Method for treating cancer with a B-RAF inhibitor

Abstract

A method for treating cancer in a subject in need of cancer treatment, the method comprising administering to the subject from about 5 mg to about 600 mg of 1-((1S,1aS,6bS)-5-((7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-4-yl)oxy)-1a,6b-dihydro-1H-cyclopropa[b]benzofuran-1-yl)-3-(2,4,5-trifluorophenyl)urea or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, isotopic molecular species, solvate, or prodrug thereof is provided.

Description

Technical Field

[0001] This specification provides a method for treating cancer in mammals. In particular, the method relates to the use of a B-Raf inhibitor for treating cancer, in particular 1-((1S,1aS,6bS)-5-((7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-4-yl)oxy)-1a,6b-dihydro-1H-cyclopropa[b]benzofuran-1-yl)-3-(2,4,5-trifluorophenyl)urea or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, isotopic molecular species, solvate, or prodrug thereof.

Background Art

[0002] Effective treatment of hyperproliferative disorders, including cancer, is an ongoing goal in the field of oncology. In general, cancer is caused by dysregulation of normal processes that control cell division, differentiation, and apoptotic cell death, and is characterized by the growth of malignant cells with the potential for unlimited proliferation, local expansion, and systemic metastasis. Dysregulation of normal processes includes abnormalities in signaling pathways and abnormal responses to factors different from those found in normal cells.

[0003] Receptor tyrosine kinases (RTKs) catalyze the phosphorylation of specific tyrosine amino acid residues in various proteins, including themselves, that govern cell growth, proliferation, and differentiation.

[0004] Downstream of several RTKs, there are several signaling pathways, and one of them is the Ras-Raf-MEK-ERK kinase pathway. Currently, it is understood that the activation of Ras GTPase proteins in response to growth factors, hormones, cytokines, etc. stimulates the phosphorylation and activation of Raf kinase. These kinases then phosphorylate and activate the intracellular protein kinases MEK1 and MEK2, and the intracellular protein kinases MEK1 and MEK2, in turn, phosphorylate and activate other protein kinases, ERK1 and 2. This signaling pathway is also known as the mitogen-activated protein kinase (MAPK) pathway or cytoplasmic cascade and mediates the cellular response to growth signals. This final function is to link the activity of receptors at the cell membrane to the modification of cytoplasmic or nuclear targets that govern cell growth, differentiation, and survival.

[0005] The constitutive activation of this pathway is sufficient to induce cell transformation. Dysregulation of the activation of the MAP kinase pathway by aberrant activation of receptor tyrosine kinases, Ras mutations or Raf mutations is frequently found in human cancers and represents a major factor in determining abnormal growth control. Ras mutations are common in human malignancies and have been identified in approximately 30% of cancers. The Ras family of GTPase proteins (proteins that convert guanosine triphosphate to guanosine diphosphate) relays signals from activated growth factor receptors to intracellular partners downstream. Among the targets mobilized by active membrane-bound Ras, prominent are the Raf family of serine / threonine protein kinases. The Raf family consists of three related kinases (A-, B-, and C-Raf) that act as downstream effectors of Ras. Ras-mediated Raf activation sequentially causes the activation of MEK1 and MEK2 (MAP / ERK kinase 1 and 2), which in turn phosphorylate ERK1 and ERK2 (extracellular signal-regulated kinases 1 and 2) on tyrosine-185 and threonine-183. Activated ERK1 and ERK2 translocate to and accumulate in the nucleus, where they can phosphorylate various substrates, including transcription factors that control cell growth and survival. Considering the importance of the Ras / Raf / MEK / ERK pathway in the development of human cancers, the kinase components of the signaling cascade are integrated as potentially important targets for the regulation of disease progression in cancer and other proliferative disorders.

[0006] Mutations in various Ras GTPases and B-Raf kinases have been identified, which result in constitutive and persistent activation of the MAPK pathway and can ultimately increase cell division and survival. As a result, these mutations are strongly associated with the establishment, development, and progression of a wide range of human cancers. The biological role of Raf kinases in signal transduction, particularly that of B-Raf, is described in Davies, H., et al., Nature (2002) 9:1-6; Garnett, M.J. & Marais, R., Cancer Cell (2004) 6:313-319; Zebisch, A. & Troppmair, J., Cell Mol. Life Sci. (2006) 63:1314-1330; Midgley, R.S. & Kerr, D.J., Crit. Rev. Onc / Hematol. (2002) 44:109-120; Smith, R.A., et al., Curr. Top. Med. Chem. (2006) 6:1071-1089; and Downward, J., Nat. Rev. Cancer (2003) 3:11-22.

[0007] Naturally occurring mutations in the B-Raf kinase that activate signaling in the MAPK pathway are present at high rates in human melanoma (Davies (2002) supra), thyroid cancer (Cohen et al J. Nat. Cancer Inst. (2003) 95(8) 625-627 and Kimura et al Cancer Res. (2003) 63(7) 1454-1457), and, although at lower rates, still at significant frequencies, in Barrett's adenocarcinoma (Garnett et al., Cancer Cell (2004) 6 313-319 and Sommerer et al Oncogene (2004) 23(2) 554-558), cholangiocarcinoma (Zebisch et al., Cell. Mol. Life Sci. (2006) 63 1314-1330), breast cancer (Davies (2002) supra), cervical cancer (Moreno-Bueno et al Clin. Cancer Res. (2006) 12(12) 3865-3866), cholangiocarcinoma (Tannapfel et al Gut (2003) 52(5) 706-712), central nervous system tumors including primary CNS tumors such as glioblastoma, astrocytoma, and ependymoma (Knobbe et al., Acta Neuropathol. (Berl.) (2004) 108(6) 467-470, Davies (2002) supra, and Garnett et al., Cancer Cell (2004) supra) and secondary CNS tumors (i.e., metastases of tumors that originated outside the central nervous system to the central nervous system), colorectal cancer including colon cancer (Yuen et al Cancer Res. (2002) 62(22) 6451-6455, Davies (2002) supra, and Zebisch et al., Cell. Mol. Life Sci. (2006)), gastric cancer (Lee et al Oncogene (2003) 22(44) 6942-6945), head and neck cancers including squamous cell carcinoma of the head and neck (Cohen et al J. Nat. Cancer Inst. (2003) 95(8) 625-627 and Weber et al Oncogene (2003) 22(30) 4757-4759), hematological cancers including leukemia (Garnett et al., *Cancer Cell* (2004), supra, particularly acute lymphoblastic leukemia (Garnett et al., *Cancer Cell* (2004) supra and Gustafsson et al *Leukemia* (2005) 19(2) 310-312), acute myeloid leukemia (AML) (Lee et al *Leukemia* (2004) 18(1) 170-172, and Christiansen et al *Leukemia* (2005) 19(12) 2232-2240), myelodysplastic syndromes (Christiansen et al *Leukemia* (2005) supra), and chronic myeloid leukemia (Mizuchi et al Biochem. Biophys.Res.Commun. (2005) 326(3) 645-651), Hodgkin lymphoma (Figl et al Arch. Dermatol. (2007) 143(4) 495-499), non-Hodgkin lymphoma (Lee et al Br. J.Cancer (2003) 89(10) 1958-1960), megakaryoblastic leukemia (Eychene et al Oncogene (1995) 10(6) 1159-1165), and multiple myeloma (Ng et al Br. J.Haematol. (2003) 123(4) 637-645), hepatocellular carcinoma (Garnett et al., *Cancer Cell* (2004), lung cancer (Bose et al Cancer Res. (2002) 62(23) 6997-7000, Cohen et al J. Nat.Cancer Inst. (2003) supra and Davies (2002) supra), small cell lung cancer (Pardo et al EMBO J. (2006) 25(13) 3078-3088) and non-small cell lung cancer (Davies (2002) supra), ovarian cancer (Russell&McCluggage J.Pathol. (2004) 203(2) 617-619 and Davies (2002) supra), endometrial cancer (Garnett et al., *Cancer Cell* (2004) supra, and Moreno-Bueno et al Clin.Cancer Res. (2006) supra), pancreatic cancer (Ishimura et al Cancer Lett.(2003)199(2)169 - 173), pituitary adenoma (De Martino et al J.Endocrinol. Invest.(2007)30(1)RC1 - 3), prostate cancer (Cho et al Int.J.Cancer(2006)119(8)1858 - 1862), renal cancer (Nagy et al Int.J.Cancer(2003)106(6)980 - 981), sarcoma (Davies(2002) supra), and skin cancer (Rodriguez - Viciana et al.,Science(2006)311(5765)1287 - 1290 and Davies(2002) supra). Overexpression of c - Raf has been associated with AML (Zebisch et al.,Cancer Res.(2006)66(7)3401 - 3408, and Zebisch(Cell. Mol.Life Sci.(2006)) and erythremia (Zebisch et la.,Cell. Mol.Life Sci.(2006).

[0008] According to exploratory studies using various pre - clinical and therapeutic agents, including those that target the role played by Raf family kinases in these cancers and inhibitors that selectively inhibit B - Raf kinase activity (King A.J.,et al.,(2006)Cancer Res.66:11100 - 11105), it is generally recognized that inhibitors of one or more Raf family kinases are useful for the treatment of such cancers or other conditions associated with Raf kinases.

[0009] B - Raf mutations are also involved in other conditions, including cardio - cutaneous syndrome (Rodriguez - Viciana et al Science(2006)311(5765)1287 - 1290) and autosomal dominant polycystic kidney disease (Nagao et al Kidney Int.(2003)63(2)427 - 437).

[0010] Although there have been many recent advances in the treatment of cancer with compounds such as B-Raf inhibitors, there is still a need for more effective and / or enhanced treatment in individuals affected by cancer, particularly cancers with MAPK pathway abnormalities.

[0011] The citation or identification of any reference in this section should not be construed as an admission that the reference is prior art to this application.

Summary of the Invention

[0012] This specification describes a method for treating cancer in a subject in need thereof, comprising administering to the subject a compound A having the name 1-((1S,1aS,6bS)-5-((7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-4-yl)oxy)-1a,6b-dihydro-1H-cyclopropa[b]benzofuran-1-yl)-3-(2,4,5-trifluorophenyl)urea, or a structure of formula (I):

Chemical formula

[0013] In one embodiment, the cancer is selected from the group consisting of colorectal cancer, pancreatic cancer, melanoma, non-small cell lung cancer, brain tumor, lung cancer, kidney cancer, bone cancer, liver cancer, bladder cancer, breast cancer, head and neck cancer, ovarian cancer, skin cancer, adrenal cancer, cervical cancer, lymphoma, thyroid tumor; preferably melanoma, ovarian cancer, and non-small cell lung cancer. In one embodiment, the cancer is melanoma. In one embodiment, the melanoma is cutaneous melanoma. In one embodiment, the melanoma is metastatic melanoma. In one embodiment, the cancer is ovarian cancer. In one embodiment, the cancer is non-small cell lung cancer.

[0014] In one embodiment, the cancer is characterized by a mutation in a gene selected from the group consisting of RAS, NRAS, KRAS, RAF, BRAF, CRAF, ARAF, and combinations thereof, preferably RAS, NRAS, KRAS, RAF, BRAF, and combinations thereof, more preferably NRAS, KRAS, BRAF, and combinations thereof. In one embodiment, the cancer is characterized by a mutation selected from the group consisting of NRAS Q61R, NRAS Q61K, NRAS Q61L, NRAS G12S, NRAS G13R, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12V, BRAF V600E, BRAF fusion, and combinations thereof; preferably NRAS Q61R, NRAS Q61K, NRAS Q61L, KRAS G12D, KRAS G12V, BRAF V600E, BRAF fusion, and combinations thereof; more preferably NRAS Q61R, NRAS Q61K, NRAS Q61L, KRAS G12D, KRAS G12V, and combinations thereof. In one embodiment, the cancer is characterized by genomic abnormalities in other MAPK pathways. In one embodiment, the other MAPK pathway genomic abnormality is the RASA1 splice isoform.

[0015] In one embodiment, compound A is administered 1 to 3 times a day. In one embodiment, compound A is administered once a day. In one embodiment, compound A is administered at about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, or about 160 mg per day. In one embodiment, compound A is administered at about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, or about 100 mg per day. In one embodiment, compound A is administered at about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, or about 60 mg per day. In one embodiment, compound A is administered at about 5 mg, about 10 mg, about 15 mg, about 25 mg, about 40 mg, or about 60 mg per day. In one embodiment, compound A is administered at about 40 mg, or about 60 mg per day. In one embodiment, compound A is administered at about 40 mg per day. In one embodiment, compound A is administered at about 60 mg per day.

[0016] In one embodiment, the method described herein provides an AUC of compound A in plasma in a subject of about 2,128 ng*h / ml to about 3,192 ng*h / ml 8h In one embodiment, the method described herein provides an AUC of compound A in plasma in a subject of about 4,576 ng*h / ml to about 6,864 ng*h / ml 8h In one embodiment, the method described herein provides an AUC of compound A in plasma in a subject of about 7,944 ng*h / ml to about 11,916 ng*h / ml 8h In one embodiment, the method described herein provides an AUC of compound A in plasma in a subject of about 9,840 ng*h / ml to about 14,760 ng*h / ml 8hTo provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 12,640 ng*h / ml to about 18,960 ng*h / ml in a subject 8h To provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 30,000 ng*h / ml to about 45,000 ng*h / ml in a subject 8h To provide.

[0017] In one embodiment, the subject achieves stable, partial response, or complete response. In one embodiment, the subject does not experience a progressive disease.

Brief Description of the Drawings

[0018]

Figure 1

Figure 2

Figure 3

Figure 4

Modes for Carrying Out the Invention

[0019] Definitions As used herein, "Compound A" refers to the name 1-((1S,1aS,6bS)-5-((7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-4-yl)oxy)-1a,6b-dihydro-1H-cyclopropa[b]benzofuran-1-yl)-3-(2,4,5-trifluorophenyl)urea, or the structure of formula (I):

Chemical

[0020] In one embodiment, a solid form of Compound A is used in the treatment provided herein. In one embodiment, a crystalline form of Compound A is used in the treatment provided herein. In one embodiment, an amorphous form of Compound A is used in the treatment provided herein. In one embodiment, the free base of Compound A is used in the treatment provided herein. In one embodiment, the hydrochloride salt of Compound A is used in the treatment provided herein. In one embodiment, Form I of Compound A described in Example 10 of WO2020151756 is used in the treatment provided herein.

[0021] BRAF inhibitors are chemical substances or drugs that inhibit the mitogen-activated protein kinase enzyme BRAF. They can be used to affect the MAPK / ERK pathway. For example, BRAF inhibitors include, but are not limited to, Compound A, lifirafenib, dabrafenib, vemurafenib, encorafenib, LY3009120, PLX8394, LXH254, MLN2480, Raf709, TAK632, and PLX7904.

[0022] As used herein, the term "neoplasm" refers to abnormal growth of cells or tissues and is understood to include benign, i.e., non-cancerous growth, and malignant, i.e., cancerous growth. The term "neoplastic" means of or related to a neoplasm.

[0023] As used herein, the term "agent" is understood to mean a substance that produces a desired effect in a tissue, system, animal, mammal, human, or other subject. Thus, the term "antineoplastic agent" is understood to mean a substance that produces an antineoplastic effect in a tissue, system, animal, mammal, human, or other subject. It should also be understood that an "agent" can be a single compound, or a combination or composition of two or more compounds.

[0024] As used herein, the term "treating" and derivatives thereof mean a therapeutic treatment. With respect to a particular condition, treating means (1) ameliorating the condition or one or more of the biological symptoms of the condition, (2) interfering with (a) one or more points in the biological cascade that leads to or causes the condition or (b) one or more of the biological symptoms of the condition, (3) alleviating one or more symptoms, effects, or side effects associated with the condition, or one or more symptoms, effects, or side effects associated with the condition or its treatment, and (4) delaying the progression of the condition or one or more of the biological symptoms of the condition.

[0025] As used herein, "prevention" refers to the prophylactic administration of a drug to substantially reduce the likelihood or severity of a condition or its biological symptoms, or to delay the onset of such a condition or its biological symptoms. One of ordinary skill in the art will understand that "prevention" is not an absolute term. Preventive therapies are appropriate, for example, when a subject is considered to be at high risk of developing cancer, e.g., when a subject has a strong family history of cancer, or when a subject has been exposed to a carcinogen.

[0026] As used herein, the term "therapeutically effective amount" means an amount of a drug or agent that elicits a biological or medical response in, for example, a tissue, system, animal, or human as determined by a researcher or clinician. Further, "therapeutically effective dose" means a dose that, compared to a corresponding control that has not received such a dose, results in an improved treatment, cure, prevention, or remission of a disease, disorder, or side effect, or a decrease in the incidence of a disease or disorder. This term also includes, within its scope, doses effective to enhance normal physiological functions.

[0027] Compound A disclosed herein may contain one or more chiral atoms and may exist as enantiomers. Accordingly, the compounds of the present invention include mixtures of enantiomers, as well as purified enantiomers or enantiomer-rich mixtures. It is also understood that all tautomers and mixtures of tautomers are included within the scope of Compound A.

[0028] As used herein, the term "solvate" refers to a variable stoichiometric complex formed by a solute (in the present invention, a compound of formula (I) or a salt thereof and a solvent). It is also understood that Compound A may be presented separately or both as solvates. Such solvents for the purposes of the present invention may be those that do not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, dimethyl sulfoxide, ethanol, and acetic acid. In one embodiment, the solvent used is pharmaceutically acceptable. Examples of suitable pharmaceutically acceptable solvents include, but are not limited to, water, ethanol, and acetic acid. In another embodiment, the solvent used is water (i.e., a hydrate).

[0029] Compound A may have the ability to crystallize in multiple forms, and the characteristics are known as polymorphs, and it is understood that such polymorphic forms ("polymorphs") are within the scope of Compound A. Polymorphs generally occur as a reaction to changes in temperature or pressure, or both, and may also occur from variations in the crystallization process. Polymorphs can be distinguished by various physical properties known in the art, such as X-ray diffraction patterns, solubility, and melting point.

[0030] As used herein, in this specification and the appended claims, the indefinite articles "a" and "an" and the definite article "the" include a single referent along with plural referents unless the context clearly dictates otherwise.

[0031] As used herein and unless otherwise specified, the terms "about" and "approximately" when used in connection with the dosage, amount, or weight percentage of a component of a composition or dosage form mean a dosage, amount, or weight percentage recognized by one of ordinary skill in the art to provide a pharmacological effect equivalent to that obtained from the specified dosage, amount, or weight percentage. In certain embodiments, the terms "about" and "approximately" when used in this context contemplate a dosage, amount, or weight percentage within 30%, 20%, 15%, 10%, or 5% of the specified dosage, amount, or weight percentage.

[0032] As used herein, unless otherwise specified, the terms “about” and “approximately” are used in connection with a numerical value or values provided to characterize a particular solid form, e.g., a numerical value or values that describe a particular temperature or temperature range (such as those that describe a melting, dehydration, desolvation, or glass transition temperature); a mass change (such as a mass change as a function of temperature or humidity); a solvent or water content (from the perspective of mass or percentage); or a peak position (such as in an analysis by IR or Raman spectroscopy or XRPD), and indicate that the value or values can deviate to an extent that would be considered reasonable to one of ordinary skill in the art, but still describe the solid form. Techniques for characterizing crystalline forms and amorphous solids include, but are not limited to, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), single crystal X-ray diffraction, vibrational spectroscopy such as infrared (IR) and Raman spectroscopy, solid and solution nuclear magnetic resonance (NMR) spectroscopy, optical microscopy, hot stage optical microscopy, scanning electron microscopy (SEM), electron crystallography and quantitative analysis, particle size analysis (PSA), surface area analysis, solubility studies, and dissolution studies. In certain embodiments, the terms “about” and “substantially” when used in this context indicate that a numerical value or range of values can vary within 30%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1.5%, 1%, 0.5%, or 0.25% of the recited value or range of values. For example, in some embodiments, the value of an XRPD peak position can vary by up to ±0.2° 2θ (or ±0.2 degrees 2θ) and still describe a particular XRPD peak.

[0033] As used herein, the term “pharmaceutically acceptable salt(s)” refers to salts prepared from pharmaceutically acceptable non-toxic acids or bases, including inorganic acids and inorganic bases, as well as organic acids and organic bases. Suitable pharmaceutically acceptable base addition salts of the compound include, but are not limited to, those well known in the art, e.g., Remington’s Pharmaceutical Sciences, 18 theds., Mack Publishing, Easton PA (1990) or Remington: The Science and Practice of Pharmacy, 19 th eds., Mack Publishing, Easton PA (1995). See also

[0034] As used herein and unless otherwise indicated, the terms “stereoisomer” or “stereoisomerically pure” mean one stereoisomer of a compound that is substantially free of other stereoisomers of that compound. For example, a stereoisomerically pure compound having one chiral center is substantially free of the opposite enantiomer of that compound. A stereoisomerically pure compound having two chiral centers is substantially free of other diastereomers of that compound. A typical stereoisomerically pure compound contains greater than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of other stereoisomers of the compound, greater than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of other stereoisomers of the compound, greater than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of other stereoisomers of the compound, or greater than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of other stereoisomers of the compound. Compounds can have chiral centers and can exist as racemic compounds, individual enantiomers or diastereomers, and mixtures thereof. All such isomeric forms, including mixtures thereof, are included in the embodiments disclosed herein.

[0035] The use of such compounds in stereoisomerically pure form, as well as the use of mixtures of those forms, are encompassed by the embodiments disclosed herein. For example, mixtures containing equal or unequal amounts of enantiomers of a particular compound can be used in the methods and compositions disclosed herein. These isomers may be synthesized asymmetrically or resolved using standard techniques such as chiral columns or chiral resolving agents. See, for example, Jacques, J., et al., Enantiomers, Racemates and Resolutions (Wiley-Interscience, New York, 1981); Wilen, S.H., et al., Tetrahedron 33:2725 (1977); Eliel, E.L., Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, S.H., Tables of Resolving Agents and Optical Resolutions p.268 (E.L. Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN, 1972).

[0036] It should also be noted that a compound can include E isomers and Z isomers, or mixtures thereof, as well as cis isomers and trans isomers, or mixtures thereof. In certain embodiments, the compound is isolated as either the E isomer or the Z isomer. In other embodiments, the compound is a mixture of the E isomer and the Z isomer.

[0037] "Tautomers" refer to isomers of a compound that are in equilibrium with each other. The concentration of the isomers depends on the environment in which the compound is found and can vary, for example, depending on whether the compound is a solid or in an organic or aqueous solution. For example, in an aqueous solution, pyrazole can exhibit the following isomeric forms, which are called tautomers of each other:

Chemical formula

[0038] As will be readily appreciated by those skilled in the art, a wide variety of functional groups and other structures may exhibit tautomerism, and all tautomers of a compound are within the scope of the present invention.

[0039] It should also be noted that a compound can contain unnatural proportions of atomic isotopes in one or more of its atoms. For example, a compound can be radiolabeled with a radioactive isotope such as tritium ( 3 H), iodine-125( 125 I), sulfur-35( 35 S), or carbon-14( 14 C), or can be isotopically enriched with an isotope such as deuterium ( 2 H), carbon-13( 13 C), or nitrogen-15( 15 N). As used herein, an "isotope molecular species" is a compound that is isotopically enriched. The term "isotopically enriched" refers to an atom having an isotope composition other than the natural isotope composition of that atom. "Isotope enriched" can also refer to a compound containing at least one atom, where that atom has an isotope composition different from the natural isotope composition of that atom. The term "isotope composition" refers to the amount of each isotope present for a given atom. Radiolabeled compounds and isotopically enriched compounds are useful as therapeutic agents, such as cancer and anti-inflammatory agents, research reagents, such as binding assay reagents, and diagnostic agents, such as in vivo contrast agents. All isotope variations of the compounds described herein are intended to be encompassed within the scope of the embodiments provided herein, whether radioactive or not. In some embodiments, isotope substituents of the compound are provided, for example, the isotope substituent is a deuterium, carbon-13, or nitrogen-15 enriched compound.

[0040] The term "subject" includes animals such as cows, monkeys, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits, or guinea pigs, and in one embodiment includes mammals, and in another embodiment includes humans, but is not limited thereto.

[0041] When used for treatment, Compound A may be administered as the raw chemical substance, but it is also possible to provide the active ingredient as a pharmaceutical composition. Accordingly, the present invention further provides a pharmaceutical composition comprising Compound A and one or more pharmaceutically acceptable carriers, diluents, or excipients. Compound A is as described above. The carrier, diluent, or excipient must be acceptable in the sense that it enables a pharmaceutical formulation and is compatible with the other components of the formulation and not harmful to its recipient. According to another aspect of the present invention, a process for the preparation of a pharmaceutical composition is also provided, which comprises mixing Compound A with one or more pharmaceutically acceptable carriers, diluents, or excipients. Such elements of the pharmaceutical composition utilized may be provided in separate pharmaceutical combinations or formulated together in one pharmaceutical composition. Accordingly, the present invention further provides a pharmaceutical composition comprising Compound A and one or more pharmaceutically acceptable carriers, diluents, or excipients. The above-described Compound A can be utilized in any of the above-described compositions.

[0042] The pharmaceutical composition can be provided in unit dosage form containing a predetermined amount of the active ingredient per unit dose. As is well known to those skilled in the art, the amount of the active ingredient per dose depends on the condition being treated, the route of administration, and the age, weight, and condition of the patient. Preferred unit dosage formulations contain the daily dose or daily unit sub-dose of the active ingredient described herein, or an appropriate fraction thereof. Further, such pharmaceutical compositions can be prepared by any of the methods well known in the art of pharmacy.

[0043] Compound A can be administered by any suitable route. Suitable routes include oral, rectal, nasal, topical (including buccal and sublingual), intravaginal, and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal, and epidural). It will be understood that the preferred route may vary, for example, depending on the condition of the person receiving combination therapy and the state of the cancer being treated. It will also be understood that each of the agents to be administered may be administered by the same or different routes, and that Compound A may be formulated together in a pharmaceutical composition.

[0044] Pharmaceutical compositions adapted for oral administration can be presented as individual units, such as capsules or tablets; powders or granules; solutions or suspensions in aqueous or non-aqueous liquids; edible foams or edible froths; or water-in-oil liquid emulsions or oil-in-water liquid emulsions.

[0045] Unless otherwise defined, in all dosing protocols described herein, the regimen of the compound administered need not start at the beginning of treatment and end at the end of treatment, but only the number of consecutive days on which both compounds are administered is required. Optional consecutive days on which only one of the component compounds is administered, or the indicated dosing protocol including the amount of the compound administered, occur at some point during the course of treatment.

[0046] Compound A may be used in combination in accordance with the present disclosure by being administered simultaneously in a single pharmaceutical composition containing both compounds.

[0047] Furthermore, it does not matter whether the compounds are administered in the same dosage form. For example, one compound may be administered topically and the other orally. Preferably, both compounds are administered orally.

[0048] Unless otherwise defined, in all dosing protocols described herein, the regimen of the compound to be administered need not start at the beginning of treatment and end at the end of treatment, but only the number of consecutive days on which both compounds are administered is required. Optional consecutive days on which only one of the component compounds is administered, or an indicated dosing protocol that includes the amount of the compound to be administered, occur at some point during the course of treatment.

[0049] As used herein, the term "kit" or "kit of parts" means a pharmaceutical composition(s) for use in administering compound A according to the present disclosure. In one embodiment, the kit may contain compound A in a single pharmaceutical composition, such as a tablet, or in separate pharmaceutical compositions. In one aspect, a kit is provided that includes compound A and components such as pharmaceutically acceptable excipients, diluents, or carriers. The kit may also be provided with instructions such as dosage and administration instructions. Such dosage and administration instructions may be of the type provided to a physician, for example, by a pharmaceutical label, or they may be of the type provided by a physician, such as instructions to a patient.

[0050] As used herein, the term "dosage" is understood to mean a dosage intended to either slowly increase the plasma or blood concentration level of a compound to a therapeutically effective level or to maintain such a therapeutically effective level.

[0051] In certain embodiments, the treatment of cancer can be evaluated according to the Response Evaluation Criteria in Solid Tumors (RECIST 1.1) (see Thereasse P., et al. New Guidelines to Evaluate the Response to Treatment in Solid Tumors. J. of the National Cancer Institute;2000;(92)205-216 and Eisenhauer E.A., Therasse P., Bogaerts J., et al. New response evaluation criteria in solid tumors: Revised RECIST guideline(version1.1). European J.Cancer;2009;(45)228-247). Regardless of the presence or absence of new lesions, the overall response rate in all possible combinations of tumor responses in target and non-target lesions is as follows:

Table 1

[0052] For the evaluation of target lesions, complete response (CR) means the disappearance of all target lesions, partial response (PR) means at least a 30% decrease in the sum of the longest diameters of target lesions based on the baseline sum of longest diameters, progression (PD) means at least a 20% increase in the sum of the longest diameters of target lesions based on the minimum value of the sum of longest diameters recorded since the start of treatment, or the appearance of one or more new lesions, and stable disease (SD) means no shrinkage sufficient to be considered a partial response and no increase sufficient to be considered progression based on the minimum value of the sum of longest diameters since the start of treatment.

[0053] For the evaluation of non-target lesions, complete response (CR) means the disappearance of all non-target lesions and normalization of tumor marker levels; incomplete response / stable disease (SD) means the presence of one or more non-target lesions and / or tumor marker levels remaining above the upper limit of the reference value, and progression (PD) means the appearance of one or more new lesions and / or obvious worsening of existing non-target lesions.

[0054] The following procedures, conventions, and definitions provide guidance for implementing the recommendations of the Response Assessment in Neuro-Oncology (RANO) Working Group regarding response criteria for high-grade gliomas (see Wen P., Macdonald, DR., Reardon, DA., et al. Updated response assessment criteria for high-grade gliomas: Response assessment in neuro-oncology working group. J Clin Oncol 2010;28:1963-1972). Major changes to the RANO criteria for Time Point Response (TPR) may include the addition of operational conventions to define changes in glucocorticoid dose and the removal of the subject's clinical deterioration component to focus on objective radiographic assessment. The baseline MRI examination is defined as the assessment performed at the end of the postoperative rest period before initiation or resumption of compound treatment. Baseline MRI is used as the criterion for evaluating complete response (CR) and partial response (PR). On the other hand, the minimum sum of products of diameters (SPD) obtained at either baseline or subsequent evaluations is designated as the lowest assessment and used as the criterion for determining progression. For 5 days prior to any MRI examination defined by any protocol, the subject is either not receiving glucocorticoids or is taking a stable dose of glucocorticoids. A stable dose is defined as the same daily dose for 5 consecutive days prior to the MRI examination. If the glucocorticoid dose prescribed 5 days prior to the baseline scan is changed, a new baseline scan is required with the use of glucocorticoids that meet the above criteria. The following definitions are used.

[0055] Measurable lesion: A measurable lesion is a contrast-enhancing lesion that can be measured two-dimensionally. Measurements are made with the maximum enhancing diameter (also known as the longest diameter, LD). The maximum perpendicular diameter is measured on the same image. The crosshairs for two-dimensional measurement must intersect, and the product of these diameters is calculated.

[0056] T1-weighted image with a slice thickness of 5 mm and a skip of 1 mm. The minimum LD of measurable lesions is set at 5 mm × 5 mm. Larger diameters may be required for inclusion and / or designation as target lesions. After baseline, target lesions that become smaller than the minimum measurement requirement or are not suitable for two-dimensional measurement are recorded with a default value of 5 mm for each diameter less than 5 mm. Lesions that disappear are recorded as 0 mm × 0 mm.

[0057] Multicentric lesions: Lesions considered multicentric (as opposed to contiguous) are those in which normal brain tissue intervenes between two (or more) lesions. The approach for multicentric lesions with individually worsening foci is to measure each enhancing lesion that meets the selection criteria individually. If there is no normal brain tissue between two (or more) lesions, they are considered the same lesion.

[0058] Non-measurable lesions: All lesions that do not meet the criteria for measurable disease defined above are considered non-measurable lesions, as well as all non-enhancing lesions and other truly non-measurable lesions. Non-measurable lesions include enhancing foci smaller than the specified minimum diameter (i.e., less than 5 mm × 5 mm), non-worsening lesions (e.g., post-contrast T1-weighted, T2-weighted, or fluid-attenuated inversion recovery (FLAIR) images), hemorrhagic or predominantly cystic or necrotic lesions, and meningiomas. Hemorrhagic lesions often have intrinsic T1-weighted hyperintensity that can be misinterpreted as enhancing the tumor, and for this reason, pre-contrast T1-weighted images may be examined to exclude baseline or intermittent subacute hemorrhage.

[0059] At baseline, lesions are classified as follows: Target lesions: Up to five measurable lesions can be selected as target lesions, and each measurement value is at least 10 mm × 5 mm representing the target disease; Non-target lesions: All other lesions including all non-measurable lesions (including mass effect and T2 / FLAIR findings), and any measurable lesion not selected as a target lesion. At baseline, target lesions are measured as described in the definition of measurable lesions, and the SPD of all target lesions is determined. The presence of all other lesions is documented. In all (follow-up) evaluations after treatment, the baseline classification of lesions (target lesions and non-target lesions) is maintained, and lesions are recorded and described over time in a consistent manner (e.g., recorded in the same order in source documents and eCRFs). To reduce the difficulty of interpreting changes, during the study period, all measurable and non-measurable lesions must be evaluated using the same method as at baseline (e.g., the subject needs to be imaged with the same MRI scanner or at least the same magnetic force). At each evaluation, target lesions are measured and the SPD is calculated. Non-target lesions are evaluated qualitatively, and new lesions, if any, are recorded separately. At each evaluation, the worsening status is determined based on the time-point response criteria for target lesions, non-target lesions, and new lesions. Even when only a subset of lesions is evaluated, tumor progression can be established. However, unless progression is observed, the objective status (stable, PR or CR) can only be determined when all lesions are evaluated.

[0060] The confirmatory evaluation of the overall time-point response for CR and PR is performed at the next scheduled evaluation, but if the examination interval is less than 28 days, confirmation may not be performed. The best response incorporating the confirmation requirement is derived from a series of time points.

[0061] As used herein, all amounts specified for Compound A are shown as the amount of the free or non-salt compound.

[0062] Kit Kits are provided herein that include a compound provided herein and means for monitoring a patient's response to administration of the compound provided herein. In certain embodiments, the patient has colorectal cancer, pancreatic cancer, melanoma, non-small cell lung cancer, brain tumor, lung cancer, kidney cancer, bone cancer, liver cancer, bladder cancer, breast cancer, head and neck cancer, ovarian cancer, skin cancer, adrenal cancer, cervical cancer, lymphoma, thyroid tumor, and complications thereof; preferably melanoma, ovarian cancer, and non-small cell lung cancer. In one embodiment, the measured response of the patient is inhibition of disease progression, inhibition of tumor growth, reduction of primary and / or secondary tumors, alleviation of tumor-related symptoms, improvement of quality of life, delay in the appearance of primary and / or secondary tumors, blunting of the development of primary and / or secondary tumors, reduction in the occurrence of primary and / or secondary tumors, delay in or reduction in the severity of secondary effects of the disease, arrest of tumor growth or tumor regression.

[0063] In other embodiments, kits are provided herein that include a compound provided herein and means for measuring the amount of inhibition of B-RAF, KRAS, or MEK in a patient. In certain embodiments, the kit includes means for measuring the inhibition of B-RAF or MEK in the patient's circulating plasma, blood, or tumor cells and / or skin biopsy or tumor biopsy / aspirate. In certain embodiments, kits are provided herein that include a compound provided herein and means for measuring the amount of inhibition of B-RAF, KRAS, or MEK before, during, and / or after administration of the compound provided herein. In some embodiments, the patient has colorectal cancer, pancreatic cancer, melanoma, ovarian cancer, or non-small cell lung cancer.

[0064] In certain embodiments, the kits provided herein include an amount of the compounds provided herein effective for the treatment or prevention of colorectal cancer, pancreatic cancer, melanoma, non-small cell lung cancer, brain tumors, lung cancer, kidney cancer, bone cancer, liver cancer, bladder cancer, breast cancer, head and neck cancer, ovarian cancer, skin cancer, adrenal cancer, cervical cancer, lymphoma, thyroid tumors, and complications thereof; preferably melanoma, ovarian cancer, and non-small cell lung cancer. In certain embodiments, the kits provided herein include an amount of the compounds provided herein effective for the treatment or prevention of colorectal cancer, pancreatic cancer, melanoma, ovarian cancer, or non-small cell lung cancer.

[0065] In certain embodiments, the kits provided herein include instructions for administering the compounds provided herein and / or for monitoring a patient's response to the administration of the compounds.

[0066] Method of treatment Provided herein is a method of treating cancer in a subject in need thereof, the method comprising administering Compound A to the subject.

[0067] In one embodiment, the cancer is selected from the group consisting of colorectal cancer, pancreatic cancer, melanoma, non-small cell lung cancer, brain tumors, lung cancer, kidney cancer, bone cancer, liver cancer, bladder cancer, breast cancer, head and neck cancer, ovarian cancer, skin cancer, adrenal cancer, cervical cancer, lymphoma, thyroid tumors, and complications thereof; preferably melanoma, ovarian cancer, non-small cell lung cancer.

[0068] In one embodiment, the cancer is characterized by a mutation in a gene selected from the group consisting of RAS, NRAS, KRAS, RAF, BRAF, CRAF, ARAF, and combinations thereof; preferably, RAS, NRAS, KRAS, RAF, BRAF, and combinations thereof; more preferably, NRAS, KRAS, BRAF, and combinations thereof.

[0069] In one embodiment, the cancer is characterized by a mutation selected from the group consisting of NRAS Q61R, NRAS Q61K, NRAS Q61L, NRAS G12S, NRAS G13R, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12V, BRAF V600E, BRAF fusion, and combinations thereof; preferably, NRAS Q61R, NRAS Q61K, NRAS Q61L, KRAS G12D, KRAS G12V, BRAF V600E, BRAF fusion, and combinations thereof; more preferably, NRAS Q61R, NRAS Q61K, NRAS Q61L, KRAS G12D, KRAS G12V, and combinations thereof.

[0070] In one embodiment, the cancer is characterized by genomic abnormalities in other MAPK pathways. In one embodiment, the other MAPK pathway genomic abnormality is the RAS A1 splice isoform.

[0071] In one embodiment, the cancer is characterized by a mutation in a gene selected from the group consisting of ARAF, BRAF, RAF1, KRAS, HRAS, NF1, MAP2K1, MAP2K2, MAPK1, and combinations thereof.

[0072] In one embodiment, the cancer is BRAF N20T, BRAF A33T, BRAF S36A, BRAF V47_G393del, BRAF V47_G327del, BRAF V47_D380del, BRAF V47_M438del, BRAF N49I, BRAF M53I, BRAF L64I, BRAF G69S, BRAF A81_D380del, BRAF A81_M438del, BRAF G104E, BRAF T119S, BRAF P141L, BRAF S151A, BRAF P162S, BRAF V169_G327del, BRAF V169_D380del, BRAF R188T, BRAF Q201H, BRAF G203_G393del, BRAF K205Q, BRAF V226L, BRAF E228V, BRAF R239Q, BRAF T241P, BRAF T241M, BRAF L245F, BRAF A246P, BRAF F247L, BRAF Q257R, BRAF Q257H, BRAF G258V, BRAF F259L, BRAF Q262R, BRAF H269Y, BRAF R271H, BRAF E275K, BRAF D287H, BRAF F294L, BRAF T310I, BRAF A320T, BRAF I326V, BRAF P341S, BRAF R347*, BRAF P348T, BRAF S363F, BRAF S364L, BRAF P367S, BRAF P367R, BRAF P367L, BRAF D380H, BRAF R389C, BRAF T401I, BRAF A404Cfs*9, BRAF P407L, BRAF S419Y, BRAF G421V, BRAF R444W, BRAF D448Y, BRAF D449Y, BRAF W450*, BRAF W450L, BRAF E451K, BRAF E451Q, BRAF P453T, BRAF V459L, BRAF R462E, BRAF R462K, BRAF R462I, BRAF I463T, BRAF I463S, BRAF G464I, BRAF G464R, BRAF G464E, BRAF G464A, BRAF G464V, BRAF S465D, BRAF S465E, BRAF S465A, BRAF G466R, BRAF G466E, BRAFG466A, BRAF G466V, BRAF S467A, BRAF S467L, BRAF F468C, BRAF G469L, BRAF G469del, BRAF G469S, BRAF G469R, BRAF G469E, BRAF G469A, BRAF G469V, BRAF T470K, BRAF V471I, BRAF V471F, BRAF Y472dup, BRAF Y472S, BRAF Y472C, BRAF G478C, BRAF K483E, BRAF K483M, BRAF L485_P490del, BRAF L485Y, BRAF L485_P490delinsY, BRAF L485S, BRAF L485W, BRAF L485F, BRAF L485_P490delinsF, BRAF N486_Q494del, BRAF N486del, BRAF N486_T488del, BRAF N486_T491del, BRAF N486_L495del, BRAF N486D, BRAF N486_V487del, BRAF N486_P490del, BRAF N486_A489delinsK, BRAF N486_T491delinsK, BRAF V487_P490del, BRAF V487_P492delinsA, BRAF T488_P492del, BRAF T488_Q493delinsK, BRAF A489_P490del, BRAF P490del, BRAF P490_Q494del, BRAF K499E, BRAF K499N, BRAF E501K, BRAF E501G, BRAF V504_R506dup, BRAF V504I, BRAF L505F, BRAF L505H, BRAF R509G, BRAF R509H, BRAF L514V, BRAF M517I, BRAF Q524L, BRAF L525R, BRAF T529M, BRAF T529N, BRAF T529I, BRAF W531C, BRAF G534D, BRAF Y538H, BRAF R558Q, BRAF G563D, BRAF H568D, BRAF H574N, BRAF H574Y, BRAF H574Q, BRAF N581D, BRAF N581Y, BRAF N581T, BRAF N581S, BRAF N581I, BRAFN581K, BRAF I582M, BRAF F583C, BRAF L584F, BRAF H585Y, BRAF E586K, BRAF D587A, BRAF D587G, BRAF D587E, BRAF V590I, BRAF V590G, BRAF I592V, BRAF I592M, BRAF G593D, BRAF D594N, BRAF D594H, BRAF D594Y, BRAF D594_T599dup, BRAF D594A, BRAF D594G, BRAF D594V, BRAF D594E, BRAF F595L, BRAF F595S, BRAF G596S, BRAF G596R, BRAF G596C, BRAF G596D, BRAF G596V, BRAF L597S, BRAF L597V, BRAF L597Q, BRAF L597P, BRAF L597R, BRAF A598T, BRAF A598S, BRAF A598V, BRAF A598_T599insARC, BRAF A598_T599insV, BRAF T599dup, BRAF T599A, BRAF T599K, BRAF T599R, BRAF T599I, BRAF T599_V600insTT, BRAF T599_V600insS, BRAF T599_V600insETT, BRAF T599_V600insEAT, BRAF V600_K601delinsEN, BRAF V600_S605delinsEISRWR, BRAF V600K, BRAF V600R, BRAF V600Q, BRAF V600dup, BRAF V600delinsYM, BRAF V600M, BRAF V600L, BRAF V600D, BRAF V600_K601delinsE, BRAF V600E, BRAF V600A, BRAF V600G, BRAF K601del, BRAF K601Q, BRAF K601E, BRAF K601_W604del, BRAF K601T, BRAF K601I, BRAF K601_S602delinsNT, BRAF K601N, BRAF S602T, BRAF S602Y, BRAF S602F, BRAF R603*, BRAF W604del, BRAF W604R, BRAF W604G, BRAF S605A, BRAFIncluding S605F, BRAF S605E, BRAF S605G, BRAF S605N, BRAF S605I, BRAF G606W, BRAF G606E, BRAF G606A, BRAF G606V, BRAF S607P, BRAF S607F, BRAF H608R, BRAF Q609E, BRAF Q609L, BRAF Q609H, BRAF E611D, BRAF L613F, BRAF G615R, BRAF L618F, BRAF W619R, BRAF S637*, BRAF V639I, BRAF E648Q, BRAF Y656D, BRAF R671Q, BRAF P676S, BRAF L678I, BRAF V681I, BRAF E695K, BRAF K698R, BRAF L711F, BRAF A712T, BRAF R719S, BRAF H725Y, BRAF A728V, BRAF P731T, BRAF P731S, BRAF P731L, BRAF A762E, BRAF A762V, and combinations thereof.

[0073] In one embodiment, the cancer is characterized by a mutation selected from the group consisting of KIAA1549-BRAF fusion, BCAS1-BRAF fusion, CCDC6-BRAF fusion, CDC42BPB-BRAF fusion, FAM131B-BRAF fusion, FXR1-BRAF fusion, GIT2-BRAF fusion, KLHL7-BRAF fusion, RNF130-BRAF fusion, TMEM106B-BRAF fusion, MKRN1-BRAF fusion, AGAP3-BRAF fusion, AGK-BRAF fusion, AKAP9-BRAF fusion, ARMC10-BRAF fusion, CUL1-BRAF fusion, GTF2I-BRAF fusion, PAPSS1-BRAF fusion, PCBP2-BRAF fusion, PPFIBP2-BRAF fusion, SND1-BRAF fusion, TRIM24-BRAF fusion, ZKSCAN1-BRAF fusion, SEPT3-BRAF fusion, and combinations thereof.

[0074] In one embodiment, the cancer is characterized by a mutation selected from the group consisting of NRAS G12A, NRAS G12C, NRAS G12D, NRAS G12N, NRAS G12P, NRAS G12R, NRAS G12S, NRAS G12V, NRAS G12Y, NRAS G13A, NRAS G13C, NRAS G13D, NRAS G13E, NRAS G13N, NRAS G13R, NRAS G13S, NRAS G13V, NRAS A18T, NRAS I24N, NRAS P34L, NRAS Y40*, NRAS Q43*, NRAS T50I, NRAS T58I, NRAS A59G, NRAS A59D, NRAS A59T, NRAS G60E, NRAS G60R, NRAS Q61E, NRAS Q61H, NRAS Q61H, NRAS Q61K, NRAS Q61L, NRAS Q61L, NRAS Q61P, NRAS Q61R, NRAS Q61R, NRAS Q61R, NRAS Q61*, NRAS E63K, NRAS Y64D, NRAS S65C, NRAS R68S, NRAS S89A, NRAS G115Efs*46, NRAS E132K, NRAS K135N, NRAS A146P, NRAS A146T, NRAS A146V, NRAS E162*, and combinations thereof.

[0075] In some embodiments, the cancer has the mutations described herein. In some embodiments, the subject with the cancer has the mutations described herein.

[0076] In one embodiment, the cancer is melanoma. In one embodiment, the melanoma is cutaneous melanoma. In one embodiment, the melanoma is metastatic melanoma. In one embodiment, the cancer is non-small cell lung cancer. In one embodiment, the cancer is colorectal cancer. In one embodiment, the cancer is ovarian cancer.

[0077] In one embodiment, the amount of Compound A administered per day (non-salt / non-solvated amount based on weight) is an amount selected from about 5 mg to about 600 mg. In one embodiment, the amount is selected from about 10 mg to about 500 mg. In one embodiment, the amount is selected from about 20 mg to about 400 mg. In one embodiment, the amount is selected from about 30 mg to about 200 mg. In one embodiment, the amount is selected from about 40 mg to about 100 mg. For example, Compound A is administered at about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg or about 200 mg per day. In one embodiment, Compound A is administered at about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, or about 100 mg per day. In one embodiment, Compound A is administered at about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, or about 80 mg per day. In one embodiment, Compound A is administered at about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, or about 80 mg per day. In one embodiment, Compound A is administered at about 5 mg, about 10 mg, about 15 mg, about 25 mg, about 40 mg, or about 60 mg per day. In one embodiment, Compound A is administered at about 40 mg, or about 60 mg per day. In one embodiment, Compound A is administered at about 40 mg per day. In one embodiment, Compound A is administered at about 60 mg per day.

[0078] In one embodiment, Compound A is administered 1 to 3 times a day. In one embodiment, Compound A is administered 3 times a day. In one embodiment, Compound A is administered 2 times a day. In one embodiment, Compound A is administered 1 time a day.

[0079] In one embodiment, the method described herein provides a plasma compound A AUC of about 2,000 ng*h / ml to about 3,200 ng*h / ml in a subject 8h In one embodiment, the method described herein provides a plasma compound A AUC of about 2,128 ng*h / ml to about 3,192 ng*h / ml in a subject 8h In one embodiment, the method described herein provides a plasma compound A AUC of about 2,400 ng*h / ml to about 2,900 ng*h / ml in a subject 8h In one embodiment, the method described herein provides a plasma compound A AUC of about 4,600 ng*h / ml to about 6,900 ng*h / ml in a subject

[0080] In one embodiment, the method described herein provides a plasma compound A AUC of about 4,600 ng*h / ml to about 6,900 ng*h / ml in a subject 8h In one embodiment, the method described herein provides a plasma compound A AUC of about 4,576 ng*h / ml to about 6,864 ng*h / ml in a subject 8h In one embodiment, the method described herein provides a plasma compound A AUC of about 5,100 ng*h / ml to about 6,300 ng*h / ml in a subject 8h In one embodiment, the method described herein provides a plasma compound A AUC of about 8,000 ng*h / ml to about 12,000 ng*h / ml in a subject

[0081] In one embodiment, the method described herein provides a plasma compound A AUC of about 8,000 ng*h / ml to about 12,000 ng*h / ml in a subject 8h In one embodiment, the method described herein provides a plasma compound A AUC of about 7,944 ng*h / ml to about 11,916 ng*h / ml in a subject 8h In one embodiment, the method described herein provides a plasma compound A AUC of about 8,900 ng*h / ml to about 10,900 ng*h / ml in a subject 8h In one embodiment, the method described herein provides a plasma compound A AUC of about 10,000 ng*h / ml to about 14,800 ng*h / ml in a subject

[0082] In one embodiment, the method described herein provides a plasma compound A AUC of about 10,000 ng*h / ml to about 14,800 ng*h / ml in a subject 8hTo provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 9,840 ng*h / ml to about 14,760 ng*h / ml in a subject 8h To provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 11,100 ng*h / ml to about 13,500 ng*h / ml in a subject 8h To provide.

[0083] In one embodiment, the method described herein provides a plasma compound A AUC of about 12,700 ng*h / ml to about 19,000 ng*h / ml in a subject 8h To provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 12,640 ng*h / ml to about 18,960 ng*h / ml in a subject 8h To provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 14,200 ng*h / ml to about 17,400 ng*h / ml in a subject 8h To provide.

[0084] In one embodiment, the method described herein provides a plasma compound A AUC of about 30,000 ng*h / ml to about 45,000 ng*h / ml in a subject 8h To provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 33,800 ng*h / ml to about 41,300 ng*h / ml in a subject 8h To provide.

[0085] In one embodiment, the method described herein provides a plasma compound A AUC of about 2,000 ng*h / ml to about 3,200 ng*h / ml in a subject receiving compound A treatment at about 5 mg / day 8h To provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 2,128 ng*h / ml to about 3,192 ng*h / ml in a subject receiving compound A treatment at about 5 mg / day 8hTo provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 2,400 ng*h / ml to about 2,900 ng*h / ml in a subject receiving compound A treatment at about 5 mg / day 8h To provide.

[0086] In one embodiment, the method described herein provides a plasma compound A AUC of about 4,600 ng*h / ml to about 6,900 ng*h / ml in a subject receiving compound A treatment at about 10 mg / day 8h To provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 4,576 ng*h / ml to about 6,864 ng*h / ml in a subject receiving compound A treatment at about 10 mg / day 8h To provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 5,100 ng*h / ml to about 6,300 ng*h / ml in a subject receiving compound A treatment at about 10 mg / day 8h To provide.

[0087] In one embodiment, the method described herein provides a plasma compound A AUC of about 8,000 ng*h / ml to about 12,000 ng*h / ml in a subject receiving compound A treatment at about 15 mg / day 8h To provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 7,944 ng*h / ml to about 11,916 ng*h / ml in a subject receiving compound A treatment at about 15 mg / day 8h To provide. In one embodiment, the method described herein provides a plasma compound A AUC of about 8,900 ng*h / ml to about 10,900 ng*h / ml in a subject receiving compound A treatment at about 15 mg / day 8h To provide.

[0088] In one embodiment, the method described herein provides a plasma compound A AUC of about 10,000 ng*h / ml to about 14,800 ng*h / ml in a subject receiving compound A treatment at about 25 mg / day 8his provided. In one embodiment, the method described herein provides a plasma compound A AUC of about 9,840 ng*h / ml to about 14,760 ng*h / ml in a subject receiving compound A treatment at about 25 mg / day 8h is provided. In one embodiment, the method described herein provides a plasma compound A AUC of about 11,100 ng*h / ml to about 13,500 ng*h / ml in a subject receiving compound A treatment at about 25 mg / day 8h is provided.

[0089] In one embodiment, the method described herein provides a plasma compound A AUC of about 12,700 ng*h / ml to about 19,000 ng*h / ml in a subject receiving compound A treatment at about 40 mg / day 8h is provided. In one embodiment, the method described herein provides a plasma compound A AUC of about 12,640 ng*h / ml to about 18,960 ng*h / ml in a subject receiving compound A treatment at about 40 mg / day 8h is provided. In one embodiment, the method described herein provides a plasma compound A AUC of about 14,200 ng*h / ml to about 17,400 ng*h / ml in a subject receiving compound A treatment at about 40 mg / day 8h is provided.

[0090] In one embodiment, the method described herein provides a plasma compound A AUC of about 30,000 ng*h / ml to about 45,000 ng*h / ml in a subject receiving compound A treatment at about 60 mg / day 8h is provided. In one embodiment, the method described herein provides a plasma compound A AUC of about 33,800 ng*h / ml to about 41,300 ng*h / ml in a subject receiving compound A treatment at about 60 mg / day 8h is provided.

[0091] In some embodiments, the AUC 8h is measured in the plasma of the subject. In some embodiments, the AUC 8h is measured in the blood of the subject. In some embodiments, the AUC 8his measured in the plasma or blood of the subject on day 1 of cycle 2. In some embodiments, the AUC 8h is measured in the plasma or blood of the subject on approximately day 29 of treatment with compound A.

[0092] In one embodiment, the subject receives the treatment provided herein for 1 to 12 cycles, each cycle consisting of about 28 days. In one embodiment, the subject receives the treatment provided herein for 1 to 12 cycles, each cycle consisting of about 21 days. In one embodiment, the subject receives the treatment provided herein for 1 to 12 cycles, each cycle consisting of about 14 days. In one embodiment, the subject receives the treatment provided herein for 1 to 12 cycles, each cycle consisting of about 7 days.

[0093] In one embodiment, the subject receives the treatment provided herein for 1 to 12 cycles, each cycle consisting of about 28 days. In one embodiment, the subject receives the treatment provided herein for 1 to 10 cycles, each cycle consisting of about 28 days. In one embodiment, the subject receives the treatment provided herein for 1 to 8 cycles, each cycle consisting of about 28 days. In one embodiment, the subject receives the treatment provided herein for 1 to 6 cycles, each cycle consisting of about 28 days. In one embodiment, the subject receives the treatment provided herein for 1 to 4 cycles, each cycle consisting of about 28 days. In one embodiment, the subject receives the treatment provided herein for 4 cycles, each cycle consisting of about 28 days. In one embodiment, the subject receives the treatment provided herein for 3 cycles, each cycle consisting of about 28 days. In one embodiment, the subject receives the treatment provided herein for 2 cycles, each cycle consisting of about 28 days. In one embodiment, the subject receives the treatment provided herein for 1 cycle, each cycle consisting of about 28 days.

[0094] In one embodiment, the subject achieves stable, partial, or complete response. In one embodiment, the subject achieves partial or complete response. In one embodiment, the subject achieves complete response. In one embodiment, the subject does not experience a progressive disease. In one embodiment, the subject achieves stability. In one embodiment, the subject achieves partial response. In one embodiment, the subject achieves stable, partial, or complete response at 1, 2, 3, or 4 weeks. In one embodiment, the subject achieves stable, partial, or complete response for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. In one embodiment, the subject achieves stable, partial, or complete response for 1, 2, 3, or 4 years.

[0095] Provided herein is a method of treating a subject in need of treatment for cancer, the method comprising administering to the subject an inhibitor of BRAF, wherein the cancer is characterized by a mutation selected from the group consisting of RAS, NRAS, KRAS, and combinations thereof; preferably NRAS and KRAS, and combinations thereof; more preferably KRAS. Provided herein is a method of treating a subject in need of treatment for cancer, the method comprising administering to the subject an inhibitor of BRAF, wherein the cancer is characterized by a KRAS mutation.

[0096] In one embodiment, the cancer is selected from the group consisting of colorectal cancer, pancreatic cancer, melanoma, non-small cell lung cancer, brain tumor, lung cancer, kidney cancer, bone cancer, liver cancer, bladder cancer, breast cancer, head and neck cancer, ovarian cancer, skin cancer, adrenal cancer, cervical cancer, lymphoma, thyroid tumor; and combinations thereof; preferably melanoma, ovarian cancer, and non-small cell lung cancer; more preferably melanoma and non-small cell lung cancer.

[0097] In one embodiment, the cancer is characterized by a mutation selected from the group consisting of NRAS Q61R, NRAS Q61K, NRAS Q61L, NRAS G12S, NRAS G13R, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12V, BRAF V600E, BRAF fusion, and combinations thereof; preferably NRAS Q61R, NRAS Q61K, NRAS Q61L, KRAS G12D, KRAS G12V, BRAF V600E, BRAF fusion, and combinations thereof; more preferably NRAS Q61R, NRAS Q61K, NRAS Q61L, KRAS G12D, KRAS G12V, and combinations thereof. In one embodiment, the cancer is characterized by the mutations provided herein.

[0098] Method for producing a compound This embodiment can be more fully understood by referring to the detailed description and examples provided herein for purposes of exemplifying non-limiting embodiments.

[0099] Compound A is Compound 1.49 in WO2014206343 and Compound 1 in WO2020151756. Compound A can be prepared as described in WO2014206343 and WO2020151756. In one embodiment, Compound A may be prepared according to the following method:

[0100] Step 1: Synthesis of INTQ-1

[0101]

Chemical formula

[0102] 1,4-Dioxane (1.5 volumes) was added to a 2 L four-necked round-bottom flask, and the flask was evacuated and flushed with nitrogen (3 times). Then, Pd(OAc)2 (2 wt%, 0.50 kg) and XantPhos (9 wt%, 2.25 kg) were added to the flask, and the flask was evacuated and flushed with nitrogen (3 times). The mixture was stirred at room temperature for 0.5 - 1 hour under a nitrogen atmosphere. NaOH (12.25 kg, 1.6 equivalents), H2O (1 volume, 25 L), and 1,4-dioxane (8 volumes, 200 L) were charged into a 20 L reactor. The mixture was stirred until clear, and then SM3 (26.75 kg, 1.2 equivalents) was added to the mixture. The catalyst solution was transferred to the above reactor under a nitrogen atmosphere. Then, SM1 (25.00 kg, 1.0 equivalent) was added dropwise to the reactor. The system was heated to 65 ± 5 °C and maintained at 65 ± 5 °C for at least 5 hours. The reaction was monitored using HPLC until the content of SM1 was 1.0% or less. The reaction mixture was cooled to 30 ± 5 °C, then filtered, and the cake was washed with 1,4-dioxane (1.0 volume). H2O (4 volumes) was added to the filtrate and concentrated to 5 volumes. Then, H2O (2 volumes) was added to the residue and concentrated to 5 volumes. The reaction product was cooled to room temperature and filtered. The cake was washed with H2O (2 volumes). Then, the filter cake was slurried with IPA (2 volumes) at 25 ± 5 °C for 3 hours. The mixture was filtered, and the filter cake was washed with IPA (0.5 volume). The solid was dried in an oven under reduced pressure.

[0103] Steps 2 and 3: Synthesis of INTQ-3

[0104]

Chemical Structure

[0105] THF (25 volumes), and INTQ-1 (16.00 kg, 1.0 equivalent) were charged into a reactor. The mixture was stirred and cooled to -80 to -70 °C. Then, n-BuLi (n-hexane solution, 2.5 M, 51.20 kg, 2.5 equivalents) was added dropwise to the mixture at -80 to -70 °C. After reacting at -80 to -70 °C for 1 to 2 hours, the reaction was monitored by TLC. Next, a solution of DMF (9.92 kg, 1.8 equivalents) in THF (1.4 volumes) at -80 to -70 °C was added dropwise to the reaction system. After reacting at -80 to -70 °C for 1 to 2 hours, the reaction was monitored by TLC. An AcOH solution in THF (1.4 volumes) was added dropwise to the mixture to adjust the pH value to 6 to 7 at -80 to -70 °C. Then, TEA (8.00 kg, 1.05 equivalents) was charged into the reactant at -80 to -70 °C. A solution of methyltriphenylphosphoranylidene acetate (26.4 kg, 1.05 equivalents) in DCM (19 volumes) was added dropwise into the reaction mixture. Then the reaction mixture was stirred at -80 to -70 °C for 10 hours, and the reaction was monitored by TLC. H2O (10.5 volumes) and citric acid (32.00 kg, 2.1 equivalents) were charged into another reactor. The mixture was stirred to dissolve and cooled to 0 to 5 °C. The temperature was cooled to -20 °C, and the solution was transferred to the above 3 L four-necked round-bottom flask. Then, the mixture was stirred at below 20 °C for 1 hour to confirm that the pH value was 4 to 7. The organic layer was separated and washed with 25% NaCl (17 volumes). Then, the organic phase was concentrated to 5 volumes, EtOAc (17 volumes) was added to the mixture, and it was concentrated to 5 volumes. EtOAc (17 volumes) was added to the mixture, and it was concentrated to 5 volumes. The solution was used directly in the next step.

[0106] Step 4: Synthesis of INTQ-4

[0107]

Chemical formula

[0108] The EtOAc solution of INTQ-3 was charged into a reactor. The solution was stirred and cooled to -5 to 5 °C. HCl was introduced into the mixture at -5 to 5 °C for 2 hours. The mixture was heated to 20 to 30 °C. After reacting for 5 hours until the content of INTQ-3 was less than 0.5%, HPLC was used to monitor the reaction every 2 hours. The reaction mixture was concentrated to 10 volumes and cooled to 0 to 5 °C. The residue was stirred at 0 to 5 °C for 1 hour. The mixture was filtered, and the filter cake was charged into H2O (15 volumes). The mixture was stirred at 20 to 30 °C for 2 hours. The mixture was filtered, and the filter cake was washed with H2O (3 volumes). Then, the filtrate was transferred to another reactor, Na2CO3 was charged into the mixture, and the pH value was adjusted to 8 to 9. Then, the mixture was filtered, and the filter cake was washed with H2O (4 volumes). After drying in a vacuum oven, 20.73 kg (yield: 69.0%, purity: 95.0%) of INTQ-4 was obtained.

[0109] Step 5: Synthesis of INTQ-5

[0110]

Chemical formula

[0111] INTQ-4 (10.40 kg, 1.0 equivalent), Pd / C (15 wt%, 1.25 kg), and THF (11 volumes) were charged into a reactor. The mixture was stirred and heated to 30 to 35 °C. Hydrogen was charged to a pressure of 10 atm. After reacting for 15 hours until the content of INTQ-4 was less than 0.5%, HPLC was used to monitor the reaction every 2 hours. The reaction mixture was cooled to 20 to 30 °C and filtered through celite (0.2 wt%). The filter cake was washed with THF (2 volumes). The filtrate was concentrated to 3 volumes, and EtOH (6 volumes) was added to the mixture. The solution was concentrated to 3 volumes, and EtOH (6 volumes) was charged into the mixture. The mixture was concentrated to 3 volumes and used directly in the next step.

[0112] Step 6 Synthesis of BGB-INTQ-6

[0113]

Chemical formula

[0114] A solution of INTQ-5 (from the previous step) in EtOH (3 volumes), EtOH (7 volumes), and Et3N (22 wt%, 2.29 kg) were charged into a reactor. The solution was heated to 70 - 80 °C. After reacting for 15 hours until the content of INTQ-5 was less than 1.0%, HPLC was used to monitor the reaction every 2 hours. The reaction mixture was cooled to 30 - 40 °C and concentrated to 5 volumes. The mixture was cooled to -5 - 0 °C and stirred for 2 hours. The mixture was filtered and the filter cake was washed with EtOH (1 volume). After drying in an oven at 45 ± 5 °C, 7.58 kg (yield: 87.1%, purity: 99.5%) of INTQ-6 was obtained.

[0115] Step 7: Synthesis of INTQ-7

[0116]

Chemical formula

[0117] Potassium hydroxide (49.9 Kg, 1.7 equivalents) was added to a solution of 4-methoxyphenol (65 Kg, 1.0 equivalent) in DMSO (65 L, 1 volume). The system was heated to 120 °C. While maintaining the temperature at 120 - 140 °C, bromoacetaldehyde diethyl acetal (123.8 Kg, 1.2 equivalents) was added dropwise. After completion of the reaction monitored by HPLC, the reaction mixture was cooled to 20 - 40 °C. N-heptane (2 volumes) and water (2 volumes) were added to the reaction mixture. The mixture was filtered through celite (0.2 wt%) and the filter cake was washed with n-heptane (0.5 volume). The filtrate was allowed to stand for at least 30 minutes. The organic layer was separated and the aqueous layer was extracted with n-heptane (2 volumes). The combined organic layers were washed with 2N aqueous NaOH solution (2 volumes). The organic layer was washed twice with 15% aqueous NaCl solution (2 volumes). The organic layer was concentrated to 3 volumes. Toluene (3 volumes) was added and the concentration was continued to 3 volumes. The toluene solution of INTQ-7 was used directly in the next step.

[0118] Step 8: Synthesis of INTQ-8

[0119]

Chemical formula

[0120] Amberlyst-15 (3.8 Kg, 0.1 wt%) was added to toluene (760 L, 20 vol). The system was heated to 110 °C under N2 protection. While maintaining the temperature at 105 - 110 °C, a solution of INTQ-7 (38 Kg / batch, 3 batches, 1.0 eq) in toluene was added dropwise. After the reaction system was reacted at 105 - 110 °C for 1 hour under constant pressure, it was concentrated to 17 vol. Toluene (3 vol) was charged into the system. After completion of the reaction monitored by HPLC, the reaction mixture was cooled to 20 - 40 °C. The mixture was filtered through celite (0.1 wt%), and the filter cake was washed with toluene (0.5 vol). The filtrate was washed with 2N aqueous NaOH solution (2 vol). The organic layer was washed twice with 20% aqueous NaCl solution (2 vol). The organic layer was concentrated to 2 vol. The crude product was distilled at a temperature below 110 °C to obtain INTQ-8 as an off-white solid (43 Kg, yield = 61.2%, purity ≥ 98.0%).

[0121] Step 9: Synthesis of INTQ-9

[0122]

Chemical formula

[0123] 1-Dodecanethiol (147.0 Kg, 3.5 equivalents) was added to a solution of INTQ-8 (43 Kg, 1.0 equivalent) in NMP (260 L, 6 volumes). The system was heated to 75 ± 5 °C. While maintaining the temperature below 120 °C, sodium ethoxide (69.0 Kg, 3.5 equivalents) was added in small portions. The reaction mixture was heated to 130 ± 5 °C. After reacting at 130 ± 5 °C for 16 hours, the mixture was sampled hourly for HPLC until the content of INTQ-8 was ≤ 3.0%. The reaction mixture was cooled to 60 ± 5 °C, and then 8 volumes of water were added to the mixture. The reaction mixture was cooled to 25 ± 5 °C, and then 3 volumes of petroleum ether were added to the mixture. The mixture was stirred for at least 30 minutes, allowed to stand for at least 30 minutes, and separated. The organic phase was temporarily stored. The aqueous phase was adjusted to pH = 1 - 2 with 6N HCl. The aqueous phase was extracted with 5 volumes and 3 volumes of ethyl acetate, respectively. The aqueous residue was combined with the temporary organic phase, and then 4 volumes of ethanol and 4 volumes of petroleum ether were added. The mixture was stirred for at least 30 minutes, allowed to stand for at least 30 minutes, and then separated. The aqueous phase was adjusted to pH = 1 - 2 with 6N HCl. The aqueous phase was extracted with ethyl acetate (5 volumes). The organic phases of ethyl acetate were combined and concentrated to 3 volumes under a pressure below 50 °C. 5 volumes of n-heptane were charged to the residue, and the mixture was adjusted to pH = 9 - 10 with 5% NaOH. The mixture was stirred for at least 30 minutes, allowed to stand for at least 30 minutes, and separated. The aqueous phase was adjusted to pH = 1 - 2 with 6N HCl. The aqueous phase was extracted with 5 volumes and 3 volumes of ethyl acetate, respectively. Then, the organic phases of ethyl acetate were combined and washed with 6 volumes of 10% H2O2 and concentrated HCl (0.15 wt). Then, the organic phase was washed with 6 volumes of 5% H2O2 and concentrated HCl (0.15 wt). The organic layer was washed with 4 volumes of 5% Na2SO3. The organic layer was washed 3 times with 3 volumes of brine. The organic layer was concentrated to 3 volumes. Dichloromethane (5 volumes) was added, and the concentration was continued until no obvious fraction remained. The crude product of INTQ-9 was used directly in the next step.

[0124] Step 10: Synthesis of INTQ-10

[0125]

Chemical formula

[0126] Et3N (48.2 Kg, 2.0 equivalents) was added to a solution of INTQ-9 (32 Kg, 1.0 equivalent) in dichloromethane (10 volumes) at a temperature below 40 °C. The mixture was cooled to -5 ± 5 °C. While maintaining the temperature at -5 ± 5 °C, TMSCl (1.3 equivalents) in dichloromethane (1 volume) was added dropwise. After reacting for 1 hour at -5 ± 5 °C, the mixture was sampled hourly for gas chromatography until the content of INTQ-9 became ≤ 2.0%. The mixture was concentrated to 3 volumes under a pressure below 40 °C. 15 volumes of n-hexane was charged to the residue and the mixture was stirred for at least 30 minutes. The mixture was filtered and the filtrate was concentrated under a pressure below 40 °C until no obvious fractions remained. The crude product was distilled at a temperature below 120 °C to obtain INTQ-10 as a pale yellow oil (40 Kg, yield = 81.4%, purity ≥ 97.5%).

[0127] Step 11: Synthesis of INTQ-11

[0128]

Chemical formula

[0129] INTQ-10 (20 Kg / batch, 2 batches, 1.0 equivalent) in dichloromethane (5 volumes) was slurried with CuI (0.1 wt%) at 25 ± 5 °C for 2 - 3 hours. Copper(I) triflate (2:1 complex with toluene, 0.11 wt%) and (S,S)-2,2-bis(4-phenyl-2-oxazolin-2-yl)propane (0.15 wt%) were stirred in dichloromethane (4 volumes) at 20 - 30 °C under N2 atmosphere for 2 - 3 hours. A solution of INTQ-10 in dichloromethane was added through a micropore filter, and a solution of ethyl diazoacetate (2.0 equivalents) in dichloromethane (10 volumes) was slowly added dropwise at 20 - 30 °C over 15 - 25 hours. The mixture was stirred at 20 - 30 °C for 30 - 60 minutes, and the mixture was washed 3 times with 4 volumes of 0.05 N aqueous sodium dihydrogen ethylenediaminetetraacetate dihydrate solution at 20 - 30 °C. The organic layer was washed twice with 3 volumes of 25% aqueous NaCl solution. The organic layer was concentrated under vacuum at a temperature below 35 °C until the system volume was 3 volumes or less. The crude product of INTQ-11 was used directly in the next step.

[0130] Steps 12 and 13: Synthesis of INTQ-13

Chemical Structure

[0131] Step 12: The crude product of INTQ-11 was dissolved in methanol (3 volumes), and an EtOH solution of 38% HCl (0.1 volume) was added to the mixture, which was then stirred at 20 - 30 °C for 2 - 3 hours. Et3N was added dropwise to the mixture to adjust the pH to 7. The mixture was concentrated under reduced pressure to 2 volumes. Ethyl acetate (2 volumes) was charged, and the concentration was continued under pressure to 2 volumes. n-Heptane (2 volumes) was charged, and the concentration was continued under pressure to 2 volumes. Dichloromethane (2 volumes) was added to completely dissolve the material. The residue was purified by silica gel chromatography (eluted with EtOAc:PE = 1:5, a total of about 100 volumes) to obtain INTQ-12 as a yellow solid.

[0132] Step 13: INTQ-12 was charged into EtOAc (1.5 volumes) and n-heptane (20 volumes), and the mixture was heated to 75 - 85 °C until it became transparent. The transparent solution was stirred at 75 - 85 °C for 1 hour and then gradually cooled to 15 - 20 °C. The mixture was filtered and washed with n-heptane (2 volumes) to obtain the product. The wet product was dried at 55 ± 5 °C for at least 16 hours to obtain INTQ-13 as a pale yellow to off-white solid.

[0133] Steps 14 and 15: Synthesis of INTQ-15

Chemical formula

[0134] Step 14: INTQ-13 (16 Kg, 1.0 equivalent) and INTQ-6 (12.7 Kg, 1.05 equivalents) were added to DMF (5 volumes). The system was heated to 55 ± 5 °C. Cesium carbonate (29.6 Kg, 1.25 equivalents) was added. The reaction mixture was heated to 110 ± 5 °C. After reacting at 110 ± 5 °C for 2 hours, the mixture was sampled hourly for HPLC until the content of INTQ-13 became ≤ 0.5%. The reaction mixture was cooled to 30 ± 5 °C and then adjusted to pH = 6 with acetic acid (5 wt%) at 30 ± 5 °C. Water (30 volumes) was added to the mixture at 25 ± 5 °C. The mixture was stirred for 1 - 2 hours and filtered to obtain the wet product. The wet product was reslurried with water (5 volumes). The filter cake was used directly in the next step.

[0135] Step 15: The wet product of INTQ-14 was added to a mixture of 1 N NaOH (10 volumes) and THF (20 volumes). The system was stirred at 25 ± 5 °C. After reacting at 25 ± 5 °C for 4 hours, sampling was performed every hour for HPLC until the content of INTQ-14 became 0.5% or less. The system was adjusted to pH = 4 - 5 with 4 N HCl at 25 ± 5 °C and stirred for 1 hour. The system was concentrated to 8 volumes under a pressure of less than 50 °C, and then filtered to obtain a wet product. The wet product was reslurried with THF (10 volumes). The mixture was stirred for 1 - 2 hours and filtered to obtain a wet product. The wet product was dried at 55 ± 5 °C for at least 30 hours to obtain INTQ-15 as a light brown to off-white solid.

[0136] Steps 16, 17, and 18: Synthesis of INTQ-18

Chemical formula

[0137] The reactor was evacuated to a pressure of -0.08 MPa or less and then filled with inert nitrogen up to atmospheric pressure. 1,4-Dioxane (10.0 volumes) and INTQ-15 (3.6 kg, 1.0 equivalent) were added to the reactor. The mixture was concentrated to 6.0 - 6.5 volumes at a temperature of less than 50 °C, and the mixture was sampled for the water content. Et3N (1.1 equivalents) was charged into the reactor. The mixture was heated to 30 ± 5 °C, and DPPA (1.1 equivalents) was added dropwise to the reactor. After reacting at 30 ± 5 °C for 2 hours, the mixture was sampled for HPLC analysis until the content of INTQ-15 became 1.0% or less. A solution of INTQ-16 was obtained.

[0138] Another reactor was evacuated to a vacuum of -0.08 MPa or less, and then filled with inert nitrogen up to atmospheric pressure. t-BuOH (20.0 volumes), (Boc)2O (0.5 equivalent), and DMAP (0.02 equivalent) were charged into the reactor. The mixture was heated to 85 ± 5 °C and stirred for 2 - 3 hours, and the mixture was sampled for water content. The criterion was KF ≤ 0.01%. A solution of INTQ-16 was added dropwise to the above-mentioned t-BuOH-based reactor at 85 ± 5 °C (for at least 3 hours). After 2 hours at 85 ± 5 °C, the mixture was sampled for HPLC analysis until the content of INTQ-16 became 1.0% or less. Then, the mixture was cooled to below 50 °C and concentrated to 3.0 - 4.0 volumes at below 50 °C.

[0139] DCM (10.0 volumes × 2) was charged into the residue, and the mixture was concentrated to 3.0 - 4.0 volumes at below 50 °C. DCM (10.0 volumes) was charged into the residue. Then, 1 wt% aqueous NaOH solution (20.0 volumes) was charged into the reactor and stirred at 25 ± 5 °C for at least 1 hour. The mixture was filtered through celite for separation. The organic phase was washed with water (5.0 volumes) and separated. The organic phase was further washed with 25 wt% brine (5.0 volumes) and separated through silica gel to remove some impurities. The organic phase was concentrated to 6.0 - 7.0 volumes at below 40 °C. DCM was charged up to 7.0 volumes. Then, the mixture was cooled to 15 °C or less, and hydrochloric acid (1.2 volumes) was added dropwise to the reactor at a temperature of 15 °C or less. After reacting at 15 ± 5 °C for 3 hours, the mixture was sampled for HPLC analysis until the content of INTQ-17 became 4.0% or less. The mixture was heated to 25 ± 5 °C, and water (3.0 volumes) was added to the reactor.

[0140] INTQ-16: 11H NMR (400 MHz, DMSO-d6) δ 10.48 (s, 1H), 7.95 (d, J = 5.6 Hz, 1H), 7.34 (d, J = 2.4 Hz, 1H), 7.05 (d, J = 8.4 Hz, 1H), 7.00 (dd, J = 8.8, 2.4 Hz, 1H), 6.25 (d, J = 5.6 Hz, 1H), 5.42 (d, J = 5.2 Hz, 1H), 3.56 (dd, J = 5.2, 2.8 Hz, 1H), 2.92 (t, J = 7.6 Hz, 2H), 2.54 (d, J = 8.0 Hz, 2H), 1.51 (d, J = 3.2 Hz, 1H). MS: M / e 364 (M+1) + 。

[0141] INTQ-17: 1 1H NMR (400 MHz, DMSO-d6) δ 10.49 (s, 1H), 7.94 (d, J = 6.0 Hz, 1H), 7.34 (s, 1H), 7.18 (s, 1H), 6.96 - 6.83 (m, 2H), 6.22 (d, J = 5.6 Hz, 1H), 4.86 (d, J = 5.6 Hz, 1H), 2.92 (t, J = 7.6 Hz, 2H), 2.86 (d, J = 4.8 Hz, 1H), 2.54 (t, J = 7.6 Hz, 2H), 2.12 (s, 1H), 1.39 (s, 9H). MS: M / e 410 (M+1) + 。

[0142] pH adjustment process: A 4 wt% aqueous NaOH solution was added dropwise to the reactor to adjust the pH value to 2.7 - 3.1. When pH > 3.1, hydrochloric acid (0.2 volume) was added, and then a 4 wt% aqueous NaOH solution was added dropwise to the reactor to adjust the pH value to 2.7 - 3.1 (precision pH test paper, range 2.7 - 4.7). The mixture was separated, and the emulsion phase was recovered as the aqueous phase. The mixture was filtered through celite, and the obtained aqueous phase was washed once with DCM (2.0 volume). To the remaining aqueous phase in the reactor, DCM (6.0 volume) and EtOH (5.0 volume) were added. A 10.0 wt% Na2CO3 solution was added dropwise to the reactants to adjust the pH value to 8 - 9 at 25 ± 5 °C. The mixture was stirred for 10 - 15 minutes and allowed to stand for 10 - 15 minutes. The mixture was separated, and the aqueous phase was extracted twice with DCM (4.0 volume). The organic phases were combined, washed with water (2.0 volume), separated, and the organic phase was washed once with 25 wt% brine (5.0 volume). The organic phase was concentrated to 3.0 - 4.0 volume at below 45 °C, and then n-heptane (4.0 volume) was added to the residue. The mixture was concentrated to 3.0 - 4.0 volume at below 45 °C, and then n-heptane (4.0 volume) was added to the residue. The mixture was concentrated to 3.0 - 4.0 volume at below 45 °C. The residue was cooled to 25 ± 5 °C and then centrifuged, and the solid was washed with n-heptane (2.0 volume). The cake was transferred to a vacuum oven, and the mixture was dried at 45 ± 5 °C (box temperature) for 4 hours. Sampling was performed for loss on drying (LOD) until LOD ≤ 1.0%. The purity of INTQ-18 (2.25 kg) was reported. The product was placed in a double LDPE plastic bag and stored at 2 - 30 °C.

[0143] INTQ-18: 1 H NMR (400 MHz, DMSO-d6) δ 10.81 (s, 1H), 8.87 (s, 3H), 8.05 (d, J = 6.0 Hz, 1H), 7.33 (t, J = 1.2 Hz, 1H), 7.07 - 6.95 (m, 2H), 6.34 (d, J = 6.0Hz, 1H), 5.24 (d, J = 6.0 Hz, 1H), 3.32 (dd, J = 6.0, 2.0 Hz, 1H), 2.97 (t, J = 7.6Hz, 2H), 2.59 (t, J = 7.6 Hz, 2H), 2.46 (s, 1H). MS: M / e 310 (M+1)+ .

[0144] Step 19: Synthesis of INTQ-19 [Chemical formula]

[0145] The reactor was evacuated to below -0.08 MPa and then filled with inert nitrogen up to atmospheric pressure. THF (6.0 volumes), H2O (3.0 volumes), 2,4,5-trifluoroaniline (1.0 equivalent), and NaHCO3 (1.2 equivalents) were charged into the reactor. The mixture was cooled to 0 °C, and phenyl chloroformate was slowly added at 0 ± 5 °C. The mixture was stirred for at least 2 hours. The mixture was sampled for LCMS until the 2,4,5-trifluoroaniline content was 0.2% or less. Then, EA (15.0 volumes) was added. The organic phase was washed with H2O (5.0 volumes), then washed twice with 5 wt% aqueous HCl solution (5.0 volumes), and washed twice with saturated NaCl (5.0 volumes). The organic phase was concentrated to 10.0 volumes at below 45 °C. n-Heptane (10.0 volumes) was added to the residue. The mixture was concentrated to 10.0 volumes, then n-heptane (10.0 volumes) was added to the residue. The mixture was concentrated to 10.0 volumes, centrifuged, and the solid was washed with n-heptane (2.0 volumes). The cake was sampled for LCMS analysis based on the criterion of INTQ-19 > 99%. Then, the cake was transferred to a vacuum oven and dried at 35 ± 5 °C (box temperature) for 10 hours, and then sampled for LOD until LOD ≤ 2.0%. The purity of INTQ-19 was reported. The product was placed in a double LDPE plastic bag and stored at 2 - 30 °C.

[0146] INTQ-19: 1 H NMR (400 MHz, DMSO-d6) δ 10.20 (s, 1H), 7.82 (dt, J = 12.0, 8.0 Hz, 1H), 7.66 (td, J = 10.8, 7.6 Hz, 1H), 7.44 (t, J = 7.6 Hz, 2H), 7.33 - 7.20 (m, 3H).

[0147] Step 20: Synthesis of the crystalline form (Form A) of Compound A

Chemical formula

[0148] The reactor was evacuated to a pressure of -0.08 MPa or less and then filled with inert nitrogen up to atmospheric pressure. DMSO (9.0 volumes), INTQ-18 (1.63 kg, 1.0 equivalent), and N-methylmorpholine (1.0 equivalent) were charged into the reactor. The mixture was stirred at 20 ± 5 °C for at least 0.5 hour. INTQ-19 (1.27 kg, 0.9 equivalent) was charged into the reactor at 20 ± 5 °C. After reacting at 20 ± 5 °C for 3 hours, the mixture was sampled for HPLC analysis until the content of INTQ-19 became 0.3% or less. After completion of the reaction, the mixture of Compound 1 was added dropwise to a 0.5% hydrochloric acid solution through a microfilter, and this was also slowly filtered through a micron filter (30.0 volumes) at 20 ± 5 °C. The mixture was stirred for at least 4 hours and centrifuged. The filter cake was washed with purified water (5.0 volumes × 2).

[0149] Slurrying procedure: Charge the reactor with DMSO (9.0 volumes) and 0.5% hydrochloric acid through a micron filter (30.0 volumes), charge the filter cake into the reactor, stir the mixture at 20 ± 5 for at least 4 hours, and then centrifuge. Wash the filter cake with purified water (5.0 volumes × 2). Sample the cake for HPLC analysis based on Compound 1 ≥ 98.0%. If Compound 1 < 98.0%, repeat the "slurry procedure". Charge purified water (40.0 volumes) and the filter cake into the reactor, stir the mixture at 20 ± 5 °C for at least 4 hours, and then centrifuge. Wash the filter cake with purified water (5.0 volumes × 2). Then, dry the cake under vacuum at 45 ± 5 °C for at least 8 hours until LOD ≤ 3.0%. If the solvent residue does not meet the standard, remove the residual solvent by slurry: Charge purified water (40.0 volumes) and the product into the reactor, stir the mixture at 20 ± 5 °C for at least 4 hours, and then centrifuge. Wash the filter cake with purified water (5.0 volumes × 2). Dry the cake under vacuum at 45 ± 5 °C for at least 8 hours until LOD ≤ 3.0%. Sample the cake for the solvent residue. If the solvent residue does not meet the standard, repeat the "removal of residual solvent by slurry" procedure until the solvent residue meets the standard. Sample the material for HPLC analysis based on the purity of Compound 1 ≥ 98.0% (2.02 kg) and the standard of Impurity-1 < 0.5%. It was determined here by HPLC analysis that the content of Impurity-1 was less than 0.1%. Package the product in a double LDPE bag with a desiccant and store it at room temperature.

[0150] Compound A: 11H NMR (400 MHz, DMSO-d6) δ 10.47 (s, 1H), 8.54 (s, 1H), 8.23 - 8.07 (m, 1H), 7.96 (d, J = 5.6 Hz, 1H), 7.65 - 7.51 (m, 1H), 7.23 (s, 1H), 7.01 (d, J = 2.0 Hz, 1H), 6.96 - 6.87 (m, 2H), 6.25 (d, J = 5.6 Hz, 1H), 4.98 (d, J = 6.0 Hz, 1H), 2.97 (dd, J = 5.6, 1.6 Hz, 1H), 2.93 (t, J = 7.6 Hz, 2H), 2.54 (t, J = 7.6 Hz, 2H), 2.26 (s, 1H).

[0151] This embodiment can be further fully understood by referring to the detailed description and examples that illustrate non-limiting embodiments for the purpose of illustration.

Example

[0152] The following examples are merely intended to be exemplary and should in no way be construed as limiting. Unless otherwise indicated, the experimental methods in the examples described below are conventional methods.

[0153]

Table 2-1

Table 2-2

Table 2-3

[0154] Example 1 The entire disclosure of NCT04249843 at ClinicalTrials.gov is incorporated herein by reference.

[0155] This study was a multicenter, open-label, two-part (dose escalation and expansion) Phase 1 trial of Compound A in patients with tumors harboring B-RAF or K-RAS / N-RAS mutations that are responsive to RAF dimer inhibitors.

[0156] Objectives and Evaluation Items

[0157] Study Objectives for the Dose Escalation Part 1a

[0158] Primary Objective:

[0159] Evaluate the safety and tolerability of Compound A in solid tumor patients.

[0160] Determine the maximum tolerated dose (MTD), if any, and the recommended Phase 2 dose (RP2D) for Compound A.

[0161] Secondary Objectives

[0162] Characterize the pharmacokinetics of Compound A after single and multiple dose administrations.

[0163] Evaluate the preliminary anti-tumor activity of Compound A.

[0164] Exploratory Objectives

[0165] Determine potential predictive biomarkers of efficacy.

[0166] Evaluate potential pharmacodynamic biomarkers of target engagement, biological activity, and mechanism of action.

[0167] Investigate the mechanisms of treatment resistance in patients who do not achieve a response or develop resistance.

[0168] Study Objectives for the Dose Expansion Part 1b

[0169] Primary Objective

[0170] To determine the objective response rate (ORR) (confirmed complete response [CR] or partial response [PR]) of Compound A when orally administered to patients with tumors selected at the RP2D, as evaluated by the Response Evaluation Criteria in Solid Tumors (RECIST version [v] 1.1).

[0171] Secondary objectives

[0172] To determine the progression-free survival (PFS); disease control rate (DCR; confirmed CR, PR, or stable disease [SD]) as evaluated by RECIST v1.1, duration of response (DOR); and overall survival of Compound A as a single agent.

[0173] To further characterize the safety and tolerability of Compound A.

[0174] To characterize the pharmacokinetics of Compound A.

[0175] Exploratory objectives

[0176] To determine potential predictive biomarkers of efficacy.

[0177] To evaluate potential pharmacodynamic biomarkers of target engagement, biological activity, and mechanism of action.

[0178] To investigate the mechanisms of treatment resistance in patients who do not achieve a response or develop resistance.

[0179] Test evaluation items for the dose-escalation Phase 1a study

[0180] Primary evaluation items

[0181] The safety and tolerability of Compound A were evaluated by the occurrence of serious adverse events (SAEs), the occurrence and severity of adverse events (AEs) (preferred terms [PTs] and system organ class [SOC] using the Medical Dictionary for Regulatory Activities [MedDRA]: Radiation), and dose-limiting toxicity (DLT) and AEs graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events version 5.0 (NCI-CTCAE v5.0), physical examinations, vital signs, electrocardiograms (ECGs), and clinical laboratory tests.

[0182] The maximum tolerated dose (MTD) was determined by the Safety Monitoring Committee (SMC) based on the occurrence of DLT, safety, and tolerability. The SMC defined the recommended phase 2 dose (RP2D) based on safety, preliminary efficacy, and other complementary data.

[0183] Secondary evaluation items

[0184] For single-dose profiles: area under the curve from zero to the last measurable concentration (AUC[last]), area under the curve from zero to infinity (AUC[0-∞]), maximum plasma concentration (Cmax), time to maximum plasma concentration (tmax), t1 / 2, apparent clearance, and apparent volume of distribution (Vz / F). Additionally, AUC[last,ss], AUC[tau], Cmax[ss], and tmax[ss] were calculated at steady state.

[0185] The objective response rate was defined as the proportion of patients with confirmed complete response (CR) or partial response (PR) as evaluated by the investigator using Response Evaluation Criteria in Solid Tumors version 1.1 (RECIST v1.1).

[0186] The duration of response was defined as the time to the first occurrence of either progression or death due to any cause from the first determination of objective response according to RECIST v1.1.

[0187] The clinical benefit rate was defined as the proportion of patients with a best overall response (BOR) of confirmed CR, PR, or stable disease (SD) lasting 16 weeks or more.

[0188] The progression-free survival period was defined as the time to the first occurrence of either the first documented disease progression according to RECIST v1.1 or death from any cause, starting from the day of the first dose of the investigational medicinal product (IMP).

[0189] The duration of stable disease (DSD) was defined as the time interval to the first occurrence of either the first documented disease progression according to RECIST v1.1 or death from any cause, starting from the day of the first dose of IMP, in the absence of either a confirmed CR or PR.

[0190] Exploratory evaluation items

[0191] Predictive biomarkers of efficacy, including but not limited to MAPK signaling, including pERK levels, the mutation status of B-RAF and K-RAS, and other abnormalities in the MAPK pathway.

[0192] Identification of potential biomarkers of resistance: Using any tumor biopsy samples taken at the time of disease progression, potential biomarkers of resistance were identified, including but not limited to the expression (protein and RNA) of MAPK signaling.

[0193] Test evaluation items for the dose escalation phase 1b

[0194] Primary evaluation items

[0195] The objective response rate (confirmed CR+PR) described in the section entitled "Secondary evaluation items" above in patients with selected tumors.

[0196] Secondary evaluation items

[0197] The above progression-free survival period, DCR (confirmed CR, PR, or SD), DOR, CBR (confirmed CR or PR or long-term SD [SD≥16 weeks]), and DSD.

[0198] The overall survival period was defined as the time from the first administration date of IMP to the date of death due to any cause.

[0199] Safety and tolerability evaluations of AE, SAE, physical examinations, ECG, and clinical laboratory values.

[0200] Single-dose and steady-state pharmacokinetics including but not limited to Ctrough.

[0201] Exploratory evaluation items

[0202] As described in Phase 1a as provided above.

[0203] Study design

[0204] Summary of the study design

[0205] This was a two-part Phase 1 trial of Compound A in patients with tumors harboring B-RAF mutations likely to respond to RAF dimer inhibitors. Compound A was a second-generation B-RAF inhibitor that showed potent inhibitory activity against the RAF family of serine / threonine kinases. In preclinical studies, Compound A was shown to inhibit tumor cell lines harboring non-V600 B-RAF mutations. It was also active against B-RAF / MEK inhibitor-resistant tumors.

[0206] Phase 1a consisted of dose escalation and dose determination components to establish the MTD and / or RP2D and to evaluate the pharmacokinetics of Compound A. Phase 1b consisted of expansion components to further evaluate the pharmacokinetics, safety, and tolerability of Compound A at the RP2D and to evaluate the preliminary antitumor activity of the compound in each of the two treatment groups of patients.

[0207] Baseline tumor tissue was essential for the analysis of mutations and biomarkers, either from stored tumor tissue or fresh tumor biopsies. For patients with easily accessible tumor lesions who had consented to biopsy, follow-up biopsies could ideally be performed from the same tumor lesion for the analysis of pharmacodynamic biomarkers. In Phase 1b, Cohort 2, paired fresh tumor biopsies were essential within 8 weeks before and after the first dose of IMP to evaluate PD biomarkers. Additionally, blood samples were collected from all patients for mutation and biomarker analysis.

[0208] Phase 1a

[0209] Dose escalation was conducted according to the modified toxicity probability interval (mTPI-2) mTPI-2 design (Yuan Ji et al 2010).

[0210] mTPI-2 uses a Bayesian statistical framework and a beta / binomial hierarchical model to calculate the posterior probabilities of three dosing intervals that reflect the relative differences between the toxicity rates at each dose level. The target toxicity rate for the MTD was φ = 0.33. The maximum sample size was 30. In this trial, patients were enrolled and treated in minimum cohorts of 3 patients. The model-based design, mTPI-2, automatically and appropriately adapts dose escalation and de-escalation decisions using an acceptable toxicity probability interval (0.28, 0.405). All dose escalation decisions were pre-calculated based on the mTPI-2 design and presented in a binary allocation table. The decision rules for "Dose Escalation" (E), "No Dose Change" (S), "Dose De-escalation" (D), or "Dose De-escalation, Unacceptable Toxicity" (DU) are described in Table 2. Patient cohorts can receive doses that have already been tested, but cannot repeat the doses associated with the decision of "Dose De-escalation, Unacceptable Toxicity", and additional patients should not be treated at this dose or higher for the remainder of the trial.

[0211]

Table 3

[0212] Sample size / dose level = 15, target toxicity probability = 0.33, epsilon1 = 0.05, epsilon2 = 0.075.

[0213] *Columns indicate the number of patients treated. Rows indicate the number of DLT patients.

[0214] *E: Escalate to the next higher dose, S: Maintain the same dose, D: De-escalate to the previous lower dose; DU: De-escalate to a lower previous dose and did not reuse the current dose during the trial.

[0215] In this trial, patients were enrolled and treated in a minimum cohort of 3 patients (see Figure 2). The design was described as follows.

[0216] Patients in the first cohort were treated at dose level 1.

[0217] To assign doses to the next cohort of patients, dose escalation / de-escalation is performed according to Table 2. When using Table 2, the following points should be noted.

[0218] "DU" means that the current and higher doses from the trial were excluded to prevent treating future patients at these doses.

[0219] If a dose is excluded, de-escalate the dose to the next lower level. If the lowest dose is excluded, end the trial for safety. In this case, the dose should not be selected as the MTD.

[0220] If the current dose is the lowest dose and the rule indicates dose de-escalation (D), continue treating new patients at the lowest dose until the number of DLTs reaches the removal boundary (i.e., DU), at which point end the trial for safety.

[0221] The current dose is the maximum dose, which may be higher than the pre-specified dose level of 60 mg. If the rule indicates a dose escalation, new patients are treated at the maximum dose (or the dose recommended by the SMC).

[0222] Repeat Step 2 until the maximum sample size of 30 is reached, or end the trial if the number of patients treated at the current dose reaches 15.

[0223] Due to the variability of small-sample binary data, the DLT may be observed occasionally at low dose levels even if the true probability (DLT) is quite low. As a result, the estimated value of the subsequent DLT may exceed the target very early in the trial and may cause early termination when few patients (e.g., 3) have been treated. To prevent early termination of the trial in such cases, it was possible to add a dose-de-escalation option at a low dose of 5 mg or the current dose level to the dose grid.

[0224] Estimation of MTD and RP2D:

[0225] Since DLT may not be frequently observed after administration of the investigational drug, the MTD may not be specified. The trial was continued until one of the following two end conditions occurred. 1. The MTD and / or RP2D are specified with sufficient accuracy: at least 6 - 12 patients have accumulated the dose presumed to be the current MTD and / or RP2D; or

[0226] 2. All doses investigated appear to be overly toxic.

[0227] The starting dose was 5 mg / day (5 mg QD). The initial treatment cycle (Cycle 1) of 30 days for each dose level cohort consisted of a single administration of Compound A on Day 1, followed by a 2-day treatment-free period (Days 2 and 3) and a 27-day repeated drug administration period (Days 4 - 30).

[0228] Continuous safety evaluations were conducted by the SMC, which consisted of the sponsor, the principal investigator of the clinical trial, and the investigator. During dose escalation, the SMC made decisions on dose levels and / or adjustments to the dosing regimen (i.e., dose level and / or schedule) based on available data from current and previous dose levels. At least three DLT-evaluable patients were required for that dose level before determining the dose level and dosing regimen for the next cohort. The RP2D and dosing regimen used in Phase 1b were recommended by the SMC and determined by the sponsor based on available safety, pharmacokinetic, preliminary antitumor activity, and other complementary data from Phase 1a. The sponsor could evaluate pre-defined or non-excluded dose levels and / or schedules as long as they did not exceed 60 mg / day or the MTD level, whichever was lower.

[0229] Phase 1b

[0230] Phase 1b was a multicenter, open-label, two-group dose-escalation study. In this Phase 1b of the trial, the RP2D was examined to investigate the potential efficacy, safety, and tolerability of Compound A in various advanced solid tumor indications. There were two treatment groups.

[0231] Group 1: Solid tumors with non-V600B-RAF mutations including RAF fusions, approximately 15 patients; and

[0232] Group 2: B-RAF V600 mutant melanoma or NSCLC progressing on B-RAF inhibitors and / or MEK inhibitors, approximately 15 patients.

[0233] Phase 1a and Phase 1b

[0234] From the date of the first administration of the IMP to 30 (+7) days after the last administration of the IMP, patients were monitored for safety, tolerability, and efficacy throughout the trial.

[0235] Patients were initially enrolled in the Phase 1a (dose escalation component of the trial). After completion of Phase 1a and determination of the RP2D for monotherapy with Compound A, patients may be enrolled in parallel in different groups of Phase 1b (advanced solid tumors).

[0236] Tumor responses were evaluated by the study physician responsible for the trial based on RECIST v1.1. In patients with ovarian cancer, tumor responses were also evaluated by the study physician responsible for the trial based on the Gynecologic Cancer InterGroup (GCIG) CA-125 criteria. For decisions regarding patient treatment and discontinuation, the RECIST criteria were prioritized.

[0237] Patients in both trial phases who continued to demonstrate clinical benefit at the end of two years of treatment were automatically rolled over to the new trial protocol and continued to be followed for treatment and safety and other clinically relevant information.

[0238] During the trial, visits, follow-up, or discontinuation

[0239] This trial consists of the following three periods.

[0240] Screening period:

[0241] Screening evaluations were performed within 21 days prior to the first administration of the IMP. Patients who consented to participate signed an informed consent form (ICF) before undergoing any screening procedures. Screening evaluations can be repeated as needed within the screening period. However, screening tumor evaluations should not be repeated unless clinically indicated. The study physician responsible for the trial was to evaluate patient eligibility according to the most recent screening evaluation results.

[0242] Rescreening under limited conditions may be permitted after consultation with the medical monitor or designee (e.g., if the patient's clinical test results slightly deviate from the clinical test criteria, which are modifiable and the condition is rapidly deteriorating or not modifiable due to PD). Rescreening was only permitted once. When rescreening a subject, a new ICF was required.

[0243] Baseline tumor tissue was essential for the analysis of mutations and biomarkers, either from the stored tumor tissue or a fresh tumor biopsy.

[0244] Registration

[0245] Before determining eligibility, all screening results and relevant medical histories must be obtained. The principal investigator of the clinical trial must confirm that all eligibility criteria are met. The site staff completed an eligibility approval packet that needed to be signed by the principal investigator of the clinical trial or a sub-investigator. Where appropriate, it was reviewed and approved by the medical monitor or designee. The approval of the eligibility approval packet and the corresponding approval date indicate the patient's registration date. Waivers of eligibility were not permitted. The site staff must ensure that confirmation of eligibility by the medical monitor has been received before the patient receives the first dose of the investigational medicinal product(s).

[0246] Treatment period

[0247] After all screening activities were completed, patients who were confirmed to be eligible and registered in the study were treated with Compound A.

[0248] The start of treatment with Compound A (i.e., Day 1 of Cycle 1) must begin within 5 days of patient registration.

[0249] The first day of Cycle 1 was the first day of the trial treatment. In Phase 1a, the first treatment cycle 1 was 30 days long, with a 2-day rest period provided on Days 2 and 3. Starting from Cycle 2 of Phase 1a, patients were treated with a repeated treatment cycle of 28-day continuous treatment cycles. (I.e., without a rest period). All patients received IMP until the completion of 2 years of treatment, when they met one of the following: 1) disease progression; 2) death, discontinuation of the trial treatment due to intolerance, or withdrawal of consent from the trial; or 3) one of the discontinuation criteria. Patients in both trial phases who continued to demonstrate a clinical benefit at the end of 2 years of treatment could continue to receive treatment according to the protocol or could participate in an expanded access protocol or compassionate use assuming the availability of the drug.

[0250] The test procedures for each clinic visit were outlined.

[0251] Unscheduled visits

[0252] Unscheduled visits can be made at any time as necessary, at the discretion of the principal investigator of the clinical trial or at the patient's request, for reasons such as additional evaluation or follow-up of AEs. If PD is suspected, tumor evaluation is necessary.

[0253] Permanent discontinuation of trial treatment

[0254] Patients can permanently discontinue trial treatment for any of the following reasons:

[0255] Disease progression

[0256] Completion of 2 years of treatment

[0257] Adverse event(s)

[0258] Death

[0259] Pregnancy

[0260] Major deviation from the protocol

[0261] The patient withdrew consent for the investigational treatment

[0262] Initiation of other anticancer therapies

[0263] Every effort must be made to encourage the patient to attend the end-of-treatment (EOT) visit and complete the appropriate safety follow-up.

[0264] Withdrawal of the patient from the study (end of study for an individual patient)

[0265] The patient may discontinue the study for reasons including, but not limited to:

[0266] Withdrawal of patient consent

[0267] Death

[0268] Loss to follow-up

[0269] Completion of all study evaluations

[0270] Visit for end of treatment

[0271] All patients attended the EOT visit within 7 days of completing all study treatment.

[0272] Patients were scheduled to attend the visit as soon as possible, but in certain circumstances (e.g., hospitalization), the EOT visit may be postponed after consultation with the medical monitor. A visit showing PD on tumor assessment may be used as the EOT visit if all necessary assessments have been performed. Tumor assessments do not need to be repeated if they were performed within 14 days of the EOT visit or at the time of a prior efficacy assessment where PD was demonstrated. Electrocardiograms do not need to be repeated if they were performed within 14 days of the EOT visit.

[0273] Safety follow-up period

[0274] The patient came to the hospital for follow-up about 30 (+7) days after the last administration of the IMP. Patients who discontinued the IMP due to drug-related AEs were followed until either resolution of the AE (Grade 1, baseline, or stabilization) or initiation of a new administration, whichever occurred first. If a new anticancer therapy was incidentally initiated before this safety follow-up (e.g., without the knowledge of the trial center team), it is necessary to schedule the safety follow-up as soon as possible.

[0275] Unable to follow up

[0276] If attempts to contact the patient by phone are unsuccessful, the following additional attempts should be made to obtain the follow-up information required by the protocol. Contact with the patient should be mailed in a way that the patient provides proof of receipt. If this fails, other contacts such as referrals from physicians or relatives should be considered. The attempts at contact should be documented in the patient's source documents. If the patient cannot be contacted despite all attempts, the patient is considered unable to be followed up, and if permitted by the local institution, death information should be obtained through a public record search.

[0277] End of the trial

[0278] The end of the trial is defined as the time when the final data of the clinical trial have been collected, which was after the final visit / follow-up for the last trial patient.

[0279] The sponsor has the right to terminate this trial at any time. Reasons for early termination of the trial include, but are not limited to:

[0280] The incidence or severity of AEs in this trial or other trials indicates a potential health risk to the patients

[0281] The overall patient enrollment was not satisfactory

[0282] When the sponsor decided to end the trial, it notified each principal investigator. If necessary, patients who had been prematurely discontinued had to return for EOT visits and safety follow-up visits.

[0283] Principal investigators may be notified of additional procedures to follow to confirm that appropriate consideration has been given to protecting the interests of the patients. The principal investigator was responsible for notifying the IRB / IEC of the early termination of the trial.

[0284] The sponsor has the right to close the facility at any time. The decision was communicated to the facility in advance. Reasons for closing the facility include, but are not limited to, the following.

[0285] Recruitment is overly slow

[0286] Failure to comply with the protocol

[0287] Inaccurate or incomplete data records

[0288] Non-compliance with GCP or applicable laws and regulations

[0289] The trial activities are completed (i.e., all trial-related activities are completed and all obligations are fulfilled)

[0290] Definition of dose-limiting toxicity

[0291] All DLTs in Phase 1a were graded. Any of the following toxicities occurring within 30 days after the first administration of Compound A and determined by the principal investigator to be related to Compound A were considered DLTs.

[0292] Hematology:

[0293] Any toxicity of grade 4 or higher.

[0294] Grade 3 febrile neutropenia (defined as an absolute neutrophil count (ANC) < 1000 / mm3, a single temperature above 38.3°C (101°F) or a sustained temperature of 38°C (100.4°F) or more for more than 1 hour).

[0295] Grade 3 neutropenia with an infection.

[0296] Grade 3 thrombocytopenia associated with clinically significant bleeding.

[0297] Non-hematological:

[0298] Grade 4 or higher unless otherwise specified below.

[0299] Grade 3 toxicities that are clinically significant and do not resolve to baseline or Grade 1 or less within 3 days of initiating optimal supportive therapy.

[0300] Grade ≥ 3 total bilirubin or liver transaminases (ALT or AST)

[0301] Note: The following AEs were not considered DLTs:

[0302] Grade 3 rash

[0303] Grade 3 or Grade 4 clinical laboratory abnormalities that are not clinically significant or do not have clinical sequelae and resolve within 48 hours.

[0304] Furthermore, clinically important or persistent toxicities not included above may be considered DLTs after review by the SMC.

[0305] Patients who received less than 80% of the assigned dose of Compound A (within 22 days of a 28-day cycle) and did not experience a DLT were not considered in the evaluation of the overall DLT rate at a particular dose level. Such patients may be replaced.

[0306] Study population

[0307] Specific eligibility criteria were provided for patient selection. The sponsor did not allow any waiver of eligibility.

[0308] Selection criteria

[0309] Each patient eligible to participate in this trial must meet all of the following criteria:

[0310] The patient has voluntarily consented to participate by providing written informed consent.

[0311] Has a histologically or cytologically confirmed progressive or solid metastatic tumor and has experienced disease progression during or after at least one previous systemic anticancer therapy, or treatment for it is not available or not tolerated by the patient. In addition, the patient must meet the following eligibility criteria at the corresponding study phase:

[0312] Phase 1a: Patients with tumors harboring a known mutation status and an oncogenic B-RAF or K-RAS / N-RAS mutation, or any other MAPK pathway aberration. In Phase 1a, enrollment in the trial was limited to approximately 1 / 3 of patients with K-RAS / N-RAS mutations, and of these patients, enrollment in the trial was limited to approximately 1 / 3 of CRC or pancreatic patients.

[0313] Phase 1b: The patient must have a known mutation status and meet one of the following criteria depending on the enrolled group:

[0314] Group 1: Patients with solid tumors with non-V600 B-RAF mutations including RAF fusions; or

[0315] Group 2: Patients with B-RAF V600 mutant melanoma or NSCLC in whom tumors resistant to B-RAF inhibitors and / or MEK inhibitors have progressed.

[0316] The patient must provide stored tumor tissue or consent to a fresh tumor biopsy for analysis of mutations and biomarkers (fresh tumor biopsies were strongly recommended at screening for patients with easily accessible tumor lesions).

[0317] The patient must have measurable disease as defined according to RECIST v1.1.

[0318] The patient must be 18 years of age or older on the date of signing the ICF.

[0319] The Eastern Cooperative Oncology Group (ECOG) performance status at screening must be 1 or less.

[0320] The life expectancy at screening must be 12 weeks or more.

[0321] Appropriate hematologic and organ function as indicated by the following laboratory values, regardless of transfusion, within 14 days of Day 1 of Cycle 1:

[0322] ANC ≥ 1500 cells / μL.

[0323] Platelets ≥ 100,000 / μL.

[0324] Hemoglobin ≥ 8 g / dL or ≥ 4.96 mmol / L.

[0325] Serum CRE ≤ 1.5 × upper limit of normal (ULN), or estimated glomerular filtration rate (GFR) ≥ 60 mL / min / 1.73 m2 according to the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation.

[0326] Serum total bilirubin ≤ 1.5 × ULN (patients with Gilbert's syndrome must have a total bilirubin less than 3 times the ULN).

[0327] AST and ALT ≤ 3 × ULN or ≤ 5 × ULN for patients with liver metastases.

[0328] Female patients were eligible to register for and participate in the study if they were in the following situations:

[0329] Not at risk of pregnancy (i.e., unable to become pregnant physiologically), including all women who:

[0330] Have had a hysterectomy.

[0331] Have had bilateral oophorectomy (ovariectomy).

[0332] Have had bilateral tubal ligation or

[0333] Are postmenopausal (complete cessation of menstruation for at least 1 year).

[0334] Are at risk of pregnancy, have a negative serum pregnancy test at screening (within 7 days of the first dose of IMP), are not breastfeeding, and use contraception before registration for the study and throughout the study until 180 days after the last dose of IMP.

[0335] Male patients are eligible to register for and participate in the study if they have had a vasectomy or have agreed to use contraception during the study treatment period and for at least 180 days after the last dose of IMP.

[0336] Exclusion Criteria

[0337] Patients who meet any of the following criteria are excluded from this study registration:

[0338] Female patients who are pregnant or breastfeeding.

[0339] Patients who are receiving cancer therapy (chemotherapy or other systemic anti-cancer therapy, immunotherapy, radiation therapy, or surgery) on Day 1 of Cycle 1.

[0340] All patients who had received prior systemic anti-cancer treatment within the following periods were excluded:

[0341] Prior to the first day of Cycle 1, cyclical chemotherapy within a period shorter than the length of the cycle used for that treatment (i.e., 6 weeks for nitrosourea, mitomycin C); and

[0342] Biological therapy (i.e., antibodies), continuous or intermittent small molecule therapy, or any other investigational drug within a period of up to 5 times the half-life of the drug or within 4 weeks (whichever is shorter) prior to the first day of Cycle 1.

[0343] Active infectious diseases requiring systemic treatment.

[0344] Any of the following cardiovascular criteria:

[0345] Current evidence of cardiac ischemia.

[0346] Ongoing symptomatic pulmonary embolism.

[0347] Acute myocardial infarction up to 6 months prior to the first day of Cycle 1.

[0348] New York Heart Association Class III or IV heart failure up to 6 months prior to the first day of Cycle 1.

[0349] Grade ≥2 ventricular arrhythmia up to 6 months prior to the first day of Cycle 1.

[0350] Cerebrovascular accident (CVA) or transient ischemic attack (TIA) up to 6 months prior to the first day of Cycle 1.

[0351] Uncontrolled hypertension: systolic blood pressure of 150 mmHg or diastolic pressure ≥100 mmHg despite the use of antihypertensive medications up to 28 days prior to the first day of Cycle 1.

[0352] Syncope or seizures up to 28 days prior to the first day of Cycle 1.

[0353] Major surgery within 28 days prior to the first day of Cycle 1.

[0354] Patients with toxicity (as a result of previous anticancer therapy) that has not recovered to baseline or is not stable (however, AEs not considered safety risks, such as alopecia, neuropathy, and certain clinical laboratory value abnormalities).

[0355] History or presence of gastrointestinal diseases or other conditions known to interfere with drug absorption, distribution, metabolism, or excretion.

[0356] Known immediate or delayed hypersensitivity reactions or specificities to drugs chemically related to Compound A (to date, there are no known US FDA-approved drugs chemically related to Compound A).

[0357] Leptomeningeal tumors or brain metastases, except for patients with previously treated brain metastases who are radiologically stable (evidence by imaging diagnosis is required), asymptomatic, and have not used steroids and anticonvulsants for more than 28 days by Day 1 of Cycle 1 are permitted.

[0358] Progressive malignancies within 2 years prior to Day 1 of Cycle 1, excluding specific cancers under investigation in this trial and any locally recurrent cancers receiving curative treatment (e.g., resected basal cell or squamous cell carcinoma, superficial bladder cancer, or cervical or breast intraepithelial carcinoma).

[0359] Unstable and existing major medical conditions for which the opinion of the principal investigator of the clinical trial contraindicates the use of the IMP, including known human immunodeficiency virus (HIV) or active hepatitis B virus (HBV) or hepatitis C virus (HCV) infections. Patients who were positive for hepatitis B surface antigen (HBsAg) or HCV antibody at the time of screening can be enrolled only if the HBV DNA titer is less than 500 IU / mL or negative by HCV RNA polymerase chain reaction test, respectively.

[0360] Psychological, familial, social, or geographical conditions that do not permit compliance with the protocol.

[0361] Unsuitable for the trial based on the opinion of the principal investigator of the clinical trial after medical interview, physical examination, or screening survey.

[0362] Taking any of the prohibited drugs or requiring any of these drugs.

[0363] Simultaneous participation in another therapeutic clinical trial.

[0364] Tumor and Efficacy Evaluation

[0365] Tumor imaging was performed within 21 days before the first test treatment. The tests or results of the standard treatment performed before obtaining informed consent and 21 days before the first administration of the IMP may be used for screening purposes instead of repeating the tests of the standard treatment. During the trial, tumor imaging was performed approximately every 8 (±1) weeks for the first year and then approximately every 12 (±1) weeks. Tumor evaluation had to be performed as scheduled regardless of whether the test treatment was administered or withheld.

[0366] Tumor evaluation must include computed tomography (CT) scans of the chest, abdomen, and pelvis (using oral / IV contrast, except in case of contraindication) or MRI (CT preferred). All measurable and evaluable lesions must be evaluated and documented at the screening visit and re-evaluated at each subsequent tumor evaluation. The same radiological procedure used to evaluate the disease site at screening should be used throughout the study (e.g., the same contrast protocol for CT scans).

[0367] Magnetic resonance images of the head at baseline (informed consent within 21 days) are required for all screened patients, except in case of contraindication, and thereafter, head CT may be sufficient.

[0368] If it is known that the patient has a contraindication to the CT contrast agent, or if a contraindication develops during the test, in addition to non-contrast CT of the chest, contrast-enhanced MRI of the abdomen and pelvis (if possible) should be performed.

[0369] When the CT scan for tumor evaluation is performed with a positron emission tomography (PET) / CT scanner, the CT acquisition must conform to the standards of a complete contrast CT scan.

[0370] If clinically indicated, a bone scan (Technetium-99m [TC-99m]) or sodium fluoride PET [NaF-PET]) should be performed at the time of screening. If bone metastases are present at the time of screening and cannot be seen on subsequent CT or MRI examinations, or if clinically indicated, if CR is suspected in the target lesion, or if bone progression is suspected, a TC-99m or NaF-PET bone scan should be repeated.

[0371] Computed tomography of the neck or extremities should also be performed if clinically indicated and if there is evidence of metastatic disease in these areas at the time of screening and followed throughout the trial. At the discretion of the principal investigator of the clinical trial, other methods of target and non-target lesion assessment according to RECIST v1.1 may be used.

[0372] Tumor response was evaluated by the principal investigator of the clinical trial using RECIST v1.1. To ensure internal consistency throughout the visit, the same evaluator should perform the evaluation if possible.

[0373] After documentation of the first response (CR or PR), confirmation of tumor response should be performed at least 4 weeks (≥4 weeks) after the first response or at the next scheduled evaluation time point.

[0374] Patients who had their test treatment discontinued early for reasons other than disease progression (e.g., toxicity) continued to receive tumor evaluations according to the original plan until any of the following occurred first: the patient initiated subsequent anticancer treatment, experienced disease progression, withdrew consent, died, or the trial ended.

[0375] All patient files and radiographic images must be made available for source verification and potential peer review.

[0376] Pharmacokinetic assessment

[0377] The following evaluations were performed by the central laboratory.

[0378] Pharmacokinetic assay: Plasma samples were assayed for the concentration of Compound A (or a potential major metabolite) using a validated chromatographic method.

[0379] Transport, storage, and handling of samples for PK assays were managed by the central laboratory. Instructions and consumable kits were provided for all central laboratory evaluations. The time points of PK sampling were documented.

[0380] Biomarker evaluation

[0381] Patients must provide stored tumor tissue or consent to a tumor biopsy for analysis of mutations and biomarkers (patients with easily accessible tumor lesions who consented to biopsy were strongly recommended to have a fresh tumor biopsy at screening). For patients with easily accessible tumor lesions who consented to biopsy, follow-up biopsies may ideally be performed from the same tumor lesion for analysis of pharmacodynamic biomarkers. In cohort 2 of phase 1b, paired fresh tumor biopsies were required within 8 weeks before the first dose and at post-dose time points to evaluate PD biomarkers. Additionally, blood samples were collected from all patients for analysis of mutations and biomarkers.

[0382] The central testing agency managed the delivery, storage, and handling of tumor tissue and blood samples for biomarker evaluation. For details on sample handling, please refer to the laboratory manual.

[0383] Stored tumor tissue (formalin-fixed paraffin-embedded blocks containing tumor tissue, or approximately 15 unstained slides) needs to be sent to the central testing agency for immunohistochemical analysis. In addition to evaluating MAPK signaling at the phosphorylated ERK level, mutations in B-RAF, K-RAS, N-RAS, and other abnormalities in the MAPK pathway, as well as other markers related to the efficacy or clinical benefit of Compound A, can also be evaluated.

[0384] For fresh biopsy specimens, acceptable samples include core needle biopsies for deep tumor tissue or excisions, incisions, punches, or forceps biopsies for skin, subcutaneous, or mucosal lesions.

[0385] Tumor tissue must be of good quality based on the tumor content of the overall survival rate. Aspiration biopsies, brushings, cell pellets from pleural effusions, and wash samples are not acceptable.

[0386] Peripheral blood samples were collected at specific time points described in the evaluation schedule for use in longitudinal evaluation of ctDNA by molecular (NGS) methods as pharmacodynamic biomarkers related to efficacy or resistance.

[0387] Hospital visit period

[0388] If the timing of the study hospital visits mandated in the clinical trial protocol conflicts with vacations, weekends, or other events, the hospital visits should be planned for the nearest available date, and subsequent hospital visits should be conducted according to the planned schedule.

[0389] Statistical methods and determination of sample size

[0390] Statistical analysis was performed by the sponsor or designee after the trial was completed, the database was locked and released. The data were enumerated and, where applicable, enumerated and summarized using SAS® version 9.4 or higher (SAS Institute, Inc., Cary, North Carolina) according to reporting criteria agreed by the sponsor. Generally, there was no missing data and the statistical methods were mainly descriptive in nature. Details of the planned analyses were documented in a separate Statistical Analysis Plan (SAP).

[0391] Statistical analysis

[0392] Randomization method

[0393] Not applicable.

[0394] Analysis population

[0395] The analysis population may include the following.

[0396] The safety population included all patients who received at least one dose of the IMP.

[0397] The evaluable population included all dosed patients with evaluable disease at baseline and at least one evaluable disease after baseline, confirmed tumor response assessments.

[0398] The DLT evaluation population included patients who received at least 80% of the dose of Compound A assigned in the dose escalation phase (Phase 1a), or patients who experienced DLT in the dose escalation phase (Phase 1a).

[0399] The PK population included all dosed patients in whom PK parameters of the active Compound A could be estimated.

[0400] The pharmacodynamic population included all patients in whom PD sampling of the active Compound A was performed after treatment with Compound A.

[0401] Patient demographics

[0402] The number of treated patients, patients who discontinued the IMP and / or the trial, and patients with major protocol deviations was counted. The main reasons for IMP and / or trial discontinuation were summarized according to the eCRF categories. The status of trial completion (survival, death, withdrawal of consent, or lost to follow-up) at the data cut-off date was summarized using data from the eCRF.

[0403] Major protocol deviations were summarized and described for each category of deviation.

[0404] Demographic and other baseline characteristics

[0405] Demographic and other baseline characteristics were summarized in the safety population using descriptive statistics. Continuous variables included age, weight, vital signs, time from first cancer diagnosis, and time from diagnosis of progressive / metastatic disease. Categorical variables included number of previous systemic treatments, tumor node metastasis classification of malignancy staging, gender, ECOG performance status, country, race, and site of metastasis.

[0406] Use of prior medications and concomitant medications

[0407] Concomitant medications were assigned an 11-digit code using the drug codes of the World Health Organization (WHO) Drug Dictionary. Concomitant medications were further coded with the appropriate anatomical therapeutic chemical (ATC) codes indicating the therapeutic classification using MedDRA version 21 or higher. Prior medications and concomitant medications were summarized and listed by drug and drug class. Prior dosing was defined as dosing completed prior to the first dose of the IMP. Concomitant medications were defined as (1) dosing that started prior to the first dose of the IMP and continued at the time of the first dose of the IMP, or (2) dosing that started on or after the first dosing day of the IMP and up to a maximum of 30 days after the patient's last dose.

[0408] Efficacy analysis

[0409] The efficacy evaluation items were analyzed using the evaluable population, except for the PFS analyzed using the safety population.

[0410] Summaries were provided for each dose level of Phase 1a and for each patient group of Phase 1b.

[0411] In the analysis of ORR, DCR, and CBR, a summary table presenting the number and percentage of responders with two-sided exact (Clopper-Pearson) 95% CI for the response rate was presented. Summaries for each response category of BOR (confirmed CR, confirmed PR, unconfirmed CR, unconfirmed PR, SD, or PD) were also presented. A waterfall plot of the maximum tumor shrinkage rate per patient was presented.

[0412] The evaluation items of time to event, including PFS, DOR, DSD, and overall survival, were analyzed by the Kaplan-Meier method. The survival functions of the evaluation items of time to event were summarized for the 25th percentile, median, and 75th percentile, and their 95% CI. The PFS, DOR, DSD, and overall survival rates at the 3rd month and every 3 months thereafter as needed, and their 95% CI, were derived based on the Kaplan-Meier estimates. Furthermore, a graph of the Kaplan-Meier estimates of the survival function was presented.

[0413] Safety analysis

[0414] The safety evaluation items (other than DLT) were summarized using the safety population. All summaries of safety were by dose level in Phase 1a and by patient group (tumor type) in Phase 1b.

[0415] Degree of exposure

[0416] The administered exposure was descriptively summarized. The measurement criteria for the degree of exposure included the number of treatment cycles received, the exposure period, and the cumulative dose during the study period. As needed, the number of patients with dose delay and the reasons for dose discontinuation were also summarized.

[0417] Adverse event

[0418] Adverse events were coded using MedDRA, and AE data were summarized by SOC and PT. The frequency and percentage of patients by reported SOC / PT were summarized, and the number of AEs was also counted. Adverse events were also summarized by the severity / grade of the worst AE and the relationship of the AE to the investigational treatment. The number of SAEs leading to discontinuation of the investigational treatment and AEs emerging from treatment (TEAEs) were also tabulated.

[0419] In Phase 1a, DLTs were summarized for the DLT evaluation population.

[0420] All AE summaries were limited to TEAEs only. AEs emerging from treatment were defined as AEs that occurred or worsened in severity after the start of IMP administration.

[0421] A patient-level AE data list was provided, including the verbatim term used by the reporter, MedDRA SOC and PT, severity, outcome, and relationship to the investigational treatment. Separate lists were generated for serious AEs and AEs leading to discontinuation of the investigational treatment.

[0422] Laboratory analysis

[0423] All hematology, chemistry, and coagulation (continuous variable) parameters were summarized using descriptive statistics for all time points scheduled in all protocols, including changes from baseline for all post-dose evaluations. The number of patients with abnormal clinical laboratory values, including clinical significance (where applicable), was also summarized. For each safety clinical laboratory parameter, a shift table from baseline to post-dose evaluation was created, and the shift categories used were "within normal range", "high", and "low" values. Laboratory values were compared to the test institution's normal range, and values outside the normal range were indicated as H (high) and L (low) in the data list.

[0424] The results of the dipstick urine tests were summarized at the time points scheduled by each protocol. Microscopic data were tabulated if available.

[0425] Vital signs

[0426] The measured values of vital signs and changes from baseline were summarized at the time points scheduled by each protocol.

[0427] Physical examination

[0428] Newly developed or worsening clinically significant abnormalities identified on physical examination were recorded as AEs and not summarized or described individually.

[0429] 12-lead electrocardiogram

[0430] For 12-lead ECG parameters, descriptive statistics including changes from baseline were calculated for all time points evaluated. Additionally, the overall interpretation of 12-lead ECG results was classified, where applicable, for all time points evaluated, into the categories of normal, clinically insignificant abnormalities, and clinically significant abnormalities, using frequencies and percentages. QT intervals corrected for heart rate (QTc) that were significantly abnormal were also summarized according to FDA Industry Guidance E14. Further details are described in the SAP.

[0431] Eastern Cooperative Oncology Group Performance Status

[0432] Shifts from baseline in ECOG Performance Status were descriptively summarized using frequencies and percentages for each dose level / treatment group at the time points scheduled by all protocols.

[0433] Pharmacokinetic analysis (PK)

[0434] Pharmacokinetic data were analyzed using PK populations. PK variables for Compound A (i.e., AUClast, AUC0-∞, Cmax, Tmax, t1 / 2, apparent clearance, and Vz / F) were calculated after the single-dose administration in Phase 1a. Further, AUClast,ss, AUCtau, Cmax,ss, and tmax,ss were calculated in Phase 1b. PK parameters were calculated using the non-compartmental method as needed and summary statistics were provided. Plasma concentration data and PK parameters of Compound A (or its potential major metabolite) were tabulated and summarized on Day 1 of Cycle 1, Day 1 of Cycle 2, and at steady state (Phase 1b). Descriptive statistics included mean, median, minimum, maximum, standard deviation, coefficient of variation (CV), geometric mean, and geometric CV as needed. Mean plasma concentrations were also plotted against time for each dose level. Additional PK analyses may be performed as needed.

[0435] When supported by the data, exposure-response (efficacy or safety assessment item) analysis may be performed. The results of such analysis can be reported separately from the clinical study report (CSR).

[0436] Other exploratory analyses

[0437] Biomarker data were analyzed using evaluable biomarkers and pharmacodynamic populations. Summary statistics were provided for pharmacodynamic biomarkers including, but not limited to, mutations, transcription, and phosphorylation profiles of MAPK pathway signaling in both paired biopsies and longitudinal blood samples. Exploratory analyses regarding potential correlations between these PD markers and dose, pharmacokinetics, safety, and antitumor activity were performed as needed. Primary predictive biomarker analysis was based on subsets of patients with abnormalities in the MAPK pathway such as oncogenic B-RAF, K-RAS, N-RAS, and NF-1 mutations. Exploratory analyses of other candidate predictive biomarkers were performed similarly. Depending on the available data, biomarker analysis was essentially descriptive. Details are provided in the SAP if applicable.

[0438] Considerations Regarding Sample Size

[0439] Formal statistical calculations for sample size were not used, and the sample size was empirical. In this trial, approximately 30 to 60 patients are planned to be enrolled.

[0440] Phase 1a: Approximately 18 to 30 patients, increasing the dose gradually until determination of MTD and / or RP2D

[0441] Phase 1b: Approximately 30 patients for expansion in two selected groups. Each group was evaluated separately and could be terminated based on statistical evaluation or futility or clinical effect due to insufficient patient enrollment. After treating all the planned patients, if promising preliminary efficacy results were observed in one of the arms (i.e., higher ORR or longer PFS), it was possible to add more patients to that arm to further evaluate the efficacy before proceeding to Phase 2 / 3 clinical development.

[0442] Interim Analysis

[0443] Formal hypothesis-related interim analyses were not conducted. However, the sponsor could request descriptive interim analyses of safety and efficacy data. The interim analysis was for the purpose of safety monitoring and future trial planning and did not affect the conduct of this trial. Efforts were made to minimize potential biases associated with the interim analysis.

[0444] The summary of all cohorts is shown in Table 3. One patient in Cohort 1 was not evaluable for DLT because of rapid disease progression.

[0445] The summary of Cohort 5 and the following results are shown in Table 4.

[0446] Four patients were enrolled in the cohort

[0447] Four patients completed the DLT period, no DLT

[0448] Dosage interruption or administration delay:

[0449] 102 - 012 interrupted, disease progression.

[0450] 201 - 010 interrupted, disease progression.

[0451] 202 - 012 administration delay, expulsion rate decreased; timing TBC.

[0452] At week 16, there was a CR target lesion in 102 - 012.

[0453] 2 SAEs (no addition from previous meeting).

[0454] Grade 2 fever, related.

[0455] Grade 3 intestinal obstruction, not related.

[0456] The administration of 2 patients was delayed (no addition from previous time).

[0457] Tumor response:

[0458] 101 - 010 PR at week 8, SD at week 16, 102 - 012 PR at week 8, CR at week 16, 202 - 012 SD at week 8.

[0459] The summary of cohort 6 and the following results are shown in Table 5.

[0460] 4 patients were enrolled in the cohort.

[0461] 2 patients completed the DLT period and 1 had DLT.

[0462] 1 patient discontinued in cycle 1, SAE (fever and angioedema).

[0463] 8 SAEs:

[0464] 201 - 011: AF, grade 3, related.

[0465] 202 - 014: Transverse striated muscle lysis × 2, Grade 2 and Grade 3 (several days later), related; Lower limb pain, Grade 3, related; Acneiform rash, Grade 2, related.

[0466] 203 - 007: Febrile syndrome x2, Grade 2 and Grade 3, related; Angioedema, Grade 2, related.

[0467] Due to AF Grade 3, the administration of 201 - 011 was delayed as a related result.

[0468] Tumor response:

[0469] 1 evaluable patient, 201 - 011: SD at week 8.

[0470] The preliminary PK parameters and data are listed in Table 6.

[0471] The parameters are described as mean (%CV)

[0472] T max is reported as the median (minimum, maximum)

[0473] Accumulation ratio R acc is based on AUC 8h

[0474] Patient 101 - 007 was excluded from C2D1 (10 mg) of Cohort 2 due to dose reduction.

[0475] ** Patient 101 - 011 was excluded from the summary statistics of C1D1 (60 mg) of Cohort 6 for the AQL sample.

[0476] ## The C2D1 parameters of Patient 201~011 were excluded from the summary statistics of C2D1 of Cohort 6 due to long - term dose interruption.

[0477] An overall summary of adverse events is provided in Table 7.

[0478] ​ *201-003 Respiratory insufficiency - Not related to IP

[0479] **Fever, Grade 1, related, recovered.

[0480] a: Update the numbers in the last safety meeting.

[0481] Summary of adverse events - All cohorts

[0482] Grade 5:

[0483] One case of respiratory insufficiency in patient 201-003 of cohort 2, not related.

[0484] Grade 4:

[0485] One case of hypercalcemia in patient 201-005 of cohort 3, not related.

[0486] Grade 3:

[0487] 55 cases in 22 patients, 18 cases related to the investigational drug.

[0488] 37 SAEs in 19 patients.

[0489] 11 non-serious cases in 9 patients.

[0490] Grade 2:

[0491] 107 cases in 23 patients, 38 cases related to the investigational drug.

[0492] 18 SAEs in 9 patients.

[0493] 86 non-serious cases in 22 patients.

[0494] Grade 1:

[0495] 241 cases occurred in 36 patients, 84 cases related to the investigational drug.

[0496] 6 SAEs in 3 patients.

[0497] 230 non - serious cases in 35 patients.

[0498] Summary of Adverse Events, Cohort 6

[0499] Grade 3:

[0500] 8 events in 3 patients.

[0501] 4 SAEs:

[0502] Related to 201 - 011AF;

[0503] Related to 202 - 014 lower limb pain;

[0504] Related to 202 - 014 rhabdomyolysis;

[0505] Related to 203 - 007 febrile syndrome.

[0506] Grade 2:

[0507] 18 events in 2 patients, 15 related to the investigational drug.

[0508] 4 SAEs in 2 patients.

[0509] 12 non - serious cases in 2 patients.

[0510] Grade 1:

[0511] 19 events in 3 patients.

[0512] 15 non - serious events in 3 patients. Patient 101 - 011 (×3), 202 - 014 (×5), 203 - 007 (×7).

[0513] 12 events are related to the investigational drug.

[0514] Note that some AEs are not designated as either severe or non-severe at the time of data extraction.

[0515] Figures 3 and 4 provide the preliminary mean PK profiles of all cohorts of C1D1 and C2D1, respectively.

[0516] [Table 4]

[0517] [Table 5]

[0518] [Table 6]

[0519] [Table 7]

[0520] [Table 8]

[0521] Compound A is a very potent RAF kinase inhibitor with equivalent potency against RAF monomers and dimers.

[0522] Compound A has the potential to target non-V600E BRAF mutations and addresses resistance to first-generation RAF / MEKi.

[0523] Compound A showed good tolerability at 5 mg, 10 mg, 15 mg, 25 mg, and 40 mg QD in cohort 5.

[0524] The results observed in Cohort 5 were as follows:

[0525] The exposure of Compound A in the entire cohort generally proportional to the increase in dose.

[0526] Two patients evaluable for tumor response showed PR at week 8.

[0527] Cohort determination and next steps:

[0528] Increase the dose of Compound A to 60 mg QD in Cohort 6 (a 50% increase from Cohort 5), and administer it as 6 capsules of 10 mg Compound A each.

[0529] Figure 1 shows the best change (%) from the baseline in the sum of diameters based on RECIST 1.1 (efficacy in a total of 22 evaluable patients). Figure 1 showed the following.

[0530] Overall response rate: 5 / 22 patients (22.7%).

[0531] 4 / 5 responders (80%) had melanoma.

[0532] 7 / 22 patients (31.8%) evaluable for efficacy had melanoma.

[0533] 7 / 7 patients (100%) with melanoma showed stable, partial response, or complete response.

[0534] One patient with melanoma showed a complete response (one CT reading).

[0535] Three patients with melanoma showed partial response.

[0536] Three patients with melanoma showed stable disease.

[0537] 2 / 2 patients (100%) with CRC showed stable disease.

[0538] Materials and Methods

[0539] The target patients were 18 years of age or older, had an ECOG of 0 - 1, and had solid tumors with changes in the MAPK pathway. Dose escalation and cohort size determination were performed using the mTPI - 2 design. The starting dose was 5 mg QD.

[0540] Adverse events (TEAEs) occurring during treatment were graded according to NCI CTCAE v5.0. Tumor response was evaluated by the principal investigator of the trial using RECIST v1.1.

[0541] Results

[0542] Forty - two patients were treated across 6 cohorts (5 - 60 mg QD). The median age was 60 years. Patients had previously received a median of 3 lines of treatment.

[0543] 98% of patients had TEAEs and 76% showed treatment - related (TR) AEs. The most frequently reported TRAEs (≥10%) were skin - related (69%) and fever and diarrhea (both 12%). Thirteen patients had Gr≥3 TRAEs (≥2 patients): decreased platelet count (3 patients), acneiform rash (2 patients), and elevated ALT and AST (2 patients each). Dose reduction occurred in 4 patients due to rhabdomyolysis (2 patients), angioedema (1 patient), and elevated AST (1 patient). Dose interruption due to AEs occurred in 36% of patients. The MTD was determined to be 40 mg QD.

[0544] Serious TEAEs occurred in 71% of patients. 29% were drug - related. DLT occurred in 6 / 42 patients (2 patients at 10 mg, 1 patient at 40 mg, and 3 patients at 60 mg).

[0545] The results of the preliminary PK showed that exposure generally increased proportionally with dose. Tmax occurred at approximately 2 hours. Compound A had a long terminal half - life and a 7.4 - fold mean exposure accumulation at steady state.

[0546] 100% of the patients were evaluable for efficacy. The disease control rate (DCR) was 33% with 1 CR, 5 cPR, 2 uPR, and 8 SD ≥ 24 weeks. Objective responders included patients with BRAF V600E melanoma who had progressed on prior BRAF / MEK and checkpoint inhibitors (2 patients, 1 CR, 1 PR), 1 patient with NRAS G12S melanoma, 1 patient with NRAS Q61K melanoma, 1 patient with BRAF V600E LGSOC, 1 patient with BRAF K601E / PIK3CA endometrial cancer, 1 patient with BRAF V600E cholangiocarcinoma, and 1 patient with KRAS G12D appendiceal cancer.

[0547] Analysis of circulating tumor DNA samples derived from a subset of melanoma patients showed that the maximum decrease in variant allele frequency (VAF) of the detected mutations was 99.3% (range: 98.7 - 99.9%) in the subjects evaluated after treatment with Compound A, which corresponded to clinical efficacy.

[0548] Conclusion

[0549] Compound A has a manageable safety profile and generally dose-proportional PK. Antitumor activity was observed in patients with no approved targeted therapy options. The safety and early efficacy profile of Compound A supported further investigation in selected MAPK-altered tumors.

[0550] Forty-two patients were treated at six dose levels (5–60 mg per day). The patients had received various prior treatments and had, on median, received three prior therapies (range 1–9), including standard-of-care immunotherapy and targeted therapy regimens. The results showed that Compound A had a manageable safety profile and that the adverse event findings were consistent with those of other MAPK pathway inhibitors. The most common treatment-related adverse events (>15%) were acneiform rash (33%), maculopapular rash (24%), and fever (17%). A dose of 40 mg once daily was determined to be the maximum tolerated dose of Compound A. Furthermore, antitumor activity was observed in patients who had received various prior treatments, and the objective response rate was 18% (6 responses were confirmed, including 1 complete response, out of 33 patients in whom efficacy was evaluable). The disease control rate was 79% and the clinical benefit rate was 42%. Objective responders included patients with tumors harboring BRAF V600E that had progressed on prior BRAF / MEK inhibitors, BRAF class II mutations, BRAF fusions, NRAS, and KRAS mutations, regardless of prior checkpoint inhibitor treatment. The median treatment duration was approximately 5 months (range: 1.9–23.6 months), and 9 patients were continuing treatment. These data supported the progression of Compound A to the dose expansion part of the 1b phase of the trial.

[0551] These data supported the investigation of Compound A in a limited cohort that included BRAF V600 tumors that had progressed after prior BRAF and / or MEK inhibitor treatment, solid tumors with BRAF class II mutations and BRAF fusions, and NRAS mutant melanomas. These patients had very limited treatment options.

[0552] Example 2 The general properties of the solid forms of Compound A used are shown in Table 8.

[0553] [Table 9]

[0554] Numerous references are cited, and their disclosures are hereby incorporated by reference in their entirety.

Claims

**Claim 1** A method for treating cancer in a subject in need of cancer treatment, the method comprising administering to the subject a compound A (1-((1S,1aS,6bS)-5-((7-oxo-5,6,7,8-tetrahydro-1,8-naphthyridin-4-yl)oxy)-1a,6b-dihydro-1H-cyclopropa[b]benzofuran-1-yl)-3-(2,4,5-trifluorophenyl)urea) having the following structure, or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, isotopic molecular species, solvate, or prodrug thereof. 【Chemical Formula 1】 **Claim 2** The method of claim 1, wherein the cancer is selected from the group consisting of colorectal cancer, pancreatic cancer, melanoma, non-small cell lung cancer, brain tumor, lung cancer, kidney cancer, bone cancer, liver cancer, bladder cancer, breast cancer, head and neck cancer, ovarian cancer, skin cancer, adrenal cancer, cervical cancer, lymphoma, thyroid tumor, and combinations thereof, preferably melanoma, and non-small cell lung cancer. **Claim 3** The method of claim 2, wherein the cancer is characterized by a mutation in a gene selected from the group consisting of RAS,NRAS,KRAS,RAF, BRAF, CRAF, ARAF, and combinations thereof. **Claim 4** The method of claim 3, wherein the cancer is characterized by a mutation in a gene selected from the group consisting of RAS,NRAS,KRAS,RAF, BRAF, and combinations thereof. **Claim 5** The method of claim 4, wherein the cancer is characterized by a mutation in a gene selected from the group consisting of NRAS,KRAS, BRAF, and combinations thereof. **Claim 6** The cancer is (i) a mutation in a gene selected from the group consisting of ARAF, BRAF, RAF1, KRAS, HRAS, NF1, MAP2K1, MAP2K2, MAPK1, and combinations thereof, ​ (ii) BRAF N20T, BRAF A33T, BRAF S36A, BRAF V47_G393del, BRAF V47_G327del, BRAF V47_D380del, BRAF V47_M438del, BRAF N49I, BRAF M53I, BRAF L64I, BRAF G69S, BRAF A81_D380del, BRAF A81_M438del, BRAF G104E, BRAF T119S, BRAF P141L, BRAF S151A, BRAF P162S, BRAF V169_G327del, BRAF V169_D380del, BRAF R188T, BRAF Q201H, BRAF G203_G393del, BRAF K205Q, BRAF V226L, BRAF E228V, BRAF R239Q, BRAF T241P, BRAF T241M, BRAF L245F, BRAF A246P, BRAF F247L, BRAF Q257R, BRAF Q257H, BRAF G258V, BRAF F259L, BRAF Q262R, BRAF H269Y, BRAF R271H, BRAF E275K, BRAF D287H, BRAF F294L, BRAF T310I, BRAF A320T, BRAF I326V, BRAF P341S, BRAF R347*, BRAF P348T, BRAF S363F, BRAF S364L, BRAF P367S, BRAF P367R, BRAF P367L, BRAF D380H, BRAF R389C, BRAF T401I, BRAF A404Cfs*9, BRAF P407L, BRAF S419Y, BRAF G421V, BRAF R444W, BRAF D448Y, BRAF D449Y, BRAF W450*, BRAF W450L, BRAF E451K, BRAF E451Q, BRAF P453T, BRAF V459L, BRAF R462E, BRAF R462K, BRAF R462I, BRAF I463T, BRAF I463S, BRAF G464I, BRAF G464R, BRAF G464E, BRAF G464A, BRAF G464V, BRAF S465D, BRAF S465E, BRAF S465A, BRAF G466R, BRAF G466E, BRAF G466A, BRAFG466V, BRAF S467A, BRAF S467L, BRAF F468C, BRAF G469L, BRAF G469del, BRAF G469S, BRAF G469R, BRAF G469E, BRAF G469A, BRAF G469V, BRAF T470K, BRAF V471I, BRAF V471F, BRAF Y472dup, BRAF Y472S, BRAF Y472C, BRAF G478C, BRAF K483E, BRAF K483M, BRAF L485_P490del, BRAF L485Y, BRAF L485_P490delinsY, BRAF L485S, BRAF L485W, BRAF L485F, BRAF L485_P490delinsF, BRAF N486_Q494del, BRAF N486del, BRAF N486_T488del, BRAF N486_T491del, BRAF N486_L495del, BRAF N486D, BRAF N486_V487del, BRAF N486_P490del, BRAF N486_A489delinsK, BRAF N486_T491delinsK, BRAF V487_P490del, BRAF V487_P492delinsA, BRAF T488_P492del, BRAF T488_Q493delinsK, BRAF A489_P490del, BRAF P490del, BRAF P490_Q494del, BRAF K499E, BRAF K499N, BRAF E501K, BRAF E501G, BRAF V504_R506dup, BRAF V504I, BRAF L505F, BRAF L505H, BRAF R509G, BRAF R509H, BRAF L514V, BRAF M517I, BRAF Q524L, BRAF L525R, BRAF T529M, BRAF T529N, BRAF T529I, BRAF W531C, BRAF G534D, BRAF Y538H, BRAF R558Q, BRAF G563D, BRAF H568D, BRAF H574N, BRAF H574Y, BRAF H574Q, BRAF N581D, BRAF N581Y, BRAF N581T, BRAF N581S, BRAF N581I, BRAF N581K, BRAFI582M, BRAF F583C, BRAF L584F, BRAF H585Y, BRAF E586K, BRAF D587A, BRAF D587G, BRAF D587E, BRAF V590I, BRAF V590G, BRAF I592V, BRAF I592M, BRAF G593D, BRAF D594N, BRAF D594H, BRAF D594Y, BRAF D594_T599dup, BRAF D594A, BRAF D594G, BRAF D594V, BRAF D594E, BRAF F595L, BRAF F595S, BRAF G596S, BRAF G596R, BRAF G596C, BRAF G596D, BRAF G596V, BRAF L597S, BRAF L597V, BRAF L597Q, BRAF L597P, BRAF L597R, BRAF A598T, BRAF A598S, BRAF A598V, BRAF A598_T599insARC, BRAF A598_T599insV, BRAF T599dup, BRAF T599A, BRAF T599K, BRAF T599R, BRAF T599I, BRAF T599_V600insTT, BRAF T599_V600insS, BRAF T599_V600insETT, BRAF T599_V600insEAT, BRAF V600_K601delinsEN, BRAF V600_S605delinsEISRWR, BRAF V600K, BRAF V600R, BRAF V600Q, BRAF V600dup, BRAF V600delinsYM, BRAF V600M, BRAF V600L, BRAF V600D, BRAF V600_K601delinsE, BRAF V600E, BRAF V600A, BRAF V600G, BRAF K601del, BRAF K601Q, BRAF K601E, BRAF K601_W604del, BRAF K601T, BRAF K601I, BRAF K601_S602delinsNT, BRAF K601N, BRAF S602T, BRAF S602Y, BRAF S602F, BRAF R603*, BRAF W604del, BRAF W604R, BRAF W604G, BRAF S605A, BRAF S605F, BRAFMutations selected from the group consisting of S605E, BRAF S605G, BRAF S605N, BRAF S605I, BRAF G606W, BRAF G606E, BRAF G606A, BRAF G606V, BRAF S607P, BRAF S607F, BRAF H608R, BRAF Q609E, BRAF Q609L, BRAF Q609H, BRAF E611D, BRAF L613F, BRAF G615R, BRAF L618F, BRAF W619R, BRAF S637*, BRAF V639I, BRAF E648Q, BRAF Y656D, BRAF R671Q, BRAF P676S, BRAF L678I, BRAF V681I, BRAF E695K, BRAF K698R, BRAF L711F, BRAF A712T, BRAF R719S, BRAF H725Y, BRAF A728V, BRAF P731T, BRAF P731S, BRAF P731L, BRAF A762E, BRAF A762V, and combinations thereof. (iii) A mutation selected from the group consisting of KIAA1549-BRAF fusion, BCAS1-BRAF fusion, CCDC6-BRAF fusion, CDC42BPB-BRAF fusion, FAM131B-BRAF fusion, FXR1-BRAF fusion, GIT2-BRAF fusion, KLHL7-BRAF fusion, RNF130-BRAF fusion, TMEM106B-BRAF fusion, MKRn1-BRAF fusion, AGAP3-BRAF fusion, AGK-BRAF fusion, AKAP9-BRAF fusion, ARMC10-BRAF fusion, CUL1-BRAF fusion, GTF2I-BRAF fusion, PAPSS1-BRAF fusion, PCBP2-BRAF fusion, PPFIBP2-BRAF fusion, SND1-BRAF fusion, TRIM24-BRAF fusion, ZKSCAN1-BRAF fusion, SEPT3-BRAF fusion, and combinations thereof, or (iv) The method according to claim 2, characterized by a mutation selected from the group consisting of NRAS G12A, NRAS G12C, NRAS G12D, NRAS G12N, NRAS G12P, NRAS G12R, NRAS G12S, NRAS G12V, NRAS G12Y, NRAS G13A, NRAS G13C, NRAS G13D, NRAS G13E, NRAS G13N, NRAS G13R, NRAS G13S, NRAS G13V, NRAS A18T, NRAS I24N, NRAS P34L, NRAS Y40*, NRAS Q43*, NRAS T50I, NRAS T58I, NRAS A59G, NRAS A59D, NRAS A59T, NRAS G60E, NRAS G60R, NRAS Q61E, NRAS Q61H, NRAS Q61H, NRAS Q61K, NRAS Q61L, NRAS Q61L, NRAS Q61P, NRAS Q61R, NRAS Q61R, NRAS Q61R, NRAS Q61*, NRAS E63K, NRAS Y64D, NRAS S65C, NRAS R68S, NRAS S89A, NRAS G115Efs*46, NRAS E132K, NRAS K135N, NRAS A146P, NRAS A146T, NRAS A146V, NRAS E162*, and combinations thereof.

7. The method according to claim 2, wherein the cancer is characterized by a mutation selected from the group consisting of NRAS Q61R, NRAS Q61K, NRAS Q61L, NRAS G12S, NRAS G13R, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12V, BRAF V600E, BRAF fusion, and combinations thereof.

8. The method according to claim 7, wherein the cancer is characterized by a mutation selected from the group consisting of NRAS Q61R, NRAS Q61K, NRAS Q61L, KRAS G12D, KRAS G12V, BRAF V600E, BRAF fusion, and combinations thereof.

9. The method according to claim 8, wherein the cancer is characterized by a mutation selected from the group consisting of NRAS Q61R, NRAS Q61K, NRAS Q61L, KRAS G12D, KRAS G12V, and combinations thereof.

10. The method according to claim 2, wherein the cancer is characterized by genomic abnormalities in other MAPK pathways.

11. The method according to claim 2, wherein the cancer is melanoma.

12. The method according to claim 11, wherein the melanoma is cutaneous melanoma.

13. The method according to claim 11, wherein the melanoma is metastatic melanoma.

14. The method according to claim 2, wherein the cancer is non-small cell lung cancer.

15. The method according to claim 2, wherein the cancer is colorectal cancer.

16. The method according to claim 2, wherein the cancer is ovarian cancer.

17. Compound A or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, isotopic molecular species, solvate, or prodrug thereof is administered 1 to 3 times a day in the method according to claim 2.

18. Compound A or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, isotopic molecular species, solvate, or prodrug thereof is administered once a day in the method according to claim 2.

19. Compound A or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, isotopic molecular species, solvate, or prodrug thereof is administered twice a day in the method according to claim 2.

20. The method according to claim 2, wherein compound A or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, isotopic molecular species, solvate, or prodrug thereof is administered three times a day.

21. The method according to claim 2, wherein compound A or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, isotopic molecular species, solvate, or prodrug thereof is administered at about 5 mg to about 600 mg per day.

22. The method according to claim 21, wherein compound A or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, isotopic molecular species, solvate, or prodrug thereof is administered at about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 110 mg, about 115 mg, or about 120 mg per day.

23. The method according to claim 22, wherein compound A or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, isotopic molecular species, solvate, or prodrug thereof is administered at about 40 mg or about 60 mg per day.

24. The method according to claim 23, wherein compound A or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, isotopic molecular species, solvate, or prodrug thereof is administered at about 40 mg per day.

25. The method according to claim 23, wherein compound A or a pharmaceutically acceptable salt, tautomer, stereoisomer, enantiomer, isotopic molecular species, solvate, or prodrug thereof is administered at about 60 mg per day.

26. The method provides a plasma compound A AUC of about 2,128 ng*h / ml to about 3,192 ng*h / ml in the subject ８ｈ The method according to claim 2

27. The method provides, in the subject, a plasma compound A AUC of about 4,576 ng*h / ml to about 6,864 ng*h / ml ８ｈ The method according to claim 2, which provides the same.

28. The method provides a plasma compound A AUC of about 7,944 ng*h / ml to about 11,916 ng*h / ml in the subject ８ｈ The method according to claim 2, which provides the same.<

29. The method provides a plasma compound A AUC of about 9,840 ng*h / ml to about 14,760 ng*h / ml in the subject ８ｈ The method according to claim 2

30. The method provides a plasma compound A AUC of about 12,640 ng*h / ml to about 18,960 ng*h / ml in the subject ８ｈ The method according to claim 2, which provides such.

31. The method provides, in the subject, a plasma compound A AUC of from about 30,000 ng*h / ml to about 45,000 ng*h / ml ８ｈ The method according to claim 2, which provides the same.<

32. The method according to claim 2, wherein the subject achieves stability, partial efficacy, or complete efficacy.

33. The method according to claim 2, wherein the subject achieves partial efficacy or complete efficacy.

34. The method according to claim 2, wherein the subject achieves complete efficacy.

35. The method according to claim 2, wherein the subject does not experience progression.

36. The method according to claim 2, wherein the subject achieves stability.

37. The method according to claim 2, wherein the subject achieves partial efficacy.

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