3,4-dihydro-2,7-naphthyridine-1,6(2h,7h)-dione as MEK inhibitor

Abstract

To provide a pharmaceutical composition, effective for treating tumors mediated by MEK, including therapies that can target tumors in the CNS by penetrating the blood-brain barrier (BBB) and / or blood-CSF barrier (BCSFB).SOLUTION: The present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising such a compound and salt. The present invention further provides a solid form of 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to novel 3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione compounds or pharmaceutically acceptable salts thereof that act as MEK inhibitors and are useful in treating abnormal cell growth, e.g., cancer, in patients. The present invention also relates to pharmaceutical compositions containing the compounds and methods of using the compounds and compositions in treating abnormal cell growth, e.g., cancer, in subjects in need thereof. The present invention also relates to solid forms of 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, pharmaceutical compositions containing the solid forms, and methods of using the solid forms and compositions in treating abnormal cell growth, e.g., cancer, in subjects in need thereof. [Background technology]

[0002] MEK kinase (mitogen-activated protein kinase kinase (MAPKK)) is a key component of the Ras-RAF-MEK-ERK cell survival pathway. The Ras pathway is activated by the binding of growth factors, cytokines, and hormones to their cognate receptors. However, this pathway is constitutively activated in cancer cells, leading to increased cancer cell survival, cell proliferation, angiogenesis, and metastasis. Tumors that exhibit constitutive activation of the pathway include, but are not limited to, tumors of the colon, pancreas, breast, brain, ovary, lung, and skin. Activation of Ras (due to upstream signaling or as a result of activating point mutations in the Ras oncogene) leads to the phosphorylation and activation of Raf kinase, which in turn phosphorylates and activates MEK1 and MEK2 (also known as MAPKK1 and MAPKK2). MEK1 and MEK2 are dual-specificity kinases that phosphorylate and activate ERK1 / 2 kinase (also known as MAP kinase), activating ERK1 and ERK2, which further phosphorylates and regulates the function of proteins such as Mcl-1, Bim, and Bad, which are involved in cell survival and apoptosis. This phosphorylation-mediated cascade leads to enhanced cell proliferation, cell survival, and reduced cell death, which are necessary for the initiation and maintenance of tumorigenic phenotypes. Inhibition of this pathway, particularly MEK activity, is known to be beneficial in treating hyperproliferative diseases. MEK inhibitors have shown variable activity in several settings, including BRAF V600-mutant melanoma, NRAS-mutant melanoma, low-grade serous ovarian cancer, plexiform neurofibroma, thyroid cancer, and low-grade glioma, but responses in KRAS-mutant pancreatic or lung cancer have been more limited.

[0003] Cancers that frequently metastasize to the brain, such as melanoma and non-small cell lung cancer, are known to have MAPK pathway-activating alterations, such as BRAF V600E and KRAS G12 mutations (Cancer Genome Atlas N., Cell 2015;161:1681-96). Activating mutations can occur at various levels in the classical pathway, but all require signaling through mitogen / extracellular signal-regulated kinase (MEK) to enhance proliferation and survival (Schubbert S, Shannon K, Bollag G., Nat Rev Cancer. 2007;7:295-308). Given the general activation of the MAPK pathway and central and downstream positions of MEK in malignant tumors, MEK inhibitors are potentially interesting for the treatment of intracranial tumors.

[0004] The blood-brain interface includes cerebral microvascular endothelium, which forms the blood-brain barrier (BBB), and choroid plexus epithelium, which forms the blood-CSF barrier (BCSFB). The blood-brain barrier (BBB) ​​is a highly selective physical transport and metabolic barrier that separates the CNS from the blood. The BBB can prevent certain drugs from entering brain tissue and therefore limits the delivery of many peripherally administered drugs to the CNS. The efficacy of many molecularly targeted drugs in central nervous system tumors is limited by penetration of the blood-brain barrier (BBB), which is composed of a single layer of endothelial cells connected by tight junctions, which act as a physical barrier protecting the brain. In addition, these endothelial cells express multidrug efflux transporters, including P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP), which are known to exclude many anticancer drugs from the brain (Ohtsuki and Terasaki, 2007, Pharm Res 24:1745-1758; Agarwal et al., 2011, Pharm Res 24:1745-1758). Similar to the blood-brain barrier, the blood-CSF barrier functions to prevent the passage of most blood-borne substances into the brain, while selectively allowing the passage of certain substances into the brain and facilitating the removal of brain metabolites and metabolic products into the blood. Summary of the Invention [Problem to be solved by the invention]

[0005] Thus, there remains a need for therapies for the treatment of MEK-mediated tumors, including therapies that can penetrate the BBB and / or BCSFB and target tumors in the CNS. [Means for solving the problem]

[0006] Provided herein, in part, are compounds of Formula I and Formula II, as well as pharmaceutically acceptable salts thereof. Such compounds can inhibit the activity of MEK, thereby effecting biological function, and can be useful in treating subjects with MEK-associated tumors. Also provided herein are solid forms of 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione. Also provided herein are pharmaceutical compositions and medicaments comprising compounds according to any of the formulas described herein, and pharmaceutically acceptable salts thereof, which can be useful alone or in combination with additional anti-cancer therapies in treating subjects with MEK-associated tumors. Also provided herein are methods for preparing compounds according to any of the formulas described herein, and pharmaceutically acceptable salts thereof, and pharmaceutical compositions, as well as methods of using the foregoing. This Summary is provided to introduce selected concepts in a simplified form that are further described in the Detailed Description below. This Summary is not intended to identify critical features or essential features of the claimed subject matter, nor is it intended to be used in isolation as an aid in determining the scope of the claimed subject matter.

[0007] In accordance with an embodiment of the present invention, provided herein is a compound of formula I

[0008] [ka] or a pharmaceutically acceptable salt thereof, wherein: R 1 is H, Br, C1-C6 alkyl or phenyl; R 2 is H, halogen or CH3-, R 3 is H, hydroxyC1-C6 alkyl-, hydroxyC1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or (C3-C6 cycloalkyl)C1-C6 alkoxy-; R 4 is phenyl substituted with 1, 2, or 3 substituents independently selected from halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(=O)-.

[0009] Also provided herein is a compound of formula II

[0010] [ka] or a pharmaceutically acceptable salt thereof, wherein: R 1 is H, Br, C1-C6 alkyl or phenyl; R 2 is H, halogen or CH3-, R 3 is H, hydroxyC1-C6 alkyl-, hydroxyC1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or (C3-C6 cycloalkyl)C1-C6 alkoxy-; R a and R bare independently selected from halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(=O)-.

[0011] In one embodiment, provided herein is a solid form of 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.

[0012] In one embodiment, provided herein is a pharmaceutical composition comprising a compound according to any of the formulas described herein or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. In some embodiments, the pharmaceutical composition comprises two or more pharmaceutically acceptable carriers and / or excipients.

[0013] In one embodiment, provided herein are methods of treatment and uses comprising administering to a subject a compound according to any of the formulas described herein, or a pharmaceutically acceptable salt thereof.

[0014] In one embodiment, provided herein is a method for treating abnormal cell growth, e.g., a tumor, e.g., a MEK-associated tumor, in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound according to any of the formulas described herein, or a pharmaceutically acceptable salt thereof. The compound according to any of the formulas described herein can be administered as a single agent or in combination with one or more anti-cancer therapies.

[0015] In one embodiment, provided herein is a method for treating abnormal cell growth, e.g., a tumor, e.g., a MEK-associated tumor, in a subject in need thereof, comprising administering to the subject an amount of a compound according to any of the formulas described herein or a pharmaceutically acceptable salt thereof, in combination with an amount of an additional anti-cancer agent, which amounts together are effective to treat said abnormal cell growth.

[0016] In one embodiment, provided herein is a compound according to any of the formulas described herein, or a pharmaceutically acceptable salt thereof, for use as a pharmaceutical.

[0017] In one embodiment, provided herein is a compound according to any of the formulas described herein, or a pharmaceutically acceptable salt thereof, for use in treating abnormal cell growth, e.g., a tumor, e.g., a MEK-associated tumor.

[0018] In one embodiment, provided herein is the use of a compound according to any of the formulas described herein, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of abnormal cell growth, e.g., a tumor, e.g., a MEK-associated tumor, in a subject.

[0019] In one embodiment, provided herein is a pharmaceutical composition comprising a compound according to any of the formulas described herein, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient.

[0020] Each embodiment of a compound according to any of the formulas described herein may be combined with one or more other embodiments of a compound according to any of the formulas described herein that are not inconsistent with the embodiment(s) with which it is combined.

[0021] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed. [Brief explanation of the drawings]

[0022] [Figure 1] 1 shows the powder X-ray diffraction pattern of crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 1. [Figure 2] 1 shows the powder X-ray diffraction pattern of crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 2. [Figure 3] 1 shows the powder X-ray diffraction pattern of crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3. [Figure 4] 1 shows the powder X-ray diffraction pattern of amorphous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 4. [Figure 5] 1 shows the sorption isotherm of crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3. DETAILED DESCRIPTION OF THE INVENTION

[0023] In one aspect, the present invention provides a compound of formula I

[0024] [ka] or a pharmaceutically acceptable salt thereof, R 1is H, Br, C1-C6 alkyl or phenyl; R 2 is H, halogen or CH3-, R 3 is H, hydroxyC1-C6 alkyl-, hydroxyC1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or (C3-C6 cycloalkyl)C1-C6 alkoxy-; R 4 is phenyl substituted with 1, 2, or 3 substituents independently selected from halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(=O)-.

[0025] As used herein, the singular forms "a," "an," and "the," when referring to substituents, include plural referents unless otherwise specified. For example, "a substituent" includes one or more substituents.

[0026] For complex chemical names used herein, a substituent is typically named before the group to which it is attached, e.g., methoxyethyl contains an ethyl skeleton bearing a methoxy substituent.

[0027] The term "halogen" refers to --F (sometimes referred to herein as "fluoro" or "fluoros"), --Cl, --Br, and --I.

[0028] The term "C1-C6 alkyl," as used herein, refers to a saturated, straight- or branched-chain monovalent hydrocarbon radical of one to six carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, 1-propyl, isopropyl, 1-butyl, isobutyl, sec-butyl, tert-butyl, 2-methyl-2-propyl, pentyl, neopentyl, and hexyl.

[0029] The term "hydroxyC1-C6 alkyl-", as used herein, refers to a C1-C6 alkyl radical, as defined herein, in which one of the hydrogen atoms has been replaced with a hydroxy group.

[0030] The term "hydroxy" refers to the group --OH.

[0031] The term "C3-C6 cycloalkyl" means a fully saturated carbocyclic ring having from 3 to 6 ring carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0032] The term "fluoroC1-C6 alkyl," as used herein, refers to a C1-C6 alkyl radical, as defined herein, in which one, two, or three hydrogen atoms are replaced with one, two, or three fluoro atoms, respectively. Examples include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 2,2-difluoroethyl, and 2,2,2-trifluoroethyl.

[0033] The term "C1-C6 alkoxy," as used herein, refers to a C1-C6 alkyl radical, as defined herein, that is single-bonded to an oxygen atom, where the radical is on the oxygen atom (i.e., C1-C6-O-). Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, and isopropoxy.

[0034] The term "fluoro C1-C6 alkoxy," as used herein, refers to C1-C6 alkoxy, as defined herein, in which one, two, or three hydrogen atoms are replaced with one, two, or three fluoro atoms, respectively. Examples include, but are not limited to, trifluoromethoxy.

[0035] The term "(C3-C6 cycloalkyl)C1-C6 alkoxy-" refers to a C1-C6 alkoxy- as defined herein, in which one of the hydrogen atoms is replaced with a C3-C6 cycloalkyl group as defined herein.

[0036] The term "C1-C6 alkylthio," as used herein, refers to a (C1-C6 alkyl)S- radical, wherein the C1-C6 alkyl moiety is as defined herein.

[0037] The term "fluoro C1-C6 alkylthio," as used herein, refers to a C1-C6 alkylthio group, as defined herein, in which one, two, or three hydrogen atoms are replaced by one, two, or three fluoro atoms, respectively.

[0038] In one embodiment of Formula I, R 1 is H.

[0039] In one embodiment of Formula I, R 1 is Br.

[0040] In one embodiment of Formula I, R 1 is C1-C6 alkyl. In one embodiment of Formula I, R 1 is methyl.

[0041] In one embodiment of Formula I, R 1 is phenyl.

[0042] In one embodiment of Formula I, R 2 is H.

[0043] In one embodiment, R 2 is a halogen.

[0044] In one embodiment of Formula I, R 2 is F.

[0045] In one embodiment of Formula I, R 2 is Cl.

[0046] In one embodiment of Formula I, R 2 is Br.

[0047] In one embodiment of Formula I, R 2 is I.

[0048] In one embodiment of Formula I, R 2 is CH3-.

[0049] In one embodiment of Formula I, R 1 is H and R 2 is H.

[0050] In one embodiment of Formula I, R 3 is H.

[0051] In one embodiment of Formula I, R 3 is hydroxyC1-C6 alkyl-. Non-limiting examples include 2-hydroxyethyl.

[0052] In one embodiment of Formula I, R 3 are hydroxyC1-C6 alkoxy-. Non-limiting examples include the structures

[0053] [ka] Examples include 2-hydroxyethoxy and 2-hydroxypropoxy having the formula:

[0054] In one embodiment of Formula I, R 3is C1-C6 alkoxy. Non-limiting examples include methoxy, ethoxy, 1-methylethoxy, and 2,2-dimethylethoxy.

[0055] In one embodiment of Formula I, R 3 is fluoroC1-C6 alkoxy. Non-limiting examples include 2,2-difluoroethoxy.

[0056] In one embodiment of Formula I, R 3 is C3-C6 cycloalkyl. A non-limiting example is cyclopropyl.

[0057] In one embodiment of Formula I, R 3 is (C3-C6 cycloalkyl)C1-C6 alkoxy-. A non-limiting example is cyclopropylmethoxy.

[0058] In one embodiment of Formula I, R 4 is phenyl substituted with 1, 2, or 3 substituents independently selected from fluoro, chloro, bromo, iodo, ethyl, propyl, isopropyl, methylthio, difluoromethylthio, trifluoromethyl, methoxy, difluoromethoxy, cyclopropyl, and C1-C6 alkyl-C(=O)-.

[0059] In one embodiment of Formula I, R 4 is phenyl substituted with 1 or 2 substituents independently selected from halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(=O)-.

[0060] In one embodiment of Formula I, R 4is phenyl substituted with 1 or 2 substituents independently selected from fluoro, chloro, bromo, iodo, ethyl, propyl, isopropyl, methylthio, difluoromethylthio, trifluoromethyl, methoxy, difluoromethoxy, cyclopropyl, and C1-C6 alkyl-C(=O)-.

[0061] In one embodiment of Formula I, R 4 is phenyl substituted with one substituent selected from halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(=O)-.

[0062] In one embodiment of Formula I, R 4 is phenyl substituted with one substituent selected from fluoro, chloro, bromo, iodo, ethyl, propyl, isopropyl, methylthio, difluoromethylthio, trifluoromethyl, methoxy, difluoromethoxy, cyclopropyl, and C1-C6 alkyl-C(=O)-.

[0063] In one embodiment of Formula I, R 4 is the structure

[0064] [ka] is selected from.

[0065] In one embodiment of Formula I, R 4 teeth,

[0066] [ka] is.

[0067] In one embodiment of Formula I, R 4 teeth,

[0068] [ka] is.

[0069] In one embodiment of Formula I, R 4 is the structure

[0070] [ka] is selected from.

[0071] In one embodiment of Formula I, R 4 is the structure

[0072] [ka] is selected from.

[0073] In one embodiment, R 4 is the structure

[0074] [ka] wherein R a and R b is independently selected from halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(=O)-. a is halogen. In one embodiment, R b is halogen, C1-C6 alkyl, C1-C6 alkylthio, or fluoroC1-C6 alkoxy. a is a halogen and R b is halogen, C1-C6 alkyl, C1-C6 alkylthio, or fluoroC1-C6 alkoxy.

[0075] In one embodiment, provided herein is a compound of formula II

[0076] [ka] or a pharmaceutically acceptable salt thereof, wherein: R 1 is H, Br, C1-C6 alkyl or phenyl; R 2 is H, halogen or CH3-, R 3 is H, hydroxyC1-C6 alkyl-, hydroxyC1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or (C3-C6 cycloalkyl)C1-C6 alkoxy-; R a and R b are independently selected from halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(=O)-.

[0077] In one embodiment of Formula II, R 1 is H.

[0078] In one embodiment of Formula II, R 1 is Br.

[0079] In one embodiment of Formula II, R 1 is C1-C6 alkyl. In one embodiment of Formula II, R 1 is methyl.

[0080] In one embodiment of Formula II, R 1 is phenyl.

[0081] In one embodiment of Formula II, R 2 is H.

[0082] In one embodiment of Formula II, R2 is a halogen.

[0083] In one embodiment of Formula II, R 2 is F.

[0084] In one embodiment of Formula II, R 2 is Cl.

[0085] In one embodiment of Formula II, R 2 is Br.

[0086] In one embodiment of Formula II, R 2 is I.

[0087] In one embodiment of Formula II, R 2 is CH3-.

[0088] In one embodiment of Formula II, R 2 is H or CH3-.

[0089] In one embodiment of Formula II, R 1 is H and R 2 is H or CH3-.

[0090] In one embodiment of Formula II, R 1 is H and R 2 is H.

[0091] In one embodiment of Formula II, R 3 is H.

[0092] In one embodiment of Formula II, R 3 is hydroxyC1-C6 alkyl-. Non-limiting examples include 2-hydroxyethyl.

[0093] In one embodiment of Formula II, R 3 are hydroxyC1-C6 alkoxy-. Non-limiting examples include the structures

[0094] [ka] Examples include 2-hydroxyethoxy and 2-hydroxypropoxy having the formula:

[0095] In one embodiment of Formula II, R 3 is C1-C6 alkoxy. Non-limiting examples include methoxy, ethoxy, 1-methylethoxy, and 2,2-dimethylethoxy.

[0096] In one embodiment of Formula II, R 3 is fluoroC1-C6 alkoxy. Non-limiting examples include 2,2-difluoroethoxy.

[0097] In one embodiment of Formula II, R 3 is C3-C6 cycloalkyl. A non-limiting example is cyclopropyl.

[0098] In one embodiment of Formula II, R 3 is (C3-C6 cycloalkyl)C1-C6 alkoxy-. A non-limiting example is cyclopropylmethoxy.

[0099] In one embodiment of Formula II, R 3 is H or hydroxyC1-C6 alkoxy-.

[0100] In one embodiment of Formula II, R a is halogen. In one embodiment of Formula II, R a is fluoro or chloro. In one embodiment of Formula II, R a is fluoro.

[0101] In one embodiment of Formula II, R b is fluoro, chloro, bromo, iodo, ethyl, propyl, isopropyl, methylthio, difluoromethylthio, trifluoromethyl, methoxy, difluoromethoxy, cyclopropyl, or CHC(=O)-.

[0102] In one embodiment of Formula II, R b is halogen, C1-C6 alkyl, C1-C6 alkylthio, or fluoroC1-C6 alkoxy.

[0103] In one embodiment of Formula II, R b is bromo, iodo, ethyl, methylthio, or difluoromethoxy. In one embodiment of Formula II, R b is methylthio.

[0104] In one embodiment of Formula II, R a is fluoro and R b is methylthio.

[0105] In one embodiment of Formula II, R a is a halogen and R b is halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or C1-C6 alkyl-C(=O)-.

[0106] In one embodiment of Formula II, R a is a halogen and R b is halogen, C1-C6 alkyl, C1-C6 alkylthio, or fluoroC1-C6 alkoxy.

[0107] In one embodiment of Formula II, R a is a halogen and R b is halogen. In one embodiment of Formula II, R a is a halogen and R b is a halogen and R 1 is H and R 2 is H.

[0108] In one embodiment of Formula II, R a is a halogen and Rb is C1-C6 alkyl. In one embodiment of Formula II, R a is a halogen and R b is a C1-C6 alkyl, and R 1 is H and R 2 is H.

[0109] In one embodiment of Formula II, R a is a halogen and R b is C1-C6 alkylthio. In one embodiment of Formula II, R a is a halogen and R b is C1-C6 alkylthio, and R 1 is H and R 2 is H.

[0110] In one embodiment of Formula II, R a is a halogen and R b is fluoroC1-C6 alkylthio. In one embodiment of Formula II, R a is a halogen and R b is fluoro C1-C6 alkylthio, and R 1 is H and R 2 is H.

[0111] In one embodiment of Formula II, R a is fluoro and R b is methylthio and R 1 is H and R 2 is H.

[0112] In one embodiment of Formula II, R a is a halogen and R b is fluoroC1-C6 alkyl. In one embodiment of Formula II, R a is a halogen and R b is fluoro C1-C6 alkyl, and R 1 is H and R 2 is H.

[0113] In one embodiment of Formula II, R a is a halogen and R b is C1-C6 alkoxy. In one embodiment of Formula II, R a is a halogen and R b is C1-C6 alkoxy, and R 1 is H and R 2 is H.

[0114] In one embodiment of Formula II, R a is a halogen and R b is fluoroC1-C6 alkoxy. In one embodiment of Formula II, R a is a halogen and R b is fluoro C1-C6 alkoxy, and R 1 is H and R 2 is H.

[0115] In one embodiment of Formula II, R a is a halogen and R b is C3-C6 cycloalkyl. In one embodiment of Formula II, R a is a halogen and R b is a C3-C6 cycloalkyl, and R 1 is H and R 2 is H.

[0116] In one embodiment of Formula II, R a is a halogen and R b is C1-C6 alkyl-C(=O)-. In one embodiment of Formula II, R a is a halogen and R b is C1-C6 alkyl-C(=O)-, and R 1 is H and R 2 is H.

[0117] In one embodiment of Formula II, R 1 is H and R 2 is H or CH3-, and R 3 is H or hydroxyC1-C6 alkoxy-, and Ra is a halogen and R b is halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or C1-C6 alkyl-C(=O)-.

[0118] In one embodiment of Formula II, R 1 is H and R 2 is H or CH3-, and R 3 is H or hydroxyC1-C6 alkoxy-, and R a is a halogen and R b is halogen, C1-C6 alkyl, C1-C6 alkylthio, or fluoroC1-C6 alkoxy.

[0119] In one embodiment of any of the above embodiments of Formula II, the group

[0120] [ka] is the structure

[0121] [ka] is selected from.

[0122] The term "compound," as used herein, is meant to include all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted. Compounds identified herein by name or structure as one particular tautomeric form are intended to include other tautomeric forms unless otherwise specified.

[0123] Compounds of the formulas provided herein may have asymmetric carbon atoms. Carbon-carbon bonds of the compounds of the invention are represented herein by solid lines (

[0124] [ka] ), solid wedge (

[0125] [ka] ), or a dotted wedge (

[0126] [ka] ). The use of a solid line to depict bonds to an asymmetric carbon atom is meant to indicate that all possible stereoisomers at that carbon atom (e.g., a specific enantiomer, a racemic mixture, etc.) are included. The use of either a solid or dotted wedge to depict bonds to an asymmetric carbon atom is meant to indicate that only the stereoisomer shown is meant to be included. It is possible that the compounds of the present invention may contain more than one asymmetric carbon atom. In those compounds, the use of a solid line to depict bonds to an asymmetric carbon atom indicates that all possible stereoisomers are meant to be included and is meant to indicate the attached stereocenter. For example, unless otherwise stated, it is intended that the compounds of the present invention can exist as enantiomers and diastereomers, or as racemates and mixtures thereof. The use of a solid line to depict bonds to one or more asymmetric carbon atoms in a compound of the present invention, and the use of a solid or dotted wedge to depict bonds to other asymmetric carbon atoms in the same compound, is meant to indicate that a diastereomeric mixture is present.

[0127] Compounds of the present invention that contain chiral centers can exist as stereoisomers, eg, racemates, enantiomers, or diastereomers.

[0128] Stereoisomers of the compounds of the formulae herein may include cis and trans isomers, optical isomers, e.g., (R) and (S) enantiomers, diastereomers, geometric isomers, rotamers, atropisomers, stereoisomers, and tautomers of the compounds of the invention, including compounds exhibiting more than one type of isomerism, and mixtures thereof (e.g., racemates and diastereomeric pairs).

[0129] Also included are acid or base addition salts wherein the counterion is optically active, for example, d-lactate or l-lysine, or racemic, for example, dl-tartrate or dl-arginine.

[0130] When any racemate crystallizes, two different types of crystals are possible. The first type is the racemate (true racemate) described above, in which one homogeneous crystalline form occurs containing both enantiomers in equimolar amounts. The second type is a racemic mixture or conglomerate, in which two crystalline forms, each containing a single enantiomer, occur in equimolar amounts.

[0131] Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from suitable optically pure precursors, or resolution of the racemate (or racemate of a salt or derivative) using, for example, chiral high-pressure liquid chromatography (HPLC) or supercritical fluid chromatography (SFC).

[0132] Alternatively, the racemate (or racemic precursor) can be reacted with a suitable optically active compound, for example, an alcohol, or, if the compound contains an acidic or basic moiety, with an acid or base such as tartaric acid or 1-phenylethylamine. The resulting diastereomeric mixture can be separated by chromatography and / or fractional crystallization, and one or both of the diastereoisomers can be converted into the corresponding pure enantiomer(s) by means well known to those skilled in the art.

[0133] The chiral compounds of the invention (and their chiral precursors) can be obtained in enantiomerically enriched form using chromatography, typically HPLC, on an asymmetric resin with a mobile phase consisting of a hydrocarbon, typically 0-50% isopropanol, typically 2-20% isopropanol, and heptane or hexane containing 0-5% alkylamine, typically 0.1% diethylamine. The eluent is concentrated to yield the enriched mixture.

[0134] Stereoisomeric conglomerates can be separated by conventional techniques known to those skilled in the art, see, for example, "Stereochemistry of Organic Compounds" by E.L. Eliel (Wiley, New York, 1994), the entire disclosure of which is incorporated herein by reference.

[0135] The enantiomeric purity of the compounds described herein can be described in units of enantiomeric excess (ee), which indicates the degree to which a sample contains one enantiomer in greater amount than the other. A racemic mixture has 0% ee, while a single, completely pure enantiomer has 100% ee. Similarly, diastereomeric purity can be described in units of diastereomeric excess (de).

[0136] The compounds of the present invention may exhibit the phenomena of tautomerism and structural isomerism. For example, the compounds may exist in several tautomeric forms, including enol and imine forms, and keto and enamine forms, as well as geometric isomers and mixtures thereof. All such tautomeric forms are included within the scope of the compounds of the present invention. Tautomers exist as a mixture of tautomeric sets in solution. In solid form, one tautomer usually predominates. Although one tautomer may be described, the present invention includes all tautomers of the compounds of the formula provided. Tautomers of the compounds of formula I may be, for example, R 3 If is hydrogen,

[0137] [ka] may occur.

[0138] In addition, some of the compounds of the present invention can form atropisomers (e.g., substituted biaryls). Atropisomers are conformational stereoisomers that arise as a result of steric interactions with other parts of a molecule when rotation around a single bond in the molecule is hindered or significantly slowed, resulting in asymmetric substituents at both ends of the single bond. The interconversion of atropisomers is slow enough to allow separation and isolation under certain conditions. The energy barrier to thermal racemization can be determined by the steric hindrance to the free rotation of one or more bonds that form the chiral axis.

[0139] The present invention also includes pharmaceutically acceptable isotopically labeled compounds identical to those recited in one of the formulas provided, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature.

[0140] Isotopically labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by methods analogous to those described herein, using an appropriate isotopically labeled reagent in place of an otherwise non-labeled reagent.

[0141] Examples of isotopes that may be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, including, but not limited to, 2 H, 3 H, 13 C. 14 C. 15 N, 18 O. 17 O. 32 P, 35 S, 18 F and 36 Certain isotopically labeled compounds of the present invention, such as 2 H, 3H or 14 Those incorporating a radioactive isotope, such as 13C, are useful in drug and / or substrate tissue distribution assays. 3 H, and carbon-14, i.e. 14 C isotopes are particularly preferred due to their ease of preparation and detectability. Additionally, deuterium, i.e. 2 Substitution with heavier isotopes, such as H, can offer certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and therefore may be preferable in some circumstances. 11 C. 18 F, 15 O and 13 Substitution with positron emitting isotopes, such as N, can be useful in positron emission tomography (PET) studies for examining substrate receptor occupancy. Isotopically labeled compounds of the present invention can generally be prepared by following the procedures disclosed in the following schemes and / or examples and preparations by substituting isotopically labeled reagents for non-isotopically labeled reagents.

[0142] Pharmaceutically acceptable solvates in accordance with the invention include those in which the solvent of crystallization may be isotopically substituted, e.g., D2O, d 6 - acetone, d 6 -Contains DMSO.

[0143] Unless otherwise specified, all references herein to compounds of the invention include references to salts, solvates, hydrates and complexes thereof, and to solvates, hydrates and complexes of salts thereof, including polymorphs, stereoisomers and isotopically labeled forms thereof.

[0144] The compounds of the present invention can exist in the form of pharmaceutically acceptable salts, such as, for example, acid addition salts and base addition salts of the compounds of one of the formulas provided herein.As used herein, the term "pharmaceutically acceptable salt" refers to a salt that retains the biological effect and properties of the parent compound.As used herein, the phrase "pharmaceutically acceptable salt(s)" includes, unless otherwise specified, the salts of acidic or basic groups that may be present in the compounds of the formulas disclosed herein.

[0145] For example, the compounds of the present invention, which are basic in nature, can form a wide variety of salts with various inorganic and organic acids. Although such salts must be pharmaceutically acceptable for administration to animals, it is often practical to first isolate the compounds of the present invention from the reaction mixture as pharmaceutically unacceptable salts, then simply convert the latter back to the free base compound by treatment with an alkaline reagent, and then convert the latter free base to a pharmaceutically acceptable acid addition salt. Acid addition salts of the base compounds of the present invention can be prepared by treating the base compound with a substantially equivalent amount of a selected inorganic or organic acid in an aqueous solvent medium or in a suitable organic solvent such as methanol or ethanol. Evaporation of the solvent yields the desired solid salt. The desired acid salt can also be precipitated from a solution of the free base in an organic solvent by adding an appropriate inorganic or organic acid to the solution.

[0146] Acids can be used to prepare pharmaceutically acceptable acid addition salts of basic compounds, including those that form non-toxic acid addition salts, i.e., salts containing a pharmacologically acceptable anion, such as hydrochloride, hydrobromide, hydroiodide, nitrate, sulfate, bisulfate, phosphate, superphosphate, isonicotinate, acetate, lactate, salicylate, citrate, acid citrate, tartrate, pantothenate, bitartrate, ascorbate, succinate, maleate, gentisate, fumarate, gluconate, glucuronate, saccharate, formate, benzoate, glutamate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate salts.

[0147] Examples of salts include acetate, acrylate, benzenesulfonate, benzoate (e.g., chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, and methoxybenzoate), bicarbonate, bisulfate, bisulfite, bitartrate, borate, bromide, butyne-1,4-dioate, calcium edetate, camsylate, carbonate, chloride, caproate, caprylate, clavulanate, and citrate. , decanoate, dihydrochloride, dihydrogen phosphate, edetate, edislyate, estolate, esylate, ethylsuccinate, formate, fumarate, gluceptate, gluconate, glutamate, glycolate, glycolylarsanilate, heptanoate, hexyne-1,6-dioate, hexylresorcinol, hydrabamine, hydrobromide, hydrochloride, gamma-hydroxybutyrate, iodide, isobutyric acid Salts include, but are not limited to, isothionate, lactate, lactobionate, laurate, malate, maleate, malonate, mandelate, mesylate, metaphosphate, methanesulfonate, methylsulfate, monohydrogenphosphate, mucate, napsylate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, nitrate, oleate, oxalate, pamoate (embonate), palmitate, pantothenate, phenylacetate, phenylbutyrate, phenylpropionate, phthalate, phosphate / diphosphate, polygalacturonate, propanesulfonate, propionate, propiolate, pyrophosphate, pyrosulfate, salicylate, stearate, subacetate, suberate, succinate, sulfate, sulfonate, sulfite, tannate, tartrate, teoclate, tosylate, and valerate salts.

[0148] Illustrative examples of suitable salts include organic salts derived from amino acids such as glycine and arginine, ammonia, primary, secondary and tertiary amines, and cyclic amines such as piperidine, morpholine and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.

[0149] The compounds of the present invention that include a basic moiety, such as an amino group, can form pharmaceutically acceptable salts with various amino acids, in addition to the acids mentioned above.

[0150] Alternatively, useful compounds that are acidic in nature can form base salts with various pharmacologically acceptable cations. Examples of such salts include alkali metal or alkaline earth metal salts, particularly sodium and potassium salts. All of these salts are prepared by conventional techniques. The chemical bases used as reagents to prepare the pharmaceutically acceptable base salts of the present invention are those that form non-toxic base salts with the acidic compounds herein. These salts can be prepared by any suitable method, such as treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary, or tertiary), an alkali metal hydroxide, or an alkaline earth metal hydroxide. These salts can also be prepared by treating the corresponding acidic compound with an aqueous solution containing the desired pharmacologically acceptable cation, and then evaporating the resulting solution to dryness, preferably under reduced pressure. Alternatively, salts can be prepared by mixing lower alkanol solutions of the acidic compound with the desired alkali metal alkoxide, and then evaporating the resulting solution to dryness in the same manner as above. In either case, stoichiometric quantities of reagents are preferably used to ensure completeness of reaction and maximum yields of the desired final product.

[0151] Chemical bases that can be used as reagents to prepare pharmaceutically acceptable base salts of compounds of the present invention that are acidic in nature are those that form non-toxic base salts with such compounds, including, but not limited to, those derived from such pharmacologically acceptable cations, such as alkali metal cations (e.g., potassium and sodium) and alkaline earth metal cations (e.g., calcium and magnesium), ammonium or water-soluble amine addition salts, such as N-methylglucamine (meglumine), and lower alkanolammonium and other base salts of pharmaceutically acceptable organic amines.

[0152] Hemisalts of acids and bases may also be formed, for example, hemisulfate and hemicalcium salts.

[0153] For a review of suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley VCH, 2002). Methods for making pharmaceutically acceptable salts of the compounds of the invention and for interconverting between salt and free base forms are known to those skilled in the art.

[0154] The salts of the present invention can be prepared according to methods known to those skilled in the art.The pharmaceutically acceptable salts of the compounds of the present invention can be easily prepared by appropriately mixing the solution of the compound with the desired acid or base.The salts can precipitate from the solution and be collected by filtration, or can be recovered by evaporating the solvent.The degree of ionization in the salts can vary from completely ionized to almost non-ionized.

[0155] Those skilled in the art will recognize that free base forms of compounds of the present invention bearing a basic functional group can be converted to acid addition salts by treatment with a stoichiometric excess of an appropriate acid. Acid addition salts of compounds of the present invention can be reconverted to the corresponding free base by treatment with a stoichiometric excess of a suitable base, such as potassium carbonate or sodium hydroxide, typically in the presence of an aqueous solvent at temperatures between about 0°C and 100°C. The free base form can be isolated by conventional means, such as extraction with an organic solvent. Additionally, acid addition salts of compounds of the present invention can be exchanged by taking advantage of the differential solubility of the salt, the volatility or acidity of the acid, or by treatment with an appropriately charged ion exchange resin. For example, exchange can be effected by reacting a salt of a compound of the present invention with a slight stoichiometric excess of an acid having a lower pK than the acid component of the starting salt. This conversion is typically carried out at temperatures between about 0°C and the boiling point of the solvent used as the medium for the procedure. Similar exchanges are possible using base addition salts, typically via the free base form.

[0156] The compounds of the present invention can exist in both unsolvated and solvated forms. When the solvent or water is tightly bound, the complex will have a well-defined stoichiometry that is independent of humidity. However, when the solvent or water is weakly bound, such as in channel solvates and hygroscopic compounds, the water / solvent content will depend on humidity and drying conditions. In such cases, non-stoichiometry is the norm. The term "solvate" is used herein to describe a molecular complex containing a compound of the present invention and one or more pharmaceutically acceptable solvent molecules, such as ethanol. The term "hydrate" is used when the solvent is water. Pharmaceutically acceptable solvates according to the present invention include hydrates and solvates in which the solvent of crystallization may be isotopically substituted, such as DO, d6-acetone, and d6-DMSO.

[0157] The present invention also relates to prodrugs of the compounds of the formulas provided herein. Thus, certain derivatives of the compounds of the present invention, which themselves may have little or no pharmacological activity, can be converted to the compounds of the present invention when administered to a patient, for example, by hydrolytic cleavage. Such derivatives are called "prodrugs." Further information on the use of prodrugs can be found in "Prodrugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series (T Higuchi and W Stella); "Bioreversible Carriers in Drug Design," Pergamon Press, 1987 (ed. EB Roche, American Pharmaceutical Association); Guarino, VR; Stella, VJ: Biotech Pharm. Aspects 2007 5(Pt2) 133-187; and J. Rautio et al., Nature Reviews Drug Discovery, 17, 559-587 (2018), the disclosures of which are incorporated herein by reference in their entirety.

[0158] Prodrugs according to the present invention can be generated, for example, by replacing appropriate functional groups present in the compounds of the present invention with certain moieties known to those skilled in the art as "promoieties," for example, as described in "Design of Prodrugs" by H. Bundgaard (Elsevier, 1985), the disclosure of which is incorporated herein by reference in its entirety.

[0159] Some non-limiting examples of prodrugs according to the present invention include: (i) if the compound contains a carboxylic acid functional group (-COOH), its ester, e.g., replacement of the hydrogen with a (C1-C6) alkyl; (ii) if the compound contains an alcohol functional group (—OH), replacement of the hydrogen with an ether thereof, e.g., a (C1-C6) alkanoyloxymethyl or phosphate ether group; and (iii) if the compound contains a primary or secondary amino functional group (NH or NHR, where R is not H), replacement of one or both hydrogens by a preferably metabolically labile group such as an amide, e.g., amide, carbamate, urea, phosphonate, sulfonate, etc. Examples include:

[0160] Further examples of substituent groups following the above examples and examples of other prodrug types can be found in the references mentioned above.

[0161] Finally, certain compounds of the present invention may themselves act as prodrugs of other compounds of the present invention.

[0162] Also included within the scope of this invention are metabolites of the compounds of the formulae described herein, i.e., compounds that are formed in vivo upon administration of the drug, often by oxidation or dealkylation. Some examples of metabolites in accordance with the present invention include: (i) When the compound of the present invention contains an alkyl group, its hydroxyalkyl derivative (-CH→-COH), (ii) When the compound of the invention contains an alkoxy group, its hydroxy derivative (-OR→-OH); (iii) When the compound of the present invention contains a tertiary amino group, its secondary amino derivative (-NRR' → -NHR or -NHR'), (iv) When the compound of the present invention contains a secondary amino group, its primary derivative (-NHR → -NH2), (v) When the compound of the invention contains a phenyl moiety, its phenol derivative (-Ph→-PhOH); (vi) When the compound of the invention contains an amide group, its carboxylic acid derivative (-CONH2 → COOH), and (vii) If the compound contains a hydroxy or carboxylic acid group, the compound can be metabolized, for example, by conjugation with glucuronic acid, to form a glucuronide. Other metabolic pathways through conjugates exist, including, but not limited to, those described below. These pathways are often known as phase 2 metabolism, and include, for example, sulfation or acetylation. Other functional groups, such as NH groups, can also undergo conjugation.

[0163] In one embodiment, provided herein is a solid form of 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione. In one embodiment, the solid form is a crystalline form. In one embodiment, the solid form is an amorphous form.

[0164] The term "crystalline" as used herein means having a regularly repeating arrangement of molecules or external surface planes. A single compound can produce various crystalline forms, where each form has different and distinct solid-state physical properties, such as different solubility profiles, dissolution rates, melting point temperatures, fluidity, and / or different X-ray diffraction peaks. Differences in physical properties can affect pharmaceutical parameters, such as storage stability, compressibility, and density (which can be important in formulation and product manufacturing), and dissolution rate (which can be an important factor in bioavailability).

[0165] The term "amorphous" refers to a state in which a material lacks long-range order at the molecular level and may exhibit the physical properties of a solid or a liquid, depending on temperature. Typically, such materials do not give distinctive X-ray diffraction patterns and are more formally described as liquids, even though they exhibit the properties of a solid. Heating causes a change from the properties of a solid to that of a liquid, which is typically characterized by a second-order change of state (a "glass transition").

[0166] There are several analytical methods that those skilled in the art of solid state chemistry can use to analyze solid forms. Powder X-ray diffraction (PXRD) can also be suitable for quantifying the amount of one crystalline solid form (or forms) in a mixture. In powder X-ray diffraction, X-rays are directed at a crystalline powder, and the intensity of the diffracted X-rays is measured as a function of the angle between the X-ray source and the beam diffracted by the sample. The intensities of these diffracted X-rays can be plotted as peaks on a graph, where the x-axis is the angle between the X-ray source and the diffracted X-rays (known as the "2-theta" angle) and the y-axis is the intensity of the diffracted X-rays. This graph is called a powder X-ray diffraction pattern or powder pattern. The location of the peaks on the x-axis is characteristic of the solid-state structure of the crystal, so different crystalline solid forms will exhibit different powder patterns.

[0167] Those skilled in the art will recognize that the typical accuracy of the 2-theta x-axis values ​​of peaks in a powder pattern is approximately plus or minus 0.2 degrees 2-theta (±0.2 degrees 2-theta). Thus, for example, a diffraction peak appearing at "about 18.0 degrees 2-theta" means that the peak appears at 18.0 ±0.2 degrees 2-theta, i.e., it may be between 17.8 degrees 2-theta and 18.2 degrees 2-theta when measured on most X-ray diffractometers under most conditions. Furthermore, those skilled in the art will recognize that relative peak intensities exhibit instrument-to-instrument variability, as well as variability due to crystallinity, preferred orientation, prepared sample surface, and other factors known to those skilled in the art, and should be interpreted solely as a qualitative measure. Thus, as used herein, the term "essentially the same" in reference to powder X-ray diffraction peak positions means that the typical degree of variability in peak positions and intensities is approximately ±0.2 degrees 2-theta.

[0168] Powder X-ray diffraction is only one of several analytical techniques that can be used to characterize and / or identify crystalline solid forms. Spectroscopic techniques, such as Raman (including microscopic Raman), infrared, and solid-state NMR spectroscopy, can be used to characterize and / or identify crystalline solid forms. These techniques can also be used to quantify the amount of one or more crystalline solid forms in a mixture, and peak values ​​can be reported with the modifier "about" before the peak value.

[0169] The term "anhydrous," as used herein, refers to a crystalline form that does not contain any solvent or water molecules in the crystal lattice.

[0170] The term "hydrate" refers to a solvate containing a compound and a stoichiometric or non-stoichiometric amount of water. The term "monohydrate" refers to a hydrate containing one water molecule per compound molecule (i.e., a 1:1 stoichiometry of water and compound).

[0171] In one embodiment, the present invention provides crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 1.

[0172] In one embodiment, crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 1, is characterized by powder X-ray diffraction (PXRD) (2-theta).

[0173] Table X provides the PXRD peak listing in degrees 2-theta (±0.2 degrees 2-theta) for crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 1.

[0174] [Table 1]

[0175] In one embodiment, the invention provides crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 1, having a PXRD pattern including characteristic peaks at 5.0, 8.7, 9.3, 10.8, 14.5, 15.3, 18.8, and 20.5 degrees two-theta (±0.2 degrees two-theta).

[0176] In one embodiment, the present invention provides crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 1, having a PXRD pattern comprising peaks at 2-theta values ​​essentially the same as those shown in FIG.

[0177] In one embodiment, the present invention provides crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 2.

[0178] In one embodiment, crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 2 is characterized by powder X-ray diffraction (PXRD) (2 theta). In one embodiment, PXRD analysis of crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 2 is performed at 25° C. and a relative humidity below 10%, e.g., as described in Example 77.

[0179] Table Y provides the PXRD peak listing in degrees 2-theta (±0.2 degrees 2-theta) for crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 2.

[0180] [Table 2]

[0181] In one embodiment, the invention provides crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 2, having a PXRD pattern including characteristic peaks at 7.1, 9.4, 12.4, 12.8, 14.3, 15.6, 16.4, 17.4, 18.5, 18.9, 19.5, 19.9, 21.1, 21.4, 23.2, 23.7, 24.8, 25.6, 27.6, 30.3, 33.2, 33.5, and 37.5 degrees two-theta (±0.2 degrees two-theta).

[0182] In one embodiment, the present invention provides crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 2, having a PXRD pattern comprising peaks at 2-theta values ​​essentially the same as those shown in FIG.

[0183] In one embodiment, the present invention provides crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3.

[0184] In one embodiment, crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3 is characterized by powder X-ray diffraction (PXRD) (2 theta). In one embodiment, PXRD analysis of crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3 is performed at 25° C. and greater than 30% relative humidity.

[0185] Table Z provides the PXRD peak listing in degrees 2-theta (±0.2 degrees 2-theta) for crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3.

[0186] [Table 3]

[0187] In one embodiment, the present invention provides crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3, having a PXRD pattern including characteristic peaks at 13.7, 18.0, and 18.3 degrees two-theta (±0.2 degrees two-theta).

[0188] In one embodiment, the invention provides crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3, having a PXRD pattern including characteristic peaks at 6.9, 9.1, 13.7, 18.0, and 18.3 degrees two-theta (±0.2 degrees two-theta).

[0189] In one embodiment, the invention provides crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3, having a PXRD pattern including characteristic peaks at 6.9, 9.1, 11.8, 12.0, 13.7, 14.0, 15.2, 15.8, 18.0, 18.3, 19.0, 19.3, 20.2, 20.9, 21.6, 22.6, 23.6, 24.0, 24.9, 25.2, 25.8, 27.5, 28.1, 28.4, 29.8, 30.9, 31.7, 32.3, and 36.5 degrees two-theta (±0.2 degrees two-theta).

[0190] In one embodiment, the present invention provides crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3, having a PXRD pattern comprising peaks at 2-theta values ​​essentially the same as those shown in FIG. 3.

[0191] In one embodiment, the present invention provides amorphous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 4.

[0192] In one embodiment, amorphous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 4, is characterized by powder X-ray diffraction (PXRD) (2-theta).

[0193] In one embodiment, the present invention provides amorphous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 4, having a PXRD pattern comprising peaks at 2-theta values ​​essentially the same as those shown in FIG.

[0194] The present invention further provides methods of treatment and uses which comprise administering a compound of the present invention, or a pharmaceutically acceptable salt thereof, alone or in combination with other therapeutic or palliative agents.

[0195] The compounds of Formula I and Formula II, and pharmaceutically acceptable salts thereof, are useful for treating diseases and disorders treatable with MEK kinase inhibitors, such as MEK-associated diseases and disorders, and are useful for treating abnormal cell growth, such as tumors, e.g., MEK-associated tumors. The ability of the compounds of Formula I and Formula II, and pharmaceutically acceptable salts thereof, to act as MEK inhibitors can be demonstrated by the assay described in Example A. IC 50 The values ​​are shown in Table A.

[0196] Thus, in one embodiment, provided herein is a method of treating a tumor, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt thereof, or a compound of Formula II, or a pharmaceutically acceptable salt thereof. In one embodiment, the tumor is a MEK-associated tumor.

[0197] As used herein, the terms "MEK kinase inhibitor" and "MEK inhibitor" are used interchangeably and refer to compounds that inhibit the mitogen-activated protein kinase kinase enzymes MEK1 and / or MEK2.

[0198] The terms "MEK-related" and "MEK-mediated" are used interchangeably and refer to diseases or disorders that have constitutive activation of MEK kinase, which can be treated with a MEK inhibitor. Examples include MEK-related abnormal cell growth, such as MEK-related tumors, e.g., MEK-related cancers. In one embodiment, the term "MEK-related" refers to diseases or disorders that have dysregulation of MEK kinase expression or activity, or dysregulation of the BRAF gene or BRAF kinase.

[0199] The phrase "dysregulation of MEK kinase expression or activity" refers to gene amplification that results in overexpression of the MEK protein, or autocrine activity that results from overexpression of the MEK gene in a cell, which results in a pathogenic increase in activity of the kinase domain of the MEK protein (e.g., a constitutively active kinase domain of the MEK protein) in the cell.

[0200] The phrase "dysregulation of the BRAF gene or BRAF kinase" refers to a genetic mutation (e.g., a BRAF gene translocation resulting in the expression of a fusion protein, a deletion of the BRAF gene resulting in the expression of a BRAF protein containing at least one amino acid deletion compared to the wild-type BRAF protein, or a mutation of the BRAF gene resulting in the expression of a BRAF protein having one or more point mutations compared to the wild-type BRAF protein). As another example, dysregulation of the expression, activity, or level of the BRAF gene, BRAF protein, or either thereof can be a mutation of the BRAF gene encoding a BRAF protein that is constitutively active or has increased activity compared to a protein encoded by a BRAF gene that does not contain a mutation. For example, dysregulation of the expression, activity, or level of the BRAF gene, BRAF protein, or either thereof can be the result of a genetic or chromosomal translocation that results in the expression of a fusion protein containing a first portion of BRAF that includes a functional kinase domain and a second portion of a partner protein (i.e., not BRAF).

[0201] In one embodiment, MEK-related disease or disorder has activating BRAF mutation. In one embodiment, MEK-related disease or disorder is MEK-related cancer with activating BRAF mutation. Non-limiting examples of BRAF mutation include BRAF V600 mutation, such as V600E, V600D, V600K, V600R and V600S. In one embodiment, BRAF mutation is V600E mutation. In one embodiment, BRAF mutation is V600K mutation.

[0202] In one embodiment, the MEK-associated disease or disorder is KIAA11549-BRAF, MKRN1-BRAF, TRIM24-BRAF, AGAP3-BRAF, ZC3HAV1-BRAF, AKAP9-BRAF, CCDC6-BRAF, AGK-BRAF, EPS15-BRAF, NUP214-BRAF, ARMC10-BRAF, BTF3L4-BRAF, GHR-BRAF, ZC3HAV1-BRAF, ZNF767-BRAF, CCDC91-BRAF, DYNC112-BRAF, ZKSCAN1-BRAF, GTF2I-BRAF, MEK-associated tumors harbor one or more BRAF fusions that result in constitutive kinase activation and transforming, including, but not limited to, BRAF, MZT1-BRAF, RAD18-BRAF, CUX1-BRAF, SLC12A7-BRAF, MYRIP-BRAF, SND1-BRAF, NUB1-BRAF, KLHL7-BRAF, TANK-BRAF, RBMS3-BRAF, STRN3-BRAF, STK35-BRAF, ETFA-BRAF, SVOPL-BRAF, JHDM1D-BRAF, or BCAP29-BRAF.

[0203] In one embodiment, the MEK-associated disease or disorder is a MEK-associated tumor harboring a BRAF fusion protein, and the tumor is selected from the group consisting of breast cancer (e.g., invasive ductal carcinoma), colorectal cancer (e.g., colon adenocarcinoma), esophageal cancer (e.g., esophageal adenocarcinoma), glioma (e.g., desmoplastic infantile ganglioglioma of the brain, pilocytic astrocytoma of the brain, pleomorphic xanthoastrocytoma of the brain, low-grade glioma (NOS) of the spinal cord, anaplastic oligodendroglioma, anaplastic ganglioglioma), and head and neck carcinoma (e.g., neuroendocrine carcinoma of the head and neck). , lung cancer (e.g., lung adenocarcinoma, non-small cell lung carcinoma (NOS)), melanoma (e.g., spitzoid cutaneous melanoma, non-spitzoid mucosal melanoma, spitzoid cutaneous melanoma, melanoma of unknown primary, non-spitzoid cutaneous melanoma), pancreatic cancer (e.g., adenocarcinoma, pancreatic acinar cell carcinoma), prostate cancer (e.g., prostatic acinar adenocarcinoma), sarcoma (malignant solid fibroma), thyroid cancer (papillary thyroid carcinoma), carcinoma of unknown primary (e.g., adenocarcinoma of unknown primary), pleural mesothelioma, rectal adenocarcinoma, endometrial cancer (e.g., endometrial adenocarcinoma (NOS)), or ovarian serous carcinoma.

[0204] In one embodiment, the MEK-associated cancer is selected from cancers having a BRAF fusion protein listed in Table 1 (J.S. Ross et al., Int. J. Cancer:138, 881-890 (2016)).

[0205] [Table 4-1]

[0206] [Table 4-2]

[0207] In one embodiment, the MEK-associated tumor is a BRAF wild-type tumor.

[0208] The term "wild-type" describes a nucleic acid (eg, a BRAF gene or BRAF mRNA) that is typically found in subjects who do not have the disease or disorder associated with the reference nucleic acid or protein.

[0209] The term "wild-type BRAF" describes the BRAF nucleic acid (eg, BRAF gene or BRAF mRNA) or BRAF protein found in subjects who do not have an activating BRAF mutation.

[0210] "Abnormal cell growth," as used herein, unless otherwise specified, refers to cell growth that is independent of normal regulatory mechanisms (e.g., loss of contact inhibition), e.g., tumors. Abnormal cell growth can be benign (non-cancerous) or malignant (cancerous).

[0211] The term "cancer" or "cancerous" refers to any malignant and / or invasive growth or tumor caused by abnormal cell growth. Cancer includes primary cancers that begin in a particular part of the body, metastatic cancers that spread from where the cancer began to other parts of the body, recurrences from the original primary cancer after remission, and second primary cancers, which are new primary cancers in patients with a history of a previous cancer of a different type from the second primary cancer. Cancer includes solid tumors, named for the cell type that forms them, and cancers of the blood, bone marrow, or lymphatic system. Solid tumors are abnormal growths or masses of tissue that usually do not contain cysts or fluid areas. Examples of solid tumors are sarcomas, carcinomas, and lymphomas. Leukemias (cancers of the blood) generally do not form solid tumors (National Cancer Institute, Dictionary of Cancer Terms).

[0212] As used herein, the term "treat" or "treatment" refers to a curative or palliative measure. Beneficial or desired clinical results include, but are not limited to, the total or partial alleviation of symptoms associated with a disease or disorder or condition, whether detectable or undetectable, a decrease in the extent of the disease, a stabilized (i.e., not worsening) state of the disease, a delay or slowing of disease progression, remission or alleviation of the condition (e.g., one or more symptoms of the disease), and relief (whether partial or total).

[0213] The term "treating" or "treating" cancer, as used herein, refers to administering a compound of the present invention to a subject having or diagnosed with cancer to achieve at least one positive therapeutic effect, such as reducing the number of cancer cells, reducing tumor size, reducing the rate of cancer cell invasion into surrounding organs, or reducing the rate of tumor metastasis or tumor growth, or improving, alleviating, or inhibiting the process of the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition. The term "treatment," as used herein, unless otherwise specified, refers to the act of treating as "treating," as defined immediately above. The term "treating" also includes adjuvant and neoadjuvant treatment of a subject.

[0214] For purposes of the present invention, beneficial or desired clinical results include, but are not limited to, one or more of the following: reduction in the proliferation (or destruction) of neoplastic or cancerous cells; inhibition of metastasis or neoplastic cells; reduction or decrease in tumor size; prolonged remission in a subject (e.g., compared to one or more metric(s) in a subject with a similar cancer who has not received treatment or who has received a different treatment, or compared to one or more metric(s) in the same subject prior to treatment); reduction in symptoms resulting from cancer; improved quality of life for a patient with cancer; reduced dosages of other medications required to treat the cancer; delayed progression of cancer; curing the cancer; overcoming one or more resistance mechanisms in the cancer; and / or prolonged survival of a patient with cancer. Positive therapeutic effects in cancer can be measured in several ways (W.A. Weber, Assessing tumor response to therapy, J. Nucl. Med. 50 Suppl. 1:1S-10S (2009)). For example, with regard to tumor growth inhibition (T / C), according to the National Cancer Institute (NCI) standards, a T / C of 42% or less is a minimal level of antitumor activity. A T / C of <10% is considered a high level of antitumor activity, where T / C (%) = median treated tumor volume / median control tumor volume × 100.

[0215] In one embodiment, the treatment achieved by administering the compounds of the present invention is defined by any of the following: partial response (PR), complete response (CR), overall response (OR), progression-free survival (PFS), disease-free survival (DFS), and overall survival (OS). PFS, also known as "time to tumor progression," refers to the length of time cancer does not grow during and after treatment, including the length of time a patient experiences CR or PR, and the length of time a patient experiences stable disease (SD). DFS refers to the length of time a patient remains disease-free during and after treatment. OS refers to the extension of life expectancy compared with untreated or untreated subjects or patients. In one embodiment, the response to treatment with the compounds of the present invention is any of PR, CR, OR, PFS, DFS, or OS, which is assessed using Response Evaluation Criteria in Solid Tumors (RECIST) 1.1.

[0216] Treatment regimens for the compounds of the present invention that are effective in treating cancer patients can vary according to factors such as the patient's condition, age, and weight, as well as the ability of the treatment to elicit an anti-cancer response in the subject. Any embodiment of the present invention may not be effective in achieving a positive therapeutic effect in all subjects, but should actually achieve a statistically significant number of subjects as determined by any statistical test known in the art, such as Student's t-test, chi-square test, Mann-Whitney U test, Kruskal-Wallis test (H test), Jonkheel-Tapstra test, and Wilcoxon test.

[0217] The terms "treatment regimen," "administration protocol," and "administration regimen" are used interchangeably to refer to the dosage and timing of administration of a compound of the invention, alone or in combination with another therapeutic agent.

[0218] "Ameliorating" means that one or more symptoms are reduced or improved upon treatment with a combination described herein compared to when the combination is not administered. "Ameliorating" also includes shortening or reducing the duration of symptoms.

[0219] As used herein, the term "subject" refers to any animal, including a mammal, e.g., a human. In one embodiment, the subject has experienced and / or exhibited at least one symptom of a disease or disorder to be treated and / or prevented. In one embodiment, the subject has been identified or diagnosed as having a MEK-associated tumor (e.g., as determined using a regulatory agency-approved, e.g., FDA-approved, assay or kit). In one embodiment, the subject has a MEK-associated tumor that is positive for a BRAF mutation (e.g., as determined using a regulatory agency-approved, e.g., FDA-approved, assay or kit). The subject may be one whose tumor has a MEK mutation (e.g., when the tumor is itself identified using a regulatory agency-approved, e.g., FDA-approved, kit or assay). In one embodiment, the subject is suspected of having a MEK-associated tumor. In one embodiment, the subject has clinical records indicating that the subject has a MEK-associated tumor with a BRAF mutation (the clinical records may indicate that the subject should be treated with any of the compositions provided herein). In one embodiment, the subject is a human. In one embodiment, the human subject is a pediatric subject.

[0220] The term "pediatric subject," as used herein, refers to a subject under the age of 21 at the time of diagnosis or treatment. The term "pediatric" can be further divided into various subpopulations, including neonates (birth to 1 month), infants (1 month to 2 years), children (2 to 12 years), and adolescents (12 to 21 years (up to, but not including, their 22nd birthday)). Berhman RE, Kliegman R, Arvin AM, Nelson WE, Nelson Textbook of Pediatrics, 15th ed., Philadelphia: WB Saunders Company, 1996; Rudolph AM et al., Rudolph's Pediatrics, 21st ed., New York: McGraw-Hill, 2002; and Avery MD, First LR. Pediatric Medicine, 2nd ed., Baltimore: Williams & Wilkins; 1994. In one embodiment, the pediatric subject is from birth to 28 days old, from 29 days old to under 2 years old, from 2 years old to under 12 years old, or from 12 years old to 21 years old (up to but not including the 22nd birthday). In one embodiment, the pediatric subject is from birth to 28 days old, from 29 days old to under 1 year old, from 1 month to under 4 months old, from 3 months to under 7 months old, from 6 months to under 1 year old, from 1 year to under 2 years old, from 2 years to under 3 years old, from 2 years to under 7 years old, from 3 years to under 5 years old, from 5 years to under 10 years old, from 6 years to under 13 years old, from 10 years to under 15 years old, or from 15 years to under 22 years old.

[0221] In one embodiment, provided herein is a method of treating a tumor, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the tumor is a MEK-associated tumor. In one embodiment, the MEK-associated tumor has a BRAF mutation. In one embodiment, the BRAF mutation is V600E and / or V600K and / or V600D and / or V600R and / or V600S. In one embodiment, the BRAF mutation is V600E. In one embodiment, the BRAF mutation is V600K. In one embodiment, the MEK-associated tumor has a BRAF fusion, e.g., a BRAF fusion disclosed herein. In one embodiment, the MEK-associated tumor is a BRAF wild-type tumor.

[0222] In one embodiment of any of the methods of use described herein, the tumor is a solid tumor. In one embodiment of any of the methods disclosed herein, the solid tumor is a MEK-associated tumor. In one embodiment, the tumor is intracranial. In one embodiment, the tumor is extracranial. In one embodiment of any of the methods described herein, the tumor (e.g., a MEK-associated tumor) is a malignant tumor (i.e., cancer), such as a MEK-associated cancer. In one embodiment of any of the methods of use described herein, the MEK-associated cancer is melanoma, colon cancer, colorectal cancer, lung cancer (e.g., small cell lung cancer or non-small cell lung cancer), thyroid cancer (e.g., papillary thyroid cancer, medullary thyroid cancer, differentiated thyroid cancer, recurrent thyroid cancer, or refractory differentiated thyroid cancer), breast cancer, ovarian cancer, cancer of the CNS, bone cancer, cancer of the anus, anal canal, or anorectum, eye cancer, bile duct cancer, ductal carcinoma in situ, liver cancer, cancer of the gallbladder or pleura, or oral cancer. cancer of the oral cavity, lip, oropharyngeal, nose, nasal cavity or middle ear, vulva, esophageal, cervical, gastrointestinal carcinoid tumor, hypopharyngeal, kidney, laryngeal, liver, lung, melanoma, nasopharyngeal, peripheral nervous system (e.g., neuroblastoma), ovarian, pancreatic, peritoneal, omental and mesenteric cancer, pharyngeal, prostate, kidney (e.g., renal cell carcinoma (RCC)), small intestine, soft tissue, stomach, testicular, uterine, ureter, or bladder.

[0223] In one embodiment of any of the methods of use described herein, the MEK-associated cancer is an extracranial cancer (i.e., an extracranial tumor). In one embodiment, the extracranial cancer is selected from melanoma, colorectal cancer, thyroid cancer, non-small cell lung cancer, ovarian cancer, and neuroblastoma. In one embodiment, the MEK-associated cancer is melanoma. In one embodiment, the MEK-associated cancer is colorectal cancer. In one embodiment, the MEK-associated cancer is thyroid cancer. In one embodiment, the MEK-associated cancer is non-small cell lung cancer. In one embodiment, the MEK-associated cancer is ovarian cancer. In one embodiment, the MEK-associated cancer is neuroblastoma.

[0224] In one embodiment of any of the methods of use described herein, the MEK-associated cancer is a CNS cancer.

[0225] In one embodiment of any of the methods of use described herein, the MEK-associated cancer is an intracranial cancer (brain cancer).

[0226] In one embodiment of any of the methods of use described herein, the cancer is metastatic cancer.

[0227] The term "metastasis" is a term known in the art that refers to the spread of cancer cells from the location where they first formed (primary site) to one or more other sites (secondary site(s)) in a subject. In metastasis, cancer cells break away from the original (primary) tumor, travel through the blood or lymphatic system, and form new tumors (metastatic tumors) in other organs or tissues of the body. The new metastatic tumors contain the same or similar cancer cells as the primary tumor. In the secondary site, tumor cells proliferate and may initiate the growth or establishment of a secondary tumor at this distant site.

[0228] The term "metastatic cancer" (also known as "secondary cancer"), as used herein, refers to a type of cancer that begins in one tissue type but then spreads to one or more tissues outside of the (primary) cancer origin. Metastatic brain tumor refers to a cancer within the brain, i.e., a cancer that begins in a tissue other than the brain and has metastasized to the brain.

[0229] In one embodiment, the compound of formula I or its pharmaceutically acceptable salt and the compound of formula II or its pharmaceutically acceptable salt exhibit surprising penetration into the brain and / or CNS. Such compounds can cross the BBB and inhibit MEK kinase in the brain and / or other CNS structures. Thus, in one embodiment, the compounds provided herein are useful for treating CNS tumors, such as CNS cancer.

[0230] In one embodiment, the MEK-associated tumor is a malignant CNS tumor (i.e., a MEK-associated CNS cancer). In one embodiment, the MEK-associated CNS cancer has a BRAF mutation. In one embodiment, the MEK-associated CNS cancer has a BRAF V600 mutation. In one embodiment, the BRAF mutation is V600E and / or V600K and / or V600D and / or V600R and / or V600S. In one embodiment, the MEK-associated CNS cancer has a BRAF V600E mutation. In one embodiment, the MEK-associated CNS cancer has a BRAF V600K mutation. In one embodiment, the MEK-associated tumor has a BRAF fusion. In one embodiment, the MEK-associated tumor is a BRAF wild-type tumor.

[0231] The terms "CNS cancer" or "cancer of the CNS," used interchangeably herein, refer to cancers of the CNS (i.e., malignant tumors), including cancers of the brain (also known as intracranial tumors), cancers of the spinal cord, and cancers of the meninges that surround the brain and spinal cord. Brain cancers include metastatic brain tumors (i.e., metastatic intracranial cancers) and malignant primary brain tumors.

[0232] In one embodiment, the MEK-associated CNS cancer is a MEK-associated metastatic brain tumor. The MEK-associated metastatic brain tumor may be the result of any cancer described herein in which the subject has developed at least one brain metastasis. In one embodiment, the metastatic brain tumor is melanoma, colorectal cancer, thyroid cancer, non-small cell lung cancer, ovarian cancer, or neuroblastoma. In one embodiment, the MEK-associated metastatic brain tumor is metastatic melanoma, metastatic colorectal cancer, or metastatic non-small cell lung cancer. In one embodiment, the MEK-associated metastatic brain tumor is metastatic melanoma. In one embodiment, the MEK-associated metastatic brain tumor is metastatic colorectal cancer. In one embodiment, the MEK-associated metastatic brain tumor is metastatic non-small cell lung cancer. In one embodiment, the MEK-associated metastatic brain tumor is metastatic ovarian cancer. In one embodiment, the metastatic brain tumor is metastatic thyroid cancer. In one embodiment, the MEK-associated metastatic brain tumor is kidney cancer. In one embodiment, the cancer is a MEK-associated metastatic cancer with at least one brain metastasis (i.e., metastatic brain tumor). In one embodiment, the cancer is a MEK-associated metastatic melanoma with at least one brain metastasis. In one embodiment, the cancer is a MEK-associated metastatic colorectal cancer with at least one brain metastasis. In one embodiment, the cancer is a MEK-associated metastatic non-small cell lung cancer with at least one brain metastasis. In one embodiment, the cancer is a MEK-associated metastatic ovarian cancer with at least one brain metastasis. In one embodiment, the cancer is a MEK-associated metastatic thyroid cancer with at least one brain metastasis. In one embodiment, the cancer is a MEK-associated neuroblastoma with at least one brain metastasis. In one embodiment of any of the above MEK-associated metastatic brain tumors, the cancer has a BRAF mutation. In one embodiment, the cancer has a BRAF V600 mutation. In one embodiment, the BRAF mutation is V600E and / or V600K and / or V600D and / or V600R and / or V600S. In one embodiment, the cancer has a BRAF V600E mutation. In one embodiment, the cancer has a BRAF V600K mutation. In one embodiment, the MEK-associated tumor has a BRAF fusion.In one embodiment, the MEK-associated tumor is a BRAF wild-type tumor.

[0233] In one embodiment, the MEK-associated cancer is a leptomeningeal metastasis (leptomeningeal disease (LMD)). LMD represents a subset of CNS metastases that grow in the lining of the brain or spine and / or in the cerebrospinal fluid (CSF), or leptomeningeal carcinomatosis. In mammals, the meninges are the dura mater, arachnoid mater, and pia mater. CSF is located in the subarachnoid space between the arachnoid mater and pia mater. The arachnoid mater and pia mater are sometimes collectively referred to as the leptomeninges. When LMD occurs in the leptomeninges surrounding the spinal cord and / or CSF, it may be referred to as "extracranial LMD." When LMD occurs in the leptomeninges and / or CSF of the brain, it may be referred to as "intracranial LMD." LMD cancer cells can be suspended in CSF and therefore can spread rapidly throughout the CNS. As a result, LMD has a poor prognosis, with survival typically measured in months. In one embodiment, the metastatic cancer is LMD. In one embodiment, the metastatic cancer is a MEK-associated LMD. In one embodiment, the metastatic cancer is a MEK-associated intracranial LMD. In one embodiment, the metastatic cancer is a MEK-associated extracranial LMD. In one embodiment, the MEK-associated LMD is an LMD secondary to melanoma metastasis (i.e., the LMD is metastatic melanoma). In one embodiment, the MEK-associated LMD is an LMD secondary to colorectal cancer metastasis (i.e., the LMD is metastatic colorectal cancer). In one embodiment, the MEK-associated LMD is an LMD secondary to non-small cell lung cancer metastasis (i.e., the LMD is metastatic non-small cell lung cancer). In one embodiment of any of the foregoing MEK-associated LMDs, the LMD has a BRAF mutation. In one embodiment, the MEK-associated LMD has a BRAF V600 mutation. In one embodiment, the BRAF mutation is V600E and / or V600K and / or V600D and / or V600R and / or V600S. In one embodiment, the MEK-associated LMD has a BRAF V600E mutation. In one embodiment, the MEK-associated LMD has a BRAF V600K mutation. In one embodiment, the MEK-associated LMD has a BRAF fusion. In one embodiment, the MEK-associated LMD is a BRAF wild-type tumor.

[0234] In one embodiment, the MEK-associated tumor is a cancer with a high risk of metastasis. In one embodiment, the tumor with a high risk of metastasis is a cancer with a BRAF V600E, V600D, V600K, V600R, and / or V600S mutation. In one embodiment, the cancer with a high risk of metastasis has a BRAF fusion, such as any of the BRAF fusions disclosed herein. In one embodiment, the cancer with a high risk of metastasis is a BRAF wild-type tumor. In one embodiment, the cancer with a high risk of metastasis is melanoma, colorectal cancer, thyroid cancer, non-small cell lung cancer, ovarian cancer, or neuroblastoma. In one embodiment, the cancer with a high risk of metastasis is melanoma, colorectal cancer, thyroid cancer, non-small cell lung cancer, ovarian cancer, or neuroblastoma. In one embodiment, the cancer with a high risk of metastasis is melanoma. In one embodiment, the cancer with a high risk of metastasis is melanoma with a BRAF V600E mutation or a BRAF V600K mutation. In one embodiment, the cancer having a high risk of metastasis is colorectal cancer. In one embodiment, the cancer having a high risk of metastasis is colorectal cancer with a BRAF V600E mutation or a BRAF V600K mutation. In one embodiment, the cancer having a high risk of metastasis is thyroid cancer. In one embodiment, the cancer having a high risk of metastasis is thyroid cancer with a BRAF V600E mutation or a BRAF V600K mutation. In one embodiment, the cancer having a high risk of metastasis is non-small cell lung cancer. In one embodiment, the cancer having a high risk of metastasis is non-small cell lung cancer with a BRAF V600E mutation or a BRAF V600K mutation. In one embodiment, the cancer having a high risk of metastasis is ovarian cancer. In one embodiment, the cancer having a high risk of metastasis is ovarian cancer with a BRAF V600E mutation or a BRAF V600K mutation. In one embodiment, the cancer having a high risk of metastasis is neuroblastoma. In one embodiment, the cancer with a high risk of metastasis is a neuroblastoma with a BRAF V600E mutation or a BRAF V600K mutation.In one embodiment, the cancer having a high risk of metastasis has a KIAA11549-BRAF fusion.

[0235] In one embodiment, the CNS tumor is a primary brain tumor. Primary brain tumors are tumors that begin in the brain or spine and are collectively known as gliomas. The term "glioma" is used to describe tumors that begin in glial cells present in the CNS. According to the WHO classification of brain tumors, gliomas are graded by cellular activity and invasiveness on a scale that includes grade I (benign CNS tumors) and grades II-IV (malignant CNS tumors).

[0236] Grade I gliomas (pilocytic astrocytomas): Typically occur in the cerebellum or brainstem of children, occasionally in the cerebral hemispheres, and are slow-growing. Grade I gliomas can also occur in adults. Grade I gliomas are benign (WHO Grade I), but the difficulty of curing this disease makes their growth behavior malignant, leading to high morbidity (Rostami, Acta Neurochir (Wien). 2017;159(11):2217-2221).

[0237] Grade II gliomas (low-grade gliomas): These include astrocytomas, oligodendrogliomas, and mixed oligoastrocytomas. Grade II gliomas typically occur in young adults (e.g., 20-50 years old) and are most often found in the cerebral hemispheres. Due to the invasive nature of these tumors, recurrences can occur. Some grade II gliomas recur and develop into more aggressive tumors (grade III or IV).

[0238] Grade III gliomas (malignant gliomas): Includes anaplastic astrocytoma, anaplastic oligodendroglioma, and mixed anaplastic oligoastrocytoma. Grade III tumors are aggressive, high-grade cancers that invade nearby brain tissue with tentacle-like projections, making complete surgical removal more difficult.

[0239] Grade IV gliomas include glioblastoma multiforme (GBM) and gliosarcoma. (GBM) is a malignant glioma. GBM is the most aggressive and most common primary brain tumor. Glioblastoma multiforme usually spreads rapidly, invading other parts of the brain with tentacle-like projections, making complete surgical removal more difficult. Gliosarcoma is a malignant cancer defined as a glioblastoma composed of glioma and sarcoma components.

[0240] In one embodiment, the primary brain tumor is a glioma. In one embodiment, the glioma is a low-grade glioma. In one embodiment, the glioma is a pediatric low-grade glioma.

[0241] In one embodiment, primary brain tumor is benign primary brain tumor.Benign primary brain tumor can cause severe pain, permanent brain damage and death, and in some cases, become malignant.Non-limiting examples of benign primary brain tumor include grade I glioma, papillary craniopharyngioma, meningioma (including rhabdoid meningioma), atypical teratoid / rhabdoid tumor, and dysembryoplastic neuroepithelial tumor (DNT), pilocytic astrocytoma, oligodendroglioma, mixed oligoastrocytoma, anaplastic astrocytoma, anaplastic oligodendroglioma, anaplastic mixed oligoastrocytoma, diffuse astrocytoma, ependymoma, pleomorphic xanthoastrocytoma (PXA), ganglioglioma, gliosarcoma or anaplastic ganglioglioma.

[0242] In one embodiment, the cancer is a peripheral nervous system cancer. In one embodiment, the peripheral nervous system cancer is neuroblastoma.

[0243] In one embodiment, provided herein is a method of treating a MEK-associated CNS tumor, comprising administering (e.g., orally administering) to a subject in need thereof a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the MEK-associated CNS tumor harbors a BRAF V600 mutation. In one embodiment, the MEK-associated CNS tumor harbors a BRAF V600E and / or V600K and / or V600D and / or V600R mutation and / or V600S. In one embodiment, the MEK-associated CNS tumor harbors a BRAF V600E mutation. In one embodiment, the MEK-associated CNS tumor harbors a BRAF V600K mutation. In one embodiment, the MEK-associated CNS tumor harbors a BRAF fusion, e.g., any of the BRAF fusions disclosed herein, e.g., a KIAA11549-BRAF fusion. In one embodiment, the MEK-associated CNS tumor is a BRAF wild-type tumor. In one embodiment, the subject has been treated with one or more anti-cancer therapies independently selected from anti-cancer agents, surgery, and radiation therapy prior to administration of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, e.g., as described below. In one embodiment, the subject is treated with a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, in combination with one or more anti-cancer therapies independently selected from one or more anti-cancer agents, surgery, and / or radiation therapy, e.g., as described below. In one embodiment, the subject is treated with one or more anti-cancer therapies independently selected from anti-cancer agents, surgery, and radiation therapy after administration of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, e.g., as described below. In one embodiment, the MEK-associated tumor is a CNS tumor. In one embodiment, the MEK-associated CNS tumor is a malignant CNS tumor (i.e., CNS cancer). In one embodiment, the malignant CNS tumor is a metastatic CNS cancer.In one embodiment, the metastatic CNS cancer is selected from metastatic melanoma, metastatic colorectal cancer, metastatic non-small cell lung cancer, metastatic thyroid cancer, and metastatic ovarian cancer. In one embodiment, the metastatic CNS cancer is metastatic melanoma. In one embodiment, the metastatic CNS cancer is colorectal cancer. In one embodiment, the metastatic CNS cancer is metastatic non-small cell lung cancer. In one embodiment, the metastatic CNS cancer is metastatic thyroid cancer. In one embodiment, the metastatic CNS cancer is metastatic ovarian cancer. In one embodiment, the MEK-associated CNS cancer is LMD. In one embodiment, the LMD is intracranial. In one embodiment, the LMD is extracranial. In one embodiment, the LMD is metastatic melanoma. In one embodiment, the LMD is selected from metastatic melanoma, metastatic colorectal cancer, and metastatic non-small cell lung cancer. In one embodiment, the LMD is metastatic colorectal cancer. In one embodiment, the LMD is metastatic non-small cell lung cancer. In one embodiment, the MEK-associated CNS cancer is a primary brain tumor. In one embodiment, the primary brain tumor is a grade 2 glioma. In one embodiment, the primary brain tumor is a grade 3 glioma. In one embodiment, the primary brain tumor is a grade 4 glioma. In one embodiment, the MEK-associated CNS tumor is a benign tumor. In one embodiment, the benign CNS tumor is a papillary craniopharyngioma, a meningioma (including rhabdoid meningioma), an atypical teratoid / rhabdoid tumor, or a dysembryoplastic neuroepithelial tumor (DNT). In one embodiment, the compound is a compound of Formula I or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the compound is selected from the compounds of Examples 1-69 or a pharmaceutically acceptable salt thereof.

[0244] The ability to determine whether a compound may be suitable for treating a CNS cancer can be determined, for example, by identifying whether the compound is a substrate for an efflux transporter, and / or measuring cell permeability, and / or measuring the ratio of free blood to free plasma, as described herein.

[0245] In one embodiment, the compound of Formula I or a pharmaceutically acceptable salt thereof and the compound of Formula II or a pharmaceutically acceptable salt thereof exhibit high cell permeability. The method for determining the permeability of the compounds of the present invention can be determined according to the assay described in Example B, and the permeability coefficients are provided in Table B1.

[0246] The compounds of the present invention exhibit low efflux. In vitro methods for assessing whether compounds of the present invention are substrates of the efflux transporters P-glycoprotein (P-gp or multidrug resistance 1 (MDR1) protein) and breast cancer resistance protein (BCRP) are described in Example B, and the efflux ratios of compounds of the present invention are provided in Table B3.

[0247] In one embodiment, the compounds of the present invention exhibit a moderate to high brain (unbound) / plasma (unbound) ratio (i.e., a moderate to high free brain / plasma ratio). The ability of the compounds of the present invention to penetrate the BBB of a subject (e.g., a human) can be determined in a suitable animal model (e.g., a rodent, e.g., a mouse). For example, the ability of a particular compound to penetrate the BBB in a mouse can be determined by assessing the ratio of unbound brain concentration to unbound plasma (free B / P) in the mouse, as described, for example, in Example C, and the free brain to free plasma ratio is provided in Table C2. Calculating the free brain to free plasma ratio of a compound allows prediction of the effective concentration required to achieve efficacy in the periphery and brain based on dose-dependent exposure in the animal model. These distribution data, along with relevant pharmacokinetic data, can be used in a model to predict the dose required to achieve efficacy in human patients.

[0248] Thus, in one embodiment, the methods of the present invention include a method for treating MEK-associated CNS cancer in a subject in need thereof, comprising administering a compound of Formula II or a pharmaceutically acceptable salt thereof, wherein at least a portion of the compound of Formula II penetrates the BBB as demonstrated in a suitable animal model. In one embodiment, the brain / plasma ratio of the total drug is at least about 0.3 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 0.35 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 0.4 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 0.45 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 0.5 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 0.55 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 0.6 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 0.65 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 0.7 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 0.75 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 0.8 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 0.85 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 0.9 after administration (e.g., oral or intravenous administration) to a subject. In one embodiment, the brain / plasma ratio of the total drug is at least about 0.95 after administration (e.g., oral or intravenous administration) to a subject.In one embodiment, the brain / plasma ratio of the total drug is at least about 1.0 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 1.0 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 1.1 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 1.2 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 1.3 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 1.4 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 1.5 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 1.6 after administration to a subject (e.g., oral or intravenous administration). In one embodiment, the brain / plasma ratio of the total drug is at least about 1.7 after administration to a subject (e.g., oral or intravenous administration). The percentage of a compound that penetrates the BBB is determined by the area under the concentration-time curve (AUC) in the brain relative to the plasma for a given period of time. 0-t It should be noted that the calculation is based on the AUC 0-24h ) is 20 ng / mL in the brain and 80 ng / mL in the plasma, the percentage of the compound that penetrates the BBB is 20% (20 ng / mL in the brain divided by the total concentration (20 ng / mL + 80 ng / mL)) (i.e., a brain to plasma ratio of 0.20). In one embodiment, the percentage is calculated by the area under the concentration-time curve, i.e., (AUC 0-last ) is calculated based on

[0249] Cancers that frequently metastasize to the brain are known to have MAPK pathway activating alterations, such as BRAF mutations, including the BRAF mutations disclosed herein, or BRAF fusions, including the BRAF fusions disclosed herein. Activating mutations can occur at various levels in the classical pathway, but all of them require signal transduction via mitogen / extracellular signal-regulated kinase (MEK) to increase proliferation and survival (Schubbert S, Shannon K, Bollag G., Nat Rev Cancer. 2007;7:295-308). BRAF gene mutations have been identified in malignant melanoma, papillary thyroid carcinoma, colorectal cancer, non-small cell lung cancer (NSCLC), and ovarian cancer, as well as their metastatic tumors, and primary brain tumors (Davies H. et al., Nature 417(6892):949-954, 2002). For example, BRAF mutations, such as BRAF V600 mutations, have been observed in a number of metastatic CNS tumors, including melanoma brain metastases (Flaherty KT et al., Nat Rev Cancer (2012) 12 (5): 349-61), colorectal cancer brain metastases, and non-small cell lung cancer (Berghoff, AS, Preusser M., Curr Opin Neurol (2014) 27 (6): 689-696), papillary thyroid cancer (Kim, WW et al., J Otolaryngol Head Neck Surg. 2018; 47: 4, 1-6), and ovarian cancer (Grisham RN. et al., Cancer, 2013; 119: 548-554).

[0250] BRAF mutations, such as those disclosed herein, and BRAF fusions, such as those disclosed herein, have also been observed in malignant primary brain tumors, including grade IV gliomas, e.g., glioblastomas and gliosarcoma, anaplastic astrocytomas (high-grade tumors), and WHO grade III anaplastic gangliogliomas in pediatric and adult populations (Berghoff, A.S., Preusser M., Curr Opin Neurol (2014) 27(6):689-696; Schindler et al. (Acta Neuropathol 121(3):397-405, 2011); Behling et al. (Diagn Pathol 11(1):55, 2016); K.C. Schreck et al., Cancers, 2019, 11, 1262)).

[0251] BRAF mutations, e.g., the BRAF mutations disclosed herein, and BRAF fusions, e.g., the BRAF fusions disclosed herein, have also been observed in benign primary brain tumors in pediatric and adult populations, such as WHO grade II astrocytoma, WHO grade II pleomorphic xanthoastrocytoma (PXA), pleomorphic xanthoastrocytoma with anaplasia, pilocytic astrocytoma (PA), papillary craniopharyngioma, ganglioglioma, astroblastoma, pilocytic astrocytoma, atypical teratoid / rhabdoid tumor, and rhabdoid meningioma (Berghoff, A.S., Preusser M., Curr Opin Neurol (2014) 27(6):689-696; Schindler et al. (Acta Neuropathol 121(3):397-405, 2011); Behling et al. (Diagn Pathol 11(1):55, 2016); (Behling et al., Diagn Pathol 11(1):55, 2016; Brastianos et al., Nat Genet 46(2):161-165, 2014; Dougherty et al., Neuro Oncol 12(7):621-630, 2010; Lehman et al., Neuro Oncol 19(1):31-42, 2017; Mordechai et al., Pediatr Hematol Oncol 32(3):207-211, 2015; Myung et al., Transl Oncol 5(6):430-436, 2012; Schindler et al., Acta Neuropathol 121(3):397-405, 2011).

[0252] BRAF mutations have also been detected in recurrent neuroblastoma (Eleveld, TF et al., Nat Genet 47(8):864-871, 2015). Neuroblastoma is a pediatric tumor of the peripheral nervous system. While most patients with neuroblastoma initially have tumors that respond to chemotherapy, the majority of patients will experience a treatment-resistant relapse.

[0253] Thus, also provided herein is a method for treating a subject diagnosed or identified as having a MEK-associated tumor, such as any of the exemplary MEK-associated tumors disclosed herein, comprising administering to the subject a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, wherein the subject has been identified or diagnosed as having a tumor with a BRAF mutation of a BRAF fusion, for example, by using a regulatory-approved, e.g., FDA-approved, test or assay for identifying a BRAF mutation or fusion in the subject or in a biopsy sample from the subject, or by performing any of the non-limiting examples of assays described herein. In one embodiment, the test or assay is provided as a kit. In one embodiment, the assay utilizes next-generation sequencing, pyrosequencing, immunohistochemistry, fluorescence microscopy, break-apart FISH analysis, Southern blotting, Western blotting, FACS analysis, Northern blotting, or PCR-based amplification (e.g., RT-PCR and quantitative real-time RT-PCR). In one embodiment, the assay is a regulatory-approved assay, e.g., an FDA-approved kit.

[0254] In one embodiment, the biopsy is a tumor biopsy (e.g., a tumor sample obtained during conventional surgery, or a stereotactic needle biopsy, such as a stereotactic needle biopsy guided by a CT or MRI scan).Tissue biopsy methods can be used to detect total tumor tissue burden and / or BRAF mutations and / or BRAF fusions.

[0255] In one embodiment, BRAF mutations or fusions can be identified using liquid biopsy (variously referred to as fluid biopsy or fluid-phase biopsy). See, for example, Karachialiou et al., "Real-time liquid biopsies become a reality in cancer treatment," Ann. Transl. Med., 3(3):36, 2016. Liquid biopsy methods can be used to detect total tumor burden and / or BRAF mutations. Liquid biopsies can be performed on biological samples that are relatively easy to obtain from a subject (e.g., by a simple blood draw) and are generally less invasive than traditional methods used to detect tumor burden and / or BRAF mutations. In one embodiment, liquid biopsies can be used to detect the presence of BRAF mutations at an earlier stage than traditional methods. In one embodiment, biological samples to be used in liquid biopsies can include CSF, blood, plasma, urine, saliva, sputum, bronchoalveolar lavage fluid, bile, lymph, cyst fluid, feces, ascites, and combinations thereof. In one embodiment, liquid biopsy can be used to detect circulating tumor cells (CTCs). In one embodiment, liquid biopsy can be used to detect cell-free DNA. In one embodiment, the cell-free DNA detected using liquid biopsy is circulating tumor DNA (ctDNA) derived from tumor cells. Analysis of ctDNA (for example, using highly sensitive detection technology, such as but not limited to, next-generation sequencing (NGS), conventional PCR, digital PCR, or microarray analysis) can be used to identify BRAF mutations or BRAF fusions.

[0256] In one embodiment, the BRAF mutation or BRAF fusion identified using liquid biopsy is also present in cancer cells present in a subject (e.g., tumor). In one embodiment, any type of BRAF mutation or fusion can be detected using liquid biopsy. In one embodiment, the genetic mutation identified by liquid biopsy can be used to identify a subject as a candidate for a specific treatment.

[0257] "Tumor burden," also known as "tumor mass," refers to the total amount of tumor material distributed throughout the body. Tumor burden refers to the total number of cancer cells or the total size of tumor(s) in the body, including lymph nodes and bone marrow. Tumor burden can be determined by various methods known in the art, such as by measuring the dimensions of tumor(s) when removed from a subject, for example, using calipers, or, if present in the body, using imaging techniques, such as magnetic resonance imaging (MRI) scans, computed tomography (CT), multi-detector CT (MDCT), positron emission tomography (PET), X-rays, ultrasound, or bone scans.

[0258] The term "tumor size" refers to the total size of the tumor, which can be measured as the length and width of the tumor.Tumor size can be determined by various methods known in the art, such as by measuring the dimensions of tumor(s) when removed from the subject, for example, using calipers, or when in the body, using imaging techniques, such as MRI scan, bone scan, ultrasound or CT.

[0259] Liquid biopsies can be performed at multiple time points during a diagnostic, monitoring, and / or treatment process to determine one or more clinically relevant parameters, including, but not limited to, disease progression or treatment effectiveness after administration of a treatment to a subject. For example, during a diagnostic, monitoring, and / or treatment process, a first liquid biopsy can be performed at a first time point, and a second liquid biopsy can be performed at a second time point. In one embodiment, the first time point can be a time point before the subject is diagnosed with a disease (e.g., when the subject is healthy), and the second time point can be a time point after the subject has developed a disease (e.g., the second time point can be used to diagnose the subject as having a disease). In one embodiment, the first time point can be a time point before the subject is diagnosed with a disease (e.g., when the subject is healthy), and the subject is then monitored, and the second time point can be a time point after the subject has been monitored. In one embodiment, the first time point can be a time point after the subject is diagnosed with a disease, and then a treatment is administered to the subject, and the second time point can be a time point after the treatment is administered. In such a case, the second time point can be used to assess the effectiveness of the treatment (e.g., if the genetic variation(s) detected at the first time point are significantly reduced or undetectable).

[0260] The compounds of Formula I or a pharmaceutically acceptable salt thereof or the compounds of Formula II or a pharmaceutically acceptable salt thereof can be used alone or in combination with one or more different forms of treatment to treat a subject having abnormal cell growth, e.g., a MEK-associated tumor, e.g., a MEK-associated cancer.

[0261] In one embodiment, a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, can be used in combination with one or more, e.g., one or more additional anti-cancer treatments, e.g., one or more treatments independently selected from surgery, radiation therapy, and anti-cancer agents that operate by the same or a different mechanism of action. In one embodiment, treatment of a subject having a MEK-associated cancer with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, in combination with one or more, e.g., one or more additional treatments independently selected from one or more of surgery, radiation therapy, and anti-cancer agents (e.g., any of the anti-cancer agents described herein below, wherein the anti-cancer agent is other than a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof), may have increased therapeutic efficacy compared to treatment of the same or similar subject with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, as monotherapy. When combination therapy is used, and one or more, for example, one, two, or three, anti-cancer treatments are independently selected from one or more anti-cancer agents, for example, any of the anti-cancer agents disclosed herein, the anti-cancer agent(s) can be administered simultaneously or separately, with variable intervening time limits, in any order with the compound of the present invention, and using various administration schedules in any order. In one embodiment, the anti-cancer agent(s) are administered to the subject before the administration of the compound of the present invention. In another embodiment, the anti-cancer agent(s) are administered to the subject after the administration of the compound of the present invention. In another embodiment, the anti-cancer agent(s) are administered to the subject simultaneously with the administration of the compound of the present invention. In one embodiment, the compound of Formula I or a pharmaceutically acceptable salt thereof or the compound of Formula II or a pharmaceutically acceptable salt thereof is used in combination with one additional anti-cancer treatment, which is surgery, radiation therapy, or an anti-cancer agent that acts by the same or a different mechanism of action.

[0262] Thus, in one embodiment, provided herein is a method of treating a subject having a MEK-associated tumor (e.g., any of the MEK-associated tumors described herein), comprising administering to the subject a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, in combination with one or more additional anti-cancer therapies. In one embodiment, the anti-cancer treatment is one or more anti-cancer agents other than a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the anti-cancer treatment is one anti-cancer agent other than a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the additional anti-cancer treatment is surgery. In one embodiment, the additional anti-cancer treatment is radiation therapy.

[0263] Also provided herein are compounds of Formula I or pharmaceutically acceptable salts thereof, or compounds of Formula II or pharmaceutically acceptable salts thereof, for use in combination with one or more, e.g., one or more anti-cancer treatments. In one embodiment, the additional anti-cancer treatments are selected from one or more treatments independently selected from surgery, radiation therapy, and / or one or more anti-cancer agents that operate by the same or a different mechanism of action.

[0264] Also provided herein are one or more, e.g., one or more anti-cancer therapies, for use in combination with a compound of Formula I, or a pharmaceutically acceptable salt thereof, or a compound of Formula II, or a pharmaceutically acceptable salt thereof. In one embodiment, the additional anti-cancer therapies are independently selected from one or more therapies independently selected from surgery, radiation therapy, and / or one or more anti-cancer agents that operate by the same or a different mechanism of action.

[0265] Also provided herein are compounds of Formula I or a pharmaceutically acceptable salt thereof, or compounds of Formula II or a pharmaceutically acceptable salt thereof, for use in combination with one or more, e.g., one or more additional anti-cancer therapies, to treat MEK-associated tumors.

[0266] Also provided herein are one or more, e.g., one or more additional anti-cancer therapies, for use in treating MEK-associated tumors by co-administration with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof.

[0267] In one embodiment for treating a subject having a MEK-associated tumor, the subject is administered one or more anti-cancer therapies other than the compound of Formula I or a pharmaceutically acceptable salt thereof, or the compound of Formula II or a pharmaceutically acceptable salt thereof, prior to administration of the compound of Formula I or a pharmaceutically acceptable salt thereof, or the compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the one or more anti-cancer therapies are selected from surgery, radiation therapy, and anti-cancer agents that act by the same or a different mechanism of action. For example, in one embodiment, the subject in need thereof may undergo at least partial resection of the tumor prior to administration of the compound of Formula I or a pharmaceutically acceptable salt thereof, or the compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, treatment with at least partial resection of the tumor reduces tumor size (e.g., tumor burden) prior to administration of one or more doses of the compound of Formula I or a pharmaceutically acceptable salt thereof, or the compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, a subject in need thereof may undergo radiation therapy prior to administration of the compound of Formula I, or a pharmaceutically acceptable salt thereof, or the compound of Formula II, or a pharmaceutically acceptable salt thereof. In one embodiment, a subject in need thereof may undergo treatment with one or more anti-cancer agents other than the compound of Formula I, or a pharmaceutically acceptable salt thereof, or the compound of Formula II, or a pharmaceutically acceptable salt thereof, prior to administration of the compound of Formula I, or a pharmaceutically acceptable salt thereof, or the compound of Formula II, or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has cancer that is refractory or intolerant to one or more previous treatments.

[0268] Thus, in one embodiment, provided herein is a method of treating a subject having a MEK-associated tumor, the method comprising: (i) administering one or more, e.g., one or more, anti-cancer therapies to the subject; and (ii) after (i), (a) administering a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof as monotherapy, or (b) administering a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof in combination with one or more, e.g., one or more, additional anti-cancer therapies. In one embodiment, the additional anti-cancer therapies are independently selected from one or more of surgery, radiation therapy, and / or one or more anti-cancer agents that operate via the same or a different mechanism of action. In one embodiment, the additional anti-cancer therapies are one or more anti-cancer agents that operate via the same or a different mechanism of action. In one embodiment, the additional anti-cancer therapies are a single anti-cancer agent that operates via the same or a different mechanism of action. In one embodiment, the additional anti-cancer therapies are surgery. In one embodiment, the additional anti-cancer treatment is radiation therapy.

[0269] Non-limiting examples of additional anti-cancer agents that can be used in combination with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, according to any of the combination treatment methods described herein include additional kinase inhibitors other than the compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, including MEK inhibitors, BRAF inhibitors, EGFR inhibitors, inhibitors of HER2 and / or HER3, SHP2 inhibitors, Axl inhibitors, PI3K inhibitors, SOS1 inhibitors, signal transduction pathway inhibitors, checkpoint inhibitors, modulators of apoptosis pathways, cytotoxic chemotherapeutic agents, angiogenesis-targeted therapies, and immune-targeted agents, including immunotherapies.

[0270] In one embodiment, the anti-cancer drug that can be used in combination with the compound of Formula I or its pharmaceutically acceptable salt or the compound of Formula II or its pharmaceutically acceptable salt according to any of the combination therapies described herein is a targeted therapeutic agent.As used herein, "targeted therapeutic agent" refers to a molecule that blocks the growth of cancer cells by interfering with specific targeted molecules required for carcinogenesis and tumor growth, rather than simply interfering with all rapidly dividing cells (for example, using conventional cytotoxic chemotherapy), and includes but is not limited to receptor tyrosine kinase targeted therapeutic agents, signal transduction pathway inhibitors (for example, Ras-Raf-MEK-ERK pathway inhibitors, PI3K-Akt-mTOR-S6K pathway inhibitors ("PI3K inhibitors")), and apoptosis pathway modulators.

[0271] In one embodiment, the anti-cancer drug that can be used in combination with the compound of Formula I or a pharmaceutically acceptable salt thereof or the compound of Formula II or a pharmaceutically acceptable salt thereof according to any of the combination therapies described herein is a BRAF inhibitor.Non-limiting examples of other BRAF inhibitors include encorafenib, dabrafenib, vemurafenib, N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide (PLX4720), and (3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394), and pharmaceutically acceptable salts thereof, such as: N-(3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-2,4-difluorophenyl)propane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropropane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4,5-difluorophenyl)propane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)propane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamide; N-(2-chloro-4-fluoro-3-((5-methyl-3-(methyl-d3)-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-phenyl)-3-fluoropropane-1-sulfonamide; N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}propane-1-sulfonamide; N-(3-chloro-4-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy)-5-fluoropyridin-2-yl)propane-1-sulfonamide; and N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}-3-fluoropropane-1-sulfonamide; or a pharmaceutically acceptable salt thereof, and compounds disclosed in PCT Publication WO2021 / 250521 published December 6, 2021, including, for example, N-(2-chloro-4-fluoro-3-((5-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-2-azabicyclo[2.1.1]hexane-2-sulfonamide, (R)-N-(2-chloro-4-fluoro-3-((5-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropyrrolidine-1-sulfonamide, and N-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamide, or a pharmaceutically acceptable salt thereof. Examples include:

[0272] In one embodiment, the BRAF inhibitor is selected from encorafenib or a pharmaceutically acceptable salt thereof, N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamide or a pharmaceutically acceptable salt thereof, and N-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamide or a pharmaceutically acceptable salt thereof.

[0273] In one embodiment, the BRAF inhibitor is encorafenib or a pharmaceutically acceptable salt thereof. In one embodiment, the BRAF inhibitor is N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamide or a pharmaceutically acceptable salt thereof. In one embodiment, the BRAF inhibitor is N-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamide or a pharmaceutically acceptable salt thereof.

[0274] Further examples of BRAF inhibitors are known in the art.

[0275] In one embodiment, the anti-cancer agent that may be used in combination with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, according to any of the combination treatment methods described herein is an EGFR inhibitor. Non-limiting examples of EGFR inhibitors include cetuximab (Erbitux®), panitumumab (Vectibix®), osimertinib (merelectinib, Tagrisso®), erlotinib (Tarceva®), gefitinib (Iressa®), necitumumab (Portrazza™), neratinib (Nerlynx®), lapatinib (Tykerb®), vandetanib (Caprelsa®), brigatinib (Alunbrig®), and inhibitors of EGFR disclosed in PCT Publications WO2019 / 071351 and WO2017 / 117680. Further examples of EGFR inhibitors are known in the art.

[0276] In one embodiment, the anti-cancer agent that may be used in combination with a compound of Formula I, or a pharmaceutically acceptable salt thereof, or a compound of Formula II, or a pharmaceutically acceptable salt thereof, according to any of the combination treatment methods disclosed herein is an SHP2 inhibitor. Non-limiting examples of SHP2 inhibitors include 6-(4-amino-4-methylpiperidin-1-yl)-3-(2,3-dichlorophenyl)pyrazin-2-amine (SHP099), [3-[(3S,4S)-4-amino-3-methyl-2-oxa-8-azaspiro[4.5]decan-8-yl]-6-(2,3-dichlorophenyl)-5-methylpyrazin-2-yl]methanol (RMC-4550) RMC-4630, TNO155, and compounds disclosed in WO2020 / 081848, WO2020 / 201991, WO2015 / 107493, WO2015 / 107494, WO2015 / 107495, and WO2019 / 075265. In one embodiment, the SHP2 inhibitor is a compound disclosed in WO 2020 / 201991. In one embodiment, the SHP2 inhibitor is (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine or a pharmaceutically acceptable salt thereof.

[0277] In one embodiment, the anti-cancer agent that can be used in combination with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, according to any of the combination treatment methods disclosed herein is a PI3K inhibitor.Non-limiting examples include buparlisib (BKM120), alpelisib (BYL719), samotricisib (LY3023414), 8-[(1R)-1-[(3,5-difluorophenyl)amino]ethyl]-N,N-dimethyl-2-(morpholin-4-yl)-4-oxo-4H-chromene-6-carboxamide (AZD8186), tenalisib (RP6530), voxtalisib hydrochloride (SAR-245409), gedatricisib (PF-05212384), panulisib (P-7170), taselisib (GDC-0032), trans-2-amino-8-[4-(2-hydroxyethoxy)cyclohexyl]-6-(6-methoxypyridin-3-yl)-4-methylpyrido[2,3-d]pyrimidin-7(8H)-one (PF-04691502), duvelisib (ABBV-954), N2-[4-oxo-4-[4-(4-oxo-8-phenyl-4H-1-benzopyran-2-yl)morpholin-4-ium-4-ylmethoxy]butyryl]-L-arginyl-glycyl-L-aspartate L-serine acetate (SF-1126), pictilisib (GDC-0941), 2-methyl-1-[2-methyl-3-(trifluoromethyl)benzyl]-6-(morpholin-4-yl)-1H-benzimidazole-4-carboxylic acid (GSK2636771), idelalisib (GS-1101), umbralisib tosylate (TGR-1202), pictilisib (GDC-0941), copanlisib hydrochloride (BAY84-1236), dactolisib (BEZ-235), 1-(4-[5-[5-amino-6-(5- tert-butyl-1,3,4-oxadiazol-2-yl)pyrazin-2-yl]-1-ethyl-1H-1,2,4-triazol-3-yl]piperidin-1-yl)-3-hydroxypropan-1-one (AZD-8835), 5-[6,6-dimethyl-4-(morpholin-4-yl)-8,9-dihydro-6H-[1,4]oxazino[4,3-e]purin-2-yl]pyrimidin-2-amine (GDC-0084), everolimus, rapamycin, perifosine, sirolimus, and temsirolimus.

[0278] In one embodiment, the anti-cancer agent that can be used in combination with a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof according to any of the combination therapies disclosed herein is an immunotherapy. The term "immunotherapy" refers to an agent that modulates the immune system. In one embodiment, immunotherapy can increase the expression and / or activity of immune system regulators. In one embodiment, immunotherapy can reduce the expression and / or activity of immune system regulators. In one embodiment, immunotherapy can mobilize and / or enhance the activity of immune cells.

[0279] In one embodiment, the immunotherapy that can be used in combination with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, according to any of the combination treatment methods disclosed herein is an antibody therapy (e.g., a monoclonal antibody, a conjugated antibody). In one embodiment, the antibody therapy is selected from the group consisting of bevacizumab (Mvasti™, Avastin®), trastuzumab (Herceptin®), rituximab (MabThera™, Rituxan®), edrecolomab (Panorex), daratumumab (Darzalex®), olaratumumab (Lartruvo™), ofatumumab (Arzerra®), alemtuzumab (Campath®), cetuximab (Erbitux®), oregovomab, pembrolizumab (Keytruda®), dinutuximab (Unituxin®), obinutuzumab (Gazyva®), tremelimumab (CP-675,206), ramucirumab (C yramza®), ublituximab (TG-1101), panitumumab (Vectibix®), elotuzumab (Empliciti™), necitumumab (Portrazza™), cirmtuzumab (UC-961), ibritumomab (Zevalin®), isatuximab (SAR650984), NEMO tuzumab, fresolimumab (GC1008), lirilumab (INN), mogamulizumab (Poteligeo®), ficlatuzumab (AV-299), denosumab (Xgeva®), ganitumab, urelumab, pidilizumab, amatuximab, blinatumomab (AMG103; Blincyto®), or midostaurin (Rydapt).

[0280] In one embodiment, the immunotherapy that can be used in combination with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, according to any of the combination therapies disclosed herein, is an immune checkpoint inhibitor. In one embodiment, the immunotherapy includes one or more, for example, one or two, immune checkpoint inhibitors. In one embodiment, the immune checkpoint inhibitor is a CTLA-4 inhibitor, a PD-1 inhibitor, or a PD-L1 inhibitor. In one embodiment, the CTLA-4 inhibitor is ipilimumab (Yervoy®) or tremelimumab (CP-675,206). In one embodiment, the PD-1 inhibitor is pembrolizumab (Keytruda®), nivolumab (Opdivo®), or sasanlimab (RN888). In one embodiment, the PD-L1 inhibitor is atezolizumab (Tecentriq®) or durvalumab (Imfinzi™).

[0281] In one embodiment, the anti-cancer treatment that can be used in combination with the compound of Formula I or a pharmaceutically acceptable salt thereof or the compound of Formula II or a pharmaceutically acceptable salt thereof according to any of the combination treatment methods disclosed herein is radiation therapy.Non-limiting examples of radiation therapy include external beam radiation therapy (e.g., external beam radiation therapy using kilovoltage X-rays or megavoltage X-rays) or internal radiation therapy.Internal radiation therapy (also known as brachytherapy) can include, for example, the use of low-dose internal radiation therapy or high-dose internal radiation therapy.Low-dose internal radiation therapy includes, for example, inserting small radioactive pellets into or adjacent to the cancerous tissue of the subject.High-dose internal radiation therapy includes, for example, inserting a long, thin tube (e.g., a catheter) or implant into or adjacent to the cancerous tissue of the subject and using a radiation machine to deliver a high dose of radiation to the long, thin tube or implant.Methods for administering radiation therapy to a subject with cancer are known in the art. In embodiments in which the tumor is a CNS tumor, radiation therapy may include whole brain radiation therapy (WBRT) or stereotactic radiosurgery (SRS), such as Cyberknife®, XKnife®, Gamma knife®, or ExacTrac®.

[0282] In one embodiment, the anti-cancer treatment that can be used in combination with the compound of Formula I or its pharmaceutically acceptable salt or the compound of Formula II or its pharmaceutically acceptable salt according to any of the combination treatment methods disclosed herein is surgery.Non-limiting examples of surgery include, for example, open surgery or minimally invasive surgery.Surgery can include, for example, at least partial resection of tumor, removal of the entire tumor, tumor debulking, or removal of tumor that is causing pain or pressure in the subject.Methods for performing open surgery and minimally invasive surgery on cancer-bearing subjects are known in the art.

[0283] In one embodiment, provided herein is a method of treating a MEK-associated tumor (e.g., any of the MEK-associated tumors described herein), comprising administering to a subject in need thereof, in any order, together or separately, therapeutically effective amounts of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, and a BRAF inhibitor (e.g., any of the BRAF inhibitors disclosed herein). In one embodiment, the compound of Formula I is a compound selected from Examples 1-69, or a pharmaceutically acceptable salt thereof.

[0284] In one embodiment, provided herein is a method of treating a MEK-associated tumor (e.g., any of the MEK-associated tumors described herein), comprising administering to a subject in need thereof, in any order, together or separately, therapeutically effective amounts of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, and an EGFR inhibitor (e.g., any of the EGFR inhibitors disclosed herein). In one embodiment, the compound of Formula I is a compound selected from any one of Examples 1-69, or a pharmaceutically acceptable salt thereof.

[0285] In one embodiment, provided herein is a method of treating a MEK-associated tumor (e.g., any of the MEK-associated tumors described herein), comprising administering to a subject in need thereof, in any order, together or separately, therapeutically effective amounts of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, and an SHP2 inhibitor (e.g., any of the SHP2 inhibitors disclosed herein). In one embodiment, the compound of Formula I is a compound selected from any one of Examples 1-69, or a pharmaceutically acceptable salt thereof.

[0286] In one embodiment, provided herein is a method of treating a MEK-associated tumor (e.g., any of the MEK-associated tumors described herein), comprising administering to a subject in need thereof, in any order, together or separately, therapeutically effective amounts of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, and a checkpoint inhibitor (e.g., any of the checkpoint inhibitors disclosed herein). In one embodiment, the compound of Formula I is a compound selected from any one of Examples 1-69, or a pharmaceutically acceptable salt thereof.

[0287] Also provided herein is a pharmaceutical combination for treating MEK-associated tumors in a subject in need thereof, comprising: (a) a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, and (b) at least one additional anti-cancer agent (e.g., any of the exemplary additional anti-cancer agents described herein or known in the art), wherein the compound of Formula I or a pharmaceutically acceptable salt thereof, or the compound of Formula II or a pharmaceutically acceptable salt thereof, and the one or more, e.g., one or more additional anti-cancer agents, inhibit the progression of the tumor. Provided are (ii) pharmaceutical combinations, formulated separately for simultaneous or separate use for treatment, wherein the amounts of the compound of Formula I or a pharmaceutically acceptable salt thereof, or the compound of Formula II or a pharmaceutically acceptable salt thereof, and the amounts of the additional anti-cancer agent(s) are together effective to treat a tumor; (ii) uses of such combinations for the preparation of a medicament for the treatment of a tumor; and (iii) commercial packages or products comprising such combinations, such as combined preparations, for simultaneous, separate, or sequential use, and methods of treating a tumor in a subject in need thereof.

[0288] The term "pharmaceutical combination" as used herein refers to a non-fixed combination of active ingredients. The term "non-fixed combination" means that a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof and one or more, for example, one or more additional anti-cancer drugs, can be administered to a subject in need thereof simultaneously or separately, with a variable intervening time limit, in any order, and are formulated in separate compositions or dosages so that such administration results in an effective level of two or more compounds in the subject's body. This also applies to cocktail therapy, for example, the administration of three or more active ingredients. Similarly, the term "combination" refers to a non-fixed combination when referring to a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof in combination with one or more anti-cancer drugs.

[0289] Accordingly, also provided herein is a method of treating a MEK-associated tumor, comprising administering to a subject in need thereof a pharmaceutical combination for treating said tumor comprising (a) a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof, and (b) one or more, e.g., one or more additional anti-cancer agents for simultaneous, separate, or sequential use for treating the tumor, wherein the amounts of the compound of Formula I or a pharmaceutically acceptable salt thereof or the compound of Formula II or a pharmaceutically acceptable salt thereof and the additional anti-cancer agent(s) are together effective to treat the tumor.

[0290] In one embodiment, provided herein is a method of treating MEK-associated tumors (e.g., benign, malignant, or metastatic tumors), comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, wherein the subject has not been treated with one or more anti-cancer therapies, e.g., selected from one or more anti-cancer therapies independently selected from surgery, radiation therapy, and anti-cancer agents acting by the same or different mechanisms of action, prior to administration of the compound of Formula I or a pharmaceutically acceptable salt thereof, or the compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the patient has not been treated with an anti-cancer agent prior to administration of the compound of Formula I or a pharmaceutically acceptable salt thereof, or the compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the patient has not been treated with surgery prior to administration of the compound of Formula I or a pharmaceutically acceptable salt thereof, or the compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the patient has not been treated with radiation therapy prior to administration of said compound of formula I, or a pharmaceutically acceptable salt thereof, or compound of formula II, or a pharmaceutically acceptable salt thereof.

[0291] In one embodiment, provided herein is a method of treating a subject having a MEK-associated tumor (e.g., a benign, malignant, or metastatic tumor), comprising administering a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof, wherein the subject has been treated with previous or standard therapy (e.g., treatment with one or more anti-cancer agents other than the compound of Formula I or a pharmaceutically acceptable salt thereof and / or radiation therapy and / or surgery), and the MEK-associated tumor has become refractory or intolerant to said previous therapy. In one embodiment, the subject developed brain metastases during said previous treatment.

[0292] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject having metastatic melanoma (e.g., metastatic melanoma having a BRAF V600 mutation or a BRAF fusion) has received treatment with a BRAF inhibitor prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has been previously treated with a BRAF inhibitor selected from encorafenib, dabrafenib, vemurafenib, N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide, (3R)—N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394). In one embodiment, the subject has been previously treated with a BRAF inhibitor selected from encorafenib, dabrafenib, and vemurafenib. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the subject developed brain metastases during the previous treatment.

[0293] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject having metastatic melanoma (e.g., metastatic melanoma with a BRAF V600 mutation or a BRAF fusion) has received treatment with a BRAF inhibitor and a MEK inhibitor prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject is receiving a BRAF inhibitor selected from encorafenib, dabrafenib, vemurafenib, N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide, and (3R)—N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394), as well as binimetinib, trametinib, cobimetinib, selumetinib, pimacetonitrile, thiazolinone ... The subject has previously been treated with a MEK inhibitor selected from: luteinib, refametinib, N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide (PD-325901), 2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide (CI-1040), and 3-[2(R),3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione (TAK-733). In one embodiment, the subject has previously been treated with a BRAF inhibitor selected from encorafenib, dabrafenib, and vemurafenib, and a MEK inhibitor selected from binimetinib, trametinib, and cobimetinib. In one embodiment, the subject has been previously treated with encorafenib and binimetinib. In one embodiment, the subject has been previously treated with dabrafenib and trametinib. In one embodiment, the subject has been previously treated with vemurafenib and cobimetinib. In one embodiment, the subject has become refractory to said previous treatments.In one embodiment, the subject developed brain metastases during said previous treatment.

[0294] In one embodiment of the methods disclosed herein for treating a subject with a MEK-associated tumor, the subject with metastatic melanoma (e.g., metastatic melanoma with a BRAF V600 mutation or a BRAF fusion) has received treatment with one or more, e.g., one or two, checkpoint inhibitors (e.g., any of the checkpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and / or a PD-L1 inhibitor) prior to treatment with the compound of Formula I or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously been treated with one or more, e.g., one or two, checkpoint inhibitors independently selected from ipilimumab, nivolumab, and pembrolizumab. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the subject has developed brain metastases during the previous treatment.

[0295] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject having metastatic melanoma (e.g., metastatic melanoma having a BRAF V600 mutation or a BRAF fusion) has received treatment with one or more, e.g., one or two, PI3K inhibitors prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof.In one embodiment, the subject is receiving buparlisib (BKM120), alpelisib (BYL719), samotricisib (LY3023414), 8-[(1R)-1-[(3,5-difluorophenyl)amino]ethyl]-N,N-dimethyl-2-(morpholin-4-yl)-4-oxo-4H-chromene-6-carboxamide (AZD8186), tenalisib (RP6530), voxtalisib hydrochloride (SAR-245409), gedatricisib (PF-05212384), panulisib (P-7170), taselisib (GDC-0032) , trans-2-amino-8-[4-(2-hydroxyethoxy)cyclohexyl]-6-(6-methoxypyridin-3-yl)-4-methylpyrido[2,3-d]pyrimidin-7(8H)-one (PF-04691502), duvelisib (ABBV-954), N2-[4-oxo-4-[4-(4-oxo-8-phenyl-4H-1-benzopyran-2-yl)morpholin-4-ium-4-ylmethoxy]butyryl]-L-arginyl-glycyl-L-aspartyl-L-serine acetate (SF-1126), pictilisib (GDC-0941), 2-methyl-1-[2-methyl-3-(trifluoromethyl)benzyl]-6-(morpholin-4-yl)-1H-benzimidazole-4-carboxylic acid (GSK2636771), idelalisib (GS-1101), umbralisib tosylate (TGR-1202), pictilisib (GDC-0941), copanlisib hydrochloride (BAY84-1236), dactolisib (BEZ-235), 1-(4-[5-[5-amino-6-(5-tert-butyl-1,3,4-oxadiazol-2-yl)pyrazine and have been previously treated with one or more, for example one or two, PI3K inhibitors selected from: [6,6-dimethyl-4-(morpholin-4-yl)-8,9-dihydro-6H-[1,4]oxazino[4,3-e]purin-2-yl]pyrimidin-2-amine (GDC-0084), everolimus, rapamycin, perifosine, sirolimus, and temsirolimus.In one embodiment, the subject has been previously treated with buparlisib or alpelisib, either alone or in combination. In one embodiment, the subject has become refractory to said previous treatment. In one embodiment, the subject has developed brain metastases during said previous treatment. In one embodiment, the subject has developed brain metastases during said previous treatment.

[0296] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject having metastatic melanoma (e.g., metastatic melanoma having a BRAF V600 mutation or a BRAF fusion) has received treatment with a BRAF inhibitor and one or more, e.g., one or two, checkpoint inhibitors (e.g., any of the checkpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and / or a PD-L1 inhibitor) prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has been previously treated with a BRAF inhibitor selected from encorafenib, dabrafenib, vemurafenib, N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide, and (3R)—N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394), and one or more, e.g., one or two, checkpoint inhibitors independently selected from ipilimumab, nivolumab, and pembrolizumab. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the subject has developed brain metastases during the previous treatment.

[0297] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject having metastatic melanoma (e.g., metastatic melanoma having a BRAF V600 mutation or a BRAF fusion) has received treatment with a BRAF inhibitor, a MEK inhibitor, and one or more, e.g., one or two, checkpoint inhibitors (e.g., any of the checkpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and / or a PD-L1 inhibitor) prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject is receiving a BRAF inhibitor selected from encorafenib, dabrafenib, vemurafenib, N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide (PLX4720), and (3R)—N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394), binimetinib, trametinib, cobimetinib, selumetinib, pimasertib, refametinib, N-[2(R),3-dihydroxypropoxy]-3,4-difluoro have been previously treated with a MEK inhibitor selected from 2-(2-fluoro-4-iodophenylamino)benzamide (PD-325901), 2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide (CI-1040), and 3-[2(R),3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione (TAK-733), and one or more checkpoint inhibitors (e.g., any of the checkpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and / or a PD-L1 inhibitor).In one embodiment, the subject has previously been treated with one or more, for example, one or two, checkpoint inhibitors independently selected from the BRAF inhibitor selected from encorafenib, dabrafenib and vemurafenib, the MEK inhibitor selected from binimetinib, trametinib and cobimetinib, and ipilimumab, nivolumab and pembrolizumab.In one embodiment, the subject has become refractory to the previous treatment.In one embodiment, the subject has developed brain metastasis during the previous treatment.

[0298] In one embodiment of the methods disclosed herein for treating a subject with a MEK-associated tumor, the subject with metastatic melanoma (e.g., metastatic melanoma with a BRAF V600 mutation or BRAF fusion) has been treated with one or more, for example, one or two, alkylating agents prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has been previously treated with an alkylating agent selected from temozolomide, fotemustine, lomustine, and carmustine. In one embodiment, the subject has been previously treated with temozolomide. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the subject has developed brain metastases during the previous treatment.

[0299] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject having metastatic colorectal cancer (e.g., metastatic colorectal cancer having a BRAF V600 mutation or a BRAF fusion) has received treatment with a MEK inhibitor and one or more, e.g., one or two, checkpoint inhibitors (e.g., any of the checkpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and / or a PD-L1 inhibitor) prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof or Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject is receiving treatment with binimetinib, trametinib, cobimetinib, selumetinib, pimasertib, refametinib, N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide (PD-325901), 2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide (CI-1040), and 3-[2(R) ,3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione (TAK-733), and one or more checkpoint inhibitors (e.g., any of the checkpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and / or a PD-L1 inhibitor). In one embodiment, the subject has previously been treated with a MEK inhibitor selected from binimetinib, trametinib, and cobimetinib, and one or more, e.g., one or two, checkpoint inhibitors independently selected from ipilimumab, nivolumab, and pembrolizumab. In one embodiment, the subject has previously been treated with a MEK inhibitor that is binimetinib, and a checkpoint inhibitor that is nivolumab, ipilimumab, or pembrolizumab. In one embodiment, the subject has become refractory to said previous treatment. In one embodiment, the subject has developed brain metastases during said previous treatment.

[0300] In one embodiment of the methods disclosed herein for treating a subject with a MEK-associated tumor, the subject with metastatic colorectal cancer (e.g., metastatic colorectal cancer with a BRAF V600 mutation or a BRAF fusion) has received treatment with one or more, e.g., one or two, checkpoint inhibitors (e.g., any of the checkpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and / or a PD-L1 inhibitor) prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously been treated with one or more, e.g., one or two, checkpoint inhibitors independently selected from ipilimumab, nivolumab, pembrolizumab, and sasanlimab. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the subject has developed brain metastases during the previous treatment.

[0301] In one embodiment of the methods disclosed herein for treating a subject with a MEK-associated tumor, the subject with metastatic colorectal cancer (e.g., a mutant metastatic colorectal cancer with a BRAF V600 mutation or a BRAF fusion) has received treatment with one or more cytotoxic chemotherapeutic agents prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has received treatment with oxaliplatin, irinotecan, FOLFOXIRI (oxaliplatin, irinotecan, and fluorouracil), FOLFIRI (folinic acid, fluorouracil, and irinotecan), or CAPEOX (capecitabine and oxaliplatin) prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the subject has developed brain metastases during the previous treatment.

[0302] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject having metastatic colorectal cancer (e.g., metastatic colorectal cancer having a BRAF V600 mutation or a BRAF fusion) has received treatment with an EGFR inhibitor, a BRAF inhibitor, and one or more cytotoxic chemotherapeutic agents prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject with metastatic colorectal cancer is treated with an EGFR inhibitor selected from cetuximab, panitumumab, osimertinib, erlotinib, gefitinib, necitumumab, neratinib, lapatinib, vandetanib, and brigatinib, encorafenib, dabrafenib, vemurafenib, N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl methyl]propanol, benzodiazepine, benzocaine ... The subject has previously received treatment with a BRAF inhibitor selected from (3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX4720), and (3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394), and one or more cytotoxic chemotherapeutic agents. In one embodiment, the subject has previously received treatment with an EGFR inhibitor selected from cetuximab and panitumumab, a BRAF inhibitor that is vemurafenib, and a cytotoxic chemotherapeutic agent that is irinotecan. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the subject developed brain metastases during the previous treatment.

[0303] In one embodiment of the methods disclosed herein for treating a subject with a MEK-associated tumor, the subject with metastatic colorectal cancer (e.g., metastatic colorectal cancer with a BRAF V600 mutation or a BRAF fusion) has received treatment with an EGFR inhibitor and one or more cytotoxic chemotherapeutic agents prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously received treatment with an EGFR inhibitor selected from cetuximab, panitumumab, osimertinib, erlotinib, gefitinib, necitumumab, neratinib, lapatinib, vandetanib, and brigatinib, and one or more chemotherapeutic agents. In one embodiment, the subject has previously received treatment with an EGFR inhibitor selected from cetuximab and panitumumab, and a cytotoxic chemotherapeutic agent that is irinotecan or FOLFIRI (folinic acid, fluorouracil, and irinotecan). In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the subject has developed brain metastases during the previous treatment.

[0304] In one embodiment of the method disclosed herein for treating a subject with a MEK-associated tumor, the subject with metastatic non-small cell lung cancer (e.g., metastatic non-small cell lung cancer with a BRAF V600 mutation or BRAF fusion) has been treated with one or more, for example, one or two, EGFR inhibitors before treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously been treated with one or more, for example, one or two, EGFR inhibitors independently selected from cetuximab, panitumumab, osimertinib, erlotinib, gefitinib, necitumumab, neratinib, lapatinib, vandetanib, and brigatinib. In one embodiment, the subject has previously been treated with erlotinib. In one embodiment, the subject has previously been treated with gefitinib. In one embodiment, the subject has previously been treated with erlotinib and gefitinib. In one embodiment, the subject has become refractory to said previous treatment. In one embodiment, the subject has developed brain metastases during said previous treatment.

[0305] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject having metastatic non-small cell lung cancer (e.g., metastatic non-small cell lung cancer with a BRAF mutation) is administered one or more of the following: encorafenib, dabrafenib, vemurafenib, N-[3-(5-chloro-1H-pyrrolo[2, and (3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)propane-1-sulfonamide (PLX4720), and (3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394). Lametinib, cobimetinib, selumetinib, pimasertib, refametinib, N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide (PD-325901), 2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide (CI-1040), and 3-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide (CI-1040). The subject has previously been treated with a MEK inhibitor selected from [pyr]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione (TAK-733), and an EGFR inhibitor selected from cetuximab, panitumumab, osimertinib, erlotinib, gefitinib, necitumumab, neratinib, lapatinib, vandetanib, and brigatinib. In one embodiment, the subject has previously been treated with a BRAF inhibitor selected from vemurafenib, dabrafenib, and encorafenib, and an EGFR inhibitor selected from cetuximab and panitumumab, prior to treatment with the compound of Formula I or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the subject has developed brain metastases during the previous treatment.

[0306] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject having metastatic thyroid cancer (e.g., metastatic thyroid cancer having a BRAF V600 mutation or a BRAF fusion) is administered one or more of the following: encorafenib, dabrafenib, vemurafenib, N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonyl]propan-1-yl, N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-yl]propan ... BRAF inhibitors selected from (3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX4720), and (3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394), binimetinib, trametinib, cobimetinib, selumetinib, pimasertib, and leprae Fametinib, N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide (PD-325901), 2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide (CI-1040), and 3-[2(R),3-dihydroxypropyl]-6-fluoro-5-( The subject has previously been treated with a MEK inhibitor selected from 2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione (TAK-733) and an EGFR inhibitor selected from cetuximab, panitumumab, osimertinib, erlotinib, gefitinib, necitumumab, neratinib, lapatinib, vandetanib, and brigatinib. In one embodiment, the subject has previously been treated with a BRAF inhibitor selected from vemurafenib, dabrafenib, and encorafenib before treatment with the compound of Formula I or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the subject has developed brain metastases during the previous treatment.

[0307] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject has a LMD and was previously treated with a BRAF inhibitor and one or more, e.g., one or two, checkpoint inhibitors (e.g., any of the checkpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and / or a PD-L1 inhibitor) prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has been previously treated with a BRAF inhibitor selected from encorafenib, dabrafenib, vemurafenib, N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide (PLX4720), and (3R)—N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394), and one or more, e.g., one or two, checkpoint inhibitors independently selected from ipilimumab, nivolumab, and pembrolizumab. In one embodiment, the subject has become refractory to said previous treatment.

[0308] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject has a LMD and was previously treated with a BRAF inhibitor, a MEK inhibitor, and one or more, e.g., one or two, checkpoint inhibitors (e.g., any of the checkpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and / or a PD-L1 inhibitor) prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject is receiving treatment with a BRAF inhibitor selected from encorafenib, dabrafenib, vemurafenib, N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide (PLX4720), and (3R)—N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394), binimetinib, trametinib, cobimetinib, selumetinib, pimasertib, refametinib, N-[2(R),3-dihydroxypropoxy]-3,4-di had previously been treated with a MEK inhibitor selected from fluoro-2-(2-fluoro-4-iodophenylamino)benzamide (PD-325901), 2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide (CI-1040), and 3-[2(R),3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione (TAK-733), and a checkpoint inhibitor (e.g., any of the checkpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and / or a PD-L1 inhibitor).In one embodiment, the subject has previously been treated with one or more, for example, one or two, checkpoint inhibitors independently selected from a BRAF inhibitor selected from encorafenib, dabrafenib, and vemurafenib, a MEK inhibitor selected from binimetinib, trametinib, and cobimetinib, and ipilimumab, nivolumab, and pembrolizumab. In one embodiment, the subject has become refractory to the previous treatment.

[0309] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject has LMD and has previously been treated with one or more, e.g., one or two, checkpoint inhibitors (e.g., any of the checkpoint inhibitors disclosed herein, e.g., a CTLA-4 inhibitor, a PD-1 inhibitor, and / or a PD-L1 inhibitor) prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously been treated with one or more, e.g., one or two, checkpoint inhibitors independently selected from ipilimumab, nivolumab, and pembrolizumab. In one embodiment, the subject has become refractory to the previous treatment.

[0310] In one embodiment of the methods disclosed herein for treating a subject with a MEK-associated tumor, the subject has a glioma and has previously been treated with surgery prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the glioma is a grade 2, grade 3, or grade 4 glioma.

[0311] In one embodiment of the methods disclosed herein for treating a subject with a MEK-associated tumor, the subject has a glioma and has previously been treated with radiation therapy (e.g., whole-brain radiation therapy or stereotactic radiosurgery) prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the glioma is a grade 2, grade 3, or grade 4 glioma.

[0312] In one embodiment of the method disclosed herein for treating a subject with a MEK-associated tumor, the subject has a glioma and has previously been treated with one or more cytotoxic chemotherapeutic agents prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously been treated with one or more cytotoxic chemotherapeutic agents independently selected from cisplatin, pemetrexed, vinorelbine, and paclitaxel. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the glioma is a grade 2, grade 3, or grade 4 glioma.

[0313] In one embodiment of the method disclosed herein for treating a subject with a MEK-associated tumor, the subject has a MEK-associated glioma and has previously been treated with an ornithine decarboxylase inhibitor before treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously been treated with an ornithine decarboxylase inhibitor that is eflornithine (racemate, or as the D or L enantiomer). In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the glioma is a grade 2, grade 3, or grade 4 glioma.

[0314] In one embodiment of the methods disclosed herein for treating a subject with a MEK-associated tumor, the subject has a MEK-associated glioma and has previously been treated with an alkylating agent prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously been treated with an alkylating agent selected from temozolomide, lomustine, and carmustine. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the glioma is grade 2, grade 3, or grade 4 glioma.

[0315] In one embodiment of the method disclosed herein for treating a subject with a MEK-associated tumor, the subject has a MEK-associated glioma and has previously been treated with an alkylating agent and an ornithine decarboxylase inhibitor before treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously been treated with an alkylating agent selected from temozolomide, lomustine, and carmustine, and an ornithine decarboxylase inhibitor that is eflornithine (racemic or as D or L enantiomer). In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the glioma is grade 2, grade 3, or grade 4 glioma.

[0316] In one embodiment of the methods disclosed herein for treating a subject with a MEK-associated tumor, the subject has a MEK-associated glioma and has previously been treated with radiation therapy (e.g., whole-brain radiation therapy or stereotactic radiosurgery) and an alkylating agent prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously received radiation therapy (e.g., whole-brain radiation therapy or stereotactic radiosurgery) and treatment with an alkylating agent selected from temozolomide, lomustine, and carmustine. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the glioma is a grade 2, grade 3, or grade 4 glioma.

[0317] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject has a MEK-associated glioma and has previously been treated with an antibody therapy prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously received treatment with an antibody therapy that is bevacizumab. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the glioma is a grade 2, grade 3, or grade 4 glioma.

[0318] In one embodiment of the methods disclosed herein for treating a subject with a MEK-associated tumor, the subject has a MEK-associated glioma and has previously been treated with surgery and radiation therapy prior to treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the glioma is a grade 2, grade 3, or grade 4 glioma.

[0319] In one embodiment of the methods disclosed herein for treating a subject with a MEK-associated tumor, the subject has a MEK-associated glioma and has previously been treated with surgery, radiation therapy, and an alkylating agent before treatment with a compound of Formula I or a pharmaceutically acceptable salt thereof, a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously been treated with surgery, radiation therapy (e.g., whole-brain radiation therapy or stereotactic radiosurgery), and an alkylating agent selected from temozolomide, lomustine, and carmustine. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the glioma is a grade 2, grade 3, or grade 4 glioma.

[0320] In one embodiment of the method disclosed herein for treating a subject with a MEK-associated tumor, the subject has a MEK-associated glioma and has previously been treated with a BRAF inhibitor before treatment with a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has previously been treated with a BRAF inhibitor selected from N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide (PLX4720), vemurafenib, dabrafenib, encorafenib, and (3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394). In one embodiment, the subject has become refractory to said previous treatment. In one embodiment, the glioma is a grade 2, grade 3 or grade 4 glioma.

[0321] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject has a MEK-associated glioma and was previously treated with a BRAF inhibitor and a MEK inhibitor prior to treatment with a compound of Formula I, or a pharmaceutically acceptable salt, solvate, or polymorph thereof, or a compound of Formula II, or a pharmaceutically acceptable salt, solvate, or polymorph thereof. In one embodiment, the subject is receiving a BRAF inhibitor selected from N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide (PLX4720), vemurafenib, dabrafenib, encorafenib, and (3R)—N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394), as well as binimetinib, trametinib, cobimetinib, selumetinib, pimetinib, rifa ... The patient had previously received treatment with a MEK inhibitor selected from macertib, refametinib, N-[2(R),3-dihydroxypropoxy]-3,4-difluoro-2-(2-fluoro-4-iodophenylamino)benzamide (PD-325901), 2-(2-chloro-4-iodophenylamino)-N-(cyclopropylmethoxy)-3,4-difluorobenzamide (CI-1040), and 3-[2(R),3-dihydroxypropyl]-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione (TAK-733). In one embodiment, the subject has previously been treated with a BRAF inhibitor selected from encorafenib, dabrafenib, and vemurafenib, and a MEK inhibitor selected from binimetinib, trametinib, and cobimetinib. In one embodiment, the subject has become refractory to the previous treatment. In one embodiment, the glioma is grade 2, grade 3, or grade 4 glioma.

[0322] In one embodiment of the methods disclosed herein for treating a subject having a MEK-associated tumor, the subject has a MEK-associated brainstem ganglioglioma and was previously treated with a BRAF inhibitor prior to treatment with a compound of Formula I, or a pharmaceutically acceptable salt thereof, a compound of Formula II, or a pharmaceutically acceptable salt thereof. In one embodiment, the subject has been previously treated with a BRAF inhibitor selected from encorafenib, dabrafenib, vemurafenib, N-[3-(5-chloro-1H-pyrrolo[2,3-b]pyridin-3-ylcarbonyl)-2,4-difluorophenyl]propane-1-sulfonamide (PLX4720), and (3R)-N-(3-[[5-(2-cyclopropylpyrimidin-5-yl)-1H-pyrrolo[2,3-b]pyridin-3-yl]carbonyl]-2,4-difluorophenyl)-3-fluoropyrrolidine-1-sulfonamide (PLX8394). In one embodiment, the subject has been previously treated with a BRAF inhibitor selected from encorafenib, dabrafenib, and vemurafenib. In one embodiment, the subject has become refractory to said previous treatment.

[0323] Although the genetic basis of tumorigenesis may vary among different cancer types, the cellular and molecular mechanisms required for metastasis appear to be similar across all solid tumor types. During the metastatic cascade, cancer cells lose growth inhibitory responses, undergo altered adhesive properties, and produce enzymes capable of degrading extracellular matrix components. This allows tumor cells to detach from the original tumor and invade the circulation through newly formed vasculature, allowing them to migrate and extravasate to preferred distant sites where they can establish colonies. Several genes have been identified as promoters or suppressors of metastasis.

[0324] Accordingly, also provided herein is a method for treating, inhibiting, preventing, aiding in the prevention, or reducing metastasis of MEK-associated tumors in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt thereof, or a compound of Formula II or a pharmaceutically acceptable salt thereof. In one embodiment, the compound of Formula I or a pharmaceutically acceptable salt thereof is used in combination with one or more anti-cancer therapies independently selected from surgery (e.g., at least partial removal of the tumor), radiation therapy, and an anti-cancer agent.

[0325] As used herein, the term "treating metastases" means reducing the size, progression, and / or further spread of one or more metastases.

[0326] As used herein, the term "inhibiting metastasis" means reducing the occurrence (or recurrence) of one or more metastases, preventing the occurrence (or recurrence) of one or more metastases, or reducing the spread of one or more metastases.

[0327] In one embodiment, a subject treated according to any of the methods disclosed herein may be assessed according to one or more standard response assessment criteria known in the art, including RECIST (Response Evaluation Criteria in Solid Tumors, e.g., RECIST version 1.0, RECIST version 1.1, and modified RECIST 1.1 (mRECIST 1.1)), RANO-BM (Response Assessment in Neuro-Oncology Brain Metastases), Macdonald, RANO-LMD, and NANO (Neurologic Assessment in Neuro-Oncology). In one embodiment of any of the above criteria, the tumor is assessed by imaging study (e.g., MRI, CT, MDCT, or PET). In one embodiment, treatment response is assessed according to RECIST version 1.1, where complete response (CR) is defined as the complete disappearance of all tumor lesions, partial response (PR) is defined as at least a 30% reduction in the sum of tumor measurements, progressive disease (PD) is defined as at least a 20% increase in the sum of tumor measurements (the occurrence of new lesions or substantial progression of non-target lesions is also defined as PD), an increase of at least 5 mm from baseline is assessed as PD, and stable disease (SD) is defined as insufficient reduction in the smallest sum diameter during treatment to qualify as PR and not sufficient increase to qualify as PD. In one embodiment, assessments include intracranial response (assessed according to modified RECIST using gadolinium-enhanced MRI), extracranial response, global response rate, disease control rate (DCR), duration of response (DOR), progression-free survival (PFS), and overall survival (OS).

[0328] As used herein, an "effective dosage" or "effective amount" of a drug, compound, or pharmaceutical composition is an amount sufficient to produce any one or more beneficial or desired effects, including biochemical, histological, and / or behavioral symptoms of a disease, its complications, and intermediate pathological phenotypes manifesting during the development of a disease. For therapeutic use, a "therapeutically effective amount" refers to the amount of the compound administered that alleviates to some extent one or more symptoms of the disorder being treated. A therapeutically effective amount, when referring to the treatment of cancer, refers to an amount that has the effect of (1) reducing tumor size, (2) inhibiting (i.e., slowing to some extent, preferably stopping) tumor metastasis, (3) inhibiting to some extent (i.e., slowing to some extent, preferably stopping) tumor growth or tumor invasiveness, (4) alleviating to some extent (or preferably eliminating) one or more signs or symptoms associated with cancer, (5) reducing the dose of other medications required to treat the disease, and / or (6) enhancing the effect of another medication, and / or (7) slowing the progression of the disease in a patient.

[0329] An effective dose can be administered in one or multiple doses.For the purpose of the present invention, an effective dose of a drug, compound or pharmaceutical composition is an amount that is sufficient to directly or indirectly achieve preventive or therapeutic treatment.As understood in clinical context, the effective dose of a drug, compound or pharmaceutical composition may or may not be achieved together with another drug, compound or pharmaceutical composition.

[0330] A "pharmaceutical composition" refers to a mixture of one or more compounds of the present invention as the active ingredient, or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof, with at least one pharmaceutically acceptable carrier or excipient. In one embodiment, the pharmaceutical composition comprises two or more pharmaceutically acceptable carriers and / or excipients.

[0331] In one embodiment, the present invention provides a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier or excipient, hi one embodiment, the pharmaceutical composition comprises two or more pharmaceutically acceptable carriers and / or excipients.

[0332] Accordingly, in one embodiment, the present invention provides a pharmaceutical composition for use in treating abnormal cell growth in a subject in need thereof, comprising a compound of the present invention or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient.

[0333] As used herein, "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

[0334] Pharmaceutically acceptable carriers can include any conventional pharmaceutical carrier or excipient. The choice of carrier and / or excipient will vary greatly depending on factors such as the particular mode of administration, the effect of the carrier or excipient on solubility and stability, and the nature of the dosage form.

[0335] Suitable pharmaceutical carriers include inert diluents or fillers, water and various organic solvents (e.g., hydrates and solvates). Pharmaceutical compositions can contain additional ingredients, such as flavorings, binders, excipients, and the like, as desired.

[0336] The term "excipient" is used herein to describe any ingredient other than the compound(s) of the invention. The choice of excipient will largely depend on factors such as the mode of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form.

[0337] As used herein, "excipient" includes any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, carriers, diluents, etc. Examples of excipients include one or more of water, saline, phosphate buffer solution, glucose, glycerol, ethanol, etc., and combinations thereof, and the composition may include an isotonic agent, such as sugar, sodium chloride, or a polyhydric alcohol, such as mannitol or sorbitol. Examples of excipients also include various organic solvents (e.g., hydrates and solvates). Pharmaceutical compositions can optionally contain additional excipients, such as flavoring agents, binders / binding agents, lubricants, disintegrants, sweeteners or flavoring agents, coloring substances, or dyes. For example, for oral administration, tablets containing various excipients such as citric acid can be used with various disintegrants, such as starch, alginic acid, and certain complex silicates, and binders, such as sucrose, gelatin, and acacia. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycol. Thus, for oral administration, tablets containing various excipients such as citric acid can be used with various disintegrants, such as starch, alginic acid, and certain complex silicates, and binders, such as sucrose, gelatin, and acacia. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycol. In addition, lubricants, such as magnesium stearate, sodium lauryl sulfate, and talc, are often useful for tableting. Solid compositions of a similar type may also be employed in soft and hard-filled gelatin capsules, such materials including, by way of non-limiting example, lactose or milk sugar as well as high molecular weight polyethylene glycols.When aqueous suspensions or elixirs are desired for oral administration, the active compound therein can be combined with a diluent, for example, water, ethanol, propylene glycol, glycerin, or combinations thereof, along with various sweetening or flavoring agents, coloring substances or dyes, and, if desired, emulsifying or suspending agents.

[0338] Examples of excipients include pharmaceutically acceptable substances, such as wetting agents, which enhance the shelf life or effectiveness of the compound, or minor amounts of auxiliary substances such as wetting or emulsifying agents, preservatives, or buffers.

[0339] Pharmaceutical compositions may be in the form of tablets, capsules, pills, powders, sustained-release preparations, liquid suspensions, and other suitable forms for oral administration; liquid solutions suitable for parenteral injection (e.g., injectable insoluble solutions) as sterile solutions, suspensions, or emulsions; ointments or creams suitable for topical administration; powders, liposomes, and suppositories (e.g., rectal administration as suppositories). Exemplary parenteral administration forms include solutions or suspensions of active compounds in sterile aqueous solutions, such as aqueous propylene glycol or dextrose solutions. Such dosage forms can be suitably buffered if desired. The form depends on the intended administration method and therapeutic application.

[0340] The pharmaceutical composition may be in unit dosage forms suitable for single administration of precise dosage amounts.

[0341] The compound of the present invention can be administered orally.Oral administration can involve swallowing, which allows the compound to enter the gastrointestinal tract, or can be administered by oral buccal or sublingual administration, which allows the compound to enter the bloodstream directly from the mouth.Preparations suitable for oral administration include solid preparations, such as tablets, capsules containing microparticles, liquids or powders, lozenges (including those filled with liquid), chewable preparations, multi- and nanoparticulate preparations, gels, solid solutions, liposomes, films (including those with mucoadhesives), vaginal suppositories, sprays, and liquid preparations.Such capsules or tablets can contain controlled-release preparations.For capsules, tablets, and pills, dosage forms can also contain buffering agents or be prepared with enteric coatings.

[0342] Liquid preparations include suspensions, solutions, syrups and elixirs.Such preparations can be used as fillers in soft or hard capsules, and typically include carriers or adjuvants, such as wetting agents, emulsifying agents, suspending agents, flavoring agents (e.g., sweeteners) or flavoring agents, such as water, ethanol, polyethylene glycol, propylene glycol, methylcellulose, or suitable oils, and one or more emulsifying agents and / or suspending agents.Liquid preparations can also be prepared by reconstituting a solid, for example, from a sachet.

[0343] The compounds of the present invention may also be used in fast-dissolving, fast-disintegrating dosage forms, such as those described in Liang and Chen, Expert Opinion in Therapeutic Patents, 11(6), 981-986 (2001), the disclosure of which is incorporated herein by reference in its entirety.

[0344] For tablet dosage forms, the drug can comprise 1 wt% to 80 wt% of the dosage form, more typically 5 wt% to 60 wt%, depending on the dosage. Tablets generally contain a disintegrant in addition to the drug. Examples of disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, crospovidone, polyvinylpyrrolidone, methylcellulose, microcrystalline cellulose, lower alkyl-substituted hydroxypropylcellulose, starch, pregelatinized starch, and sodium alginate. Generally, disintegrants comprise 1 wt% to 25 wt%, preferably 5 wt% to 20 wt%, of the dosage form. Binders are commonly used to impart cohesive properties to tablet formulations. Suitable binders include microcrystalline cellulose, gelatin, sugars, polyethylene glycol, natural and synthetic gums, polyvinylpyrrolidone, pregelatinized starch, hydroxypropylcellulose, and hydroxypropylmethylcellulose. Tablets may also contain diluents such as lactose (monohydrate, spray-dried monohydrate, anhydrous, etc.), mannitol, xylitol, glucose, sucrose, sorbitol, microcrystalline cellulose, starch, and calcium hydrogen phosphate dihydrate. Tablets may also contain surfactants such as sodium lauryl sulfate and polysorbate 80, and glidants such as silicon dioxide and talc. When present, surfactants are typically present in amounts of 0.2 wt% to 5 wt% of the tablet, and glidants are typically present in amounts of 0.2 wt% to 1 wt% of the tablet. Tablets also generally contain lubricants such as magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate, and mixtures of magnesium stearate and sodium lauryl sulfate. Lubricants are generally present in an amount of 0.25 wt% to 10 wt%, preferably 0.5 wt% to 3 wt% of the tablet. Other conventional ingredients include antioxidants, colorants, flavoring agents, preservatives, and taste-masking agents. Tablet blends can be compressed directly or by roller to form tablets.Alternatively, tablet blends or portions of blends can be wet-, dry-, or melt-granulated, melt-congealed, or extruded before tableting. The final formulation may comprise one or more layers, and may be coated or uncoated, or encapsulated. Solid formulations for oral administration can be formulated for immediate and / or modified release. Modified release formulations include delayed-, sustained-pulse-, controlled-, targeted-, and programmed-release.

[0345] For oral administration, the compositions can be provided in tablet or capsule form containing 0.01, 0.05, 0.1, 0.25, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 75.0, or 100 milligrams of active ingredient, allowing for symptomatic adjustment of patient dosage. The medicament typically contains about 0.01 mg to about 100 mg of active ingredient. In another embodiment, the medicament contains about 0.01 to 0.25 mg of active ingredient. In another embodiment, the medicament contains about 0.25, 0.5, 1.0, 5.0, 15, or 25 mg of active ingredient.

[0346] The compounds of the present invention can also be administered directly into the bloodstream, muscle, or an internal organ. Suitable means for parenteral administration include intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, ​​intrasternal, intracranial, intramuscular, and subcutaneous. Suitable devices for parenteral administration include needle (including microneedle) injectors, needle-free injectors, and infusion techniques. Injectable preparations (i.e., injectable sterile aqueous or oily suspensions) can be formulated according to known techniques using one or more suitable dispersing agents, wetting agents, or suspending agents. Parenteral formulations are typically aqueous solutions that may contain excipients such as salts, carbohydrates, and buffers (preferably to achieve a pH of 3 to 9); however, for some applications, they may be more suitably formulated as sterile nonaqueous solutions or as a dry form to be combined with a suitable vehicle, such as sterile, pyrogen-free water. The preparation of parenteral formulations under sterile conditions, for example, by lyophilization, can be readily accomplished using standard pharmaceutical techniques well known to those skilled in the art. The solubility of compounds of the invention used in the preparation of parenteral solutions may be increased by the use of appropriate formulation techniques, such as the incorporation of solubility-enhancing agents.

[0347] The preparation for parenteral administration can be formulated to be immediate and / or modified release.Modified release preparations include delayed, sustained, pulsed, controlled, targeted and programmed release.Therefore, the compound of the present invention can be formulated as solid, semi-solid or thixotropic liquid to be administered as an implanted depot that provides modified release of active compound.Examples of such preparations include drug-coated stents and PGLA microspheres.

[0348] The compounds of the present invention can also be administered topically to the skin or mucosa, i.e., transdermally or transdermally, for example, by transdermal patches or iontophoresis devices, intraocular administration, or intranasally or by inhalation. Topical formulations can include compounds that enhance absorption or penetration of the active ingredient through the skin or other affected areas. When the compounds of the present invention are administered via a transdermal device, administration is achieved using a patch, either of the reservoir and porous membrane type or of the solid matrix type. Typical formulations for this purpose include gels, hydrogels, lotions, solutions, creams, ointments, dusting powders, dressings, foams, films, skin patches, wafers, implants, sponges, fibers, adhesive plasters, and microemulsions. Liposomes can also be used. Typical carriers include alcohol, water, mineral oil, liquid petrolatum, white petrolatum, glycerin, polyethylene glycol, and propylene glycol. Penetration enhancers may be incorporated. Other means of topical administration include delivery by electroporation, iontophoresis, phonophoresis, sonophoresis and microneedle or needle-free (eg Powderject™, Bioject™, etc.) injection.

[0349] Suitable formulations for topical administration to the eye include, for example, eye drops in which the compounds of the present invention are dissolved or suspended in a suitable vehicle. Typical formulations suitable for administration to the eye or ear may be in the form of droplets of a micronized suspension or solution in isotonic, pH-adjusted, sterile saline. Other formulations suitable for administration to the eye or ear include ointments, biodegradable (i.e., absorbent gel sponges, collagen) and non-biodegradable (i.e., silicone) implants, wafers, lenses, and microparticles, or vesicular systems such as niosomes or liposomes. Polymers such as cross-linked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, cellulose polymers such as hydroxypropylmethylcellulose, hydroxyethylcellulose, or methylcellulose, or heteropolysaccharide polymers such as gellan gum, together with preservatives such as benzalkonium chloride, can also be incorporated. Such formulations can also be delivered by iontophoresis.

[0350] Formulations for topical administration may be formulated to be immediate and / or modified release, including delayed-, sustained-, pulsed-, controlled-, targeted-, and programmed-release.

[0351] The compounds of the present invention can also be administered intranasally or by inhalation, typically in dry powder form (alone, in admixture, e.g., with lactose, or as mixed component particles mixed with a phospholipid, e.g., phosphatidylcholine) from a dry powder inhaler, or as an aerosol spray from a pressurized container, pump, spray, atomizer (preferably, an atomizer that uses electro-magnetic hydrodynamics to produce a fine mist) or nebulizer, with or without a suitable propellant, such as 1,1,1,2 tetrafluoroethane or 1,1,1,2,3,3,3 heptafluoropropane. For intranasal use, the powder can contain a bioadhesive agent, e.g., chitosan or cyclodextrin. The pressurized container, pump, spray, atomizer or nebulizer contains a solution or suspension of the compound(s) of the present invention, for example, with ethanol, aqueous ethanol, or alternative agent suitable for dispersing, solubilizing, or extending the release of the active agent, propellant(s) as solvent, and optional surfactant, for example, sorbitan trioleate, oleic acid, or oligolactic acid.Before being used in dry powder or suspension formulation, the drug product can be micronized to a size (typically less than 5 microns) suitable for delivery by inhalation.This can be achieved by any suitable crushing method, for example, spiral jet milling, fluidized bed jet milling, supercritical fluid processing to form nanoparticles, high-pressure homogenization, or spray drying.

[0352] Capsules (e.g., made from gelatin or HPMC), blisters, and cartridges for use in inhalers or insufflators can be formulated to contain a powder mix of a compound of the present invention, a suitable powder base such as lactose or starch, and a performance modifier such as leucine, mannitol, or magnesium stearate. Lactose can be anhydrous or in the form of the monohydrate, preferably the latter. Other suitable excipients include dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose, and trehalose.

[0353] Suitable flavors, such as menthol and levomenthol, or sweeteners, such as saccharin or saccharin sodium, may be added to those formulations of the invention intended for inhaled / intranasal administration.

[0354] The compounds of the present invention can be administered rectally or vaginally, for example, in the form of a suppository, pessary, or enema. Cocoa butter is a traditional suppository base, although various alternatives may be suitably used.

[0355] Formulations for rectal / vaginal administration may be formulated to be immediate and / or modified release, including delayed-, sustained-, pulsed-, controlled-, targeted-, and programmed-release.

[0356] The compounds of the present invention can also be administered directly to the eye or ear, typically in the form of droplets of a micronized suspension or solution in isotonic, pH-adjusted, sterile saline.Other formulations suitable for administration to the eye and ear include ointments, biodegradable (i.e., absorbent gel sponges, collagen) and non-biodegradable (i.e., silicone) implants, wafers, lenses, and microparticles, or vesicular systems, such as niosomes or liposomes.Polymers such as cross-linked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, cellulose polymers, such as hydroxypropylmethylcellulose, hydroxyethylcellulose, or methylcellulose, or heteropolysaccharide polymers, such as gellan gum, can be incorporated along with preservatives, such as benzalkonium chloride.Such formulations can also be delivered by iontophoresis.

[0357] Formulations for ocular / aural administration may be formulated to be immediate and / or modified release, including delayed-, sustained-, pulsed-, controlled-, targeted-, and programmed-release.

[0358] Other excipients and administration methods known in the pharmaceutical arts can also be used.The pharmaceutical compositions of the present invention can be prepared by any of the well-known pharmaceutical techniques, such as effective formulation and administration procedures.The above-mentioned discussions regarding effective formulation and administration procedures are well known in the art and are described in standard textbooks.Drug formulations are discussed, for example, in Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania, 1975; Liberman et al. (eds.), Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Kibbe et al. (eds.), Handbook of Pharmaceutical Excipients (3rd ed.), American Pharmaceutical Association, Washington, 1999.

[0359] Acceptable excipients are nontoxic to subjects at the dosages and concentrations employed and can include one or more of the following: 1) buffers, e.g., phosphate, citrate, or other organic acids; 2) salts, e.g., sodium chloride; 3) antioxidants, e.g., ascorbic acid or methionine; 4) preservatives, e.g., octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol; 5) alkylparabens, e.g., methyl or propylparaben, catechol, resorcinol, cyclohexanol, 3-pentanol, or m-cresol; 6) low molecular weight (less than about 10 residues) polypeptides; 7) proteins, e.g., serum albumin, gelatin, or immunoglobulins; 8) hydroxybenzoates, e.g., hydroxybenzoates ... 10) monosaccharides, disaccharides, or other carbohydrates, including glucose, mannose, or dextrins; 11) chelating agents, such as EDTA; 12) sugars, such as sucrose, mannitol, trehalose, or sorbitol; 13) salt-forming counterions, such as sodium, metal complexes (e.g., Zn-protein complexes); or 14) nonionic surfactants, such as polysorbates (e.g., polysorbate 20 or polysorbate 80), poloxamers, or polyethylene glycol (PEG).

[0360] Liposomes containing the compounds of the present invention can be prepared by methods known in the art (see, for example, Chang, HI; Yeh, MK; Clinical development of liposome-based drugs: formulation, characterization, and therapeutic efficacy; Int J Nanomedicine 2012;7;49-60). Particularly useful liposomes can be generated by the reverse-phase evaporation method using a lipid composition containing phosphatidylcholine, cholesterol, and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes are extruded through filters of defined pore size to obtain liposomes with the desired diameter.

[0361] The compounds of the present invention can also be entrapped in microcapsules prepared in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or macroemulsions, for example, by coacervation techniques or interfacial polymerization, such as hydroxymethylcellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules, respectively. Such techniques are disclosed in Remington, The Science and Practice of Pharmacy, 20th Edition, Mack Publishing (2000).

[0362] Sustained-release preparations can be used.Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the compound of the present invention, and the matrices are in the form of shaped articles, for example, films or microcapsules.Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol)), polylactide, copolymers of L-glutamic acid and 7-ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers, such as those used in leuprolide acetate for depot suspensions (injectable microspheres composed of lactic acid-glycolic acid copolymers and leuprolide acetate), sucrose acetate isobutyrate, and poly-D-(-)-3-hydroxybutyric acid.

[0363] The preparations used for intravenous administration must be sterile. This can be easily achieved, for example, by filtration through a sterile filtration membrane. The compound of the present invention is generally placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper that can be pierced by a hypodermic injection needle.

[0364] Suitable emulsions can be prepared using commercially available fat emulsions, such as lipid emulsions containing soybean oil, fat emulsions for intravenous administration (e.g., safflower oil, soybean oil, egg phosphatides, and glycerin in water), emulsions containing soybean oil and medium-chain triglycerides, and cottonseed oil lipid emulsions. The active ingredient can be dissolved in a premixed emulsion composition, or alternatively, it can be dissolved in oil (e.g., soybean oil, safflower oil, cottonseed oil, sesame oil, corn oil, or almond oil), and the emulsion is formed when mixed with phospholipids (e.g., egg phospholipids, soybean phospholipids, or soybean lecithin) and water. It will be appreciated that other ingredients, such as glycerol or glucose, can be added to adjust the osmotic pressure of the emulsion. Suitable emulsions typically contain up to 20% oil, for example, between 5 and 20% oil. The fat emulsion may contain fat droplets between 0.1 and 1.0 μm, in particular between 0.1 and 0.5 μm, and have a pH in the range of 5.5 to 8.0.

[0365] For example, an emulsion composition may be prepared by mixing a compound of the present invention with a lipid emulsion containing soybean oil or its components (soybean oil, egg phospholipids, glycerol, and water).

[0366] Drug product intermediates (DPIs) are partially processed materials that must undergo further processing steps before becoming bulk drug products. The compounds of the present invention can be formulated into drug product intermediate DPIs, which contain the active ingredient in a form with a higher free energy than the crystalline form. One reason for using DPIs is to improve oral absorption characteristics due to low solubility, slow dissolution, improved transport through the mucus layer adjacent to epithelial cells, and, in some cases, limitations due to biological barriers, such as metabolism and transporters. Other reasons may include improved solid-state stability and downstream manufacturability. In one embodiment, the drug product intermediate contains a compound of the present invention isolated and stabilized in an amorphous state (e.g., an amorphous solid dispersion (ASD)). There are many techniques known in the art for producing ASDs, which produce materials suitable for incorporation into bulk drug products, such as spray-dried dispersions (SDDs), melt extrudates (often referred to as HMEs), co-precipitates, amorphous drug nanoparticles, and nanoadsorbates. In one embodiment, the amorphous solid dispersion contains a compound of the present invention and a polymeric excipient. Other excipients and concentrations of said excipients and compounds of the invention are well known in the art and are described in standard textbooks, see, for example, "Amorphous Solid Dispersions Theory and Practice" by Navnit Shah et al.

[0367] In another aspect, the present invention provides a compound of the present invention, or a pharmaceutically acceptable salt thereof, for use as a pharmaceutical, particularly as a pharmaceutical for the treatment of abnormal cell growth.

[0368] In yet another aspect, the present invention provides use of a compound of the present invention, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating abnormal cell growth in a subject, e.g., a tumor, e.g., a MEK-associated tumor, in a subject.

[0369] In yet another aspect, the present invention provides a compound according to any of the formulas described herein, or a pharmaceutically acceptable salt thereof, for use in the treatment of abnormal cell growth, e.g., a tumor, e.g., a MEK-associated tumor.

[0370] Administration of the compounds of the present invention can be by any method capable of delivering the compound to the site of action, including oral routes, intraduodenal routes, parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion), topical, and rectal administration.

[0371] Dosage regimens can be adjusted to provide the optimum desired response. For example, a single bolus can be administered, several divided doses can be administered over time, or the dose can be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. For ease of administration and uniformity of dosage, it is particularly advantageous to formulate parenteral compositions into unit dosage forms. As used herein, unit dosage form refers to a physically discrete unit suitable as a unitary dosage for the mammalian subject being treated, each unit containing a predetermined amount of active compound calculated to produce the desired therapeutic effect together with the necessary pharmaceutical carrier. The specifications for the unit dosage forms of the present invention are directly dependent on (a) the unique characteristics of the compound to be administered and the specific therapeutic or prophylactic effect to be achieved, and (b) the limitations inherent in the field of formulating such active compounds to treat individual susceptibility.

[0372] Therefore, based on the disclosure provided herein, those skilled in the art should know that dosage and administration regimen can be adjusted according to the method well known in the therapeutic field.That is, the maximum tolerated dose can be easily established, and the time requirement for administering each drug can be determined so as to bring about detectable therapeutic benefit to patients, so that the effective amount that brings about detectable therapeutic benefit to patients can also be determined.Therefore, although certain dosage and administration regimen are exemplified herein, these examples do not in any way limit the dosage and administration regimen that can be provided to patients in the implementation of the present invention.

[0373] It should be noted that dosage values ​​vary with the type and severity of the condition to be alleviated and may include single or multiple doses. Furthermore, it should be understood that specific dosage regimens for any particular subject should be adjusted over time according to the individual needs and the professional judgment of the personnel administering and supervising the administration of the compositions, and that dosage ranges set forth herein are merely exemplary and are not intended to limit the scope or practice of the claimed compositions. For example, doses can be adjusted based on pharmacokinetic or pharmacodynamic parameters, which may include clinical effects, e.g., toxic effects and / or laboratory values. Accordingly, the present invention encompasses intrapatient dose escalation as determined by those skilled in the art. It should be understood that the determination of appropriate dosages and regimens for the administration of chemotherapeutic agents is well known in the relevant art and is within the skill of those in the art once provided with the teachings disclosed herein. In one embodiment, an effective dosage typically ranges from about 0.001 to about 100 mg per kg of body weight per day, often from about 0.01 to about 35 mg / kg / day in single or divided doses. For a 70 kg human, this would be about 0.07 mg / day to about 7000 mg / day, more typically about 10 mg / day to about 1000 mg / day. Sometimes the dosage will be about 10, 20, 30, 40, 50, 60, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 750, 800, 900, or 1000 mg / day.Sometimes the dosage is about 10 mg / day to about 1000 mg / day, about 10 mg / day to about 750 mg / day, about 10 mg / day to about 600 mg / day, about 10 mg / day to about 300 mg / day, about 10 mg / day to about 150 mg / day, about 20 mg / day to about 750 mg / day, about 20 mg / day to about 600 mg / day, about 20 mg / day to about 3 ... The dosage ranges from about 50 mg / day to about 150 mg / day, about 50 mg / day to about 750 mg / day, about 50 mg / day to about 600 mg / day, about 50 mg / day to about 300 mg / day, about 50 mg / day to about 150 mg / day, about 75 mg / day to about 750 mg / day, about 75 mg / day to about 600 mg / day, about 75 mg / day to about 300 mg / day, or about 75 mg / day to about 150 mg / day. In some cases, dosage levels below the lower end of the range may be sufficient, while in other cases, even larger doses may be used without causing any adverse side effects, and such larger doses are typically divided into several smaller doses for administration throughout the day. In one embodiment, a subject is administered about 50 mg / day.

[0374] For example, it may be desirable to administer a combination of active compounds for the purpose of treating a specific disease or condition, so it is within the scope of the present invention that two or more pharmaceutical compositions, at least one of which contains a compound according to the present invention, can be conveniently combined in the form of a kit suitable for the combined administration of the compositions.Thus, the kit of the present invention comprises two or more separate pharmaceutical compositions, at least one of which contains a compound of the present invention, and a means for separately holding the compositions, such as a container, a divided bottle, or a divided foil packet.An example of such a kit is the well-known blister pack used for packaging tablets, capsules, etc.

[0375] The kits of the invention are particularly suitable for administering different dosage forms, e.g., oral and parenteral dosage forms, for administering separate compositions at different dosing intervals, or for titrating separate compositions against one another. To aid in compliance, the kits typically include dosing instructions and may be equipped with a memory aid. In some embodiments, the kit includes a compound or a pharmaceutical composition thereof and a diagnostic agent. In other embodiments, the kit includes a compound or a pharmaceutical composition thereof and one or more therapeutic agents, e.g., a BRAF inhibitor, e.g., N-(3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-2,4-difluorophenyl)propane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropropane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4,5-difluorophenyl)propane-1-sulfonamide;N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4- ... N-(2-chloro-4-fluoro-3-((5-methyl-3-(methyl-d3)-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-phenyl)-3-fluoropropane-1-sulfonamide;N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino]-phenyl)-3-fluoropropane-1-sulfonamide )oxy]-4-fluorophenyl}propane-1-sulfonamide;N-(3-chloro-4-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy)-5-fluoropyridin-2-yl)propane-1-sulfonamide;N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}-3-fluoropropane-1-sulfonamide;or a pharmaceutically acceptable salt thereof.

[0376] The following schemes and written descriptions provide general details for the preparation of compounds of the invention.

[0377] The compounds of the present invention can be prepared by any method known in the art for the preparation of compounds of similar structure. In particular, the compounds of the present invention can be prepared by the procedures described by reference to the following schemes, or by the specific methods described in the Examples, or by methods analogous to any of these.

[0378] Those skilled in the art will recognize that the experimental conditions described in the following schemes are examples of conditions suitable for effecting the transformations shown, and that it may be necessary or desirable to vary the exact conditions used for the preparation of compounds of formula I and compounds encompassed by formula I, such as compounds of formula II.

[0379] In addition, those skilled in the art will recognize that at any stage in the synthesis of the compounds of the present invention, it may be necessary or desirable to protect one or more sensitive groups to prevent undesired side reactions. In particular, it may be necessary or desirable to protect amino or alcohol groups. The protecting groups (PG) used in the preparation of the compounds of the present invention may be used in a conventional manner. For example, see "Greene's Protective Groups in Organic Synthesis," 3rd Edition, by Theodora W Greene and Peter GM Wuts (John Wiley and Sons, 1999), which also describes methods for the removal of such groups, and is incorporated herein by reference, particularly those described in Chapter 7 ("Protection of Amino Groups") and Chapter 2 ("Protection of Hydroxy Groups, Including 1,2- and 1,3-Diols").

[0380] [ka]

[0381] Scheme 1 describes a general method for preparing compound 8, which can be prepared by reacting R 1 is H and R 2 is H and R 3 is C3-C6 cycloalkyl, and R 4 is a compound of formula I, wherein R is as defined for formula I. Commercially available 2,6-dichloro-4-methylnicotinic acid (compound 1) can be converted to the ester analog 2 by treatment with (trimethylsilyl)diazomethane. Compound 2 can be converted to the dimethylaminovinyl intermediate compound 3 by treatment with N,N-dimethylformamide dimethyl acetal. Compound 3 can be converted to the aldehyde intermediate compound 4 by treatment with a suitable acid, such as hydrochloric acid, in a suitable solvent, such as ether. Cyclization of compound 4 can be carried out by reaction with a suitable reducing agent, such as sodium cyanoborohydride, in a suitable solvent, such as methanol, to give a compound of formula R 3 NH2 (in the formula, R 3is C3-C6 cycloalkyl) to provide compound 5. Compound 5 can be converted to compound 6 upon treatment with trimethylsilyl iodide in a suitable solvent, such as acetonitrile. Compound 6 can be methylated by treatment with methyl iodide in the presence of a suitable base, such as an alkali carbonate, for example potassium carbonate, in the presence of a suitable solvent, such as THF, to provide compound 7. Compound 7 can be converted to a compound of formula R 4 NH2 (in the formula, R 4 is as defined for formula I), can undergo nucleophilic aromatic substitution to provide compound 8.

[0382] [ka]

[0383] Scheme 2 describes a general method for preparing compound 17, which can be prepared by reacting R 1 is H and R 2 is H and R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy-, and R 4is a compound of formula I, wherein: is as defined for formula I. Commercially available 4-bromo-2,6-dichloropyridine can be lithiated with a reagent such as lithium diisopropylamide and trapped with carbon dioxide to provide carboxylic acid 10. Compound 10 can be converted to compound 11 by refluxing in aqueous base, such as 4 M sodium hydroxide. Compound 11 can be methylated by treatment with methyl iodide in the presence of a suitable solvent, such as DMF, in the presence of a suitable base, such as an alkali carbonate, for example potassium carbonate, to provide compound 12. Compound 12 can be converted to the vinyl ether intermediate 13 by Suzuki reaction with (E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (compound (i)) using a catalyst such as methanesulfonate(2-dicyclohexylphosphino-2',6'-di-i-propoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) and an alkali base (e.g., an alkali carbonate, e.g., aqueous potassium carbonate) in a suitable solvent such as 1,4-dioxane. Compound 13 can be converted to the vinyl ether intermediate 13 by Suzuki reaction with (E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (compound (i)) using triethylamine and HCl in a suitable solvent such as 1,4-dioxane and heating. 3a -NH2HCl (wherein, R 3a HA P 1 O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy-, and P 1 is an alcohol protecting group, e.g., tert-butyl, benzyl, or tert-butyldimethylsilyl), can undergo oximine formation to provide compound 14. Compound 14 can be reduced to the alkoxyamine intermediate 15 using a suitable reducing agent, e.g., sodium cyanoborohydride, in a suitable solvent, e.g., isopropanol. Compound 15 can be reduced to the alkoxyamine intermediate 15 using a suitable reducing agent, e.g., sodium cyanoborohydride, in a suitable solvent, e.g., THF, in the presence of a strong base, e.g., lithium hexamethyldisilazide. 4 NH2 (in the formula, R 4is as defined for formula I), undergoes nucleophilic aromatic substitution and simultaneous cyclization, followed by the formation of compound 16 1 If it contains a protecting group, optional deprotection (using standard alcohol deprotection conditions known to those skilled in the art, such as phosphoric acid, trifluoroacetic acid, or tetrabutylammonium fluoride) will yield R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy-.

[0384] [ka]

[0385] Scheme 3 describes a general method for preparing compound 20, which can be prepared by reacting R 1 is H and R 2 is CH3- and R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy-, and R 4 is as defined for Formula I. Compound 15 (wherein R 3a HA P 1 O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy-, and P 1 is an alcohol protecting group, e.g., tert-butyl, benzyl, or tert-butyldimethylsilyl, and R 4 is as defined for formula I) in a suitable solvent, such as THF, in the presence of a strong base, such as lithium hexamethyldisilazide, to form a compound of formula R 4 NH2 (in the formula, R 4is as defined for Formula I), compound 16 can be cyclized to provide compound 16. Compound 16 can be iodinated using n-iodosuccinimide and p-toluenesulfonic acid in a suitable solvent, such as 1:1 MeOH:THF, to provide compound 18. Compound 18 can undergo a Negishi coupling with methylzinc(II) chloride using a catalyst, such as a palladium catalyst, for example, bis(tri-t-butylphosphine)palladium(0), in a suitable solvent, such as THF, followed by optional deprotection if compound 19 contains a protecting group (using standard alcohol deprotection conditions known to those skilled in the art, such as phosphoric acid, trifluoroacetic acid, or tetrabutylammonium fluoride), to provide compound 19. 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy.

[0386] [ka]

[0387] Scheme 4 describes a general method for preparing compound 24, which can be prepared by reacting R 1 is H and R 2 is H and R 3 is H and R 4 is as defined for Formula I. Compound 13, prepared as described in Scheme 2, is reacted with a compound of formula R in the presence of a strong base, such as lithium hexamethyldisilazide, in a suitable solvent, such as THF, to give a compound of formula R 4 NH2 (in the formula, R 4is as defined for formula I), can undergo nucleophilic aromatic substitution to provide compound 21. Compound 21 can be hydrolyzed to the aldehyde intermediate 22 under acidic conditions using an acid, such as trifluoroacetic acid, in a suitable solvent, such as dichloromethane. Compound 22 can be converted to the aldehyde intermediate 23 using a reducing agent, such as sodium triacetoxyborohydride, in a suitable solvent, such as dichloroethane, to provide the aldehyde intermediate 24. 2- NH2 (in the formula, P 2 is an amine protecting group, e.g., benzyl, p-methoxybenzyl, or 2,4-dimethoxybenzyl), can be cyclized to provide compound 23. Deprotection of compound 23 can be achieved using standard deprotection conditions known to those skilled in the art, such as heating with trifluoroacetic acid or hydrochloric acid, to provide compound 24.

[0388] [ka]

[0389] Scheme 5 describes a general method for preparing compound 27, which can be prepared by reacting R 1 is H and R 2 is H and R 3 is hydroxy C1-C6 alkyl, and R 4 is a compound of formula I, wherein: is as defined for formula I. Compound 13, prepared as described in Scheme 2, can be hydrolyzed to the aldehyde intermediate 25 using a suitable acid, for example, trifluoroacetic acid. Compound 25 can be converted to the aldehyde intermediate of formula P using a suitable reducing agent, for example, sodium triacetoxyborohydride, and acetic acid in a suitable solvent, for example, dichloroethane. 1 O-(C1~C6 alkyl)-ONH2HCl (wherein, P 1 is an alcohol protecting group, e.g., tert-butyldimethylsilyl) at 60°C to give the cyclized, deprotected compound 26. Compound 26 can be converted to a compound of formula R in a suitable solvent, e.g., THF, in the presence of a strong base, e.g., lithium hexamethyldisilazide.4 NH2 (in the formula, R 4 When treated with a reagent having the formula (I), it can undergo nucleophilic aromatic substitution to provide compound 27.

[0390] [ka]

[0391] Scheme 6 describes an alternative general method for preparing compound 17, which can be prepared by reacting R 1 is H and R 2 is H and R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy, and R 4 is as defined for Formula I. Compound 28, prepared according to a method similar to that described for compound 13, is reacted with a compound of formula R in a suitable solvent, such as THF, in the presence of a strong base, such as lithium hexamethyldisilazide, to give a compound of formula R 4 NH2 (in the formula, R 4 is as defined for formula I), can undergo nucleophilic aromatic substitution to provide compound 29. Compound 29 can be converted to a compound of formula R using triethylamine and HCl in a suitable solvent, such as 1,4-dioxane, and heating. 3a NH2HCl (wherein, R 3a HA P 1 O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy, and P 1 is an alcohol protecting group, e.g., tert-butyl, benzyl, or tert-butyldimethylsilyl), can undergo oximine formation to provide compound 30. Compound 30 can be cyclized using a suitable reducing agent, e.g., sodium cyanoborohydride and acetic acid, in a suitable solvent, e.g., isopropanol, followed by conversion of compound 30 to P 1If it contains a protecting group, optional deprotection (using standard alcohol deprotection conditions known to those skilled in the art, such as phosphoric acid, trifluoroacetic acid, or tetrabutylammonium fluoride) will yield R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy.

[0392] [ka]

[0393] Scheme 7 describes an alternative general method for preparing compound 17, which can be prepared by reacting R 1 is H and R 2 is H and R 3 is hydroxy C1-C6 alkoxy, and R 4 is a compound of formula I, wherein is as defined for formula I. Compound 28 can be treated with a reagent of formula TBSO-(C1-C6 alkyl)-ONH2HCl using triethylamine and HCl in a suitable solvent, such as 1,4-dioxane, and heating to undergo oximine formation to provide compound 32. Compound 32 can be converted to a suitable alcohol protecting group P using tert-butyldimethylsilyl chloride and a suitable base, such as imidazole, in a suitable solvent, such as DMF. 1 , for example, with tert-butyldimethylsilyl to provide compound 14. Compound 14 can be reduced to the alkoxyamine intermediate 15 using a suitable reducing agent, for example, sodium cyanoborohydride, in a suitable solvent, for example, isopropanol. Compound 15 can be converted to an alkoxyamine intermediate of formula R by reaction with a suitable reducing agent, for example, sodium cyanoborohydride, in a suitable solvent, for example, THF, in the presence of a strong base, for example, lithium hexamethyldisilazide. 4 NH2 (in the formula, R 4is as defined for Formula I), can undergo nucleophilic aromatic substitution to provide compound 16. Compound 16 can be deprotected using standard alcohol deprotection conditions known to those skilled in the art, such as phosphoric acid, trifluoroacetic acid, or tetrabutylammonium fluoride, to provide compound 17.

[0394] [ka]

[0395] Scheme 8 describes an alternative general method for preparing compound 20, which can be prepared by reacting R 1 is H and R 2 is CH3- and R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy, and R 4 is as defined for formula I. Compound 28 is a compound of formula I, wherein compound 28 is reacted with a compound of formula R in a suitable solvent, such as THF, in the presence of a strong base, such as lithium hexamethyldisilazide. 4 NH2 (in the formula, R 4 is as defined for Formula I), can undergo nucleophilic aromatic substitution to provide compound 29. Compound 29 can be hydrolyzed to the aldehyde intermediate 22 using a suitable acid, for example, trifluoroacetic acid. Compound 22 can be converted to a compound of formula R using a suitable reducing agent, for example, sodium triacetoxyborohydride and acetic acid in a solvent, for example, dichloroethane. 3a NH2HCl (wherein, R 3a HA P 1 O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy, and P 1is an alcohol protecting group, e.g., tert-butyl, benzyl, or tert-butyldimethylsilyl) at 60° C. to give the cyclized compound 16. Compound 16 can be iodinated using n-iodosuccinimide and p-toluenesulfonic acid in a suitable solvent, e.g., 1:1 MeOH:THF, to provide compound 18. Compound 18 can undergo Negishi coupling with methylzinc(II) chloride using a catalyst, e.g., bis(tri-t-butylphosphine)palladium(0), in a suitable solvent, e.g., THF, followed by the conversion of compound 18 to P 1 If it contains a protecting group, it can be optionally deprotected (using standard alcohol deprotection conditions known to those skilled in the art, such as phosphoric acid, trifluoroacetic acid, or tetrabutylammonium fluoride) to give R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy.

[0396] [ka]

[0397] Scheme 9 describes a general method for synthesizing compound 35, which can be prepared by reacting R 1 is H and R 2 is a halogen and R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy, and R 4 is as defined for formula I. Compound 12 is a compound of formula I, wherein compound 12 is reacted with a compound of formula R in a suitable solvent, such as THF, in the presence of a strong base, such as lithium hexamethyldisilazide. 4 NH2 (in the formula, R 4is as defined for Formula I), can undergo nucleophilic aromatic substitution to provide compound 33. Compound 33 can be converted to vinyl ether intermediate 21 by Suzuki reaction with (E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane using a catalyst, for example, a palladium catalyst, for example, methanesulfonate(2-dicyclohexylphosphino-2',6'-di-i-propoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II), and an alkali base, for example, an alkali carbonate base, for example, aqueous potassium carbonate, in a suitable solvent, for example, 1,4-dioxane. Compound 21 can be converted to vinyl ether intermediate 21 by Suzuki reaction with (E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane using triethylamine and HCl in a suitable solvent, for example, 1,4-dioxane, and heating. 3a NH2HCl (wherein, R 3a HA P 1 O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy, and P 1 is an alcohol protecting group, e.g., tert-butyl, benzyl, or tert-butyldimethylsilyl), can undergo oximine formation to provide compound 30. Compound 30 can be treated with a suitable reducing agent, e.g., sodium cyanoborohydride and acetic acid in a suitable solvent, e.g., isopropanol, to yield the cyclized compound 31. Compound 31 can be halogenated using conditions such as N-iodosuccinimide and p-toluenesulfonic acid in 1:1 THF / MeOH, or N-bromosuccinimide in a suitable solvent, e.g., DMF, or N-chlorosuccinimide in a suitable solvent, e.g., DMF, or Selectfluor in a suitable solvent, e.g., acetonitrile, followed by treatment of compound 31 with P 1 If it has a protecting group, optional deprotection (e.g., with phosphoric acid, trifluoroacetic acid, or tetrabutylammonium fluoride) will result in R 2 are iodo, bromo, chloro, or fluoro, respectively, and R 3is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy.

[0398] [ka]

[0399] Scheme 10 describes an alternative general method for preparing compound 17, which can be prepared by reacting R 1 is H and R 2 is H and R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy, and R 4 is as defined for formula I. Compound 12 is a compound of formula I, wherein compound 12 is converted to a compound of formula R in a suitable solvent, such as THF, in the presence of a suitable base, such as potassium tert-butoxide. 4 NH2 (in the formula, R 4 is as defined for Formula I), can undergo nucleophilic aromatic substitution to provide compound 36. Compound 36 can be converted to vinyl ether intermediate 29 by Suzuki reaction with (Z)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane using a catalyst, for example, a palladium catalyst, for example, methanesulfonate(2-dicyclohexylphosphino-2',6'-di-i-propoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II), in a solvent, for example, 2-methyltetrahydrofuran, and a suitable base, for example, an alkali base, for example, an alkali carbonate base, for example, aqueous potassium carbonate. Compound 29 can be converted to vinyl ether intermediate 29 by Suzuki reaction with (Z)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane using triethylamine and hydrochloric acid in a suitable solvent, for example, 1,4-dioxane. 3a NH2HCl (wherein, R 3a HA P 1O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy, and P 1 is an alcohol protecting group, e.g., tert-butyl, benzyl, or tert-butyldimethylsilyl), followed by treatment with a suitable reducing agent, e.g., pyridine borane and hydrochloric acid, and heating at 60°C, followed by conversion of compound 29 to P 1 If it has a protecting group, optional deprotection (e.g., with phosphoric acid, trifluoroacetic acid, or tetrabutylammonium fluoride) will result in R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy, compound 17 can be obtained.

[0400] [ka]

[0401] Scheme 11 describes a method for preparing compound 43, which is obtained by reacting R 1 is phenyl and R 2 is hydrogen and R 3 is hydrogen and R 4is a compound of Formula I, wherein R is as described for Formula I. Commercially available 2,6-dichloro-4-iodopyridine can be lithiated with a reagent such as lithium diisopropylamide and trapped with carbon dioxide to provide compound 37. Compound 37 can be converted to compound 38 by refluxing in an aqueous base, such as 4 M sodium hydroxide. Compound 38 can be methylated by treatment with methyl iodide in the presence of a suitable solvent, such as DMF, in the presence of a suitable base, such as an alkali carbonate, such as potassium carbonate, to provide compound 39. Compound 39 can be converted to vinyl ether intermediate 40 by Suzuki reaction with 4,4,5,5-tetramethyl-2-(1-phenylvinyl)-1,3,2-dioxaborolane using a suitable catalyst, such as a palladium catalyst (e.g., Pd(dppf)Cl) and a base, such as an alkali carbonate (e.g., aqueous potassium carbonate), in a suitable solvent, such as 1,4-dioxane. Compound 40 can be prepared by reacting a compound of formula R in the presence of a strong base such as lithium hexamethyldisilazide in a suitable solvent such as THF. 4 NH2 (in the formula, R 4 where R is as defined for Formula I), can undergo nucleophilic aromatic substitution to provide compound 41. Compound 41 can be cyclized with (2,4-dimethoxyphenyl)methanamine in the presence of a Lewis acid, such as trimethylaluminum, in a suitable solvent, such as toluene, by heating to provide compound 42. Compound 42 can be deprotected by heating with a suitable acid, such as TFA, to provide compound 43.

[0402] [ka]

[0403] Scheme 12 describes a method for preparing compound 51, which is obtained by reacting R 1 is methyl and R 2 is hydrogen and R 3is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy, and R 4 is a compound of formula I, wherein is as defined for formula I. Compound 44 can be treated with NH, Fe(NO).9H0, and NaNH at low temperature, followed by treatment with methyl iodide, to provide compound 45. Compound 45 can be heated with benzo[d][1,3,2]dioxaborole in a suitable solvent, such as toluene, along with a catalyst, such as NiCl(dppe), to provide compound 46. Compound 46 can be converted to compound 47, for example, by reacting with compound 39 using Suzuki reaction conditions in a suitable solvent, such as a solvent mixture, such as toluene / THF, in the presence of a catalyst, such as a palladium catalyst, such as Pd(dppf)Cl, and a base, such as KPO, KCO, KOtBu, CsCO, NaOH, or triethylamine. Compound 47 can be converted to compound 47 using triethylamine and HCl in a suitable solvent, such as 1,4-dioxane, and heating to provide a compound of formula R. 3a NH2HCl (wherein, R 3a HA P 1 O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy, and P 1 is an alcohol protecting group, e.g., tert-butyl, benzyl, or tert-butyldimethylsilyl), can undergo oximine formation to provide compound 48. Compound 48 can be treated with a suitable reducing agent, e.g., sodium cyanoborohydride and acetic acid in a suitable solvent, e.g., isopropanol, to give the cyclized product 49. Compound 49 can be converted to a compound of formula R in the presence of a strong base, e.g., lithium hexamethyldisilazide, in a suitable solvent, e.g., THF. 4 NH2 (in the formula, R 4 is as defined for formula I), compound 49 undergoes nucleophilic aromatic substitution, and subsequently compound 49 is converted to P 1If it contains a protecting group, optional deprotection (using standard alcohol deprotection conditions known to those skilled in the art, such as phosphoric acid, trifluoroacetic acid, or tetrabutylammonium fluoride) will yield R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy.

[0404] The term "amine protecting group," as used herein, refers to derivatives of groups commonly used to block or protect amino groups while reactions are being carried out on other functional groups on a compound. Examples of protecting groups suitable for use in any of the methods described herein include carbamates, amides, alkyl and aryl groups, imines, and many N-heteroatom derivatives, which can be removed to regenerate the desired amine group. Non-limiting examples of amine protecting groups are t-butyloxycarbonyl ("Boc"), 2-trimethylsilylethoxymethyl (SEM), and p-methoxybenzyl (PMB). Further examples of these and other protecting groups can be found in T.W. Greene et al., Greene's Protective Groups in Organic Synthesis. New York: Wiley Interscience, 2006.

[0405] The term "alcohol protecting group," as used herein, refers to a derivative of a group commonly used to block a hydroxy group while reactions are carried out on other functional groups on a compound. Examples of protecting groups suitable for use in any of the methods described herein include benzyl, trityl, silyl ethers, and the like.

[0406] Intermediate compounds 7, 15, 18, 22, 26, 29, 30, and 31 are also novel intermediates useful in the preparation of compounds of formula I, as shown in the above scheme, and provide further embodiments of the present invention.

[0407] In one embodiment, provided herein is a method for preparing a compound of formula I, the method comprising: (a)R 1 is H and R 2 is H and R 3 is C3-C6 cycloalkyl, and R 4 is as defined for formula I, a compound of formula 7

[0408] [ka] , the formula R 4 NH2 (in the formula, R 4 is as defined for formula I) in the presence of a strong base; or (b)R 1 is H and R 2 is H and R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy or (C3-C6 cycloalkyl)C1-C6 alkoxy-, and R 4 is as defined for formula I, a compound of formula 15

[0409] [ka] [In the formula, R 3a HA P 1 O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy-, and P 1 is an alcohol protecting group], 4 NH2 (in the formula, R 4 is as defined for formula I), optionally followed by removal of the alcohol protecting group, if present, by cyclization in the presence of a strong base; or (c)R 1 is H and R 2 is CH3- and R 3is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy or (C3-C6 cycloalkyl)C1-C6 alkoxy-, and R 4 is as defined for formula I, a compound of formula 18

[0410] [ka] [In the formula, R 3a HA P 1 O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy, P 1 is an alcohol protecting group, and R 4 is as defined for Formula I] with methylzinc(II) chloride in the presence of a palladium catalyst, followed by removal of the alcohol protecting group, if present; or (d)R 1 is H and R 2 is H and R 3 is H and R 4 is as defined for formula I, a compound of formula 22

[0411] [ka] [In the formula, R 4 is as defined for formula I] into a compound of formula P 2- NH2 (in the formula, P 2 is an amine protecting group), followed by removal of the amine protecting group; or (e)R 1 is H and R 2 is H and R 3 is hydroxy C1-C6 alkyl, and R 4 is as defined for formula I, a compound of formula 26

[0412] [ka] [In the formula, R 4 is as defined for Formula I] in the presence of a strong base with 4 reacting in the presence of a reagent having NH2, or (f)R 1 is H and R 2 is H and R 3 is hydroxy C1-C6 alkyl, and R 4 is as defined for formula I, a compound of formula 30

[0413] [ka] [In the formula, P 1 is an alcohol protecting group, and R 4 is as defined for Formula I] in the presence of a reducing agent, followed by removal of the alcohol protecting group; or (g)R 1 is H and R 2 is a halogen and R 3 is hydroxy C1-C6 alkoxy-, C1-C6 alkoxy, fluoro C1-C6 alkoxy or (C3-C6 cycloalkyl) C1-C6 alkoxy-, and R 4 is as defined for formula I, a compound of formula 31

[0414] [ka] [In the formula, R 4 is as defined for Formula I, and R 3a HA P 1 O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy-, and P 1 is an alcohol protecting group] followed by removal of the alcohol protecting group, if present; or (h)R 1 is H and R2 is H and R 3 is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy or (C3-C6 cycloalkyl)C1-C6 alkoxy-, and R 4 is as defined for formula I, a compound of formula 29

[0415] [ka] [In the formula, R 4 is as defined for formula I] into a compound of formula P 1 O-(C1~C6 alkyl)-ONH2HCl (wherein, P 1 is an alcohol protecting group) in the presence of triethylamine and hydrochloric acid, followed by removal of the alcohol protecting group; or (i)R 1 is phenyl and R 2 is hydrogen and R 3 is hydrogen and R 4 For compounds of formula I, where is as described for formula I, compounds having formula 41

[0416] [ka] [In the formula, R 4 is as defined for Formula I] is cyclized with (2,4-dimethoxyphenyl)methanamine in the presence of a Lewis acid at elevated temperature to give a compound having formula 42

[0417] [ka] followed by treating compound 42 with an acid; or (j)R 1 is methyl and R 2 is hydrogen and R 3is hydroxy-C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy or (C3-C6 cycloalkyl)C1-C6 alkoxy, and R 4 For compounds of formula I, where is as defined for formula I, compounds having formula 49

[0418] [ka] [In the formula, R 3a HA P 1 O—C1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, or (C3-C6 cycloalkyl)C1-C6 alkoxy-, and P 1 is an alcohol protecting group], 4 NH2 (in the formula, R 4 is as defined for formula I) in the presence of a strong base, followed by reaction with P, if present. 1 removing the protecting group, and The compounds of formula I may be converted into pharmaceutically acceptable salts. Includes:

[0419] Synthetic intermediates 3, 4, 5, 6, 7, 13, 14, 15, 16, 18, 21, 22, 23, 25, 26, 28, 29, 30, 32, 33, 36, 37, 38, 39, 40, 41, 42, 47, 48, and 49 are also believed to be novel and are further embodiments of the present invention.

[0420] The present invention can be further understood by reference to the following detailed description of embodiments of the invention and examples contained herein. It is to be understood that the present invention is not limited to specific synthetic methods of preparation, which may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0421] E1. Compound of Formula I

[0422] [ka] or a pharmaceutically acceptable salt thereof, R 1 is H, Br, C1-C6 alkyl or phenyl; R 2 is H, halogen or CH3-, R 3 is H, hydroxyC1-C6 alkyl-, hydroxyC1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or (C3-C6 cycloalkyl)C1-C6 alkoxy-; R 4 is phenyl substituted with 1, 2, or 3 substituents independently selected from halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(=O)-.

[0423] E2.R 1 A compound according to embodiment E1, wherein is H; or a pharmaceutically acceptable salt thereof.

[0424] E3.R 2 A compound according to embodiment E1 or E2, wherein is H; or a pharmaceutically acceptable salt thereof.

[0425] E4.R 2 A compound according to embodiment E1 or E2, or a pharmaceutically acceptable salt thereof, wherein is CH3-.

[0426] E5.R 3 A compound according to any one of embodiments E1 to E4, or a pharmaceutically acceptable salt thereof, wherein is hydroxyC1-C6 alkyl-.

[0427] E6.R 4or a pharmaceutically acceptable salt thereof. A compound according to any one of embodiments E1-E5, wherein is phenyl substituted with 1 or 2 substituents independently selected from halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(=O)-.

[0428] E7.R 4 A compound according to any one of embodiments E1 to E6, wherein is phenyl substituted with 1 or 2 substituents independently selected from halogen and C1-C6 alkylthio; or a pharmaceutically acceptable salt thereof.

[0429] E8. Compound of Formula II

[0430] [ka] or a pharmaceutically acceptable salt thereof, R 1 is H, Br, C1-C6 alkyl or phenyl; R 2 is H, halogen or CH3-, R 3 is H, hydroxyC1-C6 alkyl-, hydroxyC1-C6 alkoxy-, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, or (C3-C6 cycloalkyl)C1-C6 alkoxy-; R a and R b are independently selected from halogen, C1-C6 alkyl, C1-C6 alkylthio, fluoroC1-C6 alkylthio, fluoroC1-C6 alkyl, C1-C6 alkoxy, fluoroC1-C6 alkoxy, C3-C6 cycloalkyl, and C1-C6 alkyl-C(=O)-.

[0431] E9.R 1A compound according to embodiment E8, wherein is H; or a pharmaceutically acceptable salt thereof.

[0432] E10.R 2 A compound according to embodiment E8 or E9, wherein is H; or a pharmaceutically acceptable salt thereof.

[0433] E11.R 2 A compound according to embodiment E8 or E9, or a pharmaceutically acceptable salt thereof, wherein is CH3-.

[0434] E12.R 3 A compound according to any one of embodiments E8 to E11, or a pharmaceutically acceptable salt thereof, wherein is hydroxyC1-C6alkyl-.

[0435] E13.R 3 A compound according to any one of embodiments E8 to E11, wherein is H; or a pharmaceutically acceptable salt thereof.

[0436] E14.R a A compound according to any one of embodiments E8 to E13, or a pharmaceutically acceptable salt thereof, wherein is halogen.

[0437] E15.R b A compound according to any one of embodiments E8 to E14, or a pharmaceutically acceptable salt thereof, wherein is halogen, C1-C6 alkyl, C1-C6 alkylthio, or fluoroC1-C6 alkoxy.

[0438] E16.R b A compound according to embodiment E15, wherein is C1-C6 alkylthio; or a pharmaceutically acceptable salt thereof.

[0439] E17. 8-((4-bromo-2-fluorophenyl)amino)-2-cyclopropyl-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-2-(cyclopropylmethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-2-ethoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 2-Cyclopropyl-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 2-Cyclopropyl-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-chloro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-chlorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2,3-difluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-3-chloro-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-(trifluoromethyl)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-ethyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-cyclopropyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-methoxyphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-((trifluoromethyl)thio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-isopropylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-chloro-4-cyclopropylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-acetyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-chloro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-chloro-4-ethylphenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-(trifluoromethoxy)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-((difluoromethyl)thio)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-2-isopropoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-iodophenyl)amino)-2-isopropoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 2-Ethoxy-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-iodophenyl)amino)-2-methoxy-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 2-(tert-butoxy)-8-((2-fluoro-4-iodophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; (S)-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; (R)-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; (S)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; (R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; (R)-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxypropoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-5-chloro-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 5-chloro-8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-5-fluoro-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-iodophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5-iodo-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 5-bromo-8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 4-Bromo-8-((2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-ethyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-cyclopropyl-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-iodophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-iodo-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-ethyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-cyclopropyl-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-propylphenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-(methylthio)phenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-ethyl-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-cyclopropyl-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-chlorophenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethyl)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-(difluoromethoxy)-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-5,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 2-(2,2-difluoroethoxy)-8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-7-methyl-4-phenyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-(methylthio)phenyl)amino)-7-methyl-4-phenyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-(methylthio)phenyl)amino)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((4-bromo-2-fluorophenyl)amino)-2-(2-hydroxyethoxy)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-4,7-dimethyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione; or a pharmaceutically acceptable salt thereof.

[0440] E18. Structure

[0441] [ka] or a pharmaceutically acceptable salt thereof.

[0442] E19. Structure

[0443] [ka] The compound is 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione having the formula:

[0444] E20. Crystals containing 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.

[0445] E21. Crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 1.

[0446] E22. Crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 1, according to embodiment E21, having a PXRD pattern including characteristic peaks at 5.0, 8.7, 9.3, 10.8, 14.5, 15.3, 18.8, and 20.5 degrees two-theta (±0.2 degrees two-theta).

[0447] E23. Crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione according to embodiment E21, Form 1, having a PXRD pattern comprising peaks at 2-theta values ​​essentially the same as those shown in Figure 1.

[0448] E24. Crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 2.

[0449] E25. Crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 2 according to embodiment E24, having a PXRD pattern including characteristic peaks at 7.1, 9.4, 12.4, 12.8, 14.3, 15.6, 16.4, 17.4, 18.5, 18.9, 19.5, 19.9, 21.1, 21.4, 23.2, 23.7, 24.8, 25.6, 27.6, 30.3, 33.2, 33.5, and 37.5 degrees two-theta (±0.2 degrees two-theta).

[0450] E26. Crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione according to embodiment E24, Form 2, having a PXRD pattern comprising peaks at 2-theta values ​​essentially the same as those shown in Figure 2.

[0451] E27. Crystalline anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 2, according to embodiment E25 or E26, wherein the PXRD pattern is obtained by PXRD analysis carried out at 25° C. and a relative humidity below 10%.

[0452] E28. Crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3.

[0453] E29. Crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3 according to embodiment E28, having PXRD peaks at 13.7, 18.0 and 18.3 degrees two-theta (±0.2 degrees two-theta).

[0454] Crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3 according to embodiment E28, having PXRD peaks at E30.6.9, 9.1, 13.7, 18.0 and 18.3 degrees two-theta (±0.2 degrees two-theta).

[0455] E31.2 Theta units are 6.9, 9.1, 11.8, 12.0, 13.7, 14.0, 15.2, 15.8, 18.0, 18.3, 19.0, 19.3, 20.2, 20.9, 21.6, 22.6, 23.6, 24.0, 24.9, 25.2, 25.8, 27.5, 28.1, 28.4, 29.8, 30.9, 31.7, and 32.3 and PXRD peaks at 36.5 degrees two-theta (±0.2 degrees two-theta).

[0456] E32. Crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3 according to embodiment E28, having a PXRD pattern comprising peaks at 2-theta values ​​essentially the same as those shown in Figure 3.

[0457] E33. Crystalline monohydrate 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 3 according to any one of embodiments E29 to E32, wherein the PXRD pattern is obtained by PXRD analysis carried out at 25° C. and a relative humidity greater than 35%.

[0458] E34. Amorphous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione, Form 4.

[0459] E35. Amorphous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione according to embodiment E34, Form 4, having a PXRD pattern including peaks at 2-theta values ​​essentially the same as those shown in FIG. 4.

[0460] E36. A pharmaceutical composition comprising a compound according to any one of embodiments E1 to E35, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0461] E37. A method of treating MEK-associated tumors, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of embodiments E1 to E35 or a pharmaceutically acceptable salt thereof.

[0462] E38. The method according to embodiment E37, wherein the tumor harbors a BRAF V600 mutation selected from V600E, V600K, V600D, V600R, and V600S.

[0463] E39. The method according to embodiment E37 or E38, wherein the tumor harbors a BRAF V600E mutation.

[0464] E40. The method according to any one of embodiments E37-E39, wherein the tumor is an extracranial tumor.

[0465] E41. The method according to embodiment E40, wherein the extracranial tumor is selected from melanoma, colorectal cancer, thyroid cancer, non-small cell lung cancer, ovarian cancer, and neuroblastoma.

[0466] E42. The method according to any one of embodiments E37-E39, wherein the tumor is a CNS tumor.

[0467] E43. The method according to embodiment E42, wherein the CNS tumor is an intracranial tumor.

[0468] E44. The method according to embodiment E43, wherein the intracranial tumor is a brain tumor.

[0469] E45. The method according to embodiment E44, wherein the brain tumor is a metastatic brain tumor.

[0470] E46. The method according to embodiment E45, wherein the metastatic brain tumor is selected from metastatic melanoma, metastatic colorectal cancer, metastatic non-small cell lung cancer, metastatic thyroid cancer, and metastatic ovarian cancer.

[0471] E47. The method according to embodiment E42, wherein the CNS tumor is an intracranial or extracranial LMD.

[0472] E48. The method according to embodiment E47, wherein the LMD is selected from metastatic melanoma, metastatic colorectal cancer, and metastatic non-small cell lung cancer.

[0473] E49. The method according to embodiment E43, wherein the intracranial tumor is a primary tumor.

[0474] E50. The method of embodiment E49, wherein the primary brain tumor is a malignant tumor.

[0475] E51. The method according to embodiment E50, wherein the primary brain tumor is a grade 2 glioma, a grade 3 glioma, or a grade 4 glioma.

[0476] E52. The method according to embodiment E49, wherein the primary brain tumor is a benign tumor.

[0477] The method according to embodiment E37, wherein the tumor harbors a BRAF fusion.

[0478] E54.The tumor is KIAA11549-BRAF, MKRN1-BRAF, TRIM24-BRAF, AGAP3-BRAF, ZC3HAV1-BRAF, AKAP9-BRAF, CCDC6-BRAF, AGK-BRAF, EPS15- BRAF, NUP214-BRAF, ARMC10-BRAF, BTF3L4-BRAF, GHR-BRAF, ZC3HAV1-BRAF, ZNF767-BRAF, CCDC91-BRAF, DYNC112-BRAF, ZKSCAN1 The method according to embodiment E53, wherein the patient has a BRAF fusion selected from -BRAF, GTF2I-BRAF, MZT1-BRAF, RAD18-BRAF, CUX1-BRAF, SLC12A7-BRAF, MYRIP-BRAF, SND1-BRAF, NUB1-BRAF, KLHL7-BRAF, TANK-BRAF, RBMS3-BRAF, STRN3-BRAF, STK35-BRAF, ETFA-BRAF, SVOPL-BRAF, and JHDM1D-BRAF.

[0479] E55. The tumor is breast cancer (e.g., invasive ductal carcinoma), colorectal cancer (e.g., colon adenocarcinoma), esophageal cancer (e.g., esophageal adenocarcinoma), glioma (e.g., desmoplastic infantile ganglioglioma of the brain, pilocytic astrocytoma of the brain, pleomorphic xanthoastrocytoma of the brain, low-grade glioma (NOS) of the spinal cord, anaplastic oligodendroglioma, anaplastic ganglioglioma), carcinoma of the head and neck (e.g., neuroendocrine carcinoma of the head and neck), lung cancer (e.g., lung adenocarcinoma, non-small cell lung carcinoma (NOS)), melanoma ( For example, the cancer is spitzoid cutaneous melanoma, non-spitzoid mucosal melanoma, spitzoid cutaneous melanoma, melanoma of unknown primary, non-spitzoid cutaneous melanoma), pancreatic cancer (e.g., adenocarcinoma, pancreatic acinar cell carcinoma), prostate cancer (e.g., prostatic acinar adenocarcinoma), sarcoma (malignant solid fibroma), thyroid cancer (papillary thyroid carcinoma), carcinoma of unknown primary (e.g., adenocarcinoma of unknown primary), pleural mesothelioma, rectal adenocarcinoma, endometrial cancer (e.g., endometrial adenocarcinoma (NOS)), or ovarian serous carcinoma.

[0480] E56. The method according to embodiment E37, wherein the tumor is a BRAF wild-type tumor.

[0481] E57. The method according to any one of embodiments E37 to E56, further comprising administering one or more additional anti-cancer therapies.

[0482] E58. The method according to embodiment E57, wherein the one or more additional anti-cancer treatments are independently selected from surgery, radiation therapy, and anti-cancer agents.

[0483] E59. The method according to embodiment E58, wherein the additional anticancer treatment is selected from one or more anticancer agents.

[0484] E60. The method according to embodiment E59, wherein the anticancer agent is selected from MEK inhibitors, BRAF inhibitors, EGFR inhibitors, inhibitors of HER2 and / or HER3, SHP2 inhibitors, Axl inhibitors, PI3K inhibitors, SOS1 inhibitors, signal transduction pathway inhibitors, checkpoint inhibitors, modulators of apoptosis pathways, cytotoxic chemotherapeutic agents, angiogenesis-targeted therapies, and immune-targeted agents, including immunotherapies.

[0485] E61. The method of embodiment E60, wherein the anticancer agent is a BRAF inhibitor.

[0486] E62. The method of embodiment E61, wherein the BRAF inhibitor is encorafenib or a pharmaceutically acceptable salt thereof.

[0487] E63.BRAF inhibitors N-(3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-2,4-difluorophenyl)propane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropropane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4,5-difluorophenyl)propane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)propane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamide; N-(2-chloro-4-fluoro-3-((5-methyl-3-(methyl-d3)-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-phenyl)-3-fluoropropane-1-sulfonamide; N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}propane-1-sulfonamide; N-(3-chloro-4-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy)-5-fluoropyridin-2-yl)propane-1-sulfonamide; and N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}-3-fluoropropane-1-sulfonamide; The method of embodiment E61, wherein said compound is selected from: or a pharmaceutically acceptable salt thereof.

[0488] E64. The method of embodiment E63, wherein the BRAF inhibitor is N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamide or a pharmaceutically acceptable salt thereof.

[0489] E65.BRAF inhibitors N-(2-chloro-4-fluoro-3-((5-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-2-azabicyclo[2.1.1]hexane-2-sulfonamide, (R)—N-(2-chloro-4-fluoro-3-((5-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropyrrolidine-1-sulfonamide, and N-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamide, The method of embodiment E61, wherein said compound is selected from: or a pharmaceutically acceptable salt thereof.

[0490] E66. The method of embodiment E65, wherein the BRAF inhibitor is N-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamide or a pharmaceutically acceptable salt thereof.

[0491] E67. The method of embodiment E60, wherein the anticancer agent is an SHP2 inhibitor.

[0492] E68. The method of embodiment E67, wherein the SHP2 inhibitor is (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine or a pharmaceutically acceptable salt thereof.

[0493] E69. The method according to any one of embodiments E37 to E68, wherein the compound is 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione and the subject is administered 50 mg of 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione once a day.

[0494] E70. A compound according to any one of embodiments E1 to E35 or a pharmaceutically acceptable salt thereof, for use as a medicament.

[0495] E71. A compound according to any one of embodiments E1 to E35 or a pharmaceutically acceptable salt thereof, for use in the treatment of MEK-associated tumors.

[0496] E72. Use of a compound according to any one of embodiments E1 to E35, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a MEK-associated tumor in a subject.

[0497] In order that this invention may be better understood, the following examples are set forth. These examples are for illustrative purposes only and are not to be construed as limiting the scope of the invention in any way.

[0498] The following compounds and intermediates were named using the naming conventions provided in ChemDraw, version 20.1.1.125 (Perkin Elmer Informatics). The naming conventions provided in ChemDraw, version 20.1.1.125 are well known by those skilled in the art, and the naming conventions provided in version 20.1.1.125 are generally considered to be in accordance with the IUPAC (International Union of Pure and Applied Chemistry) recommendations for organic chemical nomenclature and CAS indexing conventions. Unless otherwise noted, all reactants were commercially available without further purification or prepared using methods known in the literature. [Example]

[0499] Biological Examples Example A Cellular phospho-p44 / 42 MAPK (Erk1 / 2) (Thr202 / Tyr204) assay Inhibition of ERK1 / 2 (Thr202 / Tyr204) phosphorylation was determined by the following cellular assay, which involved incubating cells with compounds for 1 hour and quantifying the pERK signal by in-cell Western analysis on fixed cells, normalizing to the GAPDH signal. A375 cells were obtained from ATCC and grown in DMEM supplemented with 10% fetal bovine serum, penicillin / streptomycin, Glutamax (C), non-essential amino acids, and sodium pyruvate. Cells were plated at 30,000 cells / well in 96-well plates and allowed to adhere overnight at 37°C / 5% CO2. Cells were treated with compounds prepared as a 10-point 1:3 dilution series (range: 20 μM to 0.05 nM; maximum concentrations varied from 20 μM to 1 μM) using a final DMSO concentration of 0.5%. Control wells contained either 0.5% DMSO alone (no inhibition control) or 1 μM of a potent control compound (full inhibition control). After 1 hour of incubation, cells were fixed with 3.7% formaldehyde in dPBS (Dulbecco's Phosphate Buffered Saline) for 20 minutes at room temperature. Cells were then washed with dPBS and permeabilized in 100% MeOH for 10 minutes at room temperature. After permeabilization, cells were washed with dPBS and incubated in LI-COR blocking buffer (LI-COR Biosciences, Catalog No. 927-40000) for at least 1 hour. Plates were then incubated with antibodies specific for threonine 202 and tyrosine 204 (Cell Signaling Technologies, Catalog No. 9101), the MEK-dependent ERK1 / 2 phosphorylation sites downstream of MEK in the MAP kinase signaling pathway, and GAPDH (Millipore, Catalog No. MAB374). The pErk1 / 2 (Thr202 / Tyr204) antibody was diluted 1:250 in LI-COR blocking buffer containing 0.05% Tween-20, and GAPDH was diluted 1:2,500. The plates were incubated overnight at 4°C.After washing with PBS / 0.05% Tween-20, cells were incubated with fluorescently labeled secondary antibodies (anti-rabbit Alexa Flour680, Invitrogen catalog number A21109; anti-mouse IRDye800CW, LI-COR Biosciences catalog number 926-32210, both at a 1:1000 dilution) for 1 hour. Cells were then washed as described above and analyzed for fluorescence at both 680 nm and 800 nm wavelengths using an Odyssey CLx infrared imaging system (LI-COR Biosciences). Phosphorylated Erk1 / 2 (Thr202 / Tyr204) signals were normalized to GAPDH signals for each well. IC. 50 Values ​​were calculated from the normalized values ​​using a four parameter fit in BioAssay software and are provided in Table A.

[0500] [Table 5-1]

[0501] [Table 5-2]

[0502] Example B MDR1 LLC-PK1 and BCRP MDCKII permeability assays LLC-PK1 cells transfected with both LLC-PK1 and MDR1 were cultured and plated according to the manufacturer's recommendations, except that the passaging medium contained only 2% fetal bovine serum to extend the passaging time to 7 days.

[0503] The BCRP-transfected MDCKII canine P-gp knockout cell line was cultured and plated according to the manufacturer's recommendations.

[0504] Both positive and negative controls were used to assess the efflux function of P-gp or BCRP in the assays. Assay control and test article stock solutions were prepared in DMSO at final test concentrations of 10 μM and 1 μM, respectively. The final organic concentration in the assay was 1%. All dosing solutions contained 10 μM Lucifer Yellow to monitor the integrity of LLC-PK1 or MDCKII cell monolayers.

[0505] For apical-to-basolateral determinations (AB), 75 μL of test article in transport buffer was added to the apical side of each Transwell, and 250 μL of basolateral medium without compound or Lucifer Yellow was added to each well. For basolateral-to-apical determinations (BA), 250 μL of test article in transport buffer was added to each well, and 75 μL of transport buffer without compound or Lucifer Yellow was added to each Transwell. All experiments were performed in triplicate, and each compound was tested for both apical-to-basolateral and basolateral-to-apical transport. Plates were incubated for 2 hours at 50 rpm and 37°C in 5% CO2 on a Lab-Line Instruments titration orbital shaker (VWR, West Chester, PA). All culture plates were removed from the incubator, and 50 μL of medium was removed from the apical and basolateral portions of each well and added to 150 μL of 1 μM labetalol in 2:1 acetonitrile (acetonitrile):H O, v / v.

[0506] Plates were read using a Molecular Devices (Sunnyvale, CA) Gemini fluorometer to assess Lucifer Yellow concentration at excitation / emission wavelengths of 425 / 535 nm. These values ​​were accepted when found to be less than 2% for apical-to-basolateral flux and less than 5% for basolateral-to-apical flux across MDR1-transfected LLC-PK1 or BCRP-transfected MDCKII cell monolayers. Plates were sealed, and the contents of each well were analyzed by LC-MS / MS. Compound concentrations were determined from the ratio of compound peak area to internal standard (labetalol) compared to the dosing solution.

[0507] LC-MS analysis The LC-MS / MS system consisted of an HTS-PAL autosampler (Leap Technologies, Carrboro, NC), an HP1200 HPLC (Agilent, Palo Alto, CA), and an MDS Sciex 4000QTrap system (Applied Biosystems, Foster City, CA). Chromatographic separation of analytes and internal standards was achieved at room temperature using a C18 column (Reaction Velocity®, 50 × 300 mm, 2.6 μm particle size, Phenomenex, Torrance, CA) with gradient conditions using mobile phases A (water containing 1% isopropyl alcohol and 0.1% formic acid) and B (0.1% formic acid in acetonitrile). The total run time, including re-equilibration, for a single injection was 1.2 min. Mass spectrometric detection of analytes was achieved using ion spray positive mode. Analyte responses were measured by multiple reaction monitoring (MRM) of transitions unique to each compound (protonated precursor ions and selected product ions for each test article, and m / z 329 to m / z 162 for the internal standard, labetalol).

[0508] Permeability coefficient (P app ) is calculated from the following equation: P app =[((C d *V*(1×106 )) / (t*0.12cm 2 *C)] In the formula, C d , V, t, and C0 are the detection concentration (μM), the volume at the administration site (mL), the incubation time (sec), and the initial administration concentration (μM), respectively. app Calculations were performed for each replicate and then averaged. The permeability coefficients for compounds of Formula I are provided in Table B1. In this assay, compounds have a permeability of 8×10 -6 A compound is defined as having a high permeability if its permeability is greater than 2×10 cm / sec. -6 cm / sec~8×10 -6 A compound is defined as having moderate permeability if its permeability is greater than 2×10 cm / sec. -6 A value less than cm / sec is defined as having low permeability.

[0509] The efflux ratio is the average apical-to-basolateral (AB) P app Data and mean P from basolateral to apical (BA) app Calculate from the data. Emission ratio=P app (BA) / P app (AB)

[0510] [Table 6-1]

[0511] [Table 6-2]

[0512] The efflux ratio is the average apical-to-basolateral (AB) P app Data and mean P from basolateral to apical (BA) app Calculate from the data. Emission ratio=P app (BA) / P app (AB)

[0513] Table B2 provides the efflux ratios for compounds of formula I when used in this assay.

[0514] [Table 7-1]

[0515] [Table 7-2]

[0516] Example C PK (free brain to free plasma ratio) (mouse) The ability of representative compounds to penetrate the mouse BBB was determined by assessing the unbound brain to unbound plasma (also called free brain to free plasma) concentration ratio in male CD-1 mice.

[0517] Brain compound levels were generated from PK data of orally administered mice, with typical sampling times of 2, 4, 8, 12, and 24 h after a 10 mg / kg oral gavage dose. Brain samples were stored at -20 ± 5°C prior to analysis. Test compound concentrations in mouse brain homogenates were determined by liquid chromatography-tandem mass spectrometry (LC-MS / MS) after protein precipitation with acetonitrile. A 12-point calibration curve ranging from 0.5 to 10,000 ng / mL was prepared in duplicate. A 400 μg / mL solution of test compound in dimethyl sulfoxide (DMSO) was serially diluted (3-fold) with 100% DMSO, and then 2.5 μL of each standard solution was added to 100 μL of naive male CD-1 mouse brain homogenate. To mimic extraction in the standard curve, 2.5 μL of DMSO was added to all test samples. Both calibration and test brain homogenate samples were spiked with 10 μL of IS (1 μg / mL of structural analog). Brain homogenates were generated by adding 0.75 mL of 4:1 water:MeOH to each brain sample, followed by homogenization in a bead mill tube at 6 m / s for 1 minute using an MP Fast Prep-24®. Proteins were precipitated from 100 μL of brain homogenate sample by adding 300 μL of acetonitrile. Samples were vortex mixed for 5 minutes and spun at approximately 1,500 × g for 15 minutes at 4 °C in an Allegra X-12R centrifuge (Beckman Coulter, Fullerton, CA; SX4750A rotor). A 100 μL aliquot of each supernatant was transferred to a 96-well plate using a 550 μL Personal Pipettor (Apricot Designs, Monrovia, CA) and diluted 1:1 with HPLC-grade water. The resulting plates were sealed with aluminum for LC-MS / MS analysis.

[0518] Brain to plasma ratios were calculated using the concentration of compound measured in the brain divided by the concentration of compound measured in the plasma. Brain to plasma ratios were always generated from a single animal and time point. The free brain to free plasma ratio was calculated using the following equation: (B / P)*(B fu / P fu) was used to calculate the brain to plasma ratio by multiplying the in vitro brain homogenate free fraction divided by the in vitro plasma free fraction.

[0519] Table C provides the free brain to free plasma ratios for representative compounds of Examples 6, 7, 8, 14, 22, 36 and 46 disclosed herein.

[0520] [Table 8] Synthesis Examples Intermediate 1

[0521] [ka] Methyl (E)-2,6-dichloro-4-(2-(dimethylamino)vinyl)nicotinate Step 1. Preparation of methyl 2,6-dichloro-4-methylnicotinate. To a solution of 2,6-dichloro-4-methylnicotinic acid (1.0 g, 4.9 mmol) in 1:1 MeOH:dioxane (10 mL) at 0 °C was added (trimethylsilyl)diazomethane (3.3 mL, 2 M in hexanes, 6.6 mmol). The mixture was removed from the ice bath and stirred for 10 min, then concentrated to half volume and partitioned between water (20 mL) and EtOAc (20 mL). The aqueous layer was extracted with EtOAc (2 × 20 mL), and the combined organic phases were washed with brine (10 mL), dried over NaSO, filtered, and carefully concentrated. The residue was purified by column chromatography eluting with 0–15% EtOAc / petroleum ether to give methyl 2,6-dichloro-4-methylnicotinate (0.81 g, 76%). 1 H NMR (400 MHz, CDCl3) δ 7.2 (s, 1H), 4.0 (s, 3H), 2.3 (s, 3H) ppm.

[0522] Step 2. Preparation of methyl (E)-2,6-dichloro-4-(2-(dimethylamino)vinyl)nicotinate. To a solution of methyl 2,6-dichloro-4-methylnicotinate (810 mg, 3.68 mmol) in DMF (5 mL) was added N,N-dimethylformamide dimethyl acetal (978 mL, 7.36 mmol), and the mixture was stirred at 100° C. for 16 h. The cooled mixture was treated with water (40 mL) and stirred for 10 min, then the solid was collected by filtration, washed with water, and dried in vacuo to give methyl (E)-2,6-dichloro-4-(2-(dimethylamino)vinyl)nicotinate (764 mg, 75%). 1 H NMR (400 MHz, CDCl3) δ 7.3 (s, 1H), 7.1 (d, 1H), 4.8 (d, 1H), 3.1 (s, 3H), 2.9 (s, 6H) ppm.

[0523] Intermediate 2

[0524] [ka] Methyl 4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate Step 1. Preparation of 4-bromo-2,6-dichloronicotinic acid. A solution of 4-bromo-2,6-dichloropyridine (100 g, 440.7 mmol) in THF (1000 mL) was cooled to −78 °C. LDA (2 M in THF, 242.4 mL, 484.8 mmol) was added dropwise at −78 °C, and stirring was continued at −78 °C for 1 h. Solid CO (155.1 g, 3.53 mol) was added portionwise to the reaction, and stirring was continued at −78 °C for 2 h. The reaction was quenched by adding 1 M NaCO (1600 mL), followed by water (500 mL), and stirred for 10 min. The aqueous layer was extracted with EtOAc (300 mL). The pH of the aqueous solution was adjusted with 2 N HCl to obtain a solution with pH 2. The aqueous solution was then extracted with EtOAc (3 × 300 mL). The combined organic layers were dried over Na2SO4, filtered and concentrated to give 4-bromo-2,6-dichloronicotinic acid (540 g, 75%).1 H NMR (400 MHz, DMSO-d6) δ 15.51 - 12.87 (m, 1H), 8.13 (s, 1H) ppm.

[0525] Step 2. Preparation of 4-bromo-2-chloro-6-oxo-1,6-dihydropyridine-3-carboxylic acid. A solution of NaOH (4 M, 1.62 L, 6.46 mol) was heated to 110 °C and 4-bromo-2,6-dichloronicotinic acid (70 g, 258.4 mmol) was added in one portion. The mixture was stirred for 8 h and then cooled to 0 °C. The reaction was adjusted to pH 1 with HCl (6 M) and stirred for 30 min. The solid was collected by filtration and dried in vacuo to give 4-bromo-2-chloro-6-oxo-1,6-dihydropyridine-3-carboxylic acid (100% assumed).

[0526] Step 3. Preparation of methyl 4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate. To a mixture of 4-bromo-2-chloro-6-oxo-1,6-dihydropyridine-3-carboxylic acid (100 g, 396 mmol) in DMF (800 mL) was added methyl iodide (168.6 g, 1.19 mol, 73.98 mL) and K2CO3 (164.2 g, 1.19 mol) in one portion. The mixture was stirred at 25 °C for 3 h, then poured into saturated NH4Cl (1800 mL), and the aqueous phase was extracted with EtOAc (2 × 500 mL). The combined organic phase was washed with brine (400 mL), dried over Na2SO4, filtered, and concentrated. The combined residues (five batches) were purified by column chromatography eluting with 2-100% EtOAc / petroleum ether to give methyl 4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (141.83 g, 24.7% from five batches). 1 H NMR (400 MHz, CDCl3) δ 6.86 (s, 1H), 3.94 (s, 3H), 3.69 - 3.66 (m, 3H); MS (apci, m / z) = 280.0, 282.0 (M+H).

[0527] Intermediate 3

[0528] [ka] Methyl (Z)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate Methyl 4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (20.0 g, 71.3 mmol), methanesulfonate (2-dicyclohexylphosphino-2',6'-di-i-propoxy-1,1'-biphenyl) (2-amino-1,1'-biphenyl-2-yl)palladium(II) (5.96 g, 7.13 mmol), and (Z)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (14.8 g, 74.9 mmol) were suspended in 1,4-dioxane (700 mL), and KCO (53.5 mL, 107 mmol) (2 N aqueous solution) was added. The mixture was stirred at 60 °C for 6 h, then at ambient temperature under an argon atmosphere for 12 h. The reaction was partitioned between water (1500 mL) and EtOAc (500 mL). The aqueous layer was extracted with EtOAc (2 × 400 mL), and the combined organic layers were washed with brine (500 mL), dried over NaSO, filtered, and concentrated. The residue was purified by column chromatography eluting with 0–40–60% EtOAc / heptane to give methyl (Z)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (13.3 g, 68%). 1 H NMR (400 MHz, CDCl3) δ 7.23 (s, 1H), 6.45 (d, 1H), 4.86 (d, 1H), 4.40 (q, 2H), 3.89 (s, 3H), 3.67 (s, 3H), 1.35 (t, 3H); MS (apci, m / z) = 272.0 (M+H).

[0529] Intermediate 4

[0530] [ka] Methyl 4-(2-((2-(tert-butoxy)ethoxy)amino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate Step 1. Preparation of methyl (E / Z)-4-(2-((2-(tert-butoxy)ethoxy)imino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate. Methyl (Z)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (0.95 g, 3.50 mmol) and O-(2-(tert-butoxy)ethyl)hydroxylamine hydrochloride (593 mg, 3.50 mmol) were combined in 1,4-dioxane (10 mL). EtN (487 mL, 3.50 mmol) and HCl (1.75 mL, 6.99 mmol) (4 N / dioxane) were added. The suspension was heated to 60 °C for 1 h, then cooled and filtered. The filtrate was concentrated to give methyl (E / Z)-4-(2-((2-(tert-butoxy)ethoxy)imino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (assumed 100%) as a 1:1 mixture of isomers. MS (apci, m / z) = 359.1 (M+H).

[0531] Step 2. Preparation of methyl 4-(2-((2-(tert-butoxy)ethoxy)amino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate. To a solution of methyl (E / Z)-4-(2-((2-(tert-butoxy)ethoxy)imino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (1.25 g, 3.48 mmol) in IPA (20 mL) was added sodium cyanoborohydride (1.09 g, 17.4 mmol), followed by acetic acid (1.0 mL, 17.4 mmol). The mixture was stirred at ambient temperature for 16 h and then partitioned between saturated NaHCO3 (50 mL) and EtOAc (50 mL). The aqueous layer was extracted with EtOAc (2 × 30 mL), and the combined organic phases were washed with brine (30 mL), then dried over NaSO, filtered, and concentrated. The residue was purified by column chromatography eluting with 0–80% EtOAc / DCM to give methyl 4-(2-((2-(tert-butoxy)ethoxy)amino)ethyl)-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (0.66 g, 53%). 1 H NMR (400 MHz, CDCl3) δ 6.43 (s, 1H), 3.90 (s, 3H), 3.78 (t, 2H), 3.68 (s, 3H), 3.50 (t, 2H), 3.10 (t, 2H), 2.71 (t, 2H), 1.20 (s, 9H) ppm; MS (apci, m / z) = 361.1, 363.1 (M+H).

[0532] Intermediate 5

[0533] [ka] 2-chloro-4-ethylaniline Step 1. Preparation of N-(4-ethylphenyl)acetamide. 4-Ethylaniline (513 μL, 4.13 mmol) was dissolved in DCM (10.3 mL). Triethylamine (690 μL, 4.95 mmol) was added and cooled to 0 °C. After that, acetic anhydride (467 μL, 4.95 mmol) was added dropwise. After 30 min, the reaction was quenched by adding saturated NaHCO (50 mL). The organic layer was separated, and the aqueous layer was extracted with DCM (2 × 25 mL). The combined organic layers were dried over NaSO, filtered, and concentrated. The residue was triturated with hexane, and the solid was collected by filtration to give N-(4-ethylphenyl)acetamide (600 mg, 89%). 1 H NMR (400 MHz, CDCl3) δ 7.40 - 7.38 (d, 2H), 7.16 - 7.14 (d, 2H), 7.07 (br s, 1H), 2.64 - 2.58 (q, 2H), 2.16 (s, 3H), 1.23 - 1.20 (t, 3H) ppm.

[0534] Step 2. Preparation of N-(2-chloro-4-ethylphenyl)acetamide. N-(4-Ethylphenyl)acetamide (50 mg, 0.31 mmol) was dissolved in DMF (613 μL). After adding N-chlorosuccinimide (65 mg, 0.49 mmol), the solution was heated to 70 °C for 6 h and then allowed to cool to ambient temperature over 16 h. The reaction mixture was poured into 2 N HCl (4 mL) and stirred for 15 min. The mixture was extracted with EtOAc (10 mL). The organic layer was washed with water (3 × 10 mL), dried over NaSO, filtered, and concentrated. The residue was purified by column chromatography eluting with 0–25% EtOAc / hexane to give N-(2-chloro-4-ethylphenyl)acetamide (36 mg, 59%). 1 H NMR (400 MHz, CDCl3) δ 8.23 ​​- 8.21 (d, 1H), 7.51 (br s, 1H), 7.20 - 7.19 (d, 1H), 7.11 - 7.08 (dd, 1H), 2.63 - 2.57 (q, 2H), 2.23 (s, 3H), 1.23 - 1.20 (t, 3H) ppm.

[0535] Step 3. Preparation of 2-chloro-4-ethylaniline. N-(2-chloro-4-ethylphenyl)acetamide (36 mg, 0.18 mmol) was dissolved in EtOH (0.5 mL) and 12 N HCl (0.5 mL, 6.0 mmol) was added. The mixture was stirred at 120 °C for 2 h. The reaction mixture was cooled to ambient temperature, brought to pH 10 by the addition of 6 N NaOH, and then extracted with MTBE (3 × 25 mL). The combined organic layers were dried over NaSO, filtered, and concentrated to give 2-chloro-4-ethylaniline (26 mg, 92%). 1 H NMR (400 MHz, CDCl3) δ 7.09 - 7.08 (d, 1H), 6.91 - 6.88 (dd, 1H), 6.71 - 6.69 (d, 1H), 2.55 - 2.49 (q, 2H), 1.20 - 1.16 (t, 3H) ppm. MS (apci, m / z) = 156.1 (M+H).

[0536] Intermediate 6

[0537] [ka] 4-((difluoromethyl)thio)-2-fluoroaniline LiBF4 (196 mg, 2.10 mmol) and LiH (17.5 mg, 2.10 mmol) were combined in DMF (8.8 mL, 1.75 mmol). 4-Amino-3-fluorobenzenethiol (250 mg, 1.75 mmol) was added, and the mixture was stirred at ambient temperature for 5 minutes. (Trifluoromethyl)trimethylsilane (0.644 mL, 4.37 mmol) was added rapidly, and the solution was then stirred at the same temperature for 10 minutes. TBAF (6 mL, 1N / THF, 6.00 mmol) was added rapidly, and the solution was then stirred at the same temperature for 10 minutes. The reaction mixture was quenched with water (50 mL). The mixture was extracted with EtOAc (2 × 25 mL), and the combined organic layers were washed with water (3 × 50 mL) and brine (50 mL), then dried over Na2SO4, filtered, and concentrated. The residue was purified by column chromatography eluting with 0-15% EtOAc / hexanes to give 4-((difluoromethyl)thio)-2-fluoroaniline (56 mg, 17%). 1 H NMR (400 MHz, CDCl3) δ 7.25 - 7.22 (dd, 1H), 7.19 - 7.16 (ddd, 1H), 6.86 - 6.58 (t, 1H), 6.77 - 6.73 (dd, 1H), 3.95 (br s, 1H) ppm.

[0538] Intermediate 7

[0539] [ka] 2-chloro-4-(methylthio)aniline 2-Chloro-4-iodoaniline (250 mg, 0.986 mmol), NiBr (22 mg, 0.099 mmol), Zn powder (129 mg, 1.97 mmol), and 2,2'-bipyridine (15 mg, 0.099 mmol) were dissolved in THF (1.6 mL) under an Ar atmosphere. 1,2-Dimethyldisulfane (44 μL, 0.493 mmol) was added, and the mixture was sealed and heated to 65 °C for 16 h. The reaction mixture was diluted with EtOAc (50 mL) and washed with concentrated NHOH (50 mL), followed by 10% citric acid solution (50 mL). The combined organic layers were dried over NaSO, filtered, and concentrated. The residue was purified by column chromatography eluting with 0–15% EtOAc / hexane to give 2-chloro-4-(methylthio)aniline (116 mg, 68%). 1 H NMR (400 MHz, CDCl3) δ 7.27 - 7.26 (m, 1H), 7.09 - 7.06 (dd, 1H), 6.71 - 6.69 (d, 1H), 4.02 (br s, 2H), 2.41 (s, 3H) ppm. MS (apci, m / z) = 174.0 (M+H).

[0540] Intermediate 8

[0541] [ka] Methyl (E)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate Methyl 4-bromo-2-chloro-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (1.0 g, 3.565 mmol), methanesulfonate (2-dicyclohexylphosphino-2',6'-di-i-propoxy-1,1'-biphenyl) (2-amino-1,1'-biphenyl-2-yl)palladium(II) (0.2982 g, 0.3565 mmol), and (E)-2-(2-ethoxyvinyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (0.7414 g, 3.743 mmol) were suspended in 1,4-dioxane (35.65 mL) and potassium carbonate (2.674 mL, 2N aqueous solution, 5.35 mmol) was added. After degassing with argon, the mixture was stirred at 60 °C for 4 h. The cooled reaction was partitioned between water (150 mL) and EtOAc (50 mL). The aqueous layer was washed with EtOAc (2×40 mL). The combined organic layers were washed with brine (150 mL), dried over sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography eluting with 0–40% EtOAc / heptane to give methyl (E)-2-chloro-4-(2-ethoxyvinyl)-1-methyl-6-oxo-1,6-dihydropyridine-3-carboxylate (379 mg, 39%). 1 H NMR (400 MHz, CDCl3) δ 7.27 (s, 1H), 7.03 (d, 1H), 6.43 (s, 1H), 5.53 (d, 1H), 3.95 - 3.83 (m, 5H), 3.67 (s, 3H), 1.34 (t, 2H) ppm. MS (apci, m / z) = 272.1 (M+H).

[0542] Example 1

[0543] [ka] 8-((4-Bromo-2-fluorophenyl)amino)-2-cyclopropyl-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione Step 1. Preparation of methyl 2,6-dichloro-4-(2-oxoethyl)nicotinate. A suspension of methyl (E)-2,6-dichloro-4-(2-(dimethylamino)vinyl)nicotinate (0.797 g, 2.90 mmol) in EtO (30 mL) and 1 N HCl (30 mL) was vigorously stirred at ambient temperature for 1 h. The resulting solution was treated with brine (20 mL) and extracted with EtO (3 × 10 mL). The combined organic phase was washed with brine (10 mL), dried over MgSO, filtered, and carefully concentrated to give methyl 2,6-dichloro-4-(2-oxoethyl)nicotinate (assumed 100%), which was used immediately.

[0544] Step 2. Preparation of 8-chloro-2-cyclopropyl-6-methoxy-3,4-dihydro-2,7-naphthyridin-1(2H)-one. To a solution of methyl 2,6-dichloro-4-(2-oxoethyl)nicotinate (0.719 g, 2.90 mmol) in 1:1 IPA:MeOH (20 mL) was added cyclopropylamine (201 mL, 2.90 mmol) at 0 °C. The mixture was stirred at ambient temperature for 10 min, followed by the addition of sodium cyanoborohydride (546 mg, 8.70 mmol) and acetic acid (498 mL, 8.70 mmol). After stirring at ambient temperature for 16 h, the mixture was partitioned between saturated NaHCO (50 mL) and EtOAc (20 mL), and the aqueous layer was extracted with EtOAc (2 × 20 mL). The combined organic phase was washed with brine (10 mL), dried over NaSO, filtered, and concentrated. The residue was dissolved in methanol (10 mL), treated with 1 N NaOH (3.33 mL, 3.33 mmol), and stirred at ambient temperature for 1 h. The mixture was concentrated to half volume and then partitioned between water (20 mL) and DCM (20 mL). The aqueous layer was extracted with DCM (2 × 10 mL), and the combined organic phases were washed with brine (10 mL), dried over NaSO, filtered, and concentrated. The residue was purified by column chromatography eluting with 0–10% (20% MeOH / DCM) / DCM to give 8-chloro-2-cyclopropyl-6-methoxy-3,4-dihydro-2,7-naphthyridin-1(2H)-one (310 mg, 42%). 1H NMR (400 MHz, CDCl3) δ 6.5 (s, 1H), 4.0 (s, 3H), 3.5 (t, 2H), 2.9 (m, 3H), 0.9 (m, 2H), 0.7 (m, 2H) ppm; MS (apci, m / z) = 253.1, 255.1 (M+H).

[0545] Step 3. Preparation of 8-chloro-2-cyclopropyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione. To a solution of 8-chloro-2-cyclopropyl-6-methoxy-3,4-dihydro-2,7-naphthyridine-1(2H)-one (99 mg, 0.39 mmol) in acetonitrile (2 mL) was added trimethylsilyl iodide (2.94 mL, 1.96 mmol). The mixture was stirred at ambient temperature for 48 h and then concentrated. The residue was purified by column chromatography eluting with 0–15–20% (20% MeOH / DCM) / DCM to give 8-chloro-2-cyclopropyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione (assumed 100%), which was used immediately. MS (apci, m / z) = 239.0 (M+H).

[0546] Step 4. Preparation of 8-chloro-2-cyclopropyl-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione. To a solution of 8-chloro-2-cyclopropyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione (93 mg, 0.39 mmol) in THF (2 mL) was added KCO (110 mg, 0.78 mmol), followed by methyl iodide (29 mL, 0.47 mmol). The mixture was stirred at ambient temperature for 2 h, then treated with additional methyl iodide (50 mL, 0.84 mmol) and stirred for an additional 16 h. The mixture was filtered, washed with (4:1) DCM / MeOH (10 mL), and the filtrate was concentrated. The residue was purified by column chromatography eluting with 0-15% (20% MeOH / DCM) / DCM to give 8-chloro-2-cyclopropyl-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione (31 mg, 31%). 1 H NMR (400 MHz, CDCl3) δ 6.29 (s, 1H), 3.73 (s, 3H), 3.45 (t, 2H), 2.87-2.81 (m, 1H), 2.77 (t, 2H), 0.94-0.89 (m, 2H), 0.72-0.68 (m, 2H) ppm; MS (apci, m / z) = ...

Claims

1. structure 【Chemistry 3】 or a pharmaceutically acceptable salt thereof.

2. structure 【Chemistry 4】 A pharmaceutical composition comprising a compound which is 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione having the formula:

3. A pharmaceutical composition comprising a crystal containing 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.

4. A pharmaceutical composition comprising a crystal of anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.

5. 5. The pharmaceutical composition of claim 4, comprising a crystalline form of anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione characterized by PXRD peaks at 5.0, 8.7, 9.3, 10.8, 14.5, 15.3, 18.8, and 20.5 degrees two-theta (±0.2 degrees two-theta).

6. 5. The pharmaceutical composition of claim 4, comprising a crystalline form of anhydrous 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione having a PXRD pattern including characteristic peaks at 7.1, 9.4, 12.4, 12.8, 14.3, 15.6, 16.4, 17.4, 18.5, 18.9, 19.5, 19.9, 21.1, 21.4, 23.2, 23.7, 24.8, 25.6, 27.6, 30.3, 33.2, 33.5, and 37.5 degrees two-theta (±0.2 degrees two-theta).

7. A pharmaceutical composition comprising a crystal of 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione monohydrate.

8. A pharmaceutical composition comprising the crystalline 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione monohydrate of claim 7, characterized by having PXRD peaks at 13.7, 18.0, and 18.3 degrees two-theta (±0.2 degrees two-theta).

9. A pharmaceutical composition comprising an amorphous solid of 8-((2-fluoro-4-(methylthio)phenyl)amino)-2-(2-hydroxyethoxy)-7-methyl-3,4-dihydro-2,7-naphthyridine-1,6(2H,7H)-dione.

10. 10. The pharmaceutical composition of any one of claims 1 to 9 for use in treating MEK-associated tumors.

11. 11. The pharmaceutical composition of claim 10, wherein the tumor has a BRAF V600 mutation selected from V600E, V600K, V600D, V600R and V600S.

12. The pharmaceutical composition of claim 10, wherein the tumor has a BRAF V600E mutation.

13. The pharmaceutical composition of claim 10, wherein the tumor is a CNS tumor.

14. The pharmaceutical composition of claim 13, wherein the CNS tumor is an intracranial tumor.

15. The pharmaceutical composition of claim 14, wherein the intracranial tumor is a brain tumor.

16. The pharmaceutical composition of claim 14, wherein the brain tumor is a metastatic brain tumor.

17. 17. The pharmaceutical composition of claim 16, wherein the metastatic brain tumor is selected from metastatic melanoma, metastatic colorectal cancer, metastatic non-small cell lung cancer, metastatic thyroid cancer, and metastatic ovarian cancer.

18. 14. The pharmaceutical composition of claim 13, wherein the CNS tumor is intracranial or extracranial LMD.

19. 19. The pharmaceutical composition of claim 18, wherein the LMD is selected from metastatic melanoma, metastatic colorectal cancer, and metastatic non-small cell lung cancer.

20. The pharmaceutical composition of claim 14, wherein the intracranial tumor is a primary tumor.

21. The pharmaceutical composition of claim 20, wherein the primary brain tumor is a malignant tumor.

22. 22. The pharmaceutical composition of claim 21, wherein the primary brain tumor is a grade 2 glioma, a grade 3 glioma, or a grade 4 glioma.

23. The pharmaceutical composition of claim 20, wherein the primary brain tumor is a benign tumor.

24. The pharmaceutical composition of claim 10, wherein the tumor has a BRAF fusion.

25. The tumor is KIAA11549-BRAF, MKRN1-BRAF, TRIM24-BRAF, AGAP3-BRAF, ZC3HAV1-BRAF, AKAP9-BRAF, CCDC6-BRAF, AGK-BRAF, EPS15-BRAF F, NUP214-BRAF, ARMC10-BRAF, BTF3L4-BRAF, GHR-BRAF, ZC3HAV1-BRAF, ZNF767-BRAF, CCDC91-BRAF, DYNC112-BRAF, ZKSCAN1-BR 25. The pharmaceutical composition of claim 24, comprising a BRAF fusion selected from: BRAF, GTF2I-BRAF, MZT1-BRAF, RAD18-BRAF, CUX1-BRAF, SLC12A7-BRAF, MYRIP-BRAF, SND1-BRAF, NUB1-BRAF, KLHL7-BRAF, TANK-BRAF, RBMS3-BRAF, STRN3-BRAF, STK35-BRAF, ETFA-BRAF, SVOPL-BRAF, and JHDM1D-BRAF.

26. 26. The pharmaceutical composition of claim 25, wherein the tumor is breast cancer, colorectal cancer, esophageal cancer, glioma, head and neck carcinoma, lung cancer, melanoma, pancreatic cancer, prostate cancer, sarcoma, thyroid cancer, carcinoma of unknown primary, pleural mesothelioma, rectal adenocarcinoma, endometrial cancer, or ovarian serous carcinoma.

27. The pharmaceutical composition of claim 10, wherein the tumor is a BRAF wild-type tumor.

28. 11. The pharmaceutical composition of claim 10 for use in combination with one or more further anti-cancer treatments.

29. 29. The pharmaceutical composition of claim 28, wherein the additional anti-cancer treatment is selected from one or more anti-cancer agents.

30. 30. The pharmaceutical composition of claim 29, wherein the anticancer agent is a BRAF inhibitor.

31. 31. The pharmaceutical composition of claim 30, wherein the BRAF inhibitor is encorafenib or a pharmaceutically acceptable salt thereof.

32. BRAF inhibitors, N-(3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-2,4-difluorophenyl)propane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropropane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4,5-difluorophenyl)propane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)propane-1-sulfonamide; N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamide; N-(2-chloro-4-fluoro-3-((5-methyl-3-(methyl-d3)-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-phenyl)-3-fluoropropane-1-sulfonamide; N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}propane-1-sulfonamide; N-(3-chloro-4-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy)-5-fluoropyridin-2-yl)propane-1-sulfonamide; and N-{2-chloro-3-[(3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)oxy]-4-fluorophenyl}-3-fluoropropane-1-sulfonamide; or a pharmaceutically acceptable salt thereof.

33. 33. The pharmaceutical composition of claim 32, wherein the BRAF inhibitor is N-(2-chloro-3-((3,5-dimethyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoropropane-1-sulfonamide or a pharmaceutically acceptable salt thereof.

34. BRAF inhibitors, N-(2-chloro-4-fluoro-3-((5-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-2-azabicyclo[2.1.1]hexane-2-sulfonamide, (R)—N-(2-chloro-4-fluoro-3-((5-fluoro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-fluoropyrrolidine-1-sulfonamide, and N-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamide, or a pharmaceutically acceptable salt thereof.

35. 35. The pharmaceutical composition of claim 34, wherein the BRAF inhibitor is N-(2-chloro-3-((5-chloro-3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)-4-fluorophenyl)-3-fluoroazetidine-1-sulfonamide or a pharmaceutically acceptable salt thereof.

36. 30. The pharmaceutical composition of claim 29, wherein the anticancer agent is an SHP2 inhibitor.

37. 37. The pharmaceutical composition of claim 36, wherein the SHP2 inhibitor is (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine or a pharmaceutically acceptable salt thereof.

38. The pharmaceutical composition of claim 10 , wherein the subject is a human.

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