WO2023215256A1

WO2023215256A1 - Sos1 inhibitors and uses thereof

Info

Publication number: WO2023215256A1
Application number: PCT/US2023/020638
Authority: WO
Inventors: Andreas BUCKL; Severin THOMPSON; John E. Knox; Adrian L. Gill
Original assignee: Revolution Medicines Inc
Current assignee: Revolution Medicines Inc
Priority date: 2022-05-03
Filing date: 2023-05-02
Publication date: 2023-11-09
Anticipated expiration: 2024-11-03

Abstract

The present disclosure is directed to modulators of SOS1 and their use in the treatment of disease. Also disclosed are pharmaceutical compositions comprising the same.

Description

SOS1 INHIBITORS AND USES THEREOF CROSS REFERENCE TO RELATED APPLICATION(S) [0001] This application claims the benefit of priority to U.S. Provisional Application Serial No.63/364,083, which was filed May 3, 2022, the disclosure of which is hereby incorporated by reference as if set forth in its entirety. FIELD OF THE DISCLOSURE [0002] The present disclosure relates to inhibitors of SOS1 useful in the treatment of diseases or disorders. Specifically, the present disclosure is concerned with compounds and compositions inhibiting SOS1, methods of treating diseases associated with SOS1, and methods of synthesizing these compounds. BACKGROUND OF THE DISCLOSURE [0003] RAS-family proteins including KRAS (V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog), NRAS (neuroblastoma RAS viral oncogene homolog) and HRAS (Harvey murine sarcoma virus oncogene) and any mutants thereof are small GTPases that exist in cells in either GTP-bound or GDP-bound states (McCormick et al., J. Mol. Med. (Berl), 2016, 94(3):253-8; Nimnual et al., Sci. STKE., 2002, 2002(145):pl36). The RAS-family proteins have a weak intrinsic GTPase activity and slow nucleotide exchange rates (Hunter et al., Mol. Cancer Res., 2015, 13(9): 1325-35). Binding of GTPase activating proteins (GAPs) such as NF1 increases the GTPase activity of RAS-family proteins. The binding of guanine nucleotide exchange factors (GEFs) such as SOS1 (Son of Sevenless 1) promote release of GDP from RAS-family proteins, enabling GTP binding (Chardin et al., Science, 1993, 260(5112):1338-43). When in the GTP-bound state, RAS- family proteins are active and engage effector proteins including RAF and phosphoinositide 3-kinase (PI3K) to promote the RAF/mitogen or extracellular signal- regulated kinases (MEK/ERK). Published data indicate a critical involvement of SOS1 in mutant KRAS activation and oncogenic signaling in cancer (Jeng et al., Nat. Commun., 2012, 3:1168). Depleting SOS1 levels decreased the proliferation rate and survival of tumor cells carrying a KRAS mutation whereas no effect was observed in KRAS wild type cell lines. The effect of loss of SOS1 could not be rescued by introduction of a catalytic site mutated SOS1, demonstrating the essential role of SOS1 GEF activity in KRAS mutant cancer cells.

[0004] SOS1 is critically involved in the activation of RAS-family protein signaling in cancer via mechanisms other than mutations in RAS-family proteins. SOS1 interacts with the adaptor protein Grb2 and the resulting SOSl/Grb2 complex binds to activated/phosphorylated Receptor Tyrosine Kinases (e.g., EGFR, ErbB2, ErbB3, ErbB4, PDGFR-A/B, FGFR1/2/3, IGF1 R, INSR, ALK, ROS, TrkA, TrkB, TrkC, RET, c-MET, VEGFR1/2/3, AXL) (Pierre et al., Biochem. Pharmacol., 2011, 82(9): 1049-56). SOS1 is also recruited to other phosphorylated cell surface receptors such as the T cell Receptor (TCR), B cell Receptor (BCR) and monocyte colony-stimulating factor receptor (Salojin et al., J. Biol. Chem. 2000, 275(8):5966-75). This localization of SOS1 to the plasma membrane, proximal to RAS-family proteins, enables SOS1 to promote RAS-family protein activation. SOS 1 -activation of RAS-family proteins can also be mediated by the interaction of SOSl/Grb2 with the BCR-ABL oncoprotein commonly found in chronic myelogenous leukemia (Kardinal et al., 2001, Blood, 98:1773-81; Sini et al., Nat. Cell Biol., 2004, 6(3):268-74). Furthermore, alterations in SOS1 have been implicated in cancer. SOS1 mutations are found in embryonal rhabdomyosarcomas, Sertoli cell testis tumors, granular cell tumors of the skin (Denayer et al., Genes Chromosomes Cancer, 2010, 49(3):242-52) and lung adenocarcinoma (Cancer Genome Atlas Research Network., Nature, 2014, 511 (751 l):543-50). Meanwhile over-expression of S0S1 has been described in bladder cancer (Watanabe et al., IUBMB Life, 2000, 49(4):317-20) and prostate cancer (Timofeeva et al., Int. J. Oncol., 2009; 35(4):751-60). In addition to cancer, hereditary S0S1 mutations are implicated in the pathogenesis of RASopathies like e.g., Noonan syndrome (NS), cardio-facio-cutaneous syndrome (CFC) and hereditary gingival fibromatosis type 1 (Pierre et al., Biochem. Pharmacol., 2011, 82(9): 1049-56).

[0005] S0S1 is also a GEF for the activation of the GTPases RAC1 (Ras-related C3 botulinum toxin substrate 1) (Innocenti et al., J. Cell Biol., 2002, 156(1): 125-36). RAC1, like RAS-family proteins, is implicated in the pathogenesis of a variety of human cancers and other diseases (Bid et al., Mol. Cancer Ther. 2013, 12(10): 1925-34).

[0006] Son of Sevenless 2 (S0S2), a homolog of S0S1 in mammalian cells, also acts as a GEF for the activation of RAS-family proteins (Pierre et al., Biochem. Pharmacol., 2011, 82(9): 1049-56; Buday et al., Biochim. Biophys. Acta., 2008, 1786(2): 178-87). Published data from mouse knockout models suggests a redundant role for S0S1 and S0S2 in homeostasis in the adult mouse. Whilst germline knockout of S0S1 in mice results in lethality during mid-embryonic gestation (Qian et al., EMBO J., 2000, 19(4):642-54), systemic conditional S0S1 knockout adult mice are viable (Baltanas et al., Mol. Cell. Biol., 2013, 33(22):4562-78). S0S2 gene targeting did not result in any overt phenotype in mice (Esteban et al., Mol. Cell. Biol., 2000, 20(17):6410-3). In contrast, double S0S1 and S0S2 knockout leads to rapid lethality in adult mice (Baltanas et al., Mol. Cell. Biol., 2013, 33(22):4562-78). These published data suggest that selective targeting of individual SOS isoforms (e.g., selective S0S1 targeting) may be adequately tolerated to achieve a therapeutic index between SOSl/RAS-family protein driven cancers (or other SOSl/RAS-family protein pathologies) and normal cells and tissues.

[0007] Selective pharmacological inhibition of the binding of the catalytic site of S0S1 to RAS-family proteins is expected to prevent SOS 1 -mediated activation of RAS- family proteins to the GTP-bound form. Such SOS1 inhibitor compounds are be expected to consequently inhibit signaling in cells downstream of RAS-family proteins (e.g., ERK phosphorylation). In cancer cells associated with dependence on RAS-family proteins (e.g., KRAS mutant cancer cell lines), SOS1 inhibitor compounds are be expected to deliver anti- cancer efficacy (e.g., inhibition of proliferation, survival, metastasis, etc.). High potency towards inhibition of SOSl :RAS-family protein binding (nanomolar level IC50 values) and ERK phosphorylation in cells (nanomolar level IC50 values) are desirable characteristics for a S0S1 inhibitor compound. Furthermore, a desirable characteristic of a S0S1 inhibitor compound would be the selective inhibition of S0S1 over S0S2. This conclusion is based on the viable phenotype of S0S1 knockout mice and lethality of S0S1/S0S2 double knockout mice, as described above.

[0008] These characteristics have not been achieved in previously described S0S1 inhibitor compounds. In the last decades, the RAS family proteins-SOSl protein interaction has gained increasing recognition. Several efforts to identify and optimize binders, which target either the effector binding site of RAS or the catalytic binding site of S0S1 (for a selected review see: Lu et al., ChemMedChem. 2016, 11 (8): 814-21), have been made with limited success. [0009] Recently, small activating molecules have been identified, which bind to a lipophilic pocket of SOS1 in close proximity to the RAS binding site (Bums et al., Proc. Natl. Acad. Sci. 2014, 111(9):3401-6). However, binding of these molecules seems to lead to increased nucleotide exchange and thereby activation of RAS instead of deactivation.

[0010] In an effort to stabilize the protein-protein-interaction of RAS-family proteins with SOS1 and to prevent reloading of RAS-family proteins with GTP, several different fragments were subsequently identified (Winter et al., J. Med. Chem. 2015, 58(5):2265-74). However, reversible binding of fragments to SOS1 did not translate into a measurable effect on the nucleotide exchange and only a weak effect was observed for fragments covalently bound to RAS.

[0011] Also recently, studies have been conducted to combine rational design and screening platforms to identify small molecule inhibitors of SOS1 (Evelyn et al., Chem. Biol. 2014, 21 (12): 1618-28; Evelyn et al., J. Biol. Chem. 2015, 290(20): 12879-98; Zheng et al., WO 2016/077793), i.e., compounds which bind to SOS1 and inhibit protein-protein interaction with RAS-family proteins. Although compounds with a slight inhibitory effect on SOS1 have been identified, the effects on guanine nucleotide exchange and cellular signal transduction modulation (e.g., ERK phosphorylation) are weak.

[0012] Two recent clinical trials have been initiated to study S0S1 inhibition in different contexts: NCT04111458 (“A Study to Test Different Doses of BI 1701963 [a S0S1 inhibitor] Alone and Combined With Trametinib in Patients With Different Types of Advanced Cancer (Solid Tumours With KRAS Mutation)”) and NCT04975256 (“Adagrasib in Combination With BI 1701963 [a S0S1 inhibitor] in Patients With Cancer (KRYSTAL 14)”).

BRIEF DESCRIPTION OF THE DISCLOSURE

[0013] The present disclosure relates to compounds capable of inhibiting the activity of S0S1. The present disclosure further provides a process for the preparation of compounds, pharmaceutical preparations comprising such compounds and methods of using such compounds and compositions in the management of diseases or disorders associated with the aberrant activity of SOS1. [0014] One aspect of the present disclosure relates to compounds having a structure of Formula (I):

or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein.

[0015] R₁ is selected from the group consisting of optionally substituted 6- membered aryl and optionally substituted 5-6 membered heteroaryl.

[0016] R₂ is selected from the group consisting of H and optionally substituted C_1- ₆ alkyl.

[0017] R₃ is - OR_3a, and further wherein R_3a is optionally substituted C_1-3 alkyl.

[0018] R₄ is selected from the group consisting of H, C_1-6 alkyl, 3-14 membered cycloalkyl, 3-14 membered cycloalkenyl, 3-14 membered heterocyclyl, 6-10 membered aryl, and 5-10 membered heteroaryl; wherein each C_1-6 alkyl, 3-14 membered cycloalkyl, 3- 14 membered cycloalkenyl, 3-14 membered heterocyclyl, 6-10 membered aryl, and 5-10 membered heteroaryl is optionally substituted with C_1-6 alkyl, -R_4a, -OR_4a, -O-C_1-6 alkyl- R_4a, =O, halogen, -C(O)R_4a, -C(O)OR_4a, -C(O)NR_4bR_4c, -NR_4bC(O)R_4c, -CN, =NR_4a, - NR_4bR_4c, - SO₂R_4a, 3-6 membered cycloalkyl optionally substituted with R_4a, 3-7 membered heterocyclyl optionally substituted with R_4a, 6-10 membered aryl optionally substituted with R_4a, or 5-10 membered heteroaryl optionally substituted with R_4a.

[0019] R_4a is H, C_1-6 alkyl, C_1-6 haloalkyl, -C(O)R_4b, -C(O)NR_4bR_4c, =O, 3-6 membered cycloalkyl, 6-10 membered aryl optionally substituted with -OR_4b, -CN, =N-3- 6 membered cycloalkyl, 3-7 membered heterocyclyl, -(CH₂)_rOCH₃, or -(CH₂)_rOH wherein r is 1, 2, or 3. [0020] Each R_4b is independently H, C_1-6 alkyl. Each R_4c is independently H or C_1-6 alkyl.

[0021] Another aspect of the present disclosure relates to a pharmaceutical composition comprising a compound of Formula (I) as described herein, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, and a pharmaceutically acceptable carrier.

[0022] Another aspect of the present disclosure relates to a method of inhibiting S0S1 in a subject, comprising administering to the subject: a compound of the present disclosure, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof; or a pharmaceutical composition of the present disclosure, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof.

[0023] Another aspect of the present disclosure relates to a method of inhibiting the interaction of S0S1 and a RAS-family protein in a cell or inhibiting the interaction of S0S1 and RAC1 in a cell, comprising administering to the cell: a compound of the present disclosure, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof; or a pharmaceutical composition of the present disclosure, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof.

[0024] Another aspect of the present disclosure relates to a method of treating or preventing a disease, wherein treating or preventing the disease is characterized by inhibition of the interaction of S0S1 and a RAS-family protein or by inhibition of the interaction of S0S1 and RAC1, the method comprising administering to a subject in need thereof an effective amount of: a compound of the present disclosure, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof; or a pharmaceutical composition of the present disclosure, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof.

[0025] Another aspect of the present disclosure relates to a method of treating or preventing cancer in a subject in need thereof, comprising administering to the subject an effective amount of: a compound of the present disclosure, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof; or a pharmaceutical composition of the present disclosure, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof.

[0026] The present disclosure also provides compounds that are useful in inhibiting S0S1, in vivo or in vitro.

DETAILED DESCRIPTION OF THE DISCLOSURE

[0027] The details of the present disclosure are set forth in the accompanying description below. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, illustrative methods and materials are now described. Other features, objects, and advantages of the present disclosure will be apparent from the description and from the claims. In the specification and the appended claims, the singular forms also include the plural unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present disclosure belongs. All patents and publications cited in this specification are incorporated herein by reference in their entireties.

Terms

[0028] The articles “a” and “an” are used in this disclosure to refer to one or more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0029] The term “and/or” is used in this disclosure to mean either “and” or “or” unless indicated otherwise. The use of the term "or" is used to mean "and/or" unless explicitly indicated to refer to alternatives only or the alternative are mutually exclusive, although the disclosure supports a definition that refers to only alternatives and "and/or."

[0030] The symbol

represents the point of attachment to the compound.

[0031] As used herein, the term “about” is used to indicate that a value includes the standard deviation of error for the device or method being employed to determine the value. In certain embodiments, the term “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of a stated value, unless otherwise stated or otherwise evident from the context (e.g., where such number would exceed 100% of a possible value).

[0032] By "optional" or "optionally," it is meant that the subsequently described event or circumstance may or may not occur, and that the description includes instances where the event or circumstance occurs and instances in which it does not. For example, "optionally substituted aryl" encompasses both "aryl" and "substituted aryl" as defined herein. It will be understood by those ordinarily skilled in the art, with respect to any group containing one or more substituents, that such groups are not intended to introduce any substitution or substitution patterns that are sterically impractical, synthetically non- feasible, or inherently unstable.

[0033] The term “optionally substituted” unless otherwise specified means that a group may be unsubstituted or substituted by one or more (e.g., 0, 1, 2, 3, 4, or 5 or more, or any range derivable therein) of the substituents listed for that group in which said substituents may be the same or different. In an embodiment, an optionally substituted group has 1 substituent. In another embodiment, an optionally substituted group has 2 substituents. In another embodiment, an optionally substituted group has 3 substituents. In another embodiment, an optionally substituted group has 4 substituents. In another embodiment, an optionally substituted group has 5 substituents. For instance, an alkyl group that is optionally substituted can be a fully saturated alkyl chain (i.e., a pure hydrocarbon). Alternatively, the same optionally substituted alkyl group can have substituents different from hydrogen. For instance, it can, at any point along the chain be bonded to a halogen atom, a hydroxyl group, or any other substituent described herein. Thus the term “optionally substituted” means that a given chemical moiety has the potential to contain other functional groups, but does not necessarily have any further functional groups.

[0034] The term “alkyl,” as used herein, refers to a saturated, straight or branched monovalent hydrocarbon group containing from 1 to 20 (e.g., from 1 to 10 or from 1 to 6) carbons. In some embodiments, an alkyl group is unbranched (i.e., is linear); in some embodiments, an alkyl group is branched. Representative saturated alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, 2 -methyl-1-propyl, 2-methyl-2- propyl, 2-methyl-1-butyl, 3 -methyl-1-butyl, 2-methyl-3-butyl, 2,2-dimethyl-1-propyl, 2- m ethyl-1-pentyl, 3 -methyl-1-pentyl, 4-methyl-1-pentyl, 2-methyl-2-pentyl, 3-methyl-2- pentyl, 4-methyl-2-pentyl, 2,2-dimethyl-1-butyl, 3, 3-dimethyl-1-butyl, 2-ethyl-1-butyl, butyl, isobutyl, t-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl and the like, and longer alkyl groups, such as heptyl, and octyl and the like. An alkyl group can be unsubstituted or substituted. Alkyl groups containing three or more carbon atoms may be straight or branched. As used herein, “lower alkyl” means an alkyl having from 1 to 6 carbon atoms.

[0035] As used herein, the term “heteroalkyl” refers to an “alkyl” group (as defined herein), in which at least one carbon atom has been replaced with a heteroatom (e.g., an O, N, or S atom). The heteroatom may appear in the middle or at the end of the radical.

[0036] The term "alkenyl" means an aliphatic hydrocarbon group containing a carbon — carbon double bond and which may be straight or branched having about 2 to 20 carbons (e.g., from 2 to 6 or from 2 to 10 carbons) in the chain. Certain alkenyl groups have 2 to about 4 carbon atoms in the chain. Alkenyls include both cis and trans isomers. Branched means that one or more lower alkyl groups such as methyl, ethyl, or propyl are attached to a linear alkenyl chain. Exemplary alkenyl groups include ethenyl, propenyl, n- butenyl, and i-butenyl. A C2-C6 alkenyl group is an alkenyl group containing between 2 and 6 carbon atoms.

[0037] The term "alkynyl" means an aliphatic hydrocarbon group containing a carbon — carbon triple bond and which may be straight or branched having about 2 to 20 carbon atoms (e.g., from 2 to 4, from 2 to 6, or from 2 to 10 carbons) in the chain. Certain alkynyl groups have 2 to about 4 carbon atoms in the chain. Branched means that one or more lower alkyl groups such as methyl, ethyl, or propyl are attached to a linear alkynyl chain. Exemplary alkynyl groups include ethynyl, propynyl, n-butynyl, 2-butynyl, 3- methylbutynyl, and n-pentynyl. A C2-C6 alkynyl group is an alkynyl group containing between 2 and 6 carbon atoms.

[0038] As used herein, the term "halo" or "halogen" means a fluoro, chloro, bromo, or iodo group. [0039] As used herein, the term “haloalkyl” refers to an alkyl group substituted with at least one halogen atom. Non-limiting examples include monofluoro alkyl, difluoro alkyl, trifluoro alkyl, -CH₂F, -CHF₂, and CF₃.

[0040] The term “hydroxy,” as used herein, represents a -OH group.

[0041] The term “oxo” as used herein refers to an “=O” group. When an oxo group is bonded to a carbon atom, i.e., a carbonyl group, it can also be abbreviated herein as C(O) or as C=O. An oxo group can also be bonded to a sulfur atom (e.g., S=O and S(O)₂) or at phosphorous atom (e.g., P=O, PO₂, PO₃, PO₄, etc.).

[0042] The term “imine” as used herein refers to an “=N” group. When an imine is bonded to a carbon atom, it can also be abbreviated herein as C=N. Nitrogen can also be double bonded to sulfur, e.g., S=N, which is referred to as a thioimine.

[0043] The term “annular atoms” used in conjunction with terms relating to ring systems described herein (e.g., cycloalkyl, cycloalkenyl, aryl, heterocyclyl, and heteroaryl) refers to the total number of ring atoms present in the system. “Annular atoms” therefore does not include the atoms present in a substituent attached to the ring. Thus, the number of “annular atoms” includes all atoms present in a fused ring. For example, a 2-indolyl ring,

, is considered a 5-membered heteroaryl, but is also a heteroaryl containing 9 annular atoms. In another example, pyridine is considered a 6-membered heteroaryl, and is a heteroaryl containing 6 annular atoms.

[0044] "Cycloalkyl" refers to a single saturated all carbon ring having 3 to 20 annular carbon atoms (i.e., C₃-C₂₀ cycloalkyl), for example from 3 to 15 annular atoms, for example, from 3 to 12 annular atoms. In certain embodiments, the cycloalkyl group is either monocyclic ("monocyclic cycloalkyl") or contains a fused, bridged or spiro ring system such as a bicyclic system ("bicyclic cycloalkyl") and can be saturated.

"Cycloalkyl" includes ring systems where the cycloalkyl ring, as defined above, is fused with one or more cycloalkyl, cycloalkenyl, heterocyclyl, aryl or heteroaryl groups, wherein the point of attachment is on a cycloalkyl ring, and, in such instances, the number of carbon atoms recited continues to designate the number of carbons in the cycloalkyl ring containing the point of attachment. Examples of cycloalkyl groups include cyclohexyl, cycloheptyl, 2-adamantyl 2-(2,3-dihydro-lH-indene) and 9-fluorenyl

As noted above, cycloalkyl rings can be further characterized by the number of annular atoms. For example, a cyclohexyl ring is a C₆ cycloalkyl ring with 6 annular atoms, while 2-(2, 3 -dihydro- IH-indene) is a C₅ cycloalkyl ring with 9 annular atoms. Also, for example, 9-fluorenyl is a C₅ cycloalkyl ring with 13 annular atoms and 2- adamantyl is a C₆ cycloalkyl with 10 annular atoms.

[0045] As used herein, the term “cycloalkenyl” may refer to a partially saturated, monocyclic, fused or spiro polycyclic, all carbon ring having from 3 to 18 carbon atoms per ring and contains at least one double bond. "Cycloalkenyl" includes ring systems where the cycloalkenyl ring, as defined above, is fused with one or more cycloalkyl, cycloalkenyl, heterocyclyl, aryl or heteroaryl groups, wherein the point of attachment is on a cycloalkenyl ring, and, in such instances, the number of carbon atoms recited continues to designate the number of carbons in the cycloalkenyl ring containing the point of attachment. Cycloalkenyl rings can be further characterized by the number of annular atoms. Examples of cycloalkenyl include 1 -cyclohex- 1-enyl and cyclopent- 1-enyl.

[0046] The term "aryl" as used herein refers to a single all carbon aromatic ring or a multiple condensed all carbon ring system wherein at least one of the rings is aromatic. For example, in certain embodiments, an aryl group has 5 to 20 annular carbon atoms, 5 to 14 annular carbon atoms, or 5 to 12 annular carbon atoms. Aryl also includes multiple condensed ring systems (e.g., ring systems comprising 2, 3 or 4 rings) having about 9 to 20 carbon atoms in which at least one ring is aromatic and wherein the other rings may be aromatic or not aromatic (i.e., cycloalkyl). "Aryl" includes ring systems where the aryl ring, as defined above, is fused with one or more cycloalkyl, cycloalkenyl, heterocyclyl, aryl or heteroaryl groups, and wherein the point of attachment is on an aryl ring, and, in such instances, the number of carbon atoms recited continues to designate the number of carbon atoms in the aryl ring containing the point of attachment. Examples of aryl groups include phenyl and 5-(2,3-dihydro-lH-indene):

. As noted above, aryl rings can be further characterized by the number of annular atoms. For example, phenyl is a C₆ aryl with 6 annular atoms, while 5-(2,3-dihydro-lH-indene) is a C₆ aryl with 9 annular atoms.

[0047] "Heterocyclyl" as used herein refers to a single saturated or partially unsaturated non-aromatic ring or a non-aromatic multiple ring system (including fused and spiro polycyclic) that has at least one heteroatom in the ring (at least one annular heteroatom selected from oxygen, nitrogen, phosphorus, and sulfur). Unless otherwise specified, a heterocyclyl group has from 5 to about 20 annular atoms, for example from 5 to 15 annular atoms, for example from 5 to 10 annular atoms. Thus, the term includes single saturated or partially unsaturated rings (e.g., 3, 4, 5, 6 or 7-membered rings) having from about 1 to 6 annular carbon atoms and from about 1 to 3 annular heteroatoms selected from the group consisting of oxygen, nitrogen, phosphorus, and sulfur in the ring. The term also includes single saturated or partially unsaturated rings (e.g., 5, 6, 7, 8, 9, or 10- membered rings) having from about 4 to 9 annular carbon atoms and from about 1 to 3 annular heteroatoms selected from the group consisting of oxygen, nitrogen, phosphorus, and sulfur in the ring. “Heterocyclyl" includes ring systems where the heterocyclyl ring, as defined above, is fused with one or more cycloalkyl, cycloalkenyl, heterocyclyl, aryl or heteroaryl groups, wherein the point of attachment is on a heterocyclic ring, and, in such instances, the number of ring members recited continues to designate the number of annular atoms in the heterocyclic ring containing the point of attachment. Heterocyclic rings can be further characterized by the number of annular atoms. Examples of heterocyclic groups include piperidinyl (6-membered heterocycle with 6 annular atoms), azepanyl (7-membered heterocycle with 7 annular atoms), and 3-chromanyl (6-membered heterocycle with 10 annular atoms)

and benzofuran (9-membered heterocycle with 9 annular atoms)

[0048] The term "heteroaryl" as used herein refers to a single aromatic ring that has at least one atom other than carbon in the ring, wherein the atom is selected from the group consisting of oxygen, nitrogen and sulfur; the term also includes multiple condensed ring systems that have at least one such aromatic ring. Thus, the term includes single heteroaryl rings of from about 1 to 10 annular carbon atoms and about 1-5 annular heteroatoms selected from the group consisting of oxygen, nitrogen and sulfur in the rings. The sulfur and nitrogen atoms may also be present in an oxidized form provided the ring is aromatic. "Heteroaryl" includes ring systems where the heteroaryl ring, as defined above, is fused with one or more cycloalkyl, cycloalkenyl, heterocyclyl, aryl or heteroaryl groups, wherein the point of attachment is on a heteroaryl ring, and, in such instances, the number of ring members continues to designate the number of ring members in the heteroaryl ring containing the point of attachment. Heteroaryl rings can be further characterized by the number of annular atoms. For example, pyridine is a 6-membered heteroaryl having 6 annular atoms.

[0049] Unless otherwise stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. Exemplary isotopes that can be incorporated into compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³²P, ³³P, ³⁵S, ¹⁸F, ³⁶C1, ¹²³I and ¹²⁵I. Isotopically-labeled compounds (e.g., those labeled with ³H and ¹⁴C) can be useful in compound or substrate tissue distribution assays. Tritiated (i.e., ³H) and carbon-14 (i.e., ¹⁴C) isotopes can be useful for their ease of preparation and detectability. Further, substitution with heavier isotopes such as deuterium (i.e., ²H) may afford certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements). In some embodiments, one or more hydrogen atoms are replaced by ²H or ³H, or one or more carbon atoms are replaced by ¹³C- or ¹⁴C-enriched carbon. Positron emitting isotopes such as ¹⁵O, ¹³N, ¹¹C, and ¹⁸F are useful for positron emission tomography (PET) studies to examine substrate receptor occupancy. Preparations of isotopically labelled compounds are known to those of skill in the art. For example, isotopically labeled compounds can generally be prepared by following procedures analogous to those disclosed for compounds of the present disclosure described herein, by substituting an isotopically labeled reagent for a non-isotopically labeled reagent.

[0050] The disclosure also includes pharmaceutical compositions comprising an effective amount of a disclosed compound, or a pharmaceutically acceptable salt, solvate, stereoisomer, prodrug, or tautomer thereof, and a pharmaceutically acceptable carrier. Representative “pharmaceutically acceptable salts” include, e.g., water-soluble and water- insoluble salts, such as the acetate, amsonate (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, benzonate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium, calcium edetate, camsylate, carbonate, chloride, citrate, clavulariate, dihydrochloride, edetate, edisylate, estolate, esylate, fiunarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, sethionate, lactate, lactobionate, laurate, magnesium, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methyl sulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, 3-hydroxy- 2-naphthoate, oleate, oxalate, palmitate, pamoate (1,1-methene-bis-2-hydroxy-3- naphthoate, einbonate), pantothenate, phosphate/diphosphate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, subacetate, succinate, sulfate, sulfosalicylate, suramate, tannate, tartrate, teoclate, tosylate, triethiodide, and valerate salts.

[0051] The term “carrier”, as used in this disclosure, encompasses excipients and diluents and means a material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material, involved in carrying or transporting a pharmaceutical agent from one organ, or portion of the body, to another organ, or portion of the body of a subject.

[0052] A “pharmaceutically acceptable excipient,” as used herein, refers any inactive ingredient (for example, a vehicle capable of suspending or dissolving the active compound) having the properties of being nontoxic and non-inflammatory in a subject. Typical excipients include, for example: antiadherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colors), emollients, emulsifiers, fillers (diluents), film formers or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, sorbents, suspending or dispersing agents, sweeteners, or waters of hydration. Excipients include, but are not limited to: butylated optionally substituted hydroxyltoluene (BEIT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, optionally substituted hydroxylpropyl cellulose, optionally substituted hydroxylpropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol. Those of ordinary skill in the art are familiar with a variety of agents and materials useful as excipients. See, e.g., Ansel, et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, et al., Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. In some embodiments, a composition includes at least two different pharmaceutically acceptable excipients.

[0053] The term “prodrug,” as used in this disclosure, means a compound which is convertible in vivo by metabolic means (e.g., by hydrolysis) to a disclosed compound. Furthermore, as used herein a prodrug is a drug which is inactive in the body, but is transformed in the body typically either during absorption or after absorption from the gastrointestinal tract into the active compound. The conversion of the prodrug into the active compound in the body may be done chemically or biologically (i.e., using an enzyme).

[0054] The term "solvate" refers to a complex of variable stoichiometry formed by a solute and solvent. Such solvents for the purpose of the present disclosure may not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, MeOH, EtOH, and AcOH. Solvates wherein water is the solvent molecule are typically referred to as hydrates. Hydrates include compositions containing stoichiometric amounts of water, as well as compositions containing variable amounts of water.

[0055] The term “tautomers” refers to a set of compounds that have the same number and type of atoms, but differ in bond connectivity and are in equilibrium with one another. A “tautomer” is a single member of this set of compounds. Typically, a single tautomer is drawn but it is understood that this single structure is meant to represent all possible tautomers that might exist. Examples include enol-ketone tautomerism. When a ketone is drawn it is understood that both the enol and ketone forms are part of the present disclosure. [0056] The term "isomer" refers to compounds that have the same composition and molecular weight but differ in physical or chemical properties. The structural difference may be in constitution (geometric isomers) or in the ability to rotate the plane of polarized light (stereoisomers). It is recognized that the compounds of the disclosure can have one or more chiral centers or double bonds and, therefore, exist as stereoisomers, such as double-bond isomers (i.e., geometric E/Z isomers) or diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis/trans isomers). With regard to stereoisomers, the compounds herein may have one or more asymmetric carbon atom and may occur as racemates, racemic mixtures and as individual enantiomers or diastereomers. According to the disclosure, the chemical structures depicted herein, and therefore the compounds of the disclosure, encompass all the corresponding stereoisomers, that is, both the stereomerically pure form (e.g., geometrically pure, enantiomerically pure, or diastereomerically pure) and enantiomeric and stereoisomeric mixtures, e.g., racemates. Enantiomeric and stereoisomeric mixtures of compounds of the disclosure can typically be resolved into their component enantiomers or stereoisomers by well-known methods, such as chiral-phase gas chromatography, chiral-phase high performance liquid chromatography, crystallizing the compound as a chiral salt complex, or crystallizing the compound in a chiral solvent. Enantiomers and stereoisomers can also be obtained from stereomerically or enantiomerically pure intermediates, reagents, and catalysts by well-known asymmetric synthetic methods.

[0057] The term "stereoisomers" refers to the set of compounds which have the same number and type of atoms and share the same bond connectivity between those atoms, but differ in three dimensional structure. The term "stereoisomer" refers to any member of this set of compounds. For instance, a stereoisomer may be an enantiomer or a diastereomer.

[0058] The term "enantiomers" refers to a pair of stereoisomers which are non- superimposable mirror images of one another. The term "enantiomer” refers to a single member of this pair of stereoisomers. The term "racemic" refers to a 1 : 1 mixture of a pair of enantiomers.

[0059] The term "diastereomers" refers to the set of stereoisomers which cannot be made superimposable by rotation around single bonds. For example, cis- and trans- double bonds, endo- and exo- substitution on bicyclic ring systems, and compounds containing multiple stereogenic centers with different relative configurations are considered to be diastereomers. The term "diastereomer" refers to any member of this set of compounds. In some examples presented, the synthetic route may produce a single diastereomer or a mixture of diastereomers.

[0060] An “effective amount” when used in connection with a compound is an amount effective for treating or preventing a disease in a subject as described herein.

[0061] A “therapeutic agent” is any substance, e.g., a compound or composition, capable of treating a disease or disorder. In some embodiments, therapeutic agents that are useful in connection with the present disclosure include RAS inhibitors and cancer chemotherapeutics. Many such therapeutic agents are known in the art and are disclosed herein.

[0062] The term “therapeutically effective amount” means an amount that is sufficient, when administered to a population suffering from or susceptible to a disease, disorder, or condition in accordance with a therapeutic dosing regimen, to treat the disease, disorder, or condition. In some embodiments, a therapeutically effective amount is one that reduces the incidence or severity of, or delays onset of, one or more symptoms of the disease, disorder, or condition. Those of ordinary skill in the art will appreciate that the term “therapeutically effective amount” does not in fact require successful treatment be achieved in a particular individual. Rather, a therapeutically effective amount may be that amount that provides a particular desired pharmacological response in a significant number of subjects when administered to patients in need of such treatment. It is specifically understood that particular subjects may, in fact, be “refractory” to a “therapeutically effective amount.” In some embodiments, reference to a therapeutically effective amount may be a reference to an amount as measured in one or more specific tissues (e.g., a tissue affected by the disease, disorder or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine). Those of ordinary skill in the art will appreciate that, in some embodiments, a therapeutically effective amount may be formulated or administered in a single dose. In some embodiments, a therapeutically effective amount may be formulated or administered in a plurality of doses, for example, as part of a dosing regimen. [0063] A “therapeutic regimen” refers to a dosing regimen whose administration across a relevant population is correlated with a desired or beneficial therapeutic outcome.

[0064] The term “treatment” (also “treat” or “treating”), in its broadest sense, refers to any administration of a substance (e.g., a compound of the present disclosure) that partially or completely alleviates, ameliorates, relieves, inhibits, delays onset of, reduces severity of, or reduces incidence of one or more symptoms, features, or causes of a particular disease, disorder, or condition. In some embodiments, such treatment may be administered to a subject who does not exhibit signs of the relevant disease, disorder or condition or of a subject who exhibits only early signs of the disease, disorder, or condition. Alternatively, or additionally, in some embodiments, treatment may be administered to a subject who exhibits one or more established signs of the relevant disease, disorder or condition. In some embodiments, treatment may be of a subject who has been diagnosed as suffering from the relevant disease, disorder, or condition. In some embodiments, treatment may be of a subject known to have one or more susceptibility factors that are statistically correlated with increased risk of development of the relevant disease, disorder, or condition.

[0065] The term “prevent” or “preventing” with regard to a subject refers to keeping a disease or disorder from afflicting the subject. Preventing includes prophylactic treatment. For instance, preventing can include administering to the subject a compound disclosed herein before a subject is afflicted with a disease and the administration will keep the subject from being afflicted with the disease.

[0066] The terms “inhibiting” and “reducing,” or any variation of these terms, includes any measurable or complete inhibition to achieve a desired result. For example, there may be a decrease of about, at most about, or at least about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99%, or more, or any range derivable therein, reduction of activity (e.g., SOSl :Ras-family protein binding activity) compared to normal.

[0067] The term “disorder” is used in this disclosure to mean, and is used interchangeably with, the terms disease, condition, or illness, unless otherwise indicated. [0068] The term “administer”, “administering”, or “administration” as used in this disclosure refers to either directly administering a disclosed compound or pharmaceutically acceptable salt of the disclosed compound or a composition to a subject, or administering a prodrug derivative or analog of the compound or pharmaceutically acceptable salt of the compound or composition to the subject, which can form an equivalent amount of active compound within the subject’s body. Administration to an animal subject (e.g., to a human) may be by any appropriate route. For example, in some embodiments, administration may be bronchial (including by bronchial instillation), buccal, enteral, interdermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (including by intratracheal instillation), transdermal, vaginal or vitreal.

[0069] As used herein, the term “dosage form” refers to a physically discrete unit of a compound (e.g., a compound of the present disclosure) for administration to a subject. Each unit contains a predetermined quantity of compound. In some embodiments, such quantity is a unit dosage amount (or a whole fraction thereof) appropriate for administration in accordance with a dosing regimen that has been determined to correlate with a desired or beneficial outcome when administered to a relevant population (i.e., with a therapeutic dosing regimen). Those of ordinary skill in the art appreciate that the total amount of a therapeutic composition or compound administered to a particular subject is determined by one or more attending physicians and may involve administration of multiple dosage forms.

[0070] As used herein, the term “dosing regimen” refers to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time. In some embodiments, a given therapeutic compound (e.g., a compound of the present disclosure) has a recommended dosing regimen, which may involve one or more doses. In some embodiments, a dosing regimen includes a plurality of doses each of which are separated from one another by a time period of the same length; in some embodiments, a dosing regimen includes a plurality of doses and at least two different time periods separating individual doses. In some embodiments, all doses within a dosing regimen are of the same unit dose amount. In some embodiments, different doses within a dosing regimen are of different amounts. In some embodiments, a dosing regimen includes a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount. In some embodiments, a dosing regimen includes a first dose in a first dose amount, followed by one or more additional doses in a second dose amount same as the first dose amount. In some embodiments, a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (i.e., is a therapeutic dosing regimen).

[0071] The term “disorder” is used in this disclosure to mean, and is used interchangeably with, the terms disease, condition, or illness, unless otherwise indicated.

[0072] A "patient" or “subject” is a mammal, e.g., a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, or non-human primate, such as a monkey, chimpanzee, baboon or rhesus.

[0073] The term “monotherapy” refers to a method of treatment comprising administering to a subject a single therapeutic agent, optionally as a pharmaceutical composition. For example, a monotherapy may comprise administration of a pharmaceutical composition comprising a therapeutic agent and one or more pharmaceutically acceptable carrier, excipient, diluent, and/or surfactant. The therapeutic agent may be administered in an effective amount. The therapeutic agent may be administered in a therapeutically effective amount.

[0074] The term “combination therapy” refers to a method of treatment comprising administering to a subject at least two therapeutic agents, optionally as one or more pharmaceutical compositions. For example, a combination therapy may comprise administration of a single pharmaceutical composition comprising at least two therapeutic agents and one or more pharmaceutically acceptable carrier, excipient, diluent, and/or surfactant. A combination therapy may comprise administration of two or more pharmaceutical compositions, each composition comprising one or more therapeutic agent and one or more pharmaceutically acceptable carrier, excipient, diluent, and/or surfactant. In various embodiments, at least one of the therapeutic agents is a S0S1 inhibitor. In various embodiments, at least one of the therapeutic agents is a RAS inhibitor. The two agents may optionally be administered simultaneously (as a single or as separate compositions) or sequentially (as separate compositions). The therapeutic agents may be administered in an effective amount. The therapeutic agent may be administered in a therapeutically effective amount. In some embodiments, the effective amount of one or more of the therapeutic agents may be lower when used in a combination therapy than the therapeutic amount of the same therapeutic agent when it is used as a monotherapy, e.g., due an additive or synergistic effect of combining the two or more therapeutics.

[0075] The term "SOS" (e.g., a "SOS mutation") refers to SOS genes, which are known in the art to include RAS guanine nucleotide exchange factor proteins that are activated by receptor tyrosine kinases to promote GTP loading of RAS and signaling. The term SOS includes all SOS homologs that promotes the exchange of Ras-bound GDP by GTP. In particular embodiments, SOS refers specifically to "son of sevenless homolog 1" ("S0S1"). S0S1 is critically involved in the activation of RAS-family protein signaling in cancer via mechanisms other than mutations in RAS-family proteins. S0S1 interacts with the adaptor protein Grb2 and the resulting SOSl/Grb2 complex binds to activated/phosphorylated Receptor Tyrosine Kinases (e.g., EGFR, ErbB2, ErbB3, ErbB4, PDGFR-A/B, FGFR1/2/3, IGF1 R, INSR, ALK, ROS, TrkA, TrkB, TrkC, RET, c-MET, VEGFR1/2/3, AXL) (Pierre et al., Biochem. Pharmacol., 2011, 82(9): 1049-56). S0S1 is also recruited to other phosphorylated cell surface receptors such as the T cell Receptor (TCR), B cell Receptor (BCR) and monocyte colony-stimulating factor receptor (Salojin et al., J. Biol. Chem. 2000, 275(8):5966-75). This localization of S0S1 to the plasma membrane, proximal to RAS-family proteins, enables S0S1 to promote RAS-family protein activation. SOS 1 -activation of RAS-family proteins can also be mediated by the interaction of SOSl/Grb2 with the BCR-ABL oncoprotein commonly found in chronic myelogenous leukemia (Kardinal et al., 2001, Blood, 98: 1773-81; Sini et al., Nat. Cell Biol., 2004, 6(3):268-74). S0S1 is also a GEF for the activation of the GTPases RAC1 (Ras-related C3 botulinum toxin substrate 1) (Innocenti et al., J. Cell Biol., 2002, 156(1): 125-36). RAC1, like RAS-family proteins, is implicated in the pathogenesis of a variety of human cancers and other diseases (Bid et al., Mol. Cancer Ther. 2013, 12(10): 1925-34). Son of sevenless 2 (S0S2), a homolog of S0S1 in mammalian cells, also acts as a GEF for the activation of RAS-family proteins (Pierre et al., Biochem. Pharmacol., 2011, 82(9): 1049-56; Buday et al., Biochim. Biophys. Acta., 2008, 1786(2): 178-87). Published data from mouse knockout models suggests a redundant role for S0S1 and S0S2 in homeostasis in the adult mouse. Whilst germline knockout of S0S1 in mice results in lethality during mid-embryonic gestation (Qian et al., EMBO J., 2000, 19(4):642-54), systemic conditional S0S1 knockout adult mice are viable (Baltanas et al., Mol. Cell. Biol., 2013, 33(22):4562-78). S0S2 gene targeting did not result in any overt phenotype in mice (Esteban et al., Mol. Cell. Biol., 2000, 20(17):6410-3). In contrast, double S0S1 and S0S2 knockout leads to rapid lethality in adult mice (Baltanas et al., Mol. Cell. Biol., 2013, 33(22):4562-78). These published data suggest that selective targeting of individual SOS isoforms (e.g., selective S0S1 targeting) may be adequately tolerated to achieve a therapeutic index between SOSl/RAS-family protein driven cancers (or other SOSl/RAS-family protein pathologies) and normal cells and tissues. Selective pharmacological inhibition of the binding of the catalytic site of S0S1 to RAS-family proteins is expected to prevent SOS 1 -mediated activation of RAS-family proteins to the GTP-bound form. Such S0S1 inhibitor compounds are be expected to consequently inhibit signaling in cells downstream of RAS-family proteins (e.g., ERK phosphorylation). In cancer cells associated with dependence on RAS-family proteins (e.g., KRAS mutant cancer cell lines), S0S1 inhibitor compounds are be expected to deliver anti-cancer efficacy (e.g., inhibition of proliferation, survival, metastasis, etc.). High potency towards inhibition of S0S1 :RAS-family protein binding (nanomolar level IC50 values) and ERK phosphorylation in cells (nanomolar level IC50 values) are desirable characteristics for a S0S1 inhibitor compound. Furthermore, a desirable characteristic of a S0S1 inhibitor compound would be the selective inhibition of S0S1 over S0S2. This conclusion is based on the viable phenotype of S0S1 knockout mice and lethality of S0S1/S0S2 double knockout mice, as described above.

[0076] As used herein, a “S0S1 inhibitor” refers to any agent, (e.g., a small molecule (e.g., less than 750 Da)) capable of inhibiting S0S1. S0S1 inhibitors can include selective S0S1 inhibitors and inhibitors that also inhibit other proteins. In some embodiments, S0S1 inhibitors may also inhibit S0S2, with a selectivity ratio less than 10- fold for inhibition of S0S1 relative to S0S2. In some embodiments, S0S1 inhibitors will selectively inhibit S0S1, with a selectivity ratio greater of at least about 10-fold, such as greater than at least about 30-fold, for inhibition of S0S1 relative to S0S2.

Compounds of Disclosed Formulae

[0077] In some embodiments, the present disclosure relates to compounds having a structure of Formula (I):

or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof.

[0078] In some embodiments, Ri is optionally substituted 3-6 membered cycloalkyl, optionally substituted 3-6 membered heterocyclyl, optionally substituted 6- membered aryl, and optionally substituted 5-6 membered heteroaryl.

[0079] In some embodiments, Ri is selected from the group consisting of optionally substituted 6-membered aryl and optionally substituted 5-6 membered heteroaryl.

[0080] In some embodiments, Ri is of the following structure

and the associated compound is of Formula (II)

or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof. In such formula (II) embodiments, R₂, R₃, and R₄ are as defined herein. [0081] In some embodiments, R₅, R₆, R₇, R_8, and R₉ are independently selected from H, D, C_1-6 alkyl, C_2-6 alkenyl, 4-8 membered cycloalkenyl, C_2-6 alkynyl, 3-8 membered cycloalkyl, 3-14 membered heterocyclyl, -OH, halogen, -NO₂, -CN, - NR₁₁R₁₂, -SR₁₀, -S(O)₂NR₁₁ R₁₂, -S(O)₂R₁₀, -NR₁₀S(O)₂NR₁₁ R₁₂, -NR₁₀S(O)₂R₁₁, - S(O)NR₁₁R₁₂, -S(O)R₁₀, -NR₁₀S(O)NR₁₁R₁₂, -NR₁₀S(O)R₁₁ , -C(O)R₁₀, -CO₂R₁₀, 6-10 membered aryl, and 5-10 membered heteroaryl, wherein each C_1-6 alkyl, C_2-6 alkenyl, 4-8 membered cycloalkenyl, C_2-6 alkynyl, 3-8 membered cycloalkyl, 3-14 membered heterocyclyl, 6-10 membered aryl, and 5-10 membered heteroaryl is optionally substituted with -OH, C_1-6 alkyl optionally substituted with -R₁₀, halogen, -NO₂, oxo, -CN, -R₁₀, - OR₁₀, -NR₁₁R₁₂, — SR₁₀, -S(O)₂NR₁₁R₁₂,, -S(O)2R₁₀, -NR₁₀S(O)₂NR₁₁R₁₂, -NR₁₀S(O)₂R₁₁, -S(O)NR₁₁R₁₂, -S(O)R₁₀, -NRioS(O)NR₁₁R₁₂, -NR₁₀S(O)R₁₁, 3-8 membered cycloalkyl, 3-14 membered heterocyclyl optionally substituted with R₁₀, 6-10 membered aryl, or 5-10 membered heteroaryl.

[0082] In some embodiments, R₅, R₆, R₇, R₈, and R₉ are each independently selected from H, C_1-6 alkyl, C_1-6 haloalkyl, halogen, and -NH₂, wherein each C_1-6 alkyl is optionally substituted with halogen.

[0083] In some embodiments, R₅, R₆, R₇, R₈, and R₉ are independently selected from H, C_1-6 alkyl, halogen, and -NH₂, wherein each C_1-6 alkyl is optionally substituted with halogen.

[0084] Rw, R₁₁, and R₁₂ are at each occurrence independently selected from H, D, C_1-6 alkyl, C_2-6 alkenyl, 4-8 membered cycloalkenyl, C_2-6 alkynyl, 3-8 membered cycloalkyl, 3-14 membered heterocyclyl, -OR₁₃, -SR₁₃, halogen, -NR₁₃R₁₄, -NO₂, and - CN.

[0085] R₁₃ and R₁₄ are at each occurrence independently selected from H, D, C_1-6 alkyl, C2-6 alkenyl, 4-8 membered cycloalkenyl, C_2-6 alkynyl, 3-8 membered cycloalkyl, and 3-14 membered heterocyclyl, wherein each C_1-6 alkyl, C2-6 alkenyl, 4-8 membered cycloalkenyl, C_2-6 alkynyl, 3-8 membered cycloalkyl, and 3-14 membered heterocyclyl are independently optionally substituted with -OH, -SH, -NH₂, -NO₂, or -CN.

[0086] In some embodiments of the compounds of Formula (II), one to three of R₅, R₆, R₇, R₈, and R₉ is C_1-6 alkyl, wherein the alkyl is optionally substituted with halogen. [0087] In some embodiments of the compounds of Formula (II), one to three of R₅, R₆, R₇, R₈, and R₉ is C_1-6 alkyl, wherein the alkyl is optionally substituted with halogen or -OH.

[0088] In some embodiments of the compounds of Formula (II), one to three of R₅, R₆, R₇, R₈, and R₉ is C_1-6 alkyl, and one to three of R₅, R₆, R₇, R₈, and R₉ is C_1-6 alkyl optionally substituted with halogen.

[0089] In some embodiments of the compounds of Formula (II), one to three of R₅, R₆, R₇, R₈, and R₉ is halogen, and one to three of R₅, R₆, R₇, R₈, and R₉ is C_1-6 alkyl optionally substituted with halogen.

[0090] In some embodiments of the compounds of Formula (II), one to three of R₅, R₆, R₇, R₈, and R₉ is -NH₂.

[0091] In some embodiments of the compounds of Formula (II), one of R₅, R₆, R₇, R₈, and R₉ is -NH₂; and one of R₅, R₆, R₇, R₈, and R₉ is C_1-6 alkyl optionally substituted with halogen.

[0092] In some embodiments of the compound of Formula (II), one or more of R₅, R₆, R₇, R₈, and R₉ is selected from among -CHF₂, -CF₃, -NH₂, -F, and substituted aryl. In some embodiments of compounds of Formula (II), one of R₅, R₆, R₇, R₈, and R₉ is -CHF2 and one of R₅, R₆, R₇, R₈, and R₉ is -F. In some embodiments of compounds of Formula (II), one of R₅, R₆, R₇, R₈, and R₉ is -CF3 and one of R₅, R₆, R₇, R₈, and R₉ is - NH₂.

[0093] In some embodiments, Ri is optionally substituted 6-membered aryl.

[0094] In some embodiments, Ri is

[0095] In some embodiments of Formulae (I) and (II), R₂ is H or optionally substituted C_1-6 alkyl. In some embodiments, R₂ is H. In some embodiments, R₂ is optionally substituted C_1-3 alkyl, wherein the optional substituent is one or more halogens, or one or more fluoro. In some embodiments, R₂ is -CH₃.

[0096] In some embodiments of Formulae (I) and (II), R₃ is -OR_3a, wherein R_3a is optionally substituted C_1-3 alkyl. In some embodiments, R_3a is C_1-3 alkyl. In some embodiments, R₃ is -OCH₃.

[0097] In some embodiments of Formulae (I) and (II), R₄ is H, C_1-6 alkyl, C_1-6 haloalkyl, 3-14 membered cycloalkyl, 3-14 membered cycloalkenyl, 3-14 membered heterocyclyl, 6-10 membered aryl, or 5-10 membered heteroaryl. In some embodiments, each C_1-6 alkyl, C_1-6 haloalkyl, 3-14 membered cycloalkyl, 3-14 membered cycloalkenyl, 3-14 membered heterocyclyl, 6-10 membered aryl, and 5-10 membered heteroaryl is optionally substituted with C_1-6 alkyl, -R_4a, -OR_4a, -O-C_1-6 alkyl-R_4a, =O, halogen, - C(O)R_4a, -C(O)OR_4a, -C(O)NR_4bR_4c, -NR_4bC(O)R_4c, -CN, =NR_4a, -NR_4bR_4c, -SO₂R_4a, 3- 6 membered cycloalkyl optionally substituted with R_4a, 3-7 membered heterocyclyl optionally substituted with R_4a, 6-10 membered aryl optionally substituted with R_4a, or 5- 10 membered heteroaryl optionally substituted with R_4a.

[0098] In some embodiments, optionally substituted C_1-6 alkyl is optionally substituted C_1-3 alkyl or optionally substituted Ci alkyl. In some embodiments, optionally substituted C_1-6 haloalkyl is optionally substituted C_1-3 haloalkyl or optionally substituted Ci haloalkyl.

[0099] In some embodiments, each R_4a is independently H, C_1-6 alkyl, C_1-6 haloalkyl, -C(O)R_4b, -C(O)NR_4bR_4c, =O, 3-6 membered cycloalkyl, 6-10 membered aryl optionally substituted with -OR_4b, -CN, =N-3-6 membered cycloalkyl, 3-7 membered heterocyclyl, -(CH₂)_rOCH₃, or -(CH2)_rOH. In such embodiments, r is 1, 2, or 3.

[0100] In some embodiments, each R_4b is independently H or C_1-6 alkyl. In some embodiments, each R_4b is independently H or C_1-3 alkyl. In some embodiments, each R_4b is independently H or -CH₃.

[0101] In some embodiments, each R_4c is independently H or C_1-6 alkyl. In some embodiments, each R_4c is independently H or C_1-3 alkyl. In some embodiments, each R_4c is independently H or -CH₃. [0102] In some embodiments, R₄ is optionally substituted 3-14 membered heterocyclyl. In some embodiments, R₄ is 3-14 membered heterocyclyl optionally substituted with halogen, C_1-6 alkyl, -CN, -OR_4a, =O, -C(O)R_4a, -NR_4bR_4c, or =NR_4a. wherein: each R_4a is independently H, C_1-6 alkyl, C_3-8 cycloalkyl, or -CN; each R_4b is independently H or C_1-6 alkyl; and each R_4c is independently H or C_1-6 alkyl.

[0103] In some embodiments, R₄ is selected from the group consisting of optionally substituted piperidinyl, optionally substituted piperazinyl, optionally substituted tetrahydropyridinyl, optionally substituted dihydrothiopyranyl, and optionally substituted tetrahydrothiopyranyl .

[0104] In some embodiments, R₄ is selected from

[0105] In some aspects, the present disclosure provides a compound, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof, selected from the group consisting of compounds of Table A: Table A.

Methods of Synthesizing the Disclosed Compounds

[0106] The compounds of the present disclosure may be made by a variety of methods, including standard chemistry. Suitable synthetic routes are depicted in the examples given below.

[0107] The compounds of any of the formulae described herein may be prepared by methods known in the art of organic synthesis as set forth in part by the examples described below. Reference is also made to synthetic descriptions in WO 2020/180768, WO 2020/180770, and WO 2021/092115. [0108] It is well understood that protecting groups for sensitive or reactive groups are employed where necessary in accordance with general principles or chemistry. Protecting groups are manipulated according to standard methods of organic synthesis (T. W. Greene and P. G. M. Wuts, "Protective Groups in Organic Synthesis", Third edition, Wiley, New York 1999). These groups are removed at a convenient stage of the compound synthesis using methods that are readily apparent to those skilled in the art. The selection processes, as well as the reaction conditions and order of their execution, shall be consistent with the preparation of compounds of any formula disclosed herein.

[0109] Those skilled in the art will recognize if a stereocenter exists in any of the compounds of the present disclosure. Accordingly, the present disclosure includes both possible stereoisomers (unless specified in the synthesis) and includes not only racemic compounds but the individual enantiomers or diastereomers as well. When a compound is desired as a single enantiomer or diastereomer, it may be obtained by stereospecific synthesis or by resolution of the final product or any convenient intermediate. Resolution of the final product, an intermediate, or a starting material may be affected by any suitable method known in the art. See, for example, "Stereochemistry of Organic Compounds" by E. L. Eliel, S. H. Wilen, and L. N. Mander (Wiley-lnterscience, 1994).

Therapeutic Use

[0110] Due to their biological properties the compounds of the present disclosure, their tautomers, racemates, enantiomers, diastereomers, mixtures thereof and the salts of all the above-mentioned forms may be suitable for treating diseases characterized by excessive or abnormal cell proliferation such as cancer.

[0111] For example, the following cancers, tumors and other proliferative diseases may be treated with compounds of the present disclosure, without being restricted thereto: cancers/tumors/carcinomas of the head and neck: e.g., tumors/carcinomas/cancers of the nasal cavity, paranasal sinuses, nasopharynx, oral cavity (including lip, gum, alveolar ridge, retromolar trigone, floor of mouth, tongue, hard palate, buccal mucosa), oropharynx (including base of tongue, tonsil, tonsillar pilar, soft palate, tonsillar fossa, pharyngeal wall), middle ear, larynx (including supraglottis, glottis, subglottis, vocal cords), hypopharynx, salivary glands (including minor salivary glands); intraocular cancers (e.g., uveal melanoma), and orbital and adnexal cancers; cancers/tumors/carcinomas of the lung: e.g., non-small cell lung cancer (NSCLC) (squamous cell carcinoma, spindle cell carcinoma, adenocarcinoma, large cell carcinoma, clear cell carcinoma, bronchioalveolar), small cell lung cancer (SCLC) (oat cell cancer, intermediate cell cancer, combined oat cell cancer); neoplasms of the mediastinum: e.g., neurogenic tumors (including neurofibroma, neurilemoma, malignant schwannoma, neurosarcoma, ganglioneuroblastoma, ganglioneuroma, neuroblastoma, pheochromocytoma, paraganglioma), germ cell tumors (including seminoma, teratoma, non-seminoma), thymic tumors (including thymoma, thymolipoma, thymic carcinoma, thymic carcinoid), mesenchymal tumors (including fibroma, fibrosarcoma, lipoma, liposarcoma, myxoma, mesothelioma, leiomyoma, leiomyosarcoma, rhabdomyosarcoma, xanthogranuloma, mesenchymoma, hemangioma, hemangioendothelioma, hemangiopericytoma, lymphangioma, lymphangiopericytoma, lymphangiomyoma), astrocytoma (cerebral, cerebellar, diffuse, fibrillary, anaplastic, pilocytic, protoplasmic, gemistocytary), glioblastoma, gliomas, oligodendrogliomas, oligoastrocytomas, ependymomas, ependymoblastomas, choroid plexus tumors, medulloblastomas, meningiomas, schwannomas, hemangioblastomas, hemangiomas, hemangiopericytomas, neuromas, ganglioneuromas, neuroblastomas, retinoblastomas, neurinomas (e.g., acoustic), spinal axis tumors; cancers/tumors/carcinomas of the gastrointestinal (GI) tract: e.g., tumors/carcinomas/ cancers of the esophagus, stomach (gastric cancer), pancreas, liver and biliary tree (including hepatocellular carcinoma (HCC), e.g., childhood HCC, fibrolamellar HCC, combined HCC, spindle cell HCC, clear cell HCC, giant cell HCC, carcinosarcoma HCC, sclerosing HCC; hepatoblastoma; cholangiocarcinoma; cholangiocellular carcinoma; hepatic cystadenocarcinoma; angiosarcoma, hemangioendothelioma, leiomyosarcoma, malignant schwannoma, fibrosarcoma, Klatskin tumor), gall bladder, extrahepatic bile ducts, small intestine (including duodenumjejunum, ileum), large intestine (including cecum, colon, rectum, anus; colorectal cancer, gastrointestinal stroma tumor (GIST)), genitourinary system (including kidney, e.g., renal pelvis, renal cell carcinoma (RCC), nephroblastoma (Wilms' tumor), hypernephroma, Grawitz tumor; ureter; urinary bladder, e.g., urachal cancer, urothelial cancer; urethra, e.g., distal, bulbomembranous, prostatic; prostate (androgen dependent, androgen independent, castration resistant, hormone independent, hormone refractory), penis); cancers/tumors/carcinomas of the testis: e.g., seminomas, non-seminomas; gynecologic cancers/tumors/carcinomas: e.g., tumors/carcinomas/cancers of the ovary, fallopian tube, peritoneum, cervix, vulva, vagina, uterine body (including endometrium, fundus); cancers/tumors/carcinomas of the breast: e.g., mammary carcinoma (infiltrating ductal, colloid, lobular invasive, tubular, adenocystic, papillary, medullary, mucinous), hormone receptor positive breast cancer (estrogen receptor positive breast cancer, progesterone receptor positive breast cancer), HER2 positive breast cancer, triple negative breast cancer, Paget's disease of the breast; cancers/tumors/carcinomas of the endocrine system: e.g., tumors/carcinomas/cancers of the endocrine glands, thyroid gland (thyroid carcinomas/tumors; papillary, follicular, anaplastic, medullary), parathyroid gland (parathyroid carcinoma/tumor), adrenal cortex (adrenal cortical carcinoma/tumors), pituitary gland (including prolactinoma, craniopharyngioma), thymus, adrenal glands, pineal gland, carotid body, islet cell tumors, paraganglion, pancreatic endocrine tumors (PET; non-functional PET, PPoma, gastrinoma, insulinoma, VIPoma, glucagonoma, somatostatinoma, GRFoma, ACTHoma), carcinoid tumors; sarcomas of the soft tissues: e.g., fibrosarcoma, fibrous histiocytoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma, angiosarcoma, lymphangiosarcoma, Kaposi s sarcoma, glomus tumor, hemangiopericytoma, synovial sarcoma, giant cell tumor of tendon sheath, solitary fibrous tumor of pleura and peritoneum, diffuse mesothelioma, malignant peripheral nerve sheath tumor (MPNST), granular cell tumor, clear cell sarcoma, melanocytic schwannoma, plexosarcoma, neuroblastoma, ganglioneuroblastoma, neuroepithelioma, extraskeletal Ewing's sarcoma, paraganglioma, extraskeletal chondrosarcoma, extraskeletal osteosarcoma, mesenchymoma, alveolar soft part sarcoma, epithelioid sarcoma, extrarenal rhabdoid tumor, desmoplastic small cell tumor; sarcomas of the bone: e.g., myeloma, reticulum cell sarcoma, chondrosarcoma (including central, peripheral, clear cell, mesenchymal chondrosarcoma), osteosarcoma (including parosteal, periosteal, high-grade surface, small cell, radiation-induced osteosarcoma, Paget's sarcoma), Ewing's tumor, malignant giant cell tumor, adamantinoma, (fibrous) histiocytoma, fibrosarcoma, chordoma, small round cell sarcoma, hemangioendothelioma, hemangiopericytoma, osteochondroma, osteoid osteoma, osteoblastoma, eosinophilic granuloma, chondroblastoma; mesothelioma: e.g., pleural mesothelioma, peritoneal mesothelioma; cancers of the skin: e.g., basal cell carcinoma, squamous cell carcinoma, Merkel's cell carcinoma, melanoma (including cutaneous, superficial spreading, lentigo maligna, acral lentiginous, nodular, intraocular melanoma), actinic keratosis, eyelid cancer; neoplasms of the peripheral and central nervous system and brain: e.g., astrocytoma (cerebral, cerebellar, diffuse, fibrillary, anaplastic, pilocytic, protoplasmic, gemistocytary), glioblastoma, gliomas, oligodendrogliomas, oligoastrocytomas, ependymomas, ependymoblastomas, choroid plexus tumors, medulloblastomas, meningiomas, schwannomas, hemangioblastomas, hemangiomas, hemangiopericytomas, neuromas, ganglioneuromas, neuroblastomas, retinoblastomas, neurinomas (e.g., acoustic), spinal axis tumors, neurogenic tumors (including neurofibroma, neurilemoma, malignant schwannoma, neurosarcoma, ganglioneuroblastoma, ganglioneuroma, neuroblastoma, pheochromocytoma, paraganglioma), germ cell tumors (including seminoma, teratoma, non-seminoma), thymic tumors (including thymoma, thymolipoma, thymic carcinoma, thymic carcinoid), mesenchymal tumors (including fibroma, fibrosarcoma, lipoma, liposarcoma, myxoma, mesothelioma, leiomyoma, leiomyosarcoma, rhabdomyosarcoma, xanthogranuloma, mesenchymoma, hemangioma, hemangioendothelioma, hemangiopericytoma, lymphangioma, lymphangiopericytoma, lymphangiomyoma); lymphomas and leukemias: e.g., B-cell non-Hodgkin lymphomas (NHL) (including small lymphocytic lymphoma (SLL), lymphoplasmacytoid lymphoma (LPL), mantle cell lymphoma (MCL), follicular lymphoma (FL), diffuse large cell lymphoma (DLCL), Burkitt s lymphoma (BL)), Burkitt leukemia, T-cell non-Hodgkin lymphomas (including anaplastic large cell lymphoma (ALCL), adult T-cell leukemia/lymphoma (ATLL), cutaneous T-cell lymphoma (CTCL), peripheral T-cell lymphoma (PTCL)), lymphoblastic T-cell lymphoma (T-LBL), adult T-cell lymphoma, lymphoblastic B-cell lymphoma (B-LBL), immunocytoma, chronic B-cell lymphocytic leukemia (B-CLL), chronic T-cell lymphocytic leukemia (T-CLL) B-cell small lymphocytic lymphoma (B- SLL), cutaneous T-cell lymphoma (CTLC), primary central nervous system lymphoma (PCNSL), immunoblastoma, Hodgkin's disease (HD) (including nodular lymphocyte predominance HD (NLPHD), nodular sclerosis HD (NSHD), mixed-cellularity HD (MCHD), lymphocyte-rich classic HD, lymphocyte-depleted HD (LDHD)), large granular lymphocyte leukemia (LGL), chronic myelogenous leukemia (CML), acute myelogenous/myeloid leukemia (AML), acute lymphatic/lymphoblastic leukemia (ALL), acute promyelocytic leukemia (APL), chronic lymphocytic/lymphatic leukemia (CLL), prolymphocytic leukemia (PLL), hairy cell leukemia, chronic myelogenous/myeloid leukemia (CML), myeloma, plasmacytoma, multiple myeloma (MM), plasmacytoma, myelodysplastic syndromes (MDS), chronic myelomonocytic leukemia (CMML), JMML (juvenile myelomonocytic leukemia), acute leukemia of ambiguous lineage, myeloproliferative neoplasms, blastic plasmacytoid dendritic cell neoplasm, early T-cell precursor leukemia, natural killer cell leukemia/lymphoma, myeloid/lymphoid neoplasms with eosinophilia, myeloid sarcoma, transient abnormal myelopoiesis; and cancers of unknown primary site (CUP).

[0112] All cancers/tumors/carcinomas mentioned above which are characterized by their specific location/origin in the body are meant to include both the primary tumors and the metastatic tumors derived therefrom.

[0113] All cancers/tumors/carcinomas mentioned above may be further differentiated by their histopathological classification: epithelial cancers, e.g., squamous cell carcinoma (SCC) (carcinoma in situ, superficially invasive, verrucous carcinoma, pseudosarcoma, anaplastic, transitional cell, lymphoepithelial), adenocarcinoma (AC) (well-differentiated, mucinous, papillary, pleomorphic giant cell, ductal, small cell, signet-ring cell, spindle cell, clear cell, oat cell, colloid, adenosquamous, mucoepidermoid, adenoid cystic), mucinous cystadenocarcinoma, acinar cell carcinoma, large cell carcinoma, small cell carcinoma, neuroendocrine tumors (small cell carcinoma, paraganglioma, carcinoid); oncocytic carcinoma; and nonepithilial and mesenchymal cancers, e.g., sarcomas (fibrosarcoma, chondrosarcoma, rhabdomyosarcoma, leiomyosarcoma, hemangiosarcoma, giant cell sarcoma, lymphosarcoma, fibrous histiocytoma, liposarcoma, angiosarcoma, lymphangiosarcoma, neurofibrosarcoma), lymphoma, melanoma, germ cell tumors, hematological neoplasms, mixed and undifferentiated carcinomas.

[0114] The compounds of the present disclosure may be used in therapeutic regimens in the context of first line, second line, or any further line treatments.

[0115] The compounds of the disclosure may be used for the prevention, short- term or long-term treatment of the above-mentioned diseases, optionally also in combination with radiotherapy or surgery or other compounds.

[0116] Of course, the above also includes the use of the compounds of the present disclosure in various methods of treating the above diseases by administering a therapeutically effective dose to a patient in need thereof, as well as the use of these compounds for the manufacture of medicaments for the treatment of such diseases, as well as pharmaceutical compositions including such compounds of the disclosure, as well as the preparation or manufacture of medicaments including such compounds of the disclosure, and the like.

Additional Methods of Using the Disclosed Compounds

[0117] One aspect of the present disclosure relates to a method of inhibiting S0S1 in a subject in need thereof, the method comprising administering to the subject a S0S1 inhibitor of the present disclosure or a pharmaceutical composition comprising a S0S1 inhibitor of the present disclosure, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof.

[0118] Another aspect of the present disclosure relates to a method of treating or preventing a disease that is effected or characterized by modification (including inhibition) of the interaction of S0S1 and a RAS-family protein or RAC1 in a subject in need thereof. The method comprises administering to a subject or patient in need of treatment for diseases or disorders associated with S0S1 modulation an effective amount of a compound or a pharmaceutical composition disclosed herein, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof.

[0119] In certain embodiments, a method is provided of inhibiting the interaction of S0S1 and a RAS-family protein in a cell or inhibiting the interaction of S0S1 and RAC1 in a cell, the method comprising administering to the cell a compound or a pharmaceutical composition disclosed herein, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof, and a pharmaceutically acceptable carrier.

[0120] In certain embodiments, a method is provided of treating or preventing cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound or a pharmaceutical composition disclosed herein, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof.

[0121] In certain embodiments, the disease can be, but is not limited to, cancer. In certain embodiments, the disease or cancer is selected from the group consisting of pancreatic cancer, lung cancer, colorectal cancer, hematological cancer, cholangiocarcinoma, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, JMML (juvenile myelomonocytic leukemia), acute lymphoblastic leukemia/lymphoma, lymphomas, tumors of the central and peripheral nervous system, epithelial and nonepithelial tumors and mesenchymal tumor, bladder cancer, urothelial cancer, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular cancer, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer and sarcomas. In some embodiments, the cancer is colorectal cancer or pancreatic cancer.

[0122] In certain embodiments, the disease can be, but is not limited to, cancer. In certain embodiments, the disease or cancer is selected from the group consisting of pancreatic cancer, lung cancer, colorectal cancer, hematological cancer, cholangiocarcinoma, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, bladder cancer, urothelial cancer, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular cancer, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer and sarcomas.

[0123] In certain embodiments, the cancer comprises a Ras^MUT mutation or a NF1 ^L0F mutati on . [0124] In certain embodiments, the disease can be, but is not limited to, a RASopathy. In certain embodiments, the RASopathy is selected from the group consisting of Neurofibromatosis type 1 (NF1), Noonan Syndrome (NS), Noonan Syndrome with Multiple Lentigines (NSML), Capillary Malformation- Arteriovenous Malformation Syndrome (CM-AVM), Costello Syndrome (CS), Cardio-Facio-Cutaneous Syndrome (CFC), Legius Syndrome, and Hereditary gingival fibromatosis.

[0125] Another aspect of the present disclosure is directed to a method of inhibiting S0S1. The method involves administering to a patient in need thereof an effective amount of a compound or a pharmaceutical composition disclosed herein, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof.

[0126] The present disclosure relates to compounds and pharmaceutical compositions thereof capable of modulating the activity of (e.g., inhibiting) S0S1. The present disclosure also relates to the therapeutic use of such compounds or pharmaceutical compositions comprising such compounds, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof.

[0127] The disclosed compounds and pharmaceutical compositions can be administered in effective amounts to treat or prevent a disorder or prevent the development thereof in subjects.

[0128] Another aspect of the present disclosure relates to a compound of any formula, or a pharmaceutical composition thereof, disclosed herein, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof, for use in treating or preventing a disease that is affected by modification of the interaction of S0S1 and a RAS-family protein or RAC1. Another aspect of the present disclosure relates to a compound of any formula disclosed herein, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof, for use in treating or preventing a disease that is characterized by inhibition of the interaction of S0S1 with a RAS-family protein or the interaction of S0S1 with RAC1.

[0129] Another aspect of the present disclosure relates to a compound of any formula, or a pharmaceutical composition thereof, disclosed herein, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof, for use in treating or preventing a disease, wherein the treating or preventing is effected or characterized by inhibition of the interaction of S0S1 and a RAS-family protein or by inhibition of the interaction of SOS1 and RA.

[0130] Another aspect of the present disclosure relates to a compound of any formula, or a pharmaceutical composition thereof, disclosed herein, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof, for use inhibiting the binding of hSOSl to H- or N- or K-RAS including their clinically known mutations and which inhibits the nucleotide exchange reaction catalyzed by hSOSl in the presence of a concentration of 20 pM or lower, but which are substantially inactive against EGFR-kinase at concentrations of 20 pM or lower for the preparation of a medicament for the treatment or prophylaxis of a hyperproliferative disorder.

[0131] Another aspect of the present disclosure relates to a compound of any formula, or a pharmaceutical composition thereof, disclosed herein, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof, for the manufacture of a medicament for use inhibiting the binding of hSOSl specifically to K- RAS G12C protein or another Ras mutant, as described herein, and which inhibits the nucleotide exchange reaction catalyzed by hSOSl in the presence of a concentration of 20 pM or lower, but which are substantially inactive against EGFR-kinase at concentrations of 20 pM or lower for the preparation of a medicament for the treatment or prophylaxis of a hyperproliferative disorder.

[0132] In another aspect, the present disclosure relates to the use of a compound of any formula, or a pharmaceutical composition thereof, disclosed herein, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof, in the manufacture of a medicament for treating or preventing a disease.

[0133] Administration of the disclosed compounds and pharmaceutical composition thereof can be accomplished via any mode of administration for therapeutic agents. These modes include systemic or local administration such as oral, nasal, parenteral, intravenous, transdermal, subcutaneous, vaginal, buccal, rectal or topical administration modes. Likewise, they can also be administered in intravenous (both bolus and infusion), intraperitoneal, subcutaneous or intramuscular form, and all using forms well known to those skilled in the pharmaceutical arts.

[0134] Depending on the intended mode of administration, the disclosed compounds and pharmaceutical compositions thereof can be in solid, semi-solid or liquid dosage form, such as, for example, injectables, tablets, suppositories, pills, time-release capsules, elixirs, tinctures, emulsions, syrups, powders, liquids, suspensions, or the like, sometimes in unit dosages and consistent with conventional pharmaceutical practices.

[0135] Illustrative pharmaceutical compositions are tablets and gelatin capsules comprising a compound of the disclosure and a pharmaceutically acceptable carrier, such as a) a diluent, e.g., purified water, triglyceride oils, such as hydrogenated or partially hydrogenated vegetable oil, or mixtures thereof, corn oil, olive oil, sunflower oil, safflower oil, fish oils, such as EPA or DHA, or their esters or triglycerides or mixtures thereof, omega-3 fatty acids or derivatives thereof, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, sodium, saccharin, glucose or glycine; b) a lubricant, e.g., silica, talcum, stearic acid, its magnesium or calcium salt, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride or polyethylene glycol; for tablets also; c) a binder, e.g., magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, magnesium carbonate, natural sugars such as glucose or beta-lactose, com sweeteners, natural and synthetic gums such as acacia, tragacanth or sodium alginate, waxes or polyvinylpyrrolidone, if desired; d) a disintegrant, e.g., starches, agar, methyl cellulose, bentonite, xanthan gum, alginic acid or its sodium salt, or effervescent mixtures; e) absorbent, colorant, flavorant and sweetener; f) an emulsifier or dispersing agent, such as Tween 80, Labrasol, HPMC, DOSS, caproyl 909, labrafac, labrafil, peceol, transcutol, capmul MCM, capmul PG-12, captex 355, gelucire, vitamin E TGPS or other acceptable emulsifier; or g) an agent that enhances absorption of the compound such as cyclodextrin, hydroxypropyl-cyclodextrin, PEG400, PEG200.

[0136] Liquid, particularly injectable, compositions can, for example, be prepared by dissolution, dispersion, etc. For example, the disclosed compound or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof is dissolved in or mixed with a pharmaceutically acceptable solvent such as, for example, water, saline, aqueous dextrose, glycerol, ethanol, and the like, to thereby form an injectable isotonic solution or suspension. Proteins such as albumin, chylomicron particles, or serum proteins can be used to solubilize the disclosed compounds.

[0137] The disclosed compounds can be also formulated as a suppository that can be prepared from fatty emulsions or suspensions; using polyalkylene glycols such as propylene glycol, as the carrier.

[0138] The disclosed compounds can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles and multilamellar vesicles. Liposomes can be formed from a variety of phospholipids, containing cholesterol, stearylamine or phosphatidylcholines. In some embodiments, a film of lipid components is hydrated with an aqueous solution of drug to a form lipid layer encapsulating the drug, as described for instance in U.S. Pat. No. 5,262,564, the contents of which are hereby incorporated by reference.

[0139] Disclosed compounds or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof can also be delivered by the use of monoclonal antibodies as individual carriers to which the disclosed compounds are coupled. The disclosed compounds or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof can also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer, polyhydroxypropylmethacrylamide-phenol, polyhydroxyethylaspanamidephenol, or polyethyleneoxidepolylysine substituted with palmitoyl residues. Furthermore, the disclosed compounds can be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, polylactic acid, polyepsilon caprolactone, polyhydroxy butyric acid, polyorthoesters, polyacetals, polydihydropyrans, polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogels. In one embodiment, disclosed compounds are not covalently bound to a polymer, e.g., a polycarboxylic acid polymer, or a polyacrylate.

[0140] Parental injectable administration is generally used for subcutaneous, intramuscular or intravenous injections and infusions. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions or solid forms suitable for dissolving in liquid prior to injection. [0141] Another aspect of the present disclosure relates to a pharmaceutical composition comprising a compound of the present disclosure or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof and a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier can further include an excipient, diluent, or surfactant.

[0142] Pharmaceutical compositions can be prepared according to conventional mixing, granulating or coating methods, respectively, and the present pharmaceutical compositions can contain from about 0.1% to about 99%, from about 5% to about 90%, or from about 1% to about 20% of the disclosed compound by weight or volume.

[0143] The dosage regimen utilizing the disclosed compound is selected in accordance with a variety of factors including type, species, age, weight, sex, and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal or hepatic function of the patient; and the particular disclosed compound employed. A physician or veterinarian of ordinary skill in the art can readily determine and prescribe the effective amount of the drug required to prevent, counter or arrest the progress of the condition.

[0144] Effective dosage amounts of the disclosed compounds or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof, when used for the indicated effects, range from about 0.5 mg to about 5000 mg of the disclosed compound as needed to treat the condition. Compositions for in vivo or in vitro use can contain about 0.5, 5, 20, 50, 75, 100, 150, 250, 500, 750, 1000, 1250, 2500, 3500, or 5000 mg of the disclosed compound or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof, or, in a range of from one amount to another amount in the list of doses. In one embodiment, the compositions are in the form of a tablet that can be scored.

Combination Therapy

[0145] T he methods of the present disclosure may include a compound of the disclosure used alone or in combination with one or more additional therapies (e g., non- drug treatments or therapeutic agents). The dosages of one or more of the additional therapies (e.g., non-drug treatments or therapeutic agents) may be reduced from standard dosages when administered alone. For example, doses may be determined empirically from drug combinations and permutations or may be deduced by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6 (2005)).

[0146] A compound of the present disclosure may be administered before, after, or concurrently with one or more of such additional therapies. When combined, dosages of a compound of the disclosure and dosages of the one or more additional therapies (e.g., non-drug treatment or therapeutic agent) provide a therapeutic effect (e.g., synergistic or additive therapeutic effect). A compound of the present disclosure and an additional therapy, such as an anti-cancer agent, may be administered together, such as in a unitary pharmaceutical composition, or separately and, when administered separately, this may occur simultaneously or sequentially. Such sequential administration may be close or remote in time.

[0147] In some embodiments, the additional therapy is the administration of side- effect limiting agents (e.g., agents intended to lessen the occurrence or severity of side effects of treatment. For example, in some embodiments, the compounds of the present disclosure can also be used in combination with a therapeutic agent that treats nausea. Examples of agents that can be used to treat nausea include: dronabinol, granisetron, metoclopramide, ondansetron, and prochlorperazine, or pharmaceutically acceptable salts thereof.

[0148] In some embodiments, the one or more additional therapies includes a non-drug treatment (e.g., surgery or radiation therapy). In some embodiments, the one or more additional therapies includes a therapeutic agent (e.g., a compound or biologic that is an anti-angiogenic agent, signal transduction inhibitor, antiproliferative agent, glycolysis inhibitor, or autophagy inhibitor). In some embodiments, the one or more additional therapies includes a non-drug treatment (e.g., surgery or radiation therapy) and a therapeutic agent (e.g., a compound or biologic that is an anti-angiogenic agent, signal transduction inhibitor, antiproliferative agent, glycolysis inhibitor, or autophagy inhibitor). In other embodiments, the one or more additional therapies includes two therapeutic agents. In still other embodiments, the one or more additional therapies includes three therapeutic agents. In some embodiments, the one or more additional therapies includes four or more therapeutic agents. [0149] In this Combination Therapy section, all references are incorporated by reference for the agents described, whether explicitly stated as such or not.

Non-drug therapies

[0150] Examples of non-drug treatments include, but are not limited to, radiation therapy, cryotherapy, hyperthermia, surgery (e.g., surgical excision of tumor tissue), and T cell adoptive transfer (ACT) therapy.

[0151] In some embodiments, the compounds of the disclosure may be used as an adjuvant therapy after surgery. In some embodiments, the compounds of the disclosure may be used as a neo-adjuvant therapy prior to surgery.

[0152] Radiation therapy may be used for inhibiting abnormal cell growth or treating a hyperproliferative disorder, such as cancer, in a subject (e.g., mammal (e.g., human)). Techniques for administering radiation therapy are known in the art. Radiation therapy can be administered through one of several methods, or a combination of methods, including, without limitation, external-beam therapy, internal radiation therapy, implant radiation, stereotactic radiosurgery, systemic radiation therapy, radiotherapy, and permanent or temporary interstitial brachy therapy. The term "brachy therapy," as used herein, refers to radiation therapy delivered by a spatially confined radioactive material inserted into the body at or near a tumor or other proliferative tissue disease site. The term is intended, without limitation, to include exposure to radioactive isotopes (e.g., At-21 1, I- 131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and radioactive isotopes of Lu). Suitable radiation sources for use as a cell conditioner include both solids and liquids. By way of non-limiting example, the radiation source can be a radionuclide, such as 1-125, I- 131, Yb-169, Ir-192 as a solid source, I-125 as a solid source, or other radionuclides that emit photons, beta particles, gamma radiation, or other therapeutic rays. The radioactive material can also be a fluid made from any solution of radionuclide(s), e.g., a solution of I- 125 or I-131, or a radioactive fluid can be produced using a slurry of a suitable fluid containing small particles of solid radionuclides, such as Au-198, or Y-90. Moreover, the radionuclide(s) can be embodied in a gel or radioactive micro spheres.

[0153] In some embodiments, the compounds of the present disclosure can render abnormal cells more sensitive to treatment with radiation for purposes of killing or inhibiting the growth of such cells. Accordingly, this disclosure further relates to a method for sensitizing abnormal cells in a mammal to treatment with radiation which comprises administering to the mammal an amount of a compound of the present disclosure, which amount is effective to sensitize abnormal cells to treatment with radiation. The amount of the compound in this method can be determined according to the means for ascertaining effective amounts of such compounds described herein. In some embodiments, the compounds of the present disclosure may be used as an adjuvant therapy after radiation therapy or as a neo-adjuvant therapy prior to radiation therapy.

[0154] In some embodiments, the non-drug treatment is a T cell adoptive transfer (ACT) therapy. In some embodiments, the T cell is an activated T cell. The T cell may be modified to express a chimeric antigen receptor (CAR). CAR modified T (CAR-T) cells can be generated by any method known in the art. For example, the CAR-T cells can be generated by introducing a suitable expression vector encoding the CAR to a T cell. Prior to expansion and genetic modification of the T cells, a source of T cells is obtained from a subject. T cells can be obtained from a number of sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, cord blood, thymus tissue, tissue from a site of infection, ascites, pleural effusion, spleen tissue, and tumors. In certain embodiments of the present disclosure, any number of T cell lines available in the art may be used. In some embodiments, the T cell is an autologous T cell. Whether prior to or after genetic modification of the T cells to express a desirable protein (e.g., a CAR), the T cells can be activated and expanded generally using methods as described, for example, in U.S. Patents 6,352,694; 6,534,055; 6,905,680; 6,692,964; 5,858,358; 6,887,466; 6,905,681; 7,144,575; 7,067,318; 7,172,869; 7,232,566; 7,175,843; 7,572,631; 5,883,223; 6,905,874; 6,797,514; and 6,867,041.

Therapeutic agents

[0155] A therapeutic agent may be a compound used in the treatment of cancer or symptoms associated therewith. A compound of the present disclosure may be combined with a second, third, or fourth therapeutic agent, or more. A compound of the present disclosure may be combined with one or more therapeutic agents along with one or more non-drug therapies. [0156] For example, a therapeutic agent may be a steroid. Steroids are known in the art. Accordingly, in some embodiments, the one or more additional therapies includes a steroid. Suitable steroids may include, but are not limited to, 21 -acetoxy pregnenolone, alclometasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, clocortolone, cloprednol, corticosterone, cortisone, cortivazol, deflazacort, desonide, desoximetasone, dexamethasone, diflorasone, diflucortolone, difuprednate, enoxolone, fluazacort, fiucloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, fluocortin butyl, fluocortolone, fluoromethoIone, fluperolone acetate, fluprednidene acetate, fluprednisolone, flurandrenolide, fluticasone propionate, formocortal, halcinonide, halobetasol propionate, halometasone, hydrocortisone, loteprednol etabonate, mazipredone, medrysone, meprednisone, methylprednisolone, mometasone furoate, paramethasone, predni carb ate, prednisolone, prednisolone 25-diethylaminoacetate, prednisolone sodium phosphate, prednisone, prednival, prednylidene, rimexolone, tixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexacetonide, and salts or derivatives thereof.

[0157] Further examples of therapeutic agents that may be used in combination therapy with a compound of the present disclosure include compounds described in the following patents: U.S. Patent Nos. 6,258,812, 6,630,500, 6,515,004, 6,713,485, 5,521,184, 5,770,599, 5,747,498, 5,990,141, 6,235,764, and 8,623,885, and International Patent Applications WO01/37820, WO01/32651, WO02/68406, WO02/66470, WO02/55501, WO04/05279, WO04/07481, WO04/07458, WO04/09784, WO02/59110, WO99/45009, WO00/59509, WO99/61422, WO00/12089, and WO00/02871.

[0158] A therapeutic agent may be a biologic (e.g., cytokine (e.g., interferon or an interleukin such as IL-2)) used in treatment of cancer or symptoms associated therewith. Biologies are known in the art. In some embodiments, the biologic is an immunoglobulin- based biologic, e.g., a monoclonal antibody (e.g., a humanized antibody, a fully human antibody, an Fc fusion protein, or a functional fragment thereof) that agonizes a target to stimulate an anti-cancer response or antagonizes an antigen important for cancer. Also included are antibody-drug conjugates.

[0159] A therapeutic agent may be a T-cell checkpoint inhibitor. Such checkpoint inhibitors are known in the art. In one embodiment, the checkpoint inhibitor is an inhibitory antibody (e.g., a monospecific antibody such as a monoclonal antibody). The antibody may be, e.g., humanized or fully human. In some embodiments, the checkpoint inhibitor is a fusion protein, e.g., an Fc-receptor fusion protein. In some embodiments, the checkpoint inhibitor is an agent, such as an antibody, that interacts with a checkpoint protein. In some embodiments, the checkpoint inhibitor is an agent, such as an antibody, that interacts with the ligand of a checkpoint protein. In some embodiments, the checkpoint inhibitor is an inhibitor (e.g., an inhibitory antibody or small molecule inhibitor) of CTLA-4 (e.g., an anti- CTLA-4 antibody or fusion a protein). In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor) of PD-1. In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor) of PD-L1. In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist (e.g., an inhibitory antibody or Fc fusion or small molecule inhibitor) of PD-L2 (e.g., a PD-L2/Ig fusion protein). In some embodiments, the checkpoint inhibitor is an inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor) of B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK 1, CHK2, A2aR, B-7 family ligands, or a combination thereof. In some embodiments, the checkpoint inhibitor is pembrolizumab, nivolumab, PDR001 (NVS), REGN2810 (Sanofi/Regeneron), a PD-L1 antibody such as, e.g., avelumab, durvalumab, atezolizumab, pidilizumab, JNJ-63723283 (JNJ), BGB-A317 (BeiGene & Celgene) or a checkpoint inhibitor disclosed in Preusser, M. et al. (2015) Nat. Rev. Neurol., including, without limitation, ipilimumab, tremelimumab, nivolumab, pembrolizumab, AMP224, AMP514/ MED 10680, BMS936559, MED14736, MPDL3280A, MSB0010718C, BMS986016, IMP321, lirilumab, IPH2101, 1-7F9, and KW-6002.

[0160] A therapeutic agent may be an anti-TIGIT antibody, such as MBSA43, BMS-986207, MK-7684, COM902, AB 154, MTIG7192A or OMP-313M32 (etigilimab). Other anti-TIGIT antibodies are known in the art.

[0161] A therapeutic agent may be an agent that treats cancer or symptoms associated therewith (e.g., a cytotoxic agent, non-peptide small molecules, or other compound useful in the treatment of cancer or symptoms associated therewith, collectively, an “anti-cancer agent”). Anti-cancer agents can be, e.g., chemotherapeutics or targeted therapy agents. Such agents are known in the art. [0162] Anti-cancer agents include mitotic inhibitors, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, alkylating agents, antimetabolites, folic acid analogs, pyrimidine analogs, purine analogs and related inhibitors, vinca alkaloids, epipodopyyllotoxins, antibiotics, L-Asparaginase, topoisomerase inhibitors, interferons, platinum coordination complexes, anthracenedione substituted urea, methyl hydrazine derivatives, adrenocortical suppressant, adrenocorticosteroides, progestins, estrogens, antiestrogen, androgens, antiandrogen, and gonadotropin-releasing hormone analog. Further anti-cancer agents include leucovorin (LV), irenotecan, oxaliplatin, capecitabine, paclitaxel, and doxetaxel. In some embodiments, the one or more additional therapies includes two or more anti-cancer agents. The two or more anti-cancer agents can be used in a cocktail to be administered in combination or administered separately. Suitable dosing regimens of combination anti-cancer agents are known in the art and described in, for example, Saltz et al., Proc. Am. Soc. Clin. Oncol. 18:233a (1999), and Douillard et al., Lancet 355(9209): 1041-1047 (2000).

[0163] Other non-limiting examples of anti-cancer agents include Gleevec® (Imatinib Mesylate), Kyprolis® (carfilzomib); Velcade® (bortezomib); Casodex (bicalutamide); Iressa® (gefitinib); alkylating agents such as thiotepa and cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethiylenethiophosphoramide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; cally statin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189 and CB1-TM1); eleutherobin; pancrati statin; sarcodictyin A; spongistatin; nitrogen mustards such as chlorambucil, chlomaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, such as calicheamicin gammall and calicheamicin omegall (see, e.g., Agnew, Chem. Inti. Ed Engl. 33:183-186 (1994)); dynemicin such as dynemicin A; bisphosphonates such as clodronate; an esperamicin; neocarzinostatin chromophore and related chromoprotein enediyne antibiotic chromophores, aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, calicheamicin, carabicin, caminomycin, carminomycin, carzinophilin, chromomycins, dactinomycin, daunorubicin, detorubicin, 6- diazo- 5-oxo-L-norleucine, adriamycin (doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, deoxydoxorubicin, epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti- metabolites such as methotrexate and 5 -fluorouracil (5-FU); folic acid analogues such as denopterin, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenishers such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elfomithine; elliptinium acetate; an epothilone such as epothilone B; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, OR); razoxane; rhizoxin; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2, 2', 2"-trichlorotri ethylamine; trichothecenes such as T- 2 toxin, verracurin A, roridin A and anguidine; urethane; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, e.g., Taxol® (paclitaxel), Abraxane® (cremophor-free, albumin-engineered nanoparticle formulation of paclitaxel), and Taxotere® (doxetaxel); chloranbucil; tamoxifen (Nolvadex™); raloxifene; aromatase inhibiting 4(5)-imidazoles; 4-hydroxytamoxifen; trioxifene; keoxifene; LY 1 17018; onapristone; toremifene (Fareston®); flutamide, nilutamide, bicalutamide, leuprolide, goserelin; chlorambucil, Gemzar® gemcitabine; 6-thioguanine; mercaptopurine; platinum coordination complexes such as cisplatin, oxaliplatin and carboplatin; vinblastine; platinum; etoposide (VP- 16); ifosfamide; mitoxantrone; vincristine; Navelbine® (vinorelbine); novantrone; teniposide; edatrexate; daunomycin; aminopterin; ibandronate; irinotecan (e.g., CPT-11); topoisomerase inhibitor RFS 2000; difluoromethylomithine (DMFO); retinoids such as retinoic acid; esperamicins; capecitabine (e.g., Xeloda®); and pharmaceutically acceptable salts of any of the above.

[0164] Additional non-limiting examples of anti-cancer agents include trastuzumab (Herceptin®), bevacizumab (Avastin®), cetuximab (Erbitux®), rituximab (Rituxan®), Taxol®, Arimidex®, ABVD, avicine, abagovomab, acridine carboxamide, adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, alpharadin, alvocidib, 3- aminopyridine-2-carboxaldehyde thiosemicarbazone, amonafide, anthracenedione, anti- CD22 immunotoxins, antineoplastics (e.g., cell-cycle nonspecific antineoplastic agents, and other antineoplastics described herein), antitumorigenic herbs, apaziquone, atiprimod, azathioprine, belotecan, bendamustine, BIBW 2992, biricodar, brostallicin, bryostatin, buthionine sulfoximine, CBV (chemotherapy), calyculin, di chloroacetic acid, discodermolide, elsamitrucin, enocitabine, eribulin, exatecan, exisulind, ferruginol, forodesine, fosfestrol, ICE chemotherapy regimen, IT-101, imexon, imiquimod, indolocarbazole, irofulven, laniquidar, larotaxel, lenalidomide, lucanthone, lurtotecan, mafosfamide, mitozolomide, naf oxi dine, nedaplatin, olaparib, ortataxel, PAC-1, pawpaw, pixantrone, proteasome inhibitors, rebeccamycin, resiquimod, rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V, swainsonine, talaporfm, tariquidar, tegafur- uracil, temodar, tesetaxel, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, vadimezan, vinflunine, ZD6126, and zosuquidar.

[0165] Further non-limiting examples of anti-cancer agents include natural products such as vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), epidipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycins, plicamycin (mithramycin), mitomycin, enzymes (e.g., L-asparaginase which systemically metabolizes L-asparagine and deprives cells which do not have the capacity to synthesize their own asparagine), antiplatelet agents, antiproliferative/antimitotic alkylating agents such as nitrogen mustards (e.g., mechlorethamine, cyclophosphamide and analogs, melphalan, and chlorambucil), ethylenimines and methylmelamines (e.g., hexamethylmelamine and thiotepa), CDK inhibitors (e.g., a CDK4/6 inhibitor such as abemaciclib, ribociclib, Palbociclib, seliciclib, UCN-01, P1446A-05, PD-0332991, dinaciclib, P27-00, AT-7519, RGB286638, and SCH727965), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and analogs, and streptozocin), trazenes- dacarbazinine (DTIC), antiproliferative/antimitotic antimetabolites such as folic acid analogs, pyrimidine analogs (e.g., fluorouracil, floxuridine, and cytarabine), purine analogs and related inhibitors (e.g., mercaptopurine, thioguanine, pentostatin, and 2- chlorodeoxyadenosine), aromatase inhibitors (e.g., anastrozole, exemestane, and letrozole), and platinum coordination complexes (e.g., cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apicidan, suberoyl anilide hydroamic acid, vorinostat, belinostat, LBH 589, romidepsin, ACY-1215, and panobinostat), mTOR inhibitors (e.g., vistusertib, temsirolimus, everolimus, ridaforolimus, and sirolimus), KSP(Eg5) inhibitors (e.g., Array 520), DNA binding agents (e.g., Zalypsis®), PI3K inhibitors such as PI3K delta inhibitor (e.g., GS-1101 and TGR-1202), PI3K delta and gamma inhibitor (e.g., CAL- 130), copanlisib, alpelisib and idelalisib; multi-kinase inhibitor (e.g., TG02 and sorafenib), hormones (e.g., estrogen) and hormone agonists such as leutinizing hormone releasing hormone (LHRH) agonists (e.g., goserelin, leuprolide and triptorelin), BAFF-neutralizing antibody (e.g., LY2127399), IKK inhibitors, p38MAPK inhibitors, anti -IL-6 (e.g., CNT0328), telomerase inhibitors (e.g., GRN 163L), aurora kinase inhibitors (e.g., MLN8237), cell surface monoclonal antibodies (e.g., anti-CD38 (HUMAX-CD38), anti- CS1 (e.g., elotuzumab), HSP90 inhibitors (e.g., 17 AAG and KOS 953), P13K / Akt inhibitors (e.g., perifosine), Akt inhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., enzastaurin), FTIs (e.g., Zamestra™), anti-CD138 (e.g., BT062), Torcl/2 specific kinase inhibitors (e.g., INK128), ER/UPR targeting agents (e.g., MKC-3946), cFMS inhibitors (e.g., ARRY-382), JAK1/2 inhibitors (e.g., CYT387), PARP inhibitors (e.g., olaparib and veliparib (ABT-888)), and BCL-2 antagonists.

[0166] In some embodiments, an anti-cancer agent is selected from mechlorethamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, Navelbine®, sorafenib, or any analog or derivative variant of the foregoing.

[0167] In some embodiments, the anti-cancer agent is a HER2 inhibitor. HER2 inhibitors are known in the art. Non-limiting examples of HER2 inhibitors include monoclonal antibodies such as trastuzumab (Herceptin®) and pertuzumab (Perjeta®); small molecule tyrosine kinase inhibitors such as gefitinib (Iressa®), erlotinib (Tarceva®), pilitinib, CP-654577, CP-724714, canertinib (CI 1033), HKI-272, lapatinib (GW-572016; Tykerb®), PKI-166, AEE788, BMS-599626, HKI-357, BIBW 2992, ARRY-334543, and JNJ-26483327.

[0168] In some embodiments, an anti-cancer agent is an ALK inhibitor. ALK inhibitors are known in the art. Non-limiting examples of ALK inhibitors include ceritinib, TAE-684 (NVP-TAE694), PF02341066 (crizotinib or 1066), alectinib; brigatinib; entrectinib; ensartinib (X-396); lorlatinib; ASP3026; CEP-37440; 4SC-203; TL-398; PLB1003; TSR-011; CT-707; TPX-0005, and AP26113. Additional examples of ALK kinase inhibitors are described in examples 3-39 of W005016894.

[0169] In some embodiments, an anti-cancer agent is an inhibitor of a member downstream of a Receptor Tyrosine Kinase (RTK)/Growth Factor Receptor (e.g., a SHP2 inhibitor (e.g., SHP099, TNO155, RMC-4550, RMC-4630, JAB-3068, JAB-3312, RLY- 1971, ERAS-601, SH3809, PF-07284892, or BBP-398), or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof), an S0S1 inhibitor (e.g., B 1-1701963, BI-3406, SDR5, BAY-293, MRTX-0902, or RMC-5845, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof), a Raf inhibitor, a MEK inhibitor, an ERK inhibitor, a PI3K inhibitor, a PTEN inhibitor, an AKT inhibitor, or an mTOR inhibitor (e.g., mTORCl inhibitor or mT0RC2 inhibitor). In some embodiments, the anti-cancer agent is JAB-3312.

[0170] In some embodiments, an anti-cancer agent is a Ras inhibitor or a Ras vaccine, or another therapeutic modality designed to directly or indirectly decrease the oncogenic activity of Ras. Such agents are known in the art. In some embodiments, an anti-cancer agent is a Ras inhibitor. In some embodiments, the Ras inhibitor targets Ras in its active, or GTP-bound state. In some embodiments, the Ras inhibitor targets Ras in its inactive, or GDP-bound state. In some embodiments, the Ras inhibitor is, such as an inhibitor of K-Ras G12C, such as AMG 510, MRTX1257, MRTX849, JNJ-74699157, LY3499446, ARS-1620, ARS-853, BPI-421286, LY3537982, JDQ443, JAB-3312, JAB- 21822, JAB-21000, IBI351, ERAS-3490, RMC-6291, BI 1823911, D-1553, D3S-001, HBI-2438, HS-10370, MK-1084, YL-15293 or GDC-6036. In some embodiments, the Ras inhibitor is an inhibitor of K-Ras G12D, such as MRTX1133, JAB-22000, MRTX282, ERAS-4, HRS-4642, BI-2852, ASP3082, TH-Z827, TH-7835 and KD-8. In some embodiments, the Ras inhibitor is a K-Ras G12V inhibitor, such as JAB-23000. In some embodiments, the Ras inhibitor is JAB-23400. In some embodiments, the Ras inhibitor is RMC-6236. In some embodiments, the Ras inhibitor is selected from a Ras(ON) inhibitor (that is, Ras in its GTP-bound state) disclosed in the following, incorporated herein by reference in their entireties, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof: WO 2022/235870, WO 2022/235864, WO 2022/060836, WO 2021091982, WO 2021091967, WO 2021091956, and WO 2020132597. Other examples of Ras inhibitors are known in the art, such as in the following, incorporated herein by reference in their entireties: WO 2023287896, WO

2023287730, WO 2023284881, WO 2023284730, WO 2023284537, WO 2023283933, WO 2023283213, WO 2023280960, WO 2023280280, W02023278600, WO 2023280136, WO 2023280026, WO 2023278600, WO 2023274383, WO 2023274324, WO 2023034290, WO 2023020523, WO 2023020521, WO 2023020519, WO 2023020518, WO 2023018812, WO 2023018810, WO 2023018809, WO 2023018699, WO 2023015559, WO 2023014979, WO 2023014006, WO 2023010121, WO 2023009716, WO 2023009572, WO 2023004102, WO 2023003417, WO 2023001141, WO 2023001123, WO 2022271923, WO 2022271823, WO 2022271810, WO 2022271658, WO 2022269508, WO 2022266167, WO 2022266069, WO 2022266015, WO 2022265974, WO 2022261154, WO 2022261154, WO 2022251576, WO 2022251296, WO 2022237815, WO 2022232332, WO 2022232331, WO 2022232320, WO 2022232318, WO 2022223037, WO 2022221739, WO 2022221528, WO 2022221386, WO 2022216762, WO 2022192794, WO 2022192790, WO 2022188729, WO 2022187411, WO 2022184178, WO 2022173870, WO 2022173678, WO 2022135346, WO 2022133731, WO 2022133038, WO 2022133345, WO 2022132200, WO 2022119748, WO 2022109485, WO 2022109487, WO 2022066805, WO 2022002102, WO 2022002018, WO 2021259331, WO 2021257828, WO 2021252339, WO 2021248095, WO 2021248090, WO 2021248083, WO 2021248082, WO 2021248079, WO 2021248055, WO 2021245051, WO 2021244603, WO 2021239058, WO 2021231526, WO 2021228161, WO 2021219090, WO 2021219090, WO 2021219072, WO 2021218939, WO 2021217019, WO 2021216770, WO 2021215545, WO 2021215544, WO 2021211864, WO 2021190467, WO 2021185233, WO 2021180181, WO 2021175199, 2021173923, WO 2021169990, WO 2021169963, WO 2021168193, WO 2021158071, WO 2021155716, WO 2021152149, WO 2021150613, WO 2021147967, WO 2021147965, WO 2021143693, WO 2021142252, WO 2021141628, WO 2021139748, WO 2021139678, WO 2021129824, WO 2021129820, WO 2021127404, WO 2021126816, WO 2021126799, WO 2021124222, WO 2021121371, WO 2021121367, WO 2021121330, WO 2020050890, WO 2020047192, WO 2020035031, WO 2020028706, WO 2019241157, WO 2019232419, WO 2019217691, WO 2019217307, WO 2019215203, WO 2019213526, WO 2019213516, WO 2019155399, WO 2019150305, WO 2019110751, WO 2019099524, WO 2019051291, WO 2018218070, WO 2018217651, WO 2018218071, WO 2018218069, WO 2018206539, WO 2018143315, WO 2018140600, WO 2018140599, WO 2018140598, WO 2018140514, WO 2018140513, WO 2018140512, WO 2018119183, WO 2018112420, WO 2018068017, WO 2018064510, WO 2017201161, WO 2017172979, WO 2017100546, WO 2017087528, WO 2017058807, WO 2017058805, WO 2017058728, WO 2017058902, WO 2017058792, WO 2017058768, WO 2017058915, WO 2017015562, WO 2016168540, WO 2016164675, WO 2016049568, WO 2016049524, WO 2015054572, WO 2014152588, WO 2014143659 and WO 2013155223, CN 114195804, CN 114195788, CN 114057776, CN 114057744, CN 114057743, CN

113999226, CN 113980032, CN 113980014, CN 113929676, CN 113754653, CN

113683616, CN 113563323, CN 113527299, CN 113527294, CN 113527293, CN

113493440, CN 113429405, CN 113248521, CN 113087700, CN 113024544, CN

113004269, CN 112920183, CN 112778284, CN 112390818, CN 112390788, CN

112300196, CN 112300194, CN 112300173, CN 112225734, CN 112142735, CN

112110918, CN 112094269, CN 112047937, and CN 109574871, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof.

[0171] In some embodiments, the cancer comprises a SHP2 mutation (see, e.g., WO 2022/060583). In some embodiments, a cancer comprises a NF1^LOF mutation. In some embodiments of methods herein, the cancer comprises a Ras^MUT mutation and a compound of the present disclosure is administered to, e.g., a patient in need thereof, in combination with an additional therapeutic agent, e.g., a MEK inhibitor, such as a MEK inhibitor described herein. In some embodiments of methods herein, the cancer is colorectal cancer and a compound of the present disclosure is administered to, e.g., a patient in need thereof, in combination with an additional therapeutic agent, such as a topoisomerase I inhibitor (e.g., irinotecan). In some embodiments of methods herein, the cancer is non-small cell lung cancer and a compound of the present disclosure is administered to, e.g., a patient in need thereof, in combination with an additional therapeutic agent, e.g., a MEK inhibitor, such as a MEK inhibitor described herein (e.g., trametinib). In some embodiments of methods herein, the cancer is non-small cell lung cancer or colorectal cancer, and a compound of the present disclosure is administered to, e.g., a patient in need thereof, in combination with a Ras inhibitor, such as a Ras inhibitor described herein (e.g., AMG 510, MRTX1257, LY349946, MRTX849, ARS-3248 (JNJ- 74699157), MRTX1133, ARS-853, BPI-421286, LY3537982, JDQ443, JAB-21000, RMC-6291, RMC-6236, RMC-9805, RMC-8839, GDC-6036, ERAS-3490, ERAS-4, JAB- 22000, JAB-23000, or ARS-1620).

[0172] In some embodiments, a therapeutic agent that may be combined with a compound of the present disclosure is an inhibitor of the MAP kinase (MAPK) pathway (or “MAPK inhibitor”). Such agents are known in the art. MAPK inhibitors include, but are not limited to, one or more MAPK inhibitor described in Cancers (Basel) 2015 Sep; 7(3): 1758-1784. For example, the MAPK inhibitor may be selected from one or more of trametinib, binimetinib, selumetinib, cobimetinib, LErafAON (NeoPharm), ISIS 5132; vemurafenib, pimasertib, TAK733, RO4987655 (CH4987655); CI-1040; PD-0325901; CH5126766; MAP855; AZD6244; refametinib (RDEA 119/BAY 86-9766); GDC- 0973/XL581; AZD8330 (ARRY-424704/ARRY-704); RO5126766 (Roche, described in PLoS One. 2014 Nov 25;9(11)); and GSK1120212 (or JTP-74057, described in Clin Cancer Res. 2011 Mar l;17(5):989-1000). The MAPK inhibitor may be PLX8394, LXH254, GDC-5573, or LY3009120.

[0173] In some embodiments, an anti-cancer agent is a disrupter or inhibitor of the RAS-RAF-ERK or PI3K-AKT-TOR or PI3K-AKT signaling pathways. Such agents are known in the art. The PI3K/AKT inhibitor may include, but is not limited to, one or more PI3K/AKT inhibitor described in Cancers (Basel) 2015 Sep; 7(3): 1758-1784. For example, the PI3K/AKT inhibitor may be selected from one or more of NVP-BEZ235; BGT226; XL765/SAR245409; SF1126; GDC-0980; PI-103; PF-04691502; PKI-587; GSK2126458. [0174] In some embodiments, an anti-cancer agent is a PD-1 or PD-L1 antagonist. Such agents are known in the art.

[0175] In some embodiments, additional therapeutic agents include ALK inhibitors, HER2 inhibitors, EGFR inhibitors, IGF-1R inhibitors, MEK inhibitors, PI3K inhibitors, AKT inhibitors, TOR inhibitors, MCL-1 inhibitors, BCL-2 inhibitors, SHP2 inhibitors, proteasome inhibitors, and immune therapies. In some embodiments, additional therapeutic agents include FGFR inhibitors, PARP inhibitors, BET inhibitors, PRMT5i inhibitors, MAT2A inhibitors, VEGF inhibitors, and HD AC inhibitors. In some embodiments, a therapeutic agent may be a pan-RTK inhibitor, such as afatinib.

[0176] IGF-1R inhibitors are known in the art and include linsitinib, or a pharmaceutically acceptable salt thereof.

[0177] EGFR inhibitors are known in the art and include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotide or siRNA. Useful antibody inhibitors of EGFR include cetuximab (Erbitux®), panitumumab (Vectibix®), zalutumumab, nimotuzumab, and matuzumab. Further antibody -based EGFR inhibitors include any anti-EGFR antibody or antibody fragment that can partially or completely block EGFR activation by its natural ligand. Non-limiting examples of antibody-based EGFR inhibitors include those described in Modjtahedi et al., Br. J. Cancer 1993, 67:247-253; Teramoto et al., Cancer 1996, 77:639-645; Goldstein et al., Clin. Cancer Res. 1995, 1 : 1311-1318; Huang et al., 1999, Cancer Res. 15:59(8): 1935-40; and Yang et al., Cancer Res.1999, 59: 1236-1243. The EGFR inhibitor can be monoclonal antibody Mab E7.6.3 (Yang, 1999 supra), or Mab C225 (ATCC Accession No. HB-8508), or an antibody or antibody fragment having the binding specificity thereof.

[0178] Small molecule antagonists of EGFR include gefitinib (Iressa®), erlotinib (Tarceva®), and lapatinib (TykerB®). See, e.g., Yan et al., Pharmacogenetics and Pharmacogenomics in Oncology Therapeutic Antibody Development, BioTechniques 2005, 39(4):565-8; and Paez et al., EGFR Mutations in Lung Cancer Correlation with Clinical Response to Gefitinib Therapy, Science 2004, 304(5676): 1497-500. In some embodiments, the EGFR inhibitor is osimertinib (Tagrisso®). Further non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in the following patent publications, and all pharmaceutically acceptable salts of such EGFR inhibitors: EP 0520722; EP 0566226; WO96/33980; U.S. Pat. No. 5,747,498; WO96/30347; EP 0787772; W097/30034; W097/30044; WO97/38994; WO97/49688; EP 837063; WO98/02434; WO97/38983; WO95/19774; WO95/19970; WO97/13771; WO98/02437; WO98/02438; WO97/32881; DE 19629652; WO98/33798; WO97/32880; WO97/32880; EP 682027; WO97/02266; WO97/27199; WO98/07726; WO97/34895; WO96/31510; WO98/14449; WO98/14450; WO98/14451; WO95/09847; WO97/19065; WO98/17662; U.S. Pat. No. 5,789,427; U.S. Pat. No. 5,650,415; U.S. Pat. No. 5,656,643; WO99/35146; WO99/35132; W099/07701; and WO92/20642. Additional non-limiting examples of small molecule EGFR inhibitors include any of the EGFR inhibitors described in Traxler et al., Exp. Opin. Ther. Patents 1998, 8(12): 1599-1625. In some embodiments, an EGFR inhibitor is an ERBB inhibitor. In humans, the ERBB family contains HER1 (EGFR, ERBB1), HER.2 (NEU, ERBB2), HER3 (ERBB3), and HER (ERBB4).

[0179] MEK inhibitors are known in the art and include, but are not limited to, pimasertib, selumetinib, cobimetinib (Cotellic®), trametinib (Mekinist®), and binimetinib (Mektovi®). In some embodiments, a MEK inhibitor targets a MEK mutation that is a Class I MEK1 mutation selected from D67N; P124L; P124S; and L177V. In some embodiments, the MEK mutation is a Class II MEK1 mutation selected from AE51-Q58; AF53-Q58; E203K; L177M; C121S; F53L; K57E; Q56P; and K57N.

[0180] PI3K inhibitors are known in the art and include, but are not limited to, wortmannin; 17-hydroxywortmannin analogs described in WO06/044453; 4-[2-(lH- Indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-l-yl]methyl]thieno[3,2-d]pyrimidin-4- yl]morpholine (also known as pictilisib or GDC-0941 and described in W009/036082 and W009/055730); 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5- c]quinolin-l-yl]phenyl]propionitrile (also known as BEZ 235 or NVP-BEZ 235, and described in W006/122806); (S)-l-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4- morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)piperazin-l-yl)-2-hydroxypropan-l-one (described in W008/070740); LY294002 (2-(4-morpholinyl)-8-phenyl-4H-l-benzopyran-4- one (available from Axon Medchem); PI 103 hydrochloride (3-[4-(4-morpholinylpyrido- [3',2':4,5]furo[3,2-d]pyrimidin-2-yl] phenol hydrochloride (available from Axon Medchem); PIK 75 (2-methyl-5-nitro-2-[(6-bromoimidazo[l,2-a]pyridin-3-yl)methylene]- 1 -methylhydrazide-benzenesulfonic acid, monohydrochloride) (available from Axon Medchem); PIK 90 (N-(7,8-dimethoxy-2,3-dihydro-imidazo[l,2-c]quinazolin-5-yl)- nicotinamide (available from Axon Medchem); AS-252424 (5-[l-[5-(4-fluoro-2-hydroxy- phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidine-2,4-dione (available from Axon Medchem); TGX-221 (7-methyl-2-(4-morpholinyl)-9-[l-(phenylamino)ethyl]-4H-pyrido- [1,2-a]pyrimidin-4-one (available from Axon Medchem); XL-765; and XL-147. Other PI3K inhibitors include demethoxyviridin, perifosine, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, Palomid 529, GSK1059615, ZSTK474, PWT33597, IC87114, TGI 00-115, CAL263, PI-103, GNE-477, CUDC-907, and AEZS- 136.

[0181] AKT inhibitors are known in the art and include, but are not limited to, Akt-1-1 (inhibits Aktl) (Barnett et al., Biochem. J. 2005, 385(Pt. 2): 399-408); Akt-1-1,2 (inhibits Akl and 2) (Barnett et al., Biochem. J. 2005, 385(Pt. 2): 399-408); API-59CJ-Ome (e.g., Jin et al., Br. J. Cancer 2004, 91 : 1808-12); l-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO 05/011700); indole-3 -carbinol and derivatives thereof (e.g., U.S. Pat. No. 6,656,963; Sarkar and Li J Nutr. 2004, 134(12 Suppl):3493S-3498S); perifosine (e.g., interferes with Akt membrane localization; Dasmahapatra et al. Clin. Cancer Res. 2004, 10(15):5242-52); phosphatidylinositol ether lipid analogues (e.g., Gills and Dennis Expert. Opin. Investig. Drugs 2004, 13:787-97); and triciribine (TCN or API-2 or NCI identifier: NSC 154020; Yang et al., Cancer Res. 2004, 64:4394-9).

[0182] mTOR inhibitors are known in the art and include, but are not limited to, ATP-competitive mTORCl/mTORC2 inhibitors, e.g., PI-103, PP242, PP30; Torin 1; FKBP12 enhancers; 4H-l-benzopyran-4-one derivatives; and rapamycin (also known as sirolimus) and derivatives thereof, including: temsirolimus (Torisel®); everolimus (Afinitor®; W094/09010); ridaforolimus (also known as deforolimus or AP23573); rapalogs, e.g., as disclosed in WO98/02441 and WOOl/14387, e.g. AP23464 and AP23841; 40-(2-hydroxyethyl)rapamycin; 40- [3 -hydroxy (hy droxymethyl)m ethylpropanoate] - rapamycin (also known as CC1779); 40-epi-(tetrazolyt)-rapamycin (also called ABT578); 32-deoxorapamycin; 16-pentynyloxy-32(S)-dihydrorapanycin; derivatives disclosed in W005/005434; derivatives disclosed in U.S. Patent Nos. 5,258,389, 5,118,677, 5,118,678, 5,100,883, 5,151,413, 5,120,842, and 5,256,790, and in W094/090101, WO92/05179, WO93/111130, WO94/02136, WO94/02485, WO95/14023, WO94/02136, WO95/16691, WO96/41807, WO96/41807, and WO2018204416; and phosphorus-containing rapamycin derivatives (e.g., WO05/016252). In some embodiments, the mTOR inhibitor is a bisteric inhibitor (see, e.g., WO2018204416, WO2019212990 and WO2019212991), such as

RMC-5552, having the structure:

[0183] BRAF inhibitors that may be used in combination with compounds of the disclosure are known in the art and include, for example, vemurafenib, dabrafenib, and encorafenib. A BRAF may comprise a Class 3 BRAF mutation. In some embodiments, the Class 3 BRAF mutation is selected from one or more of the following amino acid substitutions in human BRAF: D287H; P367R; V459L; G466V; G466E; G466A; S467L; G469E; N581S; N581I; D594N; D594G; D594A; D594H; F595L; G596D; G596R and A762E.

[0184] MCL-1 inhibitors are known in the art and include, but are not limited to, AMG-176, MIK665, and S63845. The myeloid cell leukemia-1 (MCL-1) protein is one of the key anti-apoptotic members of the B-cell lymphoma-2 (BCL-2) protein family. Over- expression of MCL-1 has been closely related to tumor progression as well as to resistance, not only to traditional chemotherapies but also to targeted therapeutics including BCL-2 inhibitors such as ABT-263.

[0185] In some embodiments, the additional therapeutic agent is a SHP2 inhibitor. SHP2 inhibitors are known in the art. SHP2 is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene that contributes to multiple cellular functions including proliferation, differentiation, cell cycle maintenance and migration. SHP2 has two N- terminal Src homology 2 domains (N-SH2 and C-SH2), a catalytic domain (PTP), and a C- terminal tail. The two SH2 domains control the subcellular localization and functional regulation of SHP2. The molecule exists in an inactive, self-inhibited conformation stabilized by a binding network involving residues from both the N-SH2 and PTP domains. Stimulation by, for example, cytokines or growth factors acting through receptor tyrosine kinases (RTKs) leads to exposure of the catalytic site resulting in enzymatic activation of SHP2.

[0186] SHP2 is involved in signaling through the RAS-mitogen-activated protein kinase (MAPK), the JAK-STAT or the phosphoinositol 3-kinase-AKT pathways. Mutations in the PTPN11 gene and subsequently in SHP2 have been identified in several human developmental diseases, such as Noonan Syndrome and Leopard Syndrome, as well as human cancers, such as juvenile myelomonocytic leukemia, neuroblastoma, melanoma, acute myeloid leukemia and cancers of the breast, lung, and colon. Some of these mutations destabilize the auto-inhibited conformation of SHP2 and promote autoactivation or enhanced growth factor driven activation of SHP2. SHP2, therefore, represents a highly attractive target for the development of novel therapies for the treatment of various diseases including cancer. A SHP2 inhibitor (e.g., RMC-4550 or SHP099) in combination with a RAS pathway inhibitor (e.g., a MEK inhibitor) have been shown to inhibit the proliferation of multiple cancer cell lines in vitro (e.g., pancreas, lung, ovarian and breast cancer).

[0187] Non-limiting examples of such SHP2 inhibitors that are known in the art, include: Chen et al. Mol Pharmacol. 2006, 70, 562; Sarver et al., J. Med. Chem. 2017, 62, 1793; Xie et al., J. Med. Chem. 2017, 60, 113734; and Igbe et al., Oncotarget, 2017, 8, 113734; and PCT applications: WO 2023282702, WO 2023280283, WO2023280237, WO 2023018155, WO 2023011513, WO 2022271966, WO 2022271964, WO 2022271911, WO 2022259157, WO 2022242767, WO 2022241975, WO 2022237676, WO 2022237367, WO 2022237178, WO 2022235822, WO 2022135568, WO 20222084008, WO 2022063190, WO 2022043865, WO 2022042331, WO 2022033430, WO 2022017444, WO 2022007869, WO 2021259077, WO 2021249449, WO 2021249057, WO 2021244659, WO 2021218755, WO 2021218752, WO 2021176072, WO 2021171261, WO 2021149817, WO 2021148010, WO 2021147879, WO 2021143823, WO 2021143701, WO 2021143680, WO 2021121397, WO 2021119525, WO 2021115286, WO 2021110796, WO 2021088945, WO 2021073439, WO 2021061706, WO 2021061515, WO 2021043077, WO 2021033153, WO 2021028362, WO 2021033153, WO 2021028362, WO 2021018287, WO 2020259679, WO 2020249079, WO 2020210384, WO 2020201991, WO 2020181283, WO 2020177653, WO 2020165734, WO 2020165733, WO 2020165732, WO 2020156243, WO 2020156242, WO 2020108590, WO 2020104635, WO 2020094104, WO 2020094018, WO 2020081848, WO 2020073949, WO 2020073945, WO 2020072656, WO 2020065453, WO 2020065452, WO 2020063760, WO 2020061103, WO 2020061101, WO 2020033828, WO 2020033286, WO 2020022323, WO 2019233810, WO 2019213318, WO 2019183367, WO 2019183364, WO 2019182960, WO 2019167000, WO 2019165073, WO 2019158019, WO 2019152454, WO 2019051469, WO 2019051084, WO 2018218133, WO 2018172984, WO 2018160731, WO 2018136265, WO 2018136264, WO 2018130928, WO 2018129402, WO 2018081091, WO 2018057884, WO 2018013597, WO 2017216706, WO 2017211303, WO 2017210134, WO 2017156397, WO 2017100279, WO 2017079723, WO 2017078499, WO 2016203406, WO 2016203405, WO 2016203404, WO 2016196591, WO 2016191328, WO 2015107495, WO 2015107494, WO 2015107493, WO 2014176488, WO 2014113584, CN 115677661, CN 115677660, CN 115611869, CN 115521305, CN 115490697, CN 115466273, CN 115394612, CN 115304613, CN 115304612, CN 115300513, CN 115197225, CN 114957162, CN 114920759, CN 114716448, CN 114671879, CN 114539223, CN 114524772, CN 114213417, CN 114195799, CN 114163457, CN 113896710, CN 113248521, CN 113248449, CN 113135924, CN 113024508, CN 112920131, CN 112823796, CN 112409334, CN 112402385, CN 112174935, 111848599, CN 111704611, CN 111393459, CN 111265529, CN 110143949, CN 108113848, US 11179397, US 11044675, US 11034705, US 11033547, US 11001561, US 10988466, US 10954243, US 10934302, or US 10858359, or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof, each of which is incorporated herein by reference.

[0188] In some embodiments, a SHP2 inhibitor binds in the active site. In some embodiments, a SHP2 inhibitor is a mixed-type irreversible inhibitor. In some embodiments, a SHP2 inhibitor binds an allosteric site e.g., a non-covalent allosteric inhibitor. In some embodiments, a SHP2 inhibitor is a covalent SHP2 inhibitor, such as an inhibitor that targets the cysteine residue (C333) that lies outside the phosphatase’s active site. In some embodiments a SHP2 inhibitor is a reversible inhibitor. In some embodiments, a SHP2 inhibitor is an irreversible inhibitor. In some embodiments, the SHP2 inhibitor is SHP099. In some embodiments, the SHP2 inhibitor is RMC-4550. In some embodiments, the SHP2 inhibitor is ERAS-601. In some embodiments, the SHP2 inhibitor is BBP-398. In some embodiments, the SHP2 inhibitor is TNO155, having the structure:

or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is RMC-4630, having the structure:

or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is JAB-3068, having the structure:

or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is JAB-3312. In some embodiments, the SHP2 inhibitor is the following compound:

or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof. In some embodiments, the SHP2 inhibitor is RLY-1971, having the structure:

or a pharmaceutically acceptable salt, solvate, isomer (e.g., stereoisomer), prodrug, or tautomer thereof.

[0189] In some embodiments, the additional therapeutic agent is selected from the group consisting of a MEK inhibitor, a HER2 inhibitor, a SHP2 inhibitor, a CDK4/6 inhibitor, an mTOR inhibitor, and a PD-L1 inhibitor. In some embodiments, the additional therapeutic agent is selected from the group consisting of a MEK inhibitor, a SHP2 inhibitor, and a PD-L1 inhibitor. See, e.g., Hallin et al., Cancer Discovery, DOI: 10.1158/2159-8290 (October 28, 2019) and Canon et al., Nature, 575:217 (2019). In some embodiments, a Ras inhibitor is used in combination with a MEK inhibitor and a S0S1 inhibitor of the present disclosure. In some embodiments, a Ras inhibitor is used in combination with a PD-L1 inhibitor and a S0S1 inhibitor of the present disclosure.

[0190] Proteasome inhibitors are known in the art and include, but are not limited to, carfilzomib (Kyprolis®), bortezomib (Velcade®), and oprozomib.

[0191] Immune therapies include, but are not limited to, monoclonal antibodies, immunomodulatory imides (IMiDs), GITR agonists, genetically engineered T-cells (e.g., CAR-T cells), bispecific antibodies (e.g., BiTEs), and anti-PD-1, anti-PD-Ll, anti-CTLA4, anti-LAGl, and anti-OX40 agents). Other immune therapies are known in the art.

[0192] Immunomodulatory agents (IMiDs) are a class of immunomodulatory drugs (drugs that adjust immune responses) containing an imide group. The IMiD class includes thalidomide and its analogues (lenalidomide, pomalidomide, and apremilast). [0193] Exemplary anti-PD-1 antibodies and methods for their use are described by Goldberg et al., Blood 2007, 110(1): 186-192; Thompson et al., Clin. Cancer Res. 2007, 13(6): 1757-1761; and WO06/121168 Al), as well as described elsewhere herein.

[0194] FGFR inhibitors are known in the art, such as pemigatinib and erdafitinib, including FGFR2 inhibitors and FGFR4 inhibitors. See, e.g., Cancers (Basel), 2021 Jun; 13(12) 2968.

[0195] BET inhibitors are known in the art, such as romidepsin, panobinostat and belinostat. See, e.g., British J. Cancer 124: 1478 (2021).

[0196] PRMT5i inhibitors are known in the art, such as PF-0693999, PJ-68 and MRTX1719. See, e.g., Biomed. Pharmacotherapy 144: 112252 (2021).

[0197] MAT2A inhibitors are known in the art, such as AG-270 and IDE397. See, e.g., Exp Opin Ther Patents (2022) DOI: 10.1080/13543776.2022.2119127.

[0198] GITR agonists include, but are not limited to, GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies), such as, a GITR fusion protein described in U.S. Pat. No. 6,111,090, U.S. Pat. No. 8,586,023, W02010/003118, and WO201 1/090754; or an anti-GITR antibody described, e.g., in U.S. Pat. No. 7,025,962, EP 1947183, U.S. Pat. No. 7,812,135, U.S. Pat. No. 8,388,967, U.S. Pat. No. 8,591,886, U.S. Pat. No. 7,618,632, EP 1866339, and WO2011/028683, WO2013/039954, WO05/007190, WO07/133822, WO05/055808, WO99/40196, WO01/03720, WO99/20758, WO06/083289, WO05/115451, and WO2011/051726.

[0199] Another example of a therapeutic agent that may be used in combination with the compounds of the disclosure is an anti-angiogenic agent. Anti -angiogenic agents are known in the art and are inclusive of, but not limited to, in vitro synthetically prepared chemical compositions, antibodies, antigen binding regions, radionuclides, and combinations and conjugates thereof. An anti-angiogenic agent can be an agonist, antagonist, allosteric modulator, toxin or, more generally, may act to inhibit or stimulate its target (e.g., receptor or enzyme activation or inhibition), and thereby promote cell death or arrest cell growth. In some embodiments, the one or more additional therapies include an anti -angiogenic agent. [0200] Anti-angiogenic agents can be MMP-2 (matrix-metalloproteinase 2) inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, and COX-II (cyclooxygenase 11) inhibitors. Non-limiting examples of anti-angiogenic agents include rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II inhibitors include alecoxib, valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors are described in WO96/33172, WO96/27583, WO98/07697, WO98/03516, WO98/34918, WO98/34915, WO98/33768, WO98/30566, W090/05719, WO99/52910, WO99/52889, WO99/29667, WO99007675, EP0606046, EP0780386, EP 1786785, EPl 181017, EP0818442, EP 1004578, and US20090012085, and U.S. Patent Nos. 5,863,949 and 5,861,510. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity inhibiting MMP-1. More preferred, are those that selectively inhibit MMP-2 or AMP-9 relative to the other matrix- metalloproteinases (i.e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP- 7, MMP- 8, MMP-10, MMP-11, MMP-12, and MMP-13). Some specific examples of MMP inhibitors are AG-3340, RO 32-3555, and RS 13-0830.

[0201] Further exemplary anti-angiogenic agents include KDR (kinase domain receptor) inhibitory agents (e.g., antibodies and antigen binding regions that specifically bind to the kinase domain receptor), anti-VEGF agents (e.g., antibodies or antigen binding regions that specifically bind VEGF (e.g., bevacizumab), or soluble VEGF receptors or a ligand binding region thereof) such as VEGF-TRAP™, and anti-VEGF receptor agents (e.g., antibodies or antigen binding regions that specifically bind thereto), VEGF inhibitors, EGFR inhibitory agents (e.g., antibodies or antigen binding regions that specifically bind thereto) such as Vectibix® (panitumumab), erlotinib (Tarceva®), anti-Angl and anti-Ang2 agents (e.g., antibodies or antigen binding regions specifically binding thereto or to their receptors, e.g., Tie2/Tek), and anti-Tie2 kinase inhibitory agents (e.g., antibodies or antigen binding regions that specifically bind thereto). Other anti-angiogenic agents include Campath, IL-8, B-FGF, Tek antagonists (US2003/0162712; US6,413,932), anti-TWEAK agents (e.g., specifically binding antibodies or antigen binding regions, or soluble TWEAK receptor antagonists; see US6, 727,225), ADAM distintegrin domain to antagonize the binding of integrin to its ligands (US 2002/0042368), specifically binding anti-eph receptor or anti-ephrin antibodies or antigen binding regions (U.S. Patent Nos. 5,981,245; 5,728,813; 5,969,110; 6,596,852; 6,232,447; 6,057,124 and patent family members thereof), and anti-PDGF-BB antagonists (e.g., specifically binding antibodies or antigen binding regions) as well as antibodies or antigen binding regions specifically binding to PDGF-BB ligands, and PDGFR kinase inhibitory agents (e.g., antibodies or antigen binding regions that specifically bind thereto). Additional anti -angiogenic agents include: SD-7784 (Pfizer, USA); cilengitide (Merck KGaA, Germany, EPO 0770622); pegaptanib octasodium, (Gilead Sciences, USA); Alphastatin, (BioActa, UK); M-PGA, (Celgene, USA, US 5712291); ilomastat, (Arriva, USA, US5892112); emaxanib, (Pfizer, USA, US 5792783); vatalanib, (Novartis, Switzerland); 2-methoxyestradiol (EntreMed, USA); TLC ELL- 12 (Elan, Ireland); anecortave acetate (Alcon, USA); alpha-D148 Mab (Amgen, USA); CEP-7055 (Cephalon, USA); anti-Vn Mab (Crucell, Netherlands), DACantiangiogenic (ConjuChem, Canada); Angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, EP 0970070); ARGENT technology (Ariad, USA); YIGSR-Stealth (Johnson & Johnson, USA); fibrinogen-E fragment (BioActa, UK); angiogenic inhibitor (Trigen, UK); TBC-1635 (Encysive Pharmaceuticals, USA); SC-236 (Pfizer, USA); ABT-567 (Abbott, USA); Metastatin (EntreMed, USA); maspin (Sosei, Japan); 2-methoxyestradiol (Oncology Sciences Corporation, USA); ER-68203-00 (IV AX, USA); BeneFin (Lane Labs, USA); Tz-93 (Tsumura, Japan); TAN-1120 (Takeda, Japan); FR-111142 (Fujisawa, Japan, JP 02233610); platelet factor 4 (RepliGen, USA, EP 407122); vascular endothelial growth factor antagonist (Borean, Denmark); bevacizumab (pINN) (Genentech, USA); angiogenic inhibitors (SUGEN, USA); XL 784 (Exelixis, USA); XL 647 (Exelixis, USA); MAb, alpha5beta3 integrin, second generation (Applied Molecular Evolution, USA and Medlmmune, USA); enzastaurin hydrochloride (Lilly, USA); CEP 7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC 1 (Genoa Institute of Cancer Research, Italy); rBPI 21 and BPI-derived anti angiogenic (XOMA, USA); PI 88 (Progen, Australia); cilengitide (Merck KGaA, German; Munich Technical University, Germany, Scripps Clinic and Research Foundation, USA); AVE 8062 (Ajinomoto, Japan); AS 1404 (Cancer Research Laboratory, New Zealand); SG 292, (Telios, USA); Endostatin (Boston Children’s Hospital, USA); ATN 161 (Attenuon, USA); 2-methoxyestradiol (Boston Children’s Hospital, USA); ZD 6474, (AstraZeneca, UK); ZD 6126, (Angiogene Pharmaceuticals, UK); PPI 2458, (Praecis, USA); AZD 9935, (AstraZeneca, UK); AZD 2171, (AstraZeneca, UK); vatalanib (pINN), (Novartis, Switzerland and Schering AG, Germany); tissue factor pathway inhibitors, (EntreMed, USA); pegaptanib (Pinn), (Gilead Sciences, USA); xanthorrhizol, (Yonsei University, South Korea); vaccine, gene-based, VEGF-2, (Scripps Clinic and Research Foundation, USA); SPV5.2, (Supratek, Canada); SDX 103, (University of California at San Diego, USA); PX 478, (ProlX, USA); METASTATIN, (EntreMed, USA); troponin I, (Harvard University, USA); SU 6668, (SUGEN, USA); OXI 4503, (OXiGENE, USA); o-guanidines, (Dimensional Pharmaceuticals, USA); motuporamine C, (British Columbia University, Canada); CDP 791, (Celltech Group, UK); atiprimod (pINN), (GlaxoSmithKline, UK); E 7820, (Eisai, Japan); CYC 381, (Harvard University, USA); AE 941, (Aetema, Canada); vaccine, angiogenic, (EntreMed, USA); urokinase plasminogen activator inhibitor, (Dendreon, USA); oglufanide (pINN), (Melmotte, USA); HIF-lalfa inhibitors, (Xenova, UK); CEP 5214, (Cephalon, USA); BAY RES 2622, (Bayer, Germany); Angiocidin, (InKine, USA); A6, (Angstrom, USA); KR 31372, (Korea Research Institute of Chemical Technology, South Korea); GW 2286, (GlaxoSmithKline, UK); EHT 0101, (ExonHit, France); CP 868596, (Pfizer, USA); CP 564959, (OSI, USA); CP 547632, (Pfizer, USA); 786034, (GlaxoSmithKline, UK); KRN 633, (Kirin Brewery, Japan); drug delivery system, intraocular, 2-methoxyestradiol; anginex (Maastricht University, Netherlands, and Minnesota University, USA); ABT 510 (Abbott, USA); AAL 993 (Novartis, Switzerland); VEGI (ProteomTech, USA); tumor necrosis factor-alpha inhibitors; SU 11248 (Pfizer, USA and SUGEN USA); ABT 518, (Abbott, USA); YH16 (Yantai Rongchang, China); S-3APG (Boston Children’s Hospital, USA and EntreMed, USA); MAb, KDR (ImClone Systems, USA); MAb, alpha5 beta (Protein Design, USA); KDR kinase inhibitor (Celltech Group, UK, and Johnson & Johnson, USA); GFB 116 (South Florida University, USA and Yale University, USA); CS 706 (Sankyo, Japan); combretastatin A4 prodrug (Arizona State University, USA); chondroitinase AC (IBEX, Canada); BAY RES 2690 (Bayer, Germany); AGM 1470 (Harvard University, USA, Takeda, Japan, and TAP, USA); AG 13925 (Agouron, USA); Tetrathiomolybdate (University of Michigan, USA); GCS 100 (Wayne State University, USA) CV 247 (Ivy Medical, UK); CKD 732 (Chong Kun Dang, South Korea); irsogladine, (Nippon Shinyaku, Japan); RG 13577 (Aventis, France); WX 360 (Wilex, Germany); squalamine, (Genaera, USA); RPI 4610 (Sima, USA); heparanase inhibitors (InSight, Israel); KL 3106 (Kolon, South Korea); Honokiol (Emory University, USA); ZK CDK (Schering AG, Germany); ZK Angio (Schering AG, Germany); ZK 229561 (Novartis, Switzerland, and Schering AG, Germany); XMP 300 (XOMA, USA); VGA 1102 (Taisho, Japan); VE-cadherin-2 antagonists(ImClone Systems, USA); Vasostatin (National Institutes of Health, USA); Flk-1 (ImClone Systems, USA); TZ 93 (Tsumura, Japan); TumStatin (Beth Israel Hospital, USA); truncated soluble FLT 1 (vascular endothelial growth factor receptor 1) (Merck & Co, USA); Tie-2 ligands (Regen eron, USA); and thrombospondin 1 inhibitor (Allegheny Health, Education and Research Foundation, USA).

[0202] Further examples of therapeutic agents that may be used in combination with compounds of the disclosure include agents (e.g., antibodies, antigen binding regions, or soluble receptors) that specifically bind and inhibit the activity of growth factors, such as antagonists of hepatocyte growth factor (HGF, also known as Scatter Factor), and antibodies or antigen binding regions that specifically bind its receptor, c-Met. Such agents are known in the art.

[0203] Another example of a therapeutic agent that may be used in combination with compounds of the disclosure is an autophagy inhibitor. Autophagy inhibitors are known in the art and include, but are not limited to chloroquine, 3- methyladenine, hydroxychloroquine (Plaquenil™), bafilomycin A1, 5-amino-4-imidazole carboxamide riboside (AICAR), okadaic acid, autophagy-suppressive algal toxins which inhibit protein phosphatases of type 2A or type 1, analogues of cAMP, and drugs which elevate cAMP levels such as adenosine, LY204002, N6-mercaptopurine riboside, and vinblastine. In addition, antisense or siRNA that inhibits expression of proteins including but not limited to ATG5 (which are implicated in autophagy), may also be used. In some embodiments, the one or more additional therapies include an autophagy inhibitor.

[0204] Another example of a therapeutic agent that may be used in combination with compounds of the disclosure is an anti -neoplastic agent, which are known in the art. In some embodiments, the one or more additional therapies include an anti-neoplastic agent. Non-limiting examples of anti -neoplastic agents include acemannan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, altretamine, amifostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ancer, ancestim, arglabin, arsenic trioxide, BAM-002 (Novelos), bexarotene, bicalutamide, broxuridine, capecitabine, celmoleukin, cetrorelix, cladribine, clotrimazole, cytarabine ocfosfate, DA 3030 (Dong- A), daclizumab, denileukin diftitox, deslorelin, dexrazoxane, dilazep, docetaxel, docosanol, doxercalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT diclofenac, interferon alfa, daunorubicin, doxorubicin, tretinoin, edelfosine, edrecolomab, eflornithine, emitefur, epirubicin, epoetin beta, etoposide phosphate, exemestane, exisulind, fadrozole, filgrastim, finasteride, fludarabine phosphate, formestane, fotemustine, gallium nitrate, gemcitabine, gemtuzumab zogamicin, gimeracil/oteracil/tegafur combination, glycopine, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha fetoprotein, ibandronic acid, idarubicin, (imiquimod, interferon alfa, interferon alfa, natural, interferon alfa-2, interferon alfa-2a, interferon alfa-2b, interferon alfa-Nl, interferon alfa-n3, interferon alfacon-1, interferon alpha, natural, interferon beta, interferon beta-la, interferon beta-lb, interferon gamma, natural interferon gamma- 1a, interferon gamma-lb, interleukin-1 beta, iobenguane, irinotecan, irsogladine, lanreotide, LC 9018 (Yakult), leflunomide, lenograstim, lentinan sulfate, letrozole, leukocyte alpha interferon, leuprorelin, levamisole + fluorouracil, liarozole, lobaplatin, lonidamine, lovastatin, masoprocol, melarsoprol, metoclopramide, mifepristone, miltefosine, mirimostim, mismatched double stranded RNA, mitoguazone, mitolactol, mitoxantrone, molgramostim, nafarelin, naloxone + pentazocine, nartograstim, nedaplatin, nilutamide, noscapine, novel erythropoiesis stimulating protein, NSC 631570 octreotide, oprelvekin, osaterone, oxaliplatin, paclitaxel, pamidronic acid, pegaspargase, peginterferon alfa-2b, pentosan polysulfate sodium, pentostatin, picibanil, pirarubicin, rabbit antithymocyte polyclonal antibody, polyethylene glycol interferon alfa-2a, porfimer sodium, raloxifene, raltitrexed, rasburi embodiment, rhenium Re 186 etidronate, RII retinamide, rituximab, romurtide, samarium (153 Sm) lexidronam, sargramostim, sizofiran, sobuzoxane, sonermin, strontium-89 chloride, suramin, tasonermin, tazarotene, tegafur, temoporfm, temozolomide, teniposide, tetrachlorodecaoxide, thalidomide, thymalfasin, thyrotropin alfa, topotecan, toremifene, tositumomab-iodine 131, trastuzumab, treosulfan, tretinoin, trilostane, trimetrexate, triptorelin, tumor necrosis factor alpha, natural, ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin, vinorelbine, virulizin, zinostatin stimalamer, or zoledronic acid; abarelix; AE 941 (Aeterna), ambamustine, antisense oligonucleotide, bcl-2 (Genta), APC 8015 (Dendreon), decitabine, dexaminoglutethimide, diaziquone, EL 532 (Elan), EM 800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, galocitabine, gastrin 17 immunogen, HLA-B7 gene therapy (Vical), granulocyte macrophage colony stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM 862 (Cytran), interleukin-2, iproxifene, LDI 200 (Milkhaus), leridistim, lintuzumab, CA 125 MAb (Biomira), cancer MAb (Japan Pharmaceutical Development), HER-2 and Fc MAb (Medarex), idiotypic 105AD7 MAb (CRC Technology), idiotypic CEA MAb (Trilex), LYM-1 -iodine 131 MAb (Techni clone), polymorphic epithelial mucin-yttrium 90 MAb (Antisoma), marimastat, menogaril, mitumomab, motexafin gadolinium, MX 6 (Galderma), nelarabine, nolatrexed, P 30 protein, pegvisomant, pemetrexed, porfiromycin, prinomastat, RL 0903 (Shire), rubitecan, satraplatin, sodium phenyl acetate, sparfosic acid, SRL 172 (SR Pharma), SU 5416 (SUGEN), TA 077 (Tanabe), tetrathiomolybdate, thaliblastine, thrombopoietin, tin ethyl etiopurpurin, tirapazamine, cancer vaccine (Biomira), melanoma vaccine (New York University), melanoma vaccine (Sloan Kettering Institute), melanoma oncolysate vaccine (New York Medical College), viral melanoma cell lysates vaccine (Royal Newcastle Hospital), or valspodar.

[0205] Additional examples of therapeutic agents that may be used in combination with compounds of the disclosure include ipilimumab (Yervoy®); tremelimumab; galiximab; nivolumab, also known as BMS-936558 (Opdivo®); pembrolizumab (Keytruda®); avelumab (Bavencio®); AMP224; BMS-936559; MPDL3280A, also known as RG7446; MEDI-570; AMG557; MGA271; IMP321; BMS- 663513; PF-05082566; CDX-1127; anti-OX40 (Providence Health Services); huMAbOX40L; atacicept; CP-870893; lucatumumab; dacetuzumab; muromonab-CD3; ipilumumab; MEDI4736 (Imfinzi®); MSB0010718C; AMP 224; adalimumab (Humira®); ado-trastuzumab emtansine (Kadcyla®); aflibercept (Eylea®); alemtuzumab (Campath®); basiliximab (Simulect®); belimumab (Benlysta®); basiliximab (Simulect®); belimumab (Benlysta®); brentuximab vedotin (Adcetris®); canakinumab (IL aris®); certolizumab pegol (Cimzia®); daclizumab (Zenapax®); daratumumab (Darzalex®); denosumab (Prolia®); eculizumab (Soliris®); efalizumab (Raptiva®); gemtuzumab ozogamicin (Mylotarg®); golimumab (Simponi®); ibritumomab tiuxetan (Zevalin®); infliximab (Remicade®); motavizumab (Numax®); natalizumab (Tysabri®); obinutuzumab (Gazyva®); ofatumumab (Arzerra®); omalizumab (Xolair®); palivizumab (Synagis®); pertuzumab (Perjeta®); pertuzumab (Perjeta®); ranibizumab (Lucentis®); raxibacumab (Abthrax®); tocilizumab (Actemra®); tositumomab; tositumomab-i-131; tositumomab and tositumomab-i-131 (Bexxar®); ustekinumab (Stelara®); AMG 102; AMG 386; AMG 479; AMG 655; AMG 706; AMG 745; and AMG 951.

[0206] In some embodiments, an additional compound used in combination therapy with a compound of the present disclosure is selected from the group consisting of a CDK4/6 inhibitor (e.g., abemaciclib, palbociclib, or ribociclib), a KRAS:GDP G12C inhibitor (e.g., AMG 510, MRTX1257, MRTX849) or other mutant Ras:GDP inhibitor, a KRAS:GTP G12C inhibitor or other mutant Ras:GTP inhibitor (e.g., a Ras inhibitor described in WO 2020/132597, WO 2021/091956, WO 2021/091982, WO 2021/091967, WO 2022/060836; RMC-6291, RMC-6236, RMC-9805 or RMC-8839), a MEK inhibitor (e.g., refametinib, selumetinib, trametinib, or cobimetinib), a SHP2 inhibitor (e.g., TNO155, RMC-4630), an ERK inhibitor, and an RTK inhibitor (e.g., an EGFR inhibitor). In some embodiments, a S0S1 inhibitor may be used in combination with a Ras inhibitor, a SHP2 inhibitor, or a MEK inhibitor. In some embodiments, a combination therapy includes a S0S1 inhibitor, a RAS inhibitor and a MEK inhibitor.

[0207] In some embodiments, an additional compound used in combination therapy with a compound of the present disclosure is selected from the group consisting of ABT-737, AT-7519, carfilzomib, cobimetinib, danusertib, dasatinib, doxorubicin, GSK- 343, JQ1, MLN-7243, NVP-ADW742, paclitaxel, palbociclib and volasertib. In some embodiments, an additional compound used in combination therapy with a compound of the present disclosure is selected from the group consisting of neratinib, acetinib and reversine.

[0208] The compounds described herein can be used in combination with the agents disclosed herein or other suitable agents, depending on the condition being treated. Hence, in some embodiments the one or more compounds of the disclosure will be co- administered with other therapies as described herein. When used in combination therapy, the compounds described herein may be administered with the second agent simultaneously or separately. This administration in combination can include simultaneous administration of the two agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, a compound described herein and any of the agents described herein can be formulated together in the same dosage form and administered simultaneously. Alternatively, a compound of the disclosure and any of the therapies described herein can be simultaneously administered, wherein both the agents are present in separate formulations. In another alternative, a compound of the present disclosure can be administered and followed by any of the therapies described herein, or vice versa. In some embodiments of the separate administration protocol, a compound of the disclosure and any of the therapies described herein are administered a few minutes apart, or a few hours apart, or a few days apart.

[0209] In some embodiments, a combination therapeutic regimen employs two therapeutic agents, one compound of the present disclosure and a second selected from the therapeutic agents described herein. In some embodiments, a combination therapeutic regimen employs three therapeutic agents, one compound of the present disclosure and two selected from the therapeutic agents described herein. In some embodiments, a combination therapeutic regimen employs four or more therapeutic agents, one compound of the present disclosure and three selected from the therapeutic agents described herein.

[0210] In some embodiments of any of the methods described herein, the first therapy (e.g., a compound of the disclosure) and one or more additional therapies are administered simultaneously or sequentially, in either order. The first therapeutic agent may be administered immediately, up to 1 hour, up to 2 hours, up to 3 hours, up to 4 hours, up to 5 hours, up to 6 hours, up to 7 hours, up to, 8 hours, up to 9 hours, up to 10 hours, up to 11 hours, up to 12 hours, up to 13 hours, 14 hours, up to hours 16, up to 17 hours, up 18 hours, up to 19 hours up to 20 hours, up to 21 hours, up to 22 hours, up to 23 hours, up to 24 hours, or up to 1-7, 1-14, 1-21 or 1-30 days before or after the one or more additional therapies.

[0211] The disclosure also features kits including (a) a pharmaceutical composition including an agent (e.g., a compound of the disclosure) described herein, and (b) a package insert with instructions to perform any of the methods described herein. In some embodiments, the kit includes (a) a pharmaceutical composition including an agent (e.g., a compound of the disclosure) described herein, (b) one or more additional therapies (e.g., non-drug treatment or therapeutic agent), and (c) a package insert with instructions to perform any of the methods described herein.

[0212] As one aspect of the present disclosure contemplates the treatment of the disease or symptoms associated therewith with a combination of pharmaceutically active compounds that may be administered separately, the disclosure further relates to combining separate pharmaceutical compositions in kit form. The kit may comprise two separate pharmaceutical compositions: a compound of the present disclosure, and one or more additional therapies. The kit may comprise a container for containing the separate compositions such as a divided bottle or a divided foil packet. Additional examples of containers include syringes, boxes, and bags. In some embodiments, the kit may comprise directions for the use of the separate components. The kit form is particularly advantageous when the separate components are preferably administered in different dosage forms (e.g., oral and parenteral), are administered at different dosage intervals, or when titration of the individual components of the combination is desired by the prescribing health care professional.

Exemplary Embodiments

[0213] The present disclosure is further illustrated by the following non-limiting embodiments.

[0214] Embodiment 1 is a compound having the structure of Formula (I),

or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein:

R₁ is selected from the group consisting of optionally substituted 6-membered aryl and optionally substituted 5-6 membered heteroaryl;

R₂ is selected from the group consisting of H and optionally substituted C_1-6 alkyl;

R₃ is — OR_3a, and further wherein R_3a is optionally substituted C_1-3 alkyl; and R₄ is selected from the group consisting of H, C_1-6 alkyl, 3-14 membered cycloalkyl, 3-14 membered cycloalkenyl, 3-14 membered heterocyclyl, 6-10 membered aryl, and 5-10 membered heteroaryl; wherein each C_1-6 alkyl, 3-14 membered cycloalkyl, 3-14 membered cycloalkenyl, 3-14 membered heterocyclyl, 6-10 membered aryl, and 5-10 membered heteroaryl is optionally substituted with C_1-6 alkyl, -R_4a, -OR_4a, -O-C_1-6 alkyl-R_4a, =O, halogen, -C(O)R_4a, -C(OO)R_4a, -C(O)NR_4bR_4c, -NR_4bC(O)R_4c, -CN, =NR_4a, -NR_4bR_4c, - SO₂R_4a,, 3-6 membered cycloalkyl optionally substituted with R_4a, 3-7 membered heterocyclyl optionally substituted with R_4a, 6-10 membered aryl optionally substituted with R_4a, or 5-10 membered heteroaryl optionally substituted with R_4a; wherein R_4a is H, C_1-6 alkyl, C_1-6 haloalkyl, -C(O)R_4b, -C(O)NR_4bR_4c, =O, 3-6 membered cycloalkyl, 6-10 membered aryl optionally substituted with -OR_4b, -

CN, =N-3-6 membered cycloalkyl, 3-7 membered heterocyclyl, -(CH₂)_rOCH₃, or - (CH₂)_rOH, wherein r is 1, 2, or 3; wherein each R_4b is independently H, C_1-6 alkyl; and wherein each R_4c is independently H or C_1-6 alkyl. [0215] Embodiment 2 is a compound of embodiment 1, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₁ is optionally substituted 6-membered aryl.

[0216] Embodiment 3 is a compound of embodiment 1, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₁ is

[0217] Embodiment 4 is a compound of embodiment 1, having the structure of

Formula (II),

or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₂, R₃, and R₄ are as defined in embodiment 1, wherein R₅, R₆, R₇, R₈, and R₉ are independently selected from the group consisting of H, C_1-6 alkyl, halogen, and -NH₂, wherein each C_1-6 alkyl is optionally substituted with halogen.

[0218] Embodiment 5 is a compound of any one of embodiments 1 through 4, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₂ is H.

[0219] Embodiment 6 is a compound of any one of embodiments 1 through 4, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₂ is optionally substituted C_1-3 alkyl, wherein the optional substituent is one or more fluoro.

[0220] Embodiment 7 is a compound of any one of embodiments 1 through 4, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₂ is -CEE.

[0221] Embodiment 8 is a compound of any one of embodiments 1 through 7, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₃ is -OR_3a, and further wherein R_3a is C_1-3 alkyl.

[0222] Embodiment 9 is a compound of any one of embodiments 1 through 7, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₃ is -OCH₃.

[0223] Embodiment 10 is a compound of any one of embodiments 1 through 9, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₄ is optionally substituted 3-14 membered heterocyclyl.

[0224] Embodiment 11 is a compound of any one of embodiments 1 through 9, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₄ is 3-14 membered heterocyclyl optionally substituted with halogen, C_1- ₆ alkyl, -CN, -OR_4a, =O, -C(O)R_4a, -NR_4bR_4c, or =NR_4a; wherein: each R_4a is independently H, C_1-6 alkyl, C_3-8 cycloalkyl, or -CN; each R_4b is independently H or C_1-6 alkyl; and each R_4c is independently H or C_1-6 alkyl.

[0225] Embodiment 12 is a compound of any one of embodiments 1 through 9, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₄ is selected from the group consisting of optionally substituted piperidinyl, optionally substituted piperazinyl, optionally substituted tetrahydropyridinyl, optionally substituted dihydrothiopyranyl, and optionally substituted tetrahydrothiopyranyl.

[0226] Embodiment 13 is a compound of any one of embodiments 1 through 9, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₄ is selected from the group consisting of

[0227] Embodiment 14 is a compound of Embodiment 1, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, selected from the group consisting of: N-[(1R )-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(1-methyl-4- piperidyl)pyrido[3,4-d]pyrimidin-4-amine; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(4- methylpiperazin-1-yl)pyrido[3,4-J]pyrimidin-4-amine; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(l-methyl-3,6- dihydro-27/-pyridin-4-yl)pyrido[3,4-d/]pyrimidin-4-amine;

(R) -4-(4-((l-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8- methoxypyrido[3,4-d]pyrimidin-6-yl)-3,6-dihydro-2H-thiopyran 1,1 -dioxide; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(l,l-dioxothian-4-yl)-8- methoxy-pyrido[3,4-d]pyrimidin-4-amine;

(R)-4 -(4-((1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8- methoxypyrido[3,4-d]pyrimidin-6-yl)-4-fluorotetrahydro-2H-thiopyran 1,1 -dioxide; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(4-methoxy- l,l-dioxo-thian-4-yl)pyrido[3,4-d]pyrimidin-4-amine;

4-[4-[[(1R )-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy- pyrido[3 ,4-d]pyrimidin-6-yl]- 1 , 1 -dioxo-thiane-4-carbonitrile;

6-(4-amino-l,l-dioxo-thian-4-yl)-N-[(1R )-1- [3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine; N-[(1R )-1- -[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(1,1-dioxo-3,6-dihydro-2H- thiopyran-4-yl)-8-methoxy-2-methyl-pyrido[3,4-J]pyrimidin-4-amine; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(4-methyl-1,1- dioxo-thian-4-yl)pyrido[3,4-d]pyrimidin-4-amine;

1-[4-[4-[[(1R )-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy- pyrido[3,4-d]pyrimidin-6-yl]-l-piperidyl]ethanone; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(4-fluoro-l-methyl-4- piperidyl)-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine; N-[(1R )-1- [3 -(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-( 1 -imino-1-oxo-3,6- dihydro-2H -thiopyran-4-yl)-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(l- methylimino-1 -oxo-3, 6-dihydro-2H -thiopyran-4-yl)pyrido[3,4-J]pyrimidin-4-amine; [4-[4-[[(1R )-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy- pyrido[3,4-d/]pyrimidin-6-yl]-1 -oxo-3, 6-dihydro-2H/-thiopyran- 1 -ylidene]cyanamide;

(R) -N-(1-(3-(difluoromethyl)-2-fluorophenyl)ethyl)-8-methoxy-6-(4- methoxypiperidin-4-yl)pyrido[3,4-d]pyrimidin-4-amine; and

1-[4-[4-[[(1R )-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy- pyrido[3,4-J]pyrimidin-6-yl]-4-methoxy-l-piperidyl]ethanone.

[0228] Embodiment 15 is a pharmaceutical composition comprising a compound of any one of embodiments 1-14, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, and a pharmaceutically acceptable carrier.

[0229] Embodiment 16 is a method of inhibiting S0S1 in a subject, comprising administering to the subject: a compound of any one of embodiments 1-14, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof; or a pharmaceutical composition of embodiment 15, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof.

[0230] Embodiment 17 is a method of inhibiting the interaction of S0S1 and a RAS-family protein in a cell or inhibiting the interaction of S0S1 and RAC1 in a cell, comprising administering to the cell: a compound of any one of embodiments 1-14, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof; or a pharmaceutical composition of embodiment 15, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof.

[0231] Embodiment 18 is a method of treating or preventing a disease, wherein treating or preventing the disease is characterized by inhibition of the interaction of S0S1 and a RAS-family protein or by inhibition of the interaction of S0S1 and RAC1, the method comprising administering to a subject in need thereof an effective amount of: a compound of any of embodiments 1-14, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof; or a pharmaceutical composition of embodiment 15, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof. [0232] Embodiment 19 is a method of treating or preventing cancer in a subject in need thereof, comprising administering to the subject an effective amount of: a compound of any of embodiments 1-14, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof; or a pharmaceutical composition of embodiment 15, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof.

[0233] Embodiment 20 is a method of embodiment 18 or embodiment 19, wherein the disease or cancer is selected from the group consisting of pancreatic cancer, lung cancer, colorectal cancer, hematological cancer, cholangiocarcinoma, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, bladder cancer, urothelial cancer, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular cancer, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer and sarcomas.

[0234] Embodiment 21 is a method of embodiment 19 or embodiment 20, wherein the cancer comprises a Ras^MUT or an NF1^LOF mutation.

[0235] Embodiment 22 is a method of embodiment 18, wherein the disease is a RASopathy.

[0236] Embodiment 23 is a method of embodiment 22, wherein the RASopathy is selected from the group consisting of Neurofibromatosis type 1 (NF1), Noonan Syndrome (NS), Noonan Syndrome with Multiple Lentigines (NSML), Capillary Malformation-Arteriovenous Malformation Syndrome (CM-AVM), Costello Syndrome (CS), Cardio-Facio-Cutaneous Syndrome (CFC), Legius Syndrome, and Hereditary gingival fibromatosis.

EXAMPLES

[0237] The disclosure is further illustrated by the following examples and synthesis examples, which are not to be construed as limiting this disclosure in scope or spirit to the specific procedures herein described. It is to be understood that the examples are provided to illustrate certain embodiments and that no limitation to the scope of the disclosure is intended thereby. It is to be further understood that resort may be had to various other embodiments, modifications, and equivalents thereof which may suggest themselves to those skilled in the art without departing from the spirit of the present disclosure or scope of the appended claims. [0238] Definitions used in the following examples and elsewhere herein are:

AcOH Acetic acid

BOP (benzotri azol4~yloxytris(dimethylamino)phosphonium hexafluorophosphate) reagent n-BuLi n-butyl lithium n-BuOH n -butanol

CH₃CN Acetonitrile

CH₃ONa, MeONa Sodium Methoxide

CS₂CO₃ Cesium Carbonate

DAST Diethylaminosulfur trifluoride

DBU Di azabi cy cl oundecene

DCM Dichloromethane

DIPEA or DIEA N,N-diisopropylethylamine

DMAP 4-Dimethylaminopyridine

DMF N,N-Dimethylformamide

Et₃N Triethylamine

EtOH Ethanol

EtOAc Ethyl acetate

H₂O Water

HPLC High performance liquid chromatography

HC1 Hydrochloric acid

K₂CO₃ Potassium Carbonate

KOH Potassium Hydroxide

LCMS Liquid chromatography/mass spectrometry

MeSO₃H Methanesulfonic acid

Me₂SO₄ Dimethyl sulfate

MeOH Methanol

NaCN Sodium Cyanide

Na₂CO₃ Sodium carbonate

NaHCO₃ Sodium Bicarbonate

NaOH Sodium Hydroxide

Na₂SO₄ Sodium Sulfate

Na₂S2O₃ Sodium thiosulfate

NaO^tBu Sodium tert-butoxide

NH₃ Ammonia

NH₂CO₂NH₄ Ammonia Carbamic Acid

NBS N-bromosuccinimide

NIS N-iodosuccinimide

Oxone Potassium Peroxymonosulfate Pd/C Palladium on carbon Pd(dppf)Cl₂ [1,1 '-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) PhI(OAc)₂ [acetoxy (pheny 1)-λ3 -i odany 1 ] acetate rt Room temperature SOCl₂ Thionyl Chloride TEA Triethylamine tert-AmOH Tert-amyl alchol TFA Trifluroacetic acid THF Tetrahydrofuran TMS Trimethyl silyl

Example 1. Synthesis of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8- methoxy-6-(l-methyl-4-piperidyl)pyrido [3,4-d] pyrimidin-4-amine formate salt

[0239] Step 1.

[0240] A mixture of methyl 3-amino-2,6-dichloro-pyridine-4-carboxylate (6.0 g, 27 mmol) in CH₃ONa, 30% solution in MeOH (30 mL) was heated to 80°C and stirred for

5 h. The mixture was concentrated under reduced pressure to give methyl 3-amino-6- chloro-2-m ethoxy -pyridine-4-carboxylate (4 g, crude) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₈H₉ClN₂O₃ 216.0; found 217.0. [0241] Step 2.

[0242] To a mixture of methyl 3-amino-6-chloro-2-methoxy-pyridine-4- carboxylate (4 g, 18.5 mmol) in EtOH (80 mL) and H2O (20 mL) was added KOH (2.07 g, 37.0 mmol). The mixture was stirred at rt for 30 min, then heated to 90°C for 2 h. The mixture was concentrated under reduced pressure, then diluted with H₂O (100 mL) and extracted with EtOAc (100 mL x 2). The aqueous layer was adjusted to pH ~2 with 2M HC1, then extracted with EtOAc (100 mL x 3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure to give 3-amino-6-chloro-2-methoxy-pyridine-4-carboxylic acid (5 g, crude) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₇H₇CIN₂O₃ 202.0; found 203.0.

[0243] Step 3.

[0244] A mixture of 3-amino-6-chloro-2-methoxy-pyridine-4-carboxylic acid (1.0 g, 4.9 mmol) and formimidamide acetic acid salt (1.03 g, 9.9 mmol) in 2-methoxy ethanol (14 mL) was heated to 170°C under microwave irradiation and stirred for 2 h. A total of 6 batches were subjected to the reaction conditions, then combined, filtered, the filtrate concentrated under reduced and the residue was purified by preparative-HPLC to give 6- chloro-8-methoxy-pyrido[3,4-d ]pyrimidin-4-ol (500 mg, 10% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₈H₆CiN₃O₂ 211.0; found 212.0; ¹H NMR (400 MHz, DMSO-d₆) δ 8.15 (s, 1H), 7.41 (s, 1H), 3.97 (s, 3H). [Note: Partial decomposition during the purification by preparative-HPLC],

[0245] Step 4.

[0246] A mixture of 6-chloro-8-methoxy-pyrido[3,4-d]pyrimidin-4-ol (150 mg, 0.8 mol), 1-methyl-4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H- pyridine (475 mg, 2.1 mmol), CS2CO3 (693 mg, 2.1 mmol), Pd(dppf)C12 (104 mg, 0.14 mmol) in 1,4-di oxane (3 mL) and H2O (0.6 mL) was degassed with N2 (x3), then heated to 100°C and stirred for 5 h. The mixture was filtered, the filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give 8-methoxy-6- (1-methyl-3,6-dihydro-2H -pyridin-4-yl)pyrido[3,4-J]pyrimidin-4-ol (80 mg, 36% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₁₄H₁₆N₄O₂ 272.1; found 273.1; 'H NMR (400 MHz, CD₃OD) δ 8.12 (s, 1H), 7.66 (s, 1H), 6.95 (s, 1H), 4.13 (s, 3H), 3.84 - 3.77 (m, 2H), 3.38 - 3.33 (m, 3H), 2.98 - 2.92 (m, 2H), 2.86 (s, 3H).

[0247] Step 5.

[0248] To a mixture of 8-methoxy-6-(l-methyl-3,6-dihydro-2H -pyridin-4- yl)pyrido[3,4-d]pyrimidin-4-ol (60 mg, 0.22 mmol) in MeOH (10 mL) was added 10% Pd/C (10 mg). The mixture was stirred under an atmosphere of H₂ (50 psi) at 40°C for 2 h, then filtered and the filtrate was concentrated under reduced pressure to give 8-methoxy-6- (1-methyl-4-piperidyl)pyrido[3,4-J]pyrimidin-4-ol (60 mg, crude) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₁₄H₁₈N₄O₂ 274.1; found 275.2.

[0249] Step 6.

[0250] To a mixture of 8-methoxy-6-(l-methyl-3,6-dihydro-2H -pyridin-4- yl)pyrido[3,4-d] pyrimidin-4-ol (25 mg, 0.09 mmol) and (1R) -1-[3-(difluoromethyl)-2- fhioro-phenyl]ethanamine (26 mg, 0.14 mmol) in DMF (0.5 mL) was added DBU (42 mg, 42 mL, 0.28 mmol) and BOP (61 mg, 0.14 mmol). The mixture was stirred at rt for 2 h, then diluted with H2O (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with aqueous NaHCO₃ (10 mL x 2) and brine (10 mL), then dried over Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give /V-[(1R)-l-[3-(difluoromethyl)-2- fluoro-phenyl]ethyl]-8-methoxy-6-(l-methyl-4-piperidyl)pyrido[3,4-J]pyrimidin-4-amine formate salt (19 mg, 23% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₂₃H₂₆F₃N₅O 445.2; found 446.2; 'H NMR (400 MHz, CD3OD) δ 8.43 (s, 1H), 7.64 (s, 1H), 7.58 (t, J= 7.3 Hz, 1H), 7.49 (t, J= 7.1 Hz, 1H), 7.23 (t, J= 7.8 Hz, 1H), 7.00 (t, J= 54.8 Hz, 1H), 5.81 (q, J= 6.8 Hz, 1H), 4.11 (s, 3H), 3.48 - 3.40 (m, 2H), 3.05 - 2.83 (m, 3H), 2.74 (s, 3H), 2.25 - 2.12 (m, 4H), 1.69 (d, J= 7.1 Hz, 3H). Example 2. Synthesis of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8- methoxy-6-(4-methylpiperazin-l-yl)pyrido [3,4- d] pyrimidin-4-amine formate salt

[0251] Step l.

[0252] A mixture of 6-chloro-8-methoxy-pyrido[3,4-d]pyrimidin-4-ol (200 mg, 0.95 mmol), 1 -methylpiperazine (189 mg, 210 mL, 1.89 mmol), [2-(2- aminophenyl)phenyl]palladium;dicyclohexyl-[2-(2,4,6- triisopropylphenyl)phenyl]phosphane methanesulfonate (96 mg, 1.1 mmol) and NaO'Bu (182 mg, 1.89 mmol) in tert-AmOH (8 mL) was degassed with N2 (x3), then the mixture was heated to 100°C and stirred for 12 h. The mixture was filtered, the filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give 8-methoxy-6-(4-methylpiperazin-l-yl) pyrido[3,4-d]pyramidin-4-ol (75 mg, 29% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₁₃H₁₇N₅O₂ 275.1; found 276.1.

[0253] Step 2.

[0254] To a mixture of 8-methoxy-6-(4-methylpiperazin-l-yl)pyrido[3,4- d]pyrimidin-4-ol (75 mg, 0.27 mmol) and (17?)-l-[3-(difluoromethyl)-2-fluoro- phenyl]ethanamine (77 mg, 0.41 mmol) in DMF (1 mL) was added BOP (181 mg, 0.41 mmol) and DBU (124 mg, 123 mL, 0.82 mmol). The mixture was stirred at rt for 2 h, then diluted with H2O (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with aqueous NaHCO₃ (5 mL x 2), brine (5 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give N-[(1R )-1- [3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-8-methoxy-6-(4-methylpiperazin-l-yl)pyrido[3,4-J]pyrimidin-4-amine formate salt (25 mg, 18% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₂₂H₂₅F₃N₆O 446.2; found 447.2; LCMS (ESI): m/z: [M +H] calculated for C₂₂H₂₅F₃N₆O: 446.5; found 447.2; 'H NMR (400 MHz, CD₃OD) δ = 8.20 (s, 1H), 7.56 (t, J= 7.2 Hz, 1H), 7.48 (t, J= 6.8 Hz, 1H), 7.19 - 7.26 (m, 1H), 6.85 - 7.16 (m, 2H), 5.78 (q, J= 7.0 Hz, 1H), 4.06 (s, 3H), 3.85 - 3.75 (m, 4H), 2.98 - 2.92 (m, 4H), 2.62 (s, 3H), 1.69 (d, J= 7.1 Hz, 3H).

Example 3. Synthesis of N-[(1R )-1- (difluoromethyl)-2-fluoro-phenyl]ethyl]-8- methoxy-6-( 1 -inetliyl-3.6-diliydro-2//-pyridin-4-yl)pyrido|3.4-7|pyrimidin -4-:imine

[0255] Step 1.

[0256] To a mixture of 8-methoxy-6-(l-methyl-3,6-dihydro-2//-pyridin-4- yl)pyrido[3,4-d] pyrimidin-4-ol (25 mg, 0.09 mmol) and (1R )-1-[3-(difluoromethyl)-2- fluoro-phenyl]ethanamine (26 mg, 0.14 mmol) in DMF (0.5 mL) was added DBU (42 mg, 42 mL 0.28 mmol) and BOP (61 mg, 0.14 mmol). The mixture was stirred at rt for 2 h, then filtered and the filtrate was concentrated under reduced pressure. The residue was purified by preparative-HPLC to give N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8- methoxy-6-(l-methyl-3,6-dihydro-2//-pyridin-4-yl)pyrido[3,4-J]pyrimidin-4-amine (10 mg, crude) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C23H24F3N5O 443.2; found 444.1; 'H NMR (400 MHz, CD3OD) δ = 8.69 (s, 1H), 8.06 (s, 1H), 7.70 - 7.66 (m, 1H), 7.58 - 7.54 (m, 1H), 7.33 - 7.29 (m, 1H), 7.14 - 6.84 (m, 2H), 6.02 (q, J= 7.0 Hz, 1H), 4.24 (s, 3H), 4.21 - 4.15 (m, 1H), 4.02 - 3.89 (m, 1H), 3.88 - 3.79 (m, 1H), 3.50 - 3.39 (m, 1H), 3.14 - 3.08 (m, 1H), 3.06 (s, 4H), 1.80 (d, J= 7.1 Hz, 3H).

Example 4. Synthesis of (l?)-4-(4-((l-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)- 8-methoxypyrido[3,4-d]pyrimidin-6-yl)-3,6-dihydro-2H-thiopyran 1,1-dioxide

[0257] Step 1.

[0258] A mixture of 6-chloro-8-methoxy-pyrido[3,4-d]pyrimidin-4-ol (400 mg, 1.89 mmol) in SOCl₂ (8 mL) and DMF (291 mL, 3.78 mmol) was heated to 100°C and stirred for 4 h. The mixture was concentrated under reduced pressure and the residue was diluted with aqueous NaHCO₃ (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered, the filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give 4,6-dichloro-8-methoxy-pyrido[3,4-d]pyrimidine (180 mg, 41% yield) as a solid. 'H NMR (400 MHz, CDCl₃) δ 9.13 (s, 1H), 7.61 (s, 1H), 4.27 (s,

3H).

[0259] Step 2.

[0260] To a mixture of 4,6-dichloro-8-methoxy-pyrido[3,4-d]pyrimidine (180 mg, 0.78 mmol) and ((1R )-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethanamine HC1 salt (212 mg, 0.94 mmol) in n-BuOH (4 mL) was added DIPEA (681 mL, 3.91 mmol). The mixture was heated to 90°C and stirred for 5 h, then cooled, diluted with H₂O (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give 6-chloro-N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine (280 mg, 94% yield) as a solid. 'H NMR (400 MHz, CDCh) 6 8.69 - 8.60 (m, 1H), 7.58 - 7.43 (m, 2H), 7.26 - 7.18 (m, 2H), 7.17 - 7.09 (m, 1H), 7.08 - 6.74 (m, 1H), 5.99 - 5.89 (m, 1H), 580 - 5.69 (m, 1H), 4.19 - 4.13 (m, 3H), 1.75 - 1.67 (m, 3H).

[0261] Step 3.

[0262] A mixture of 6-chloro-A-[(lA)-l-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-8-methoxy-pyrido[3,4-d]pyrimidin-4-amine (100 mg, 0.26 mmol), 4- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H -thiopyran 1,1-dioxide (135 mg, 0.52 mmol), CS2CO3 (255 mg, 0.78 mmol), Pd(dppf)C12 (38 mg, 0.05 mmol) in 1,4- dioxane (1 mL) and H2O (0.2 mL) was degassed with N2 (x 3), then placed under an atmosphere of N2, heated to 100°C and stirred for 2 h. [Note: two separate reactions were combined for workup]. The mixture was diluted with H2O (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give A-[(lA)-l-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-6-(l,l-dioxo-3,6-dihydro- 2H -thiopyran-4-yl)-8-methoxy-pyrido[3,4- d]pyrimidin-4-amine (60 mg, 48% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C22H21F3N4O3S 478.1; found 479.1; ¹H NMR (400 MHz, CD3OD) δ 8.43 (s, 1H), 7.89 (s, 1H), 7.59 (t, J= 7.3 Hz, 1H), 7.49 (t, J= 7.0 Hz, 1H), 7.27 - 7.21 (m, 1H), 7.15 - 6.86 (m, 2H), 5.82 (q, J= 7.0 Hz, 1H), 4.12 (s, 3H), 3.96 (d, J= 2.3 Hz, 2H), 3.40 - 3.35 (m, 5H), 1.71 (d, J= 1A Hz, 3H).

Example 5. Synthesis of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(1,1- dioxo-3.6-dihydro- 2H -thiopyran-4-yl)-8-methoxy-pyrido|3.4-7|pyrimidin-4-amine

[0263] Step 1. [0264] To a mixture of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6- (1,1 -di oxo-3, 6-dihydro-2H -thiopyran-4-yl)-8-m ethoxy -pyrido[3,4-d]pyrimidin-4-amine (100 mg, 0.21 mmol) in MeOH (10 mL) was added 10% Pd on carbon (10 mg). The mixture was stirred under an atmosphere of H2 (50 psi) at rt for 2 h, then filtered and the filtrate was concentrated under reduced pressure. The residue was purified by preparative- HPLC to give N-[(1R )-1--[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(l,l-dioxothian-4- yl)-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine (32 mg, 32% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₂₂H₂₃F₃N₄O₃S 480.1; found 481.0; ¹H NMR (400 MHz, CD3OD) δ 8.43 (s, 1H), 7.68 (s, 1H), 7.58 (t, J= 7.3 Hz, 1H), 7.49 (t, J= 7.0 Hz, 1H), 7.27 - 7.21 (m, 1H), 7.15 - 6.85 (m, 1H), 5.81 (q, J= 1A Hz, 1H), 4.11 (s, 3H), 3.39 - 3.33(m,

2H), 3.26 - 3.09 (m, 3H), 2.57 - 2.39 (m, 4H), 1.69 (d, J= 7.1 Hz, 3H).

Examples 6 and 7. Synthesis of (R) -4-(4-((l-(3-(difluoromethyl)-2- fluorophenyl)ethyl)amino)-8-methoxypyrido[3,4-d]pyrimidin-6-yl)-4- fluorotetrahydro-2H-thiopyran 1,1-dioxide; Example 6) and N-[(1R )-1- [3- (difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(4-methoxy-l,l-dioxo-thian-4- yl)pyrido[3,4-d]pyrimidin-4-amine (Example 7)

[0265] Step 1.

[0266] To a mixture of methyl 3-aminopyridine-4-carboxylate (40 g, 263 mmol) in DMF (400 mL) at rt was added NBS (117 g, 657 mmol). The mixture was stirred at rt for 30 min, then diluted with aqueous Na₂SO₃ (1.5 L) and extracted with EtOAc (1 L x 3). The combined organic layers were washed with brine (1 L), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give methyl 3-amino-2,6-dibromo-pyridine-4-carboxylate (66 g, 81% yield) as a solid. 'H NMR (400 MHz, DMSO- d₆) δ 7.73 (s, 1H), 6.80 (s, 2H), 3.87 (s, 3H).

[0267] Step 2.

[0268] To a mixture of methyl 3-amino-2,6-dibromo-pyridine-4-carboxylate (10 g, 32.3 mmol) in 1,4-dioxane (100 mL) was added NaOMe, 30% purity (20.34 g, 112.9 mmol). The mixture was heated to 120°C and stirred for 1 h, then KOH (5.43 g, 96.8 mmol) and H₂O (100 mL) were added, and the mixture was stirred at 120 °C for 4 h, then diluted with 2M HC1 (300 mL) and extracted with EtOAc (300 mL x 3). The combined organic layers were washed with brined (200 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatograph) to give 3-amino-6-bromo-2-methoxy-pyridine-4-carboxylic acid (5 g, 63% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₇H₇BrN₂O₃ 246.0; found 246.9.

[0269] Step 3.

[0270] To a mixture of 3-amino-6-bromo-2-methoxy-pyridine-4-carboxylic acid (1.0 g, 4.0 mmol) and acetic acid; methanimidamide (843 mg, 8.1 mmol) in 2-methoxy ethanol (14 mL) was heated to 170°C under microwave irradiation and stirred for 2 h. Four batches were combined and worked-up together. The mixture was filtered, the filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give 6-bromo-8-methoxy-pyrido[3,4-d ]pyrimidin-4-ol (800 mg, 77% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₈H₆BrN₃O₂ 255.0; found 256.0; 'H NMR (400 MHz, DMSO-d₆) δ 12.92 - 12.66 (m, 1H), 8.18 (s, 1H), 7.62 (s, 1H), 4.00 (s, 3H).

[0271] Step 4.

[0272] To a mixture of 6-bromo-8-methoxy-pyrido[3,4-d ]pyrimidin-4-ol (2.5 g, 9.8 mmol) and (1R)-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethanamine (2.77 g, 14.7 mmol) in DMF (10 mL) was added DBU (5.89 mL, 39.0 mmol) and BOP (6.48 g, 14.7 mmol). The reaction mixture was stirred at rt for 1 h, then diluted with H₂O (30 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give 6-bromo-N-[(1R )-1- [3- (difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine (1.3 g, 31% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₁₇H₁₄BrF₃N₄O 426.0; found 426.9; 'H NMR (400 MHz, CDCl₃) δ 8.66 (s, 1H), 7.56 - 7.46 (m, 3H), 7.21 (t, J= 7.7 Hz, 1H), 7.07 - 6.76 (m, 1H), 6.70 (d, J= 6.5 Hz, 1H), 5.81 (q, J= 6.9 Hz, 1H), 4.11 (s, 3H), 1.73 (d, J = 7.0 Hz, 3H).

[0273] Step 5.

[0274] To a mixture of 6-bromo-N-[(1R )-1- [3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-8-methoxy-pyrido[3,4-d]pyrimidin-4-amine (1.3 g, 3.0 mmol) in THF (13 mL) at 0°C was added NaH, 60% dispersion in oil (146 mg, 3.7 mmol). The mixture was stirred at 0°C for 30 min, then cooled to -78°C, n-BuLi, 2.5 M in hexanes (1.46 mL, 3.7 mmol) was added followed by tetrahydrothiopyran-4-one (371 mg, 3.2 mmol). The mixture was stirred at -78°C for 2 h, then diluted with H2O (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (50 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-TLC to give 4-[4-[[(lA)-l-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-J]pyrimidin-6-yl]tetrahydrothiopyran-4-ol (150 mg, 11% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₂₂H₂₃F₃N₄O₂S 464.2; found 465.0.

[0275] Step 6.

[0276] To a mixture of AI2O3 (33 mg, 0.32 mmol) in H2O (0.15 mL) at rt was added Oxone (199 mg, 0.32 mmol) in DCM (3.75 mL) and 4-[4-[[ (1R)-1-[3- (difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-d]pyrimidin-6- yl]tetrahydrothiopyran-4-ol (150 mg, 0.32 mmol). The mixture was heated to 60 °C and stirred for 1 h, then diluted with H₂O (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-TLC to give 4-[4-[[ (1R)-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]- 8-methoxy-pyrido[3,4-J]pyrimidin-6-yl]-l,l-dioxo-thian-4-ol (120 mg, 75% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C22H23F3N4O4S 496.1; found 497.0; 'H NMR (400 MHz, CDCl₃) δ 8.71 (s, 1H), 7.55 (q, J= 6.3 Hz, 2H), 7.26 - 7.21 (m, 1H), 7.19 - 6.77 (m, 2H), 6.13 (d, J= 7.4 Hz, 1H), 5.81 (q, J= 7.1 Hz, 1H), 4.64 (s, 1H), 4.20 (s, 3H), 3.70 (t, J= 13.1 Hz, 2H), 3.05 (d, J= 13.4 Hz, 2H), 2.76 (dt, J=13.7, 3.0 Hz, 2H), 2.19 (d, J= 12.8 Hz, 2H), 1.78 (d, J= 7.0 Hz, 3H).

[0277] Step 7.

[0278] To a mixture of 4-[4-[[(lA)-l-[3-(difhioromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-J]pyrimidin-6-yl]-l,l-dioxo-thian-4-ol (120 mg, 0.24 mmol) in DCM (1.5 mL) at -30°C was added DAST (96 mL, 0.73 mmol). The mixture was stirred at -30°C for 30 min, then adjusted to pH ~9 with Et₃N (1 mL), diluted with H2O (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (10 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give /V-[ (1R)-l-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(4-fluoro-l,l-dioxo-thian-4- yl)-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine (50 mg, 40% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C22H22F4N4O3S 498.1; found 499.1; ¹H NMR (400 MHz, DMSO- d₆) δ 8.86 (d, J= 6.1 Hz, 1H), 8.49 (s, 1H), 8.16 (s, 1H), 7.65 (t, J= 7.2 Hz, 1H), 7.51 (t, J= 7.0 Hz, 1H), 7.39 - 7.09 (m, 2H), 5.82 - 5.71 (m, 1H), 4.01 (s, 3H), 3.52 - 3.43 (m, 2H), 3.33 - 3.20 (m, 2H), 2.94 - 2.71 (m, 2H), 2.46 - 2.39 (m, 2H), 1.61 (d, J= 7.1 Hz, 3H).

[0279] Step 8.

[0280] To a mixture of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6- (4-fluoro-l,l-dioxo-thian-4-yl)-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine (100 mg, 0.2 mmol) in MeOH (1 mL), CH3CN (0.5 mL) and H2O (0.25 mL) was added CH₃ONa, 30% in MeOH (72 mg, 0.4 mmol). The mixture was heated to 60°C and stirred for 12 h, then filtered, the filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8- methoxy-6-(4-methoxy-l,l-dioxo-thian-4-yl)pyrido[3,4-J]pyrimidin-4-amine (45 mg, 43% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C23H25F3N4O4S 510.2; found 511.0; 'H NMR (400 MHz, DMSO- d₆) δ 8.80 (d, J= 1A Hz, 1H), 8.47 (s, 1H), 8.06 (s, 1H), 7.63 (t, J= 7.3 Hz, 1H), 7.51 (t, J= 6.8 Hz, 1H), 7.39 - 7.09 (m, 2H), 5.76 (q, J= 7.2 Hz, 1H), 4.01 (s, 3H), 3.31 - 3.28 (m, 2H), 3.17 - 3.08 (m, 5H), 2.60 - 2.50 (m, 4H), 1.62 (d, J= 1A Hz, 3H).

Example 8. Synthesis of 4-[4-[[ (1R)-l-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-J|pyrimidin-6-yl]-l,l-dioxo-thiane-4- carbonitrile

[0281] Step 1.

[0282] To a mixture of NaH, 60% in oil (112 mg, 2.81 mmol) in THF (10 mL) under an atmosphere of N₂ at 0°C was added 6-bromo-A-[(lA)-l-[3-(difluoromethyl)-2- fluoro-phenyl]ethyl]-8-methoxy-pyrido[3,4-J] pyrimidin-4-amine (1.0 g, 2.3 mmol). The mixture was stirred at 0°C for 30 min, then cooled to -78°C and n-BuLi, 2.5 M in hexanes (1.12 mL, 2.8 mmol) was added. Tetrahydrothiopyran-4-one (272 mg, 2.34 mmol) was added, and the mixture was stirred at -78°C for 2 h, then diluted with H₂O (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give 4-[4- [[(1R )-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy-pyrido[3,4- d]pyrimidin-6-yl]tetrahydrothiopyran-4-ol (0.4 g, 37% yield) as a solid. 'H NMR (400 MHz, DMSO-d₆) δ 8.84 (d, J= 8 Hz, 1H) 8.43 (s, 1H) 8.12 (s, 1H) 7.65 (t, J= 8.0 Hz, 1H) 7.51 (t, J= 8.0 Hz, 1H) 7.38 - 7.24 (m, 2H) 5.82 - 5.74 (m, 1H) 5.29 (s, 1H) 4.01 (s, 3H) 3.13 - 3.07 (m, 2H) 2.47 - 2.44 (m, 2H) 2.35 - 2.26 (m, 2H) 1.89 (d, J= 8.0 Hz, 2H) 1.60 (d, J= 8.0 Hz, 3H).

[0283] Step 2.

[0284] To a mixture of 4-[4-[[(lA)-l-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-d]pyrimidin-6-yl]tetrahydrothiopyran-4-ol (0.4 g, 0.86 mmol) in DAST (4 mL) at 0°C was added EtOH (4 mg, 0.09 mmol). The mixture was stirred at 0°C for 1 h, then ice cold H2O (15 mL) was added and the mixture was extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL x 2), dried with anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-TLC to give N-[(1R )-1- [3 - (difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(4-fluorotetrahydrothiopyran-4-yl)-8-methoxy- pyrido[3,4-d]pyrimidin-4-amine (0.2 g, 50% yield) as a solid. ¹H NMR (400 MHz, DMSO- d₆) δ 8.85 (d, J= 8.0 Hz, 1H) 8.47 (s, 1H) 8.07 (s, 1H) 7.64 (t, J= 8.0 Hz, 1H) 7.51 (t, J = 8.0 Hz, 1H) 7.38 - 7.11 (m, 2H) 5.78 - 5.74 (m, 1H) 4.01 (s, 3H) 3.04 - 2.97 (m, 2H) 2.70 - 2.64 (m, 2H) 2.46 - 2.32 (m, 2H) 2.24 - 2.19 (m, 2H) 1.60 (d, J= 8.0 Hz, 3H).

[0285] Step 3.

[0286] To a mixture of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6- (4-fluorotetrahydrothiopyran-4-yl)-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine (0.2 g, 0.43 mmol) in MeCN (1.2 mL), MeOH (0.6 mL) and H2O (0.2 mL) was added NaCN (126 mg, 2.57 mmol). The mixture was heated to 45°C and stirred for 24 h, then heated to 80°C and stirred for 48 h. The mixture was cooled and a further aliquot of MeCN (1.2 mL), MeOH (0.6 mL), H₂O (0.2 mL) and NaCN (126 mg, 2.57 mmol) were added. The mixture was heated to 80°C and stirred for 24 h, then heated to 90°C and stirred for 24 h and heated to 110°C and stirred for 96 h. The mixture was poured into NaHCO₃ (90 mL) and extracted with EtOAc (90 mL x 3). The combined organic layers were washed with brine (270 mL x 2), dried over anhydrous Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-TLC to give 4-[4-[[( 1R)- 1 -[3 - (difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-J]pyrimidin-6- yl]tetrahydrothiopyran-4-carbonitrile (50 mg, 25% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₂₃H₂₂F₃N₅OS 473.2; found 474.1; ¹H NMR (400 MHz, DMSO- d₆) δ 8.85 (d, J= 8.0 Hz, 1H) 8.49 (s, 1H) 8.02 (s, 1H) 7.66 (t, J= 8.0 Hz, 1H) 7.52 (d, J= 8.0 Hz, 1H) 7.38 - 7.11 (m, 2H) 5.80 - 5.73 (m, 1H) 4.04 (s, 3H) 3.05 - 2.98 (m, 2H) 2.92 - 2.85 (m, 2H) 2.60 - 2.57 (m, 2H) 2.32 - 2.23 (m, 2H) 1.63 (d, J= 8.0 Hz, 3H).

[0287] Step 4.

[0288] To a mixture of 4-[4-[[ (1R))-1-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-t/]pyrimidin-6-yl]tetrahydrothiopyran-4- carbonitrile (50 mg, 0.11 mmol) in MeOH (0.5 mL) and H2O (0.5 mL) under an atmosphere of N2 was added Oxone (260 mg, 0.42 mmol) in one portion. The mixture was stirred at rt for 1 h, then filtered, the filtrate concentrated under reduced pressure and the residue was purified by preparative-HPLC to give 4-[4-[[(lA)-l-[3-(difluoromethyl)-2- fluoro-phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-J]pyrimidin-6-yl]-l,l-dioxo-thiane-4- carbonitrile (30 mg, 56% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C23H22F3N5O3S 505.1; found 506.0; ¹H NMR (400 MHz, DMSO-d₆) δ 8.85 (d, J= 8.0 Hz, 1H) 8.52 (s, 1H) 8.08 (s, 1H) 7.64 (t, J= 8.0 Hz, 1H) 7.57 - 7.52 (m, 1H) 7.38 - 7.11 (m, 2H) 5.76 (t, J= 8.0 Hz, 1H) 4.04 (s, 3H) 3.55 - 3.45 (m, 2H) 3.40 - 3.30 (m, 2H) 2.77 - 2.74 (m, 4H) 1.64 (d, J= 4.0 Hz, 3H).

Example 9. Synthesis of 6-(4-amino-l,l-dioxo-thian-4-yl)-7V-[(ll?)-l-[3- (difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine

[0289] Step 1.

[0290] A mixture of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(4- fluoro-l,l-dioxo-thian-4-yl)-8-methoxy-pyrido[3,4-d]pyrimidin-4-amine (50 mg, 0.1 mmol) in 35% NH₃ in H₂O (5 mL) was heated to 120°C and stirred for 12 h in a pressure tube. After cooling, the mixture was diluted with H2O (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, the filtrate concentrated under reduced pressure and the residue was purified by preparative-HPLC to give 6-(4-amino-l,l-dioxo-thian-4-yl)-N-[(1R )-1- [3- (difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine (20 mg, 40% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₂₂H₂₄F₃N₅O₃S 495.2; found 496.0; 'H NMR (400 MHz, DMSO- d₆) 6 8.67 (d, J= 7.3 Hz, 1H), 8.44 (s, 1H), 8.16 (s, 1H), 8.00 (s, 1H), 7.64 (t, J= 7.3 Hz, 1H), 7.52 (t, J= 6.8 Hz, 1H), 7.39 - 7.09 (m, 2H), 5.77 (q, J= 6.9 Hz, 1H), 4.02 (s, 3H), 3.59 - 3.45 (m, 3H), 3.04 (d, J= 12.6 Hz, 3H), 2.65 - 2.52 (m, 2H), 2.13 (d, J= 13.7 Hz, 2H), 1.63 (d, J= 7.1 Hz, 3H).

Examples 10. Synthesis of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6- (l,l-dioxo-3,6-dihydro- 2H -thiopyran-4-yl)-8-methoxy-2-methyl-pyrido[3,4- d]pyrimidin-4-amine

[0291] Step 1.

[0292] A sealed tube containing a mixture of methyl 3-amino-2,6-dibromo- pyridine-4-carboxylate (1.0 g, 3.2 mmol), MeCN (10 mL) and MeSCLH (2.0 mL, 28.1 mmol) was heated to 120°C and stirred for 12 h. The mixture was adjusted to pH ~7 by addition of aqueous NaOH, then filtered and the filtrate was concentrated under reduced pressure to give 6,8-dibromo-2-methyl-pyrido[3,4-J]pyrimidin-4-ol (450 mg, 44% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₈H₄Br₂N₃O 318.9; found 319.8; ¹H NMR (400 MHz, CDCl₃) δ 8.06 (s, 1H), 2.41 (s, 3H).

[0293] Step 2. [0294] To a mixture of 6,8-dibromo-2-methyl-pyrido[3,4-J]pyrimidin-4- (200 mg, 0.63 mmol) in 1,4-dioxane (2 mL) was added CH₃ONa, 30% purity (339 mg, 1.88 mmol). The mixture was heated to 100°C and stirred for 12 h, then concentrated under reduced pressure. The residue was triturated with petroleum ether (5 mL) then filtered, and the filtrate was concentrated under reduced pressure to give 6-bromo-8-methoxy-2-methyl- pyrido[3,4-d ]pyrimidin-4-ol (169 mg) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₉H₈BrN₃O₂ 269.0; found 269.9.

[0295] Step 3.

[0296] To a mixture of 6-bromo-8-m ethoxy -2-rnethyl-pyrido[3,4-d]pyrimidin-4- ol (169 mg, 0.63 mmol) in 1,4-dioxane (2 mL) and H2O (0.4 mL) under an atmosphere of N₂ was added Cs₂CO₃ (612 mg, 1.88 mmol), 4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)-3,6-dihydro- 2H -thiopyran 1,1-dioxide (485 mg, 1.88 mmol), Pd(dppf)C12 (92 mg, 0.13 mmol). The mixture was heated to 100°C and stirred for 2 h, then diluted with DMF (8 mL) and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give 6-(1,1-dioxo-3,6-dihydro-2H -thiopyran-4-yl)-8- methoxy-2-methyl-pyrido[3,4-J]pyrimidin-4-ol (60 mg, 30% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₁₄H₁₅N₃O₄S 321.1; found 322.0.

[0297] Step 4.

[0298] To a mixture of 6-(l,l-dioxo-3,6-dihydro- 2H -thiopyran-4-yl)-8-methoxy- 2-methyl-pyrido[3,4-d]pyrimidin-4-ol (60 mg, 0.19 mmol) and (1R )-1-[3-(difluoromethyl)- 2-fluoro-phenyl]ethanamine (39 mg, 0.21 mmol) in DMF (1 mL) was added BOP (132 mg, 0.3 mmol) and DBU (84 mL, 0.56 mmol). The mixture was stirred at rt for 12 h, then diluted with MeOH (3 mL) and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give /V-[ (1R)-1-[3- (difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(1,1 -di oxo-3, 6-dihydro- 2H -thiopyran-4-yl)-8- methoxy-2-methyl-pyrido[3,4-d]pyrimidin-4-amine (7 mg, 8% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C23H23F3N4O3S 492.1; found 493.1; ¹H NMR (400 MHz, DMSO-d₆) δ 8.61 (d, J= 8.0 Hz, 1H), 7.94 (s, 1H), 7.67 ( t, J= 8.0 Hz, 1H), 7.49 ( t, J= 8.0 Hz, 1H), 7.37 - 7.10 (m, 2H), 6.81 (t, J = 4.0 Hz, 1H), 5.81 - 5.74 (m, 1H), 4.00 (s, 5H), 3.46 - 3.43 (t, J= 8.0 Hz, 2H), 3.21 (d, J= 4.0 Hz, 2H), 2.40 (s, 3H), 1.63 (d, J= 8.0 Hz, 3H). Example 11. Synthesis of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8- methoxy-6-(4-methyl-l,l-dioxo-thian-4-yl)pyrido[3,4-d|pyrimidin-4-amine

[0299] Step 1. [0300] To a mixture of 6-bromo-N-[(1R )-1- [3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-8-methoxy-pyrido[3,4-d]pyrimidin-4-amine (2.5 g, 5.85 mmol) and 2-(3,6- dihydro-2H -thiopyran-4-yl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (3.97 g, 17.56 mmol) in 1,4-dioxane (75 mL) and H₂O (15 mL) under an atmosphere of N₂ was added Cs₂C_O3 (5.72 g, 17.56 mmol) and Pd(dppf)C12 (856 mg, 1.17 mmol). The mixture was heated to 100°C and stirred for 2 h, then quenched by addition H2O (100 mL) and extracted with

EtOAc (80 mL x 3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by column chromatography on silica gel to give N-[(1R )-1- [3- (difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(3,6-dihydro-2H -thiopyran-4-yl)-8-methoxy- pyrido[3,4-d]pyrimidin-4-amine (1.6 g, 61% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C22H21F3N4OS 446.1; found 447.1; 'H NMR (400MHz, DMSO- d₆) δ 8.66 (d, J= 7.3 Hz, 1H), 8.42 (s, 1H), 7.91 (s, 1H), 7.64 (t, J= 7.5 Hz, 1H), 7.51 (t, J= 7.2 Hz, 1H), 7.44 - 7.04 (m, 3H), 5.77 (m, 1H), 4.01 (s, 3H), 3.49 - 3.39 (m, 2H), 2.97 - 2.88 (m, 2H), 2.83 (m, 2H), 1.62 (d, J= 6.8 Hz, 3H). [0301] Step 2.

[0302] To a mixture of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6- (3,6-dihydro- 2H -thiopyran-4-yl)-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine (100 mg, 0.22 mmol) in THF (1 mL) at -20°C under an atmosphere of N2 was added n-BuLi, 2.5 M in hexanes (206 mL, 0.53 mmol) dropwise over 5 min. The mixture was warmed to -15°C and stirred for 30 min, then cooled to -50°C and Me2SO₄ (23 mL, 0.25 mmol) was added dropwise. The mixture was cooled to -65°C and stirred for 30 min, then slowly transferred into a dilute solution of aqueous NH₄OH (0.5 mL) and heptane (0.5 mL). The mixture was warmed to rt and stirred for 2 h, then concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give N-[ (1R)-1-[3-(difluoromethyl)-2- fluoro-phenyl]ethyl]-8-methoxy-6-(4-methyl-2,3-dihydrothiopyran-4-yl)pyrido[3,4- d]pyrimidin-4-amine (140 mg, 34% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₂₃H₂₃F₃N₄OS 460.2; found 461.1.

[0303] Step 3.

[0304] To a mixture of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8- methoxy-6-(4-methyl-2,3-dihydrothiopyran-4-yl)pyrido[3,4-J]pyrimidin-4-amine (65 mg, 0.14 mmol) in THF (2 mL) under an atmosphere of nitrogen was added 10% Pd on carbon (6 mg). The suspension was degassed under vacuum and purged with H₂ (15 psi) several times, then stirred under an atmosphere of H₂ (15 psi) for 4 h. The mixture was filtered and the filtrate was concentrated to give N-[(1R)-l-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]- 8-methoxy-6-(4-methyltetrahydrothiopyran-4-yl)pyrido[3,4-J]pyrimidin-4-amine (60 mg, 92% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C23H25F3N4OS 462.2; found 463.2; 'H NMR (400MHz, DMSO- d₆) δ 8.70 (d, J= 7.0 Hz, 1H), 8.48 (s, 1H), 7.86 (s, 1H), 7.70 (t, J= 7.4 Hz, 1H), 7.58 (t, J= 6.9 Hz, 1H), 7.47 - 6.68 (m, 3H), 5.83 (m, 1H), 4.05 (s, 3H), 2.86 - 2.58 (m, 6H), 1.99 (m, 2H), 1.69 (d, J= 7.1 Hz, 3H), 1.36 (s, 3H).

[0305] Step 4.

[0306] To a mixture of N-[(1R)-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8- methoxy-6-(4-methyltetrahydrothiopyran-4-yl)pyrido[3,4-J]pyrimidin-4-amine (50 mg, 0.11 mmol) in H₂O (0.5 mL) and MeOH (0.5 mL) at rt under an atmosphere of N₂ was added Oxone (266 mg, 0.43 mmol) in one portion. The mixture was stirred at rt for 50 min, filtered, the filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give N-[(1R )-1- 3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-8-methoxy-6-(4-methyl-l,l-dioxo-thian-4-yl)pyrido[3,4-J]pyrimidin-4- amine (17 mg, 32% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₂₃H₂₅F₃N₄O₃S 494.2; found 495.1; ¹H NMR (400 MHz, DMSO-d₆) δ 8.62 (d, J= 7.2 Hz, 1H), 8.45 (s, 1H), 7.90 (s, 1H), 7.64 (t, J= 7.4 Hz, 1H), 7.52 (t, J= 7.2 Hz, 1H), 7.43 - 7.05 (m, 2H), 5.79 - 5.76 (m, 1H), 4.00 (s, 3H), 3.20 (dd, J= 6.2, 11.4 Hz, 2H), 3.12 - 2.96 (m, 2H), 2.84 - 2.71 (m, 2H), 2.26 - 2.13 (m, 2H), 1.64 (d, J= 7.0 Hz, 3H), 1.38 (s, 3H).

Example 12. Synthesis of l-[4-[4-[[(1R )-1-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-J]pyrimidin-6-yl]-l-piperidyl]ethanone

[0307] Step 1.

[0308] To a mixture of 6-chloro-8-methoxy-pyrido[3,4-J]pyrimidin-4-ol (300 mg, 1.42 mmol) andl-[4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H/-pyridin- l-yl]ethanone (712 mg, 2.84 mmol) in 1,4-dioxane (12 mL) and H2O (2.4 mL) under an atmosphere of N₂ was added Pd(dppf)C12 (208 mg, 0.28 mmol) and CS₂CO₃ (1.39 g, 4.25 mmol). The mixture was degassed with N2 (x3) and heated to 100°C and stirred for 5 h, then filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give 8-methoxy-6-(l-methyl-3,6-dihydro-2H -pyridin-4- yl)pyrido[3,4-d]pyrimidin-4-ol formate salt (80 mg, 36% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₁₅H₁₆N₄O₃ 300.1; found 301.0; 'H NMR (400 MHz, CD₃OD) δ 8.14 (s, 1H), 7.62 (d, J= 4.8 Hz, 1H), 6.98 (s, 1H), 4.35 - 4.25 (m, 2H), 4.14 (s, 3H), 3.88

- 3.74 (m, 2H), 2.82 - 2.62 (m, 2H), 2.18 (d, J= 16.4 Hz, 3H).

[0309] Step 2.

[0310] To a mixture of 8-methoxy-6-(l-methyl-3,6-dihydro- 2H -pyridin-4- yl)pyrido[3,4-d]pyrimidin-4-ol formate salt (240 mg, 0.8 mmol) in MeOH (10 mL) under an atmosphere of Ar was added Pd, 10% on carbon (200 mg, 0.8 mmol). The mixture was degassed and purged with H₂ (x3) then heated to 40°C under an atmosphere of H₂ (50 psi) and stirred for 2 h. The mixture was filtered and the filtrate was concentrated under reduced pressure to give l-[4-(4-hydroxy-8-methoxy-pyrido[3,4-d ]pyrimidin-6-yl)-1- piperidyl]ethanone (183 mg, 76% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₁₅H₁₈N₄O₃ 302.1; found 303.1.

[0311] Step 3.

[0312] To a mixture of (lA)-l-[3-(difluoromethyl)-2-fluoro-phenyl]ethanamine HC1 salt (169 mg, 0.75 mmol) and l-[4-(4-hydroxy-8-methoxy-pyrido[3,4-d ]pyrimidin-6- yl)-l -piperidyl] ethanone (180 mg, 0.60 mmol) in DMF (2 mL) was added BOP (395 mg, 0.89 mmol) and DBU (269 mL, 1.79 mmol). The mixture was stirred at rt for 2 h, then purified by preparative-HPLC to give 1-[4-[4-[[(1R )-1-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-J]pyrimidin-6-yl]-l-piperidyl]ethanone (8.7 mg, 5% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C24H26F3N5O2 473.2; found 474.1; 'H NMR (400 MHz, DMSO- d₆) 6 10.48 (s, 1H) 8.76 (s, 1H) 8.18 (s, 1H) 7.83 (t, J= 7.2 Hz, 1H) 7.58 (t, J= 6.9 Hz, 1H) 7.41 - 7.10 (m, 2H) 5.96 (q, J= 7.1 Hz, 1H) 4.56 (d, J= 13.2 Hz, 1H) 4.10 (s, 3H) 3.98 (d, J= 13.6 Hz, 1H) 3.26 - 3.17 (m, 2H) 3.00 (tt, J= 11.8, 3.4 Hz, 1H) 2.74 - 2.65 (m, 1H) 2.08 - 1.94 (m, 5H) 1.79 - 1.57 (m, 5H). Example 13. Synthesis of N-[ (1R)-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(4- fluoro-l-methyl-4-piperidyl)-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine

[0313] Step 1.

[0314] To a mixture of 6-bromo-N-[(1R )-1- [3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-8-methoxy-pyrido[3,4-d]pyrimidin-4-amine (500 mg, 1.17 mmol) in THF (5 mL) at 0°C was added NaH, 60% dispersion in oil (56 mg, 1.40 mmol). The mixture was stirred at 0°C for 30 min, then cooled to -78°C and n-BuLi, 2.5 M in hexanes (562 mL, 1.4 mmol) added. The mixture was stirred at -78°C for 1 h, then l-methylpiperidin-4-one (159 mg, 1.40 mmol) was added at -78°C and the mixture stirred at -78°C for 1 h. The mixture was quenched with H2O (10 mL) and extracted with EtOAc (10 mL x 3). The combined organic layers were washed with brine (10 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure to give a residue and the residue was purified by silica gel column chromatography to give 4-[4-[[(lA)-l-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-J]pyrimidin-6-yl]-l-methyl-piperidin-4-ol (130 mg, 24% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₂₃H₂₆F₃N₅O₂ 461.2; found 462.2; 'H NMR (400 MHz, CD₃OD) δ 8.44 (s, 1H), 7.97 (s, 1H), 7.59 (t, J= 7.5 Hz, 1H), 7.49 (t, J= 7.0 Hz, 1H), 7.23 (t, J= 7.8 Hz, 1H), 7.16 - 6.85 (m, 2H), 5.84 (q, J= 7.1 Hz, 1H), 4.12 (s, 3H), 3.05 - 2.95 (m, 2H), 2.90 - 2.80 (m, 2H), 2.60 - 2.40 (m, 5H), 2.01 (s, 2H), 1.86 (d, J= 14.2 Hz, 2H), 1.70 (d, J= 7.2 Hz, 3H).

[0315] Step 2.

[0316] To a mixture of 4-[4-[[(lA)-l-[3-(difhioromethyl)-2-fhioro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-J]pyrimidin-6-yl]-l-methyl-piperidin-4-ol (103 mg, 0.22 mmol) in DCM (1 mL) at 0°C was added DAST (59 mL, 0.45 mmol). The mixture was warmed to rt and stirred at for 1 h, then quenched by addition of saturated aqueous NaHCCL solution (2 mL) and extracted with DCM (1 mL x 4). The combined organic layers were washed with brine (5mL), dried over anhydrous Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(4- fhioro-l-methyl-4-piperidyl)-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine (34 mg, 33% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₂₃H₂₅F₄N₅O 463.2; found 464.2; 'H NMR (400 MHz, DMSO-d₆) δ 8.83 (d, J= 7.2 Hz, 1H), 8.46 (s, 1H), 8.18 (s, 1H), 8.09 (s, 1H), 7.64 (t, J= 7.3 Hz, 1H), 7.51 (t, J= 6.8 Hz, 1H), 7.41 - 7.07 (m, 2H), 5.76 (q, J= 7.0 Hz, 1H), 4.00 (s, 3H), 2.78 (d, J= 6.4 Hz, 2H), 2.46 - 2.28 (m, 4H), 2.27 (s, 3H), 1.97 - 1.87 (m, 2H), 1.60 (d, J= 7.0 Hz, 3H).

Example 14. Synthesis of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(1- imino-l-oxo-3,6-dihydro- 2H -thiopyran-4-yl)-8-methoxy-pyrido[3,4-J]pyrimidin-4- amine

[0317] Step 1. [0318] To a mixture of 6-bromo-N-[(1R )-1- [3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-8-methoxy-pyrido[3,4-d]pyrimidin-4-amine (1.0 g, 2.3 mmol) and 2-(3,6- dihydro- 2H -thiopyran-4-yl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (1.59 g, 7.0 mmol) in 1,4-di oxane (10 mL) and H2O (2 mL) under an atmosphere of N2 was added Pd(dppf)C1₂ (343 mg, 0.47 mmol) and Cs₂CO₃ (2.29 g, 7.0 mmol). The mixture was degassed with N2 (x3), then heated to 100°C and stirred for 1 h, then quenched by H₂O (20 mL) slowly and extracted with EtOAc (30 mL x 2). The combined organic layers were washed with brine (20 mL x 2), dried with anhydrous Na2SO4 and filtered. The filtered and concentrated under reduced pressure and the residue was purified by silica gel column chromatography to giveN-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(3,6-dihydro-27/-thiopyran-4-yl)- 8-methoxy-pyrido[3,4-J]pyrimidin-4-amine (640 mg, 61% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C22H21F3N4OS 446.1; found 447.1; ¹H NMR (400 MHz, DMSO-d₆) δ 8.67 (d, J= 4.0 Hz, 1H), 8.42 (br s, 1H), 7.91 (s, 1H), 7.65 (t, J= 8.0 Hz, 1H), 7.52 (t, J= 8.0 Hz, 1H), 7.42 - 7.11 (m, 3H), 5.78 (t, J= 8.0 Hz, 1H), 4.02 (s, 3H), 3.48 - 3.42 (m, 2H), 2.92 - 2.88 (m , 2H), 2.86 - 2.73 (m, 2H), 1.63 (d, J= 8.0 Hz, 3H).

[0319] Step 2.

[0320] To a mixture of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6- (3,6-dihydro- 2H -thiopyran-4-yl)-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine (440 mg, 0.99 mmol) in THF (6 mL) and H₂O (0.1 mL) at 0°C was added Oxone (212 mg, 0.35 mmol). The mixture was stirred at 0°C for 6 h, then quenched by addition of aqueous Na₂S₂O₃ (5 mL) and extracted with EtOAc (5 mL x 2). The combined organic layers were washed with brine (10 mL x 2), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give A-[(lA)-l-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy- 6-(l-oxo-3,6-dihydro- 2H -thiopyran-4-yl)pyrido[3,4-J]pyrimidin-4-amine (310 mg, 68% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₂₂H₂₁F₃N₄O₂S 462.1; found 463.1; 'H NMR (400 MHz, DMSO-d₆) δ 8.70 (d, J= 8.0 Hz, 1H), 8.44 (br s, 1H), 8.00 (s, 1H), 7.64 (t, J = 8.0 Hz, 1H), 7.51 (t, J= 8.0 Hz, 1H), 7.40 - 7.06 (m, 2H), 6.91 - 6.79 (m, 1H), 5.77 (t, J= 8.0 Hz, 1H), 4.02 (s, 3H), 3.77 (d, J= 16.0 Hz, 1H), 3.53 - 3.49 (m, 1H), 3.30 - 3.21 (m, 1H), 3.10 - 2.88 (m, 3H), 1.63 (d, J= 8.0 Hz, 3H).

[0321] Step 3.

[0322] A mixture of A-[(lA)-l-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8- methoxy-6-(l -oxo-3, 6-dihydro- 2H -thiopyran-4-yl)pyrido[3,4-J]pyrimidin-4-amine (310 mg, 0.67 mmol), [acetoxy(phenyl)-X3-iodanyl]acetate (648 mg, 2.01 mmol) and ammonia carbamic acid (209 mg, 2.68 mmol) in MeOH (5 mL) was stirred at rt for 3 h. The mixture was quenched by addition of H₂O (20 mL) and extracted with EtOAc (30 mL x 3). The combined organic layers were washed with brine (10 mL x 2), dried over Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography, then by preparative-HPLC to give N-[(1R )-1- (difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(l -imino- 1 -oxo-3, 6-dihy dro- 2H -thiopyran-4-yl)- 8-methoxy-pyrido[3,4-J]pyrimidin-4-amine (14 mg, 4% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calculated for C₂₂H₂₂F₃N₅O₂S 477.1; found 478.2; 'H NMR (400 MHz, DMSO-d₆) δ 8.70 (d, J= 8.0 Hz, 1H), 8.44 (br s, 1H), 7.98 (s, 1H), 7.64 (t, J= 8.0 Hz, 1H), 7.51 (t, J= 8.0 Hz, 1H), 7.41 - 7.08 (m, 2H), 6.83 (t, J= 4.0 Hz, 1H), 5.77 (t, J= 4.0 Hz, 1H), 4.02 (s, 3H), 3.97 - 3.90 (m, 2H), 3.83 (s, 1H), 3.31 - 3.27 (m, 2H), 3.12 - 3.18 (m, 2H), 1.62 (d, J= 8.0 Hz, 3H).

Example 15. Synthesis of N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8- methoxy-6-(l-methylimino-l-oxo-3,6-dihydro-2Z/-thiopyran-4-yl)pyrido[3,4- d\ pyrimidin-4-amine

[0323] Step 1.

[0324] To a mixture of A-[(lA)-l-[3-(difhioromethyl)-2-fluoro-phenyl]ethyl]-6- (1 -imino-1 -oxo-3, 6-dihydro- 2H -thiopyran-4-yl)-8-methoxy-pyrido[3,4-d]pyrimidin-4- amine (20 mg, 0.04 mmol) in MeCN (2 mL) was added TFA (13 mL, 0.18 mmol) and formaldehyde, 37% in H₂O (13 mL, 0.17 mmol). The mixture was stirred at rt for 30 min then Et₃SiH (28 mL, 0.17 mmol) was added and the mixture stirred at rt for 12 h, then concentrated under reduced pressure. The residue was partitioned between H2O (20 mL) and DCM (20 mL) with saturated NaHCO₃ added to neutralize TFA. The aqueous layer was extracted with additional DCM (20 mL) and the combined organic extracts were dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give N-[(1R )-1- [3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-8-methoxy-6-(1-methylimino-1-oxo-3,6-dihydro-2H/-thiopyran-4- yl)pyrido[3,4-d]pyrimidin-4-amine (3 mg, 14% yield) as solid. LCMS (ESI): m/z [M+H]⁺ calc’d for C₂₃H₂₄F₃N₅O₂S 491.2; found 492.2; 1¹H NMR (400 MHz, DMSO-d₆) δ 8.69 (d, J = 8.0 Hz, 1H), 8.44 (s, 1H), 7.98 (s, 1H), 7.64 (t, J= 8.0 Hz, 1H), 7.52 (t, J= 8.0 Hz, 1H), 7.40 - 7.08 (m, 2H), 6.87 - 6.79 (m, 1H), 5.80 - 5.70 (m, 1H), 4.14 - 4.06 (m, 1H), 4.02 (s, 3H), 3.97 - 3.88 (m, 1H), 3.44 - 3.38 (m, 2H), 3.20 - 3.06 (m, 2H), 2.70 (s, 3H), 1.63 (d, J= 8.0 Hz, 3H).

[0325] Step 1.

[0326] To a mixture of N-[(1R )-1- [3-(difkoromethyl)-2-fluoro-phenyl]ethyl]-6- (1 -imino-1 -oxo-3, 6-dihydro- 2H -thiopyran-4-yl)-8-methoxy-pyrido[3,4-J]pyrimidin-4- amine (30 mg, 0.06 mmol) in DCM (1 mL) at 0°C was added DMAP (8.4 mg, 0.07 mmol) and carb ononi tri die bromide (9 mL, 0.13 mmol). The mixture was warmed to rt and stirred for 4 h, then concentrated under reduced pressure and the residue was purified by preparative-HPLC to give [4-[4-[[(1R)-l-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-J]pyrimidin-6-yl]-1 -oxo-3, 6-dihydro- 2H - thiopyran- l-ylidene]cyanamide (7 mg, 21% yield, 95%) as a solid. LCMS (ESI): m/z [M+H]⁺ calc’d for C₂₃H₂₁F₃N₆O₂S 502.1; found 503.1; 'H NMR (400 MHz, DMSO-d₆) δ 8.74 ( d, J= 6.8 Hz, 1H) 8.46 (s, 1H) 8.02 (s, 1H) 7.69 - 7.61 (m, 1H) 7.57 - 7.49 (m, 1H) 7.42 - 6.77 (m, 3H) 5.79 - 5.74 (m, 1H) 4.58 (s, 2H) 4.03 (s, 5H) 3.29 (s, 2H) 1.63 (d, J= 7.2 Hz, 3H). Example 17 and 18. Synthesis of (R) -N-(l-(3-(difluoromethyl)-2-fluorophenyl)ethyl)-8- methoxy-6-(4-methoxypiperidin-4-yl)pyrido[3,4-d]pyrimidin-4-amine (Example 17) and 1-[4-[4-[[ (1R)-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy- pyrido[3,4-J|pyrimidin-6-yl]-4-methoxy-l-piperidyl]ethanone (Example 18)

[0327] Step 1.

[0328] To a mixture of 6-bromo-N-[(1R )-1- [3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-8-rnethoxy-pyrido[3,4-d]pyrimidin-4-amine (1.0 g, 2.3 mmol) in THF (10 mL) at 0°C was added NaH, 60% dispersion in oil (140 mg, 3.5 mmol). The mixture was stirred at 0°C for 30 min, then cooled to -78°C and n-BuLi, 2.5 M in hexanes (1.12 mL, 2.8 mmol) was added, followed by tert-butyl 4-oxopiperidine-l -carboxylate (490 mg, 2.5 mmol). The mixture was stirred at -78°C for 2 h, then quenched with H2O (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4 and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give tert- butyl 4-[4-[[ (1R)-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy- pyrido[3,4-d ]pyrimidin-6-yl]-4-hydroxy-piperi dine- 1 -carboxylate (664 mg, 52% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc’d for C₂₇H₃₂F₃N₅O₄ 547.2; found 548.2.

[0329] Step 2.

[0330] To a mixture of tert-butyl 4-[4-[[(lA)-l-[3-(difkoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-d ]pyrimidin-6-yl]-4-hydroxy-piperidine-l- carboxylate (556 mg, 1.02 mmol) in DCM (3 mL) at 0°C was added DAST (268 mL, 2.03 mmol). The mixture was warmed to rt and stirred for 1 h, then quenched by addition of NaHCCL (5 mL) and extracted with DCM (5mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give tert-butyl 4-[4-[[(1R )-1 - [3-(difluoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-d ]pyrimidin-6-yl]-4-fluoro-piperidine-l- carboxylate (228 mg, 41% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ cak’d for C27H31F4N5O3 549.2; found 550.3; 'H NMR (400 MHz, CD₃OD)δ 8.47 (s, 1H), 8.03 (s, 1H), 7.60 (t, J= 7.5 Hz, 1H), 7.50 (t, J= 6.9 Hz, 1H), 7.29 - 7.21 (m, 1H), 7.18 - 6.88 (m, 1H), 5.85 (q, J = 7.1 Hz, 1H), 4.19 - 4.14 (m, 2H), 4.11 (s, 3H), 3.26 - 3.10 (m, 2H), 2.49 - 2.28 (m, 2H), 2.02 - 1.93 (m, 2H), 1.71 (d, J= 7.0 Hz, 3H), 1.53 (s, 9H).

[0331] Step 3.

[0332] To a mixture of tert-butyl 4-[4-[[(lA)-l-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-d ]pyrimidin-6-yl]-4-fluoro-piperidine-1- carboxylate (226 mg, 0.41 mmol) in MeCN (1.5 mL), MeOH (0.9 mL) and H₂O (0.3 mL) was added NaOMe (200 mg, 3.7 mmol). The mixture was heated to 70°C and stirred for 12 h, them diluted with H₂O (15 mL) and extracted with EtOAc (5 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by silica gel column chromatography to give tert-butyl 4-[4-[[(lA)-l-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-J]pyrimidin-6-yl]-4-methoxy-piperidine-1- carboxylate (142 mg, 62% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ cak’d for C₂₈H₃₄F₃N₅O₄ 561.3; found 562.2; 'H NMR (400 MHz, CD3OD) δ 8.44 (s, 1H), 7.93 (s, 1H), 7.58 (t, J= 7.4 Hz, 1H), 7.49 (t, J= 7.0 Hz, 1H), 7.24 (t, J= 7.8 Hz, 1H), 7.17 - 6.86 (m, 1H), 5.84 (q, J= 7.0 Hz, 1H), 4.13 - 4.04 (m, 5H), 3.92 (d, J= 13.4 Hz, 2H), 3.16 (s, 3H), 2.22 - 2.10 (m, 4H), 1.70 (d, J= 7.0 Hz, 3H), 1.48 (s, 9H).

[0333] Step 4.

[0334] A mixture of tert-butyl 4-[4-[[(lA)-l-[3-(difluoromethyl)-2-fluoro- phenyl]ethyl]amino]-8-methoxy-pyrido[3,4-J]pyrimidin-6-yl]-4-methoxy-piperidine-l- carboxylate (110 mg, 0.2 mmol) and HC1 in MeOH (1 mL) was stirred at 0°C for 1 h. The mixture was filtered, the filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give N-[(1R )-1- [3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-8-methoxy-6-(4-methoxy-4-piperidyl)pyrido[3,4-J]pyrimidin-4-amine (78 mg, 84% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc’d for C₂₃H₂₆F₃N₅O₂ 461.2; found 462.2; 'H NMR (400 MHz, CD3OD) δ 8.81 (d, J= 7.5 Hz, 1H), 8.46 (s, 1H), 8.33 (s,lH), 8.03 (s, 1H), 7.64 (t, J= 8.0 Hz, 1H), 7.52 (t, J= 8.0 Hz, 1H), 7.38 - 7.11 (m, 2H), 5.81 - 5.74 (m, 1H), 4.01 (s, 3H), 3.12 - 2.96 (m, 7H), 2.19 - 2.10 (m, 4H), 1.67 - 1.58 (m, 3H).

[0335] Step 5.

[0336] To a mixture ofN-[(1R )-1- [3-(difhioromethyl)-2-fluoro-phenyl]ethyl]-8- methoxy-6-(4-methoxy-4-piperidyl)pyrido[3,4-J]pyrimidin-4-amine (55 mg, 0.12 mmol) in DCM (5 mL) was added TEA (100 mL, 0.72 mmol) and acetyl chloride (30 mL, 0.42 mmol). The mixture was stirred at rt for 3 h, then diluted with aqueous NaHCO₃ (1 mL), H₂O (20 mL) and extracted with DCM (20 mL x 3). The combined organic layers were washed with brine (5 mL), dried with anhydrous Na₂SO₄ and filtered. The filtrate was concentrated under reduced pressure and the residue was purified by preparative-HPLC to give l-[4-[4-[[ (1R)-1-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy- pyrido[3,4-d]pyrimidin-6-yl]-4-methoxy-l-piperidyl]ethanone (12.6 mg, 21% yield) as a solid. LCMS (ESI): m/z [M+H]⁺ calc’d for C₂₅H₂₈F₃N₅O₃ 503.2; found 504.2; ¹H NMR (400 MHz, DMSO-d₆) 8 8.78 (d, J= 7.6 Hz, 1 H), 8.45 (s, 1H), 8.43 (m, 1H), 8.01 (s, 1H), 7.68 - 7.60 (m, 1H), 7.57 - 7.48 (m, 1H), 7.41 - 7.07 (m, 2H), 5.83 - 5.69 (m, 1H), 4.21 (d, J= 12.0 Hz, 1H), 3.99 (s, 3H), 3.71 (d, J= 12.0 Hz, 1H), 3.47 - 3.37 (m, 1H), 3.07 (s, 3H), 3.03 - 2.94 (m, 1H), 2.15 - 1.94 (m, 7H), 1.62 (d, J= 7.2 Hz, 1H). Biological Examples

Potency assay: pERK

[0337] The purpose of this assay is to measure the ability of test compounds to inhibit S0S1 function in cells. S0S1 activates RAS proteins by catalyzing the conversion of RAS GDP to RAS GTP in response to receptor tyrosine kinase activation. Activation of RAS induces a sequence of cellular signaling events that results in increased phosphorylation of ERK at Threonine 202 and Tyrosine 204 (pERK). The procedure described below measures the level of cellular pERK in response to test compounds in PC- 9 cells (EGFR Exl9Del).

[0338] PC-9 cells were grown and maintained using media and procedures recommended by the ATCC. On the day prior to compound addition, cells were plated in 384-well cell culture plates (40 pL/well) and grown overnight in a 37°C, 5% CO2 incubator. Test compounds were prepared in 10, 3 -fold dilutions in DMSO, with a top concentration of 10 mM. On the day of the assay, 40 nL of test compound was added to each well of cell culture plate using an Echo550 liquid handler (LabCyte). Concentrations of test compound were tested in duplicate with highest test concentration being 10 pM. After compound addition, cells were incubated for 1 hour at 37°C, 5% CO2. Following incubation, culture medium was removed and cells were washed once with phosphate buffered saline.

[0339] Cellular pERK level was determined using the AlphaLISA SureFire Ultra p-ERKl/2 Assay Kit (PerkinElmer). Cells were lysed in 25 pL lysis buffer, with shaking at 600 RPM at room temperature for 15 minutes. Lysate (10 pL) was transferred to a 384- well Opti-plate (PerkinElmer) and 5 pL acceptor mix was added. The plate was centrifuged at 1000 RPM for 1 minute, and incubated in the dark for 2 hours. Following this incubation, 5 pL of donor mix was added, the plate was sealed and centrifuged at 1000 RPM for 1 minute, and the mixture was incubated for 2 hours at room temperature. Signal was read on an Envision plate reader (PerkinElmer) using standard AlphaLISA settings. Analysis of raw data was carried out in Excel (Microsoft) and Prism (GraphPad). Signal was plotted vs. the decadal logarithm of compound concentration, and IC50 was determined by fitting a 4-parameter sigmoidal concentration response model. [0340] SOS1 pERK IC50 Assay results are shown in Table B below.

Table B

Mode of Action Assay: Inhibition of SOS1 Nucleotide Exchange Activity [0341] The purpose of this assay was to characterize the inhibitory activity of compounds on S0S1 nucleotide exchange of KRAS. Data was reported as IC₅₀ values based on the TR-FRET signal.

[0342] Note - the following protocol describes a procedure for monitoring the inhibition of S0S1 nucleotide exchange activity of wild-type KRAS in response to a compound of the disclosure. Other KRAS mutants and RAS isoforms maybe employed.

[0343] In assay buffer containing 20 mM HEPES, pH 7.5, 150 mM NaCl, 5 mM MgCl₂, 0.05% Tween-20, 0.1% BSA, 1 mM DTT, concentration series of test compounds were generated spanning 100 pM to 1.7 nM over eleven 3-fold serial dilutions in a 384- well assay plate at a volume of 20 pL. The purified tagless catalytic domain of S0S1 (residues 564 - 1049) was first diluted in assay buffer at a concentration of 100 nM, and then 20 μL of the S0S1 containing solution was directly dispensed into compound plates. The SOSl/compound mixture was incubated at room temperature with constant mixing on an orbital shaker for 20 minutes to allow the reaction to reach equilibrium. A KRAS mixture was prepared by diluting 66.7 nM avi-tagged KRAS (residue 1 - 169), 3.33 nM Streptavidin-Tb and 333 nM EDA-GTP-DY-647P1 in assay buffer. This mixture was prepared immediately before addition to the SOSl/compound mixture to prevent intrinsic nucleotide exchange. Then 5 pL of the pre-incubated SOSl/compound mixture and 7.5 μL of the KRAS mixture were added sequentially in a 384-well low volume black round bottom plate and incubated at room temperature with constant shaking for 30 minutes.

Time-resolved fluorescence was measured on a PerkinElmer Envision plate reader. DMSO and 10 pM of compound (i) were used as negative and positive controls, respectively.

compound (i)

[0344] Three replicates were performed for each compound. Data were normalized by the following: (Positive control - Sample signal)/(Positive control - negative control)* 100. The data were fit using a four-parameter logistic fit.

[0345] S0S1 TR-FRET IC50 Assay results are shown in Table C below.

Table C

Equivalents

[0346] While the present disclosure has been described in conjunction with the specific embodiments set forth above, many alternatives, modifications and other variations thereof will be apparent to those of ordinary skill in the art. All such alternatives, modifications and variations are intended to fall within the spirit and scope of the present disclosure.

Claims

WHAT IS CLAIMED IS:

1. A compound having the structure of Formula (I),

Ri is selected from the group consisting of optionally substituted 6-membered aryl and optionally substituted 5-6 membered heteroaryl; R₂ is selected from the group consisting of H and optionally substituted C_1-6 alkyl;

R₃ is — OR_3a, and further wherein R_3a is optionally substituted C_1-3 alkyl; and R₄ is selected from the group consisting of H, C_1-6 alkyl, 3-14 membered cycloalkyl, 3-14 membered cycloalkenyl, 3-14 membered heterocyclyl, 6-10 membered aryl, and 5-10 membered heteroaryl; wherein each C_1-6 alkyl, 3-14 membered cycloalkyl, 3-14 membered cycloalkenyl, 3-14 membered heterocyclyl, 6-10 membered aryl, and 5-10 membered heteroaryl is optionally substituted with C_1-6 alkyl, -R_4a, -OR_4a, -O-C_1-6 alkyl-R_4a, =0, halogen, -C(O)R_4a, -C(O)OR_4a, -C(O)NR_4bR_4c, -NR₄bC(O)R₄c, -CN, =NR_4a, -NR_4bR_4c, - SO2R_4a, 3-6 membered cycloalkyl optionally substituted with R_4a, 3-7 membered heterocyclyl optionally substituted with R_4a, 6-10 membered aryl optionally substituted with R_4a, or 5-10 membered heteroaryl optionally substituted with R_4a; wherein R_4a is H, C_1-6 alkyl, C_1-6 haloalkyl, -C(O)R_4b, -C(O)NR_4bR_4c, =O, 3-6 membered cycloalkyl, 6-10 membered aryl optionally substituted with -OR_4b, - CN, =N-3-6 membered cycloalkyl, 3-7 membered heterocyclyl, -(CH₂)_rOCH₃, or - (CH₂)_rOH, wherein r is 1, 2, or 3; wherein each R_4b is independently H, C_1-6 alkyl; and wherein each R_4c is independently H or C_1-6 alkyl.

2. The compound of claim 1, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein Ri is optionally substituted 6- membered aryl.

3. The compound of claim 1, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein Ri is

4. The compound of claim 1, having the structure of Formula (II),

or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₂, R₃, and R₄ are as defined in claim 1, wherein. R₅, R₆, R₇, R₈, and R₉ are independently selected from the group consisting of H, C_1-6 alkyl, halogen, and -NH₂, wherein each C_1-6 alkyl is optionally substituted with halogen.

5. The compound of any one of claims 1 through 4, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₂ is H.

6. The compound of any one of claims 1 through 4, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₂ is optionally substituted C_1-3 alkyl, wherein the optional substituent is one or more fluoro.

7. The compound of any one of claims 1 through 4, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₂ is - CH₃.

8. The compound of any one of claims 1 through 7, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₃ is - OR_3a, and further wherein R_3a is C_1-3 alkyl.

9. The compound of any one of claims 1 through 7, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₃ is - OCH₃.

10. The compound of any one of claims 1 through 9, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₄ is optionally substituted 3-14 membered heterocyclyl.

11. The compound of any one of claims 1 through 9, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₄ is 3-14 membered heterocyclyl optionally substituted with halogen, C_1-6 alkyl, -CN, -OR_4a, =O, -C(O)R_4a, -NR_4bR_4c, or =NR_4a; wherein: each R_4a is independently H, C_1-6 alkyl, C_3-8 cycloalkyl, or -CN; each R_4b is independently H or C_1-6 alkyl; and each R_4c is independently H or C_1-6 alkyl.

12. The compound of any one of claims 1 through 9, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₄ is selected from the group consisting of optionally substituted piperidinyl, optionally substituted piperazinyl, optionally substituted tetrahydropyridinyl, optionally substituted dihydrothiopyranyl, and optionally substituted tetrahydrothiopyranyl.

13. The compound of any one of claims 1 through 9, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, wherein R₄ is

14. The compound of claim 1, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, selected from the group consisting of: N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(l-methyl-4- piperidyl)pyrido[3,4-J]pyrimidin-4-amine; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(4- methylpiperazin-l-yl)pyrido[3,4-J]pyrimidin-4-amine; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(l-methyl-3,6- dihydro-2J/-pyridin-4-yl)pyrido[3,4-J]pyrimidin-4-amine;

(R) -4-(4-((l-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8- methoxypyrido[3,4-d]pyrimidin-6-yl)-3,6-dihydro-2H-thiopyran 1,1 -di oxide; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(l,l-dioxothian-4-yl)-8- methoxy-pyrido[3,4-t/]pyrimidin-4-amine;

(R) -4-(4-((l-(3-(difluoromethyl)-2-fluorophenyl)ethyl)amino)-8- methoxypyrido[3,4-d]pyrimidin-6-yl)-4-fluorotetrahydro-2H-thiopyran 1,1 -di oxide; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(4-methoxy- l,l-dioxo-thian-4-yl)pyrido[3,4-d/]pyrimidin-4-amine; 4-[4-[[(17?)-l-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy- pyrido[3 ,4-d]pyrimidin-6-yl]- 1 , 1 -dioxo-thiane-4-carbonitrile;

6-(4-amino-l,l-dioxo-thian-4-yl)-N-[(1R )-1- [3-(difluoromethyl)-2-fluoro- phenyl]ethyl]-8-methoxy-pyrido[3,4-d]pyrimidin-4-amine; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(l, 1 -di oxo-3, 6-dihydro-2//- thiopyran-4-yl)-8-methoxy-2-methyl-pyrido[3,4-d]pyrimidin-4-amine; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(4-methyl-l,l- dioxo-thian-4-yl)pyrido[3,4-d]pyrimidin-4-amine;

1-[4-[4-[[ (1R)-l-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy- pyrido[3,4-d]pyrimidin-6-yl]-l-piperidyl]ethanone; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-(4-fluoro-l-methyl-4- piperidyl)-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine;

N- [ (1R)- 1 -[3 -(difluoromethyl)-2-fluoro-phenyl]ethyl]-6-( 1 -imino- 1 -oxo-3 ,6- dihydro- 2H -thiopyran-4-yl)-8-methoxy-pyrido[3,4-J]pyrimidin-4-amine; N-[(1R )-1- [3-(difluoromethyl)-2-fluoro-phenyl]ethyl]-8-methoxy-6-(l- methylimino-1 -oxo-3, 6-dihydro- 2H -thiopyran-4-yl)pyrido[3,4-J]pyrimidin-4-amine;

[4-[4-[[ (1R)-l-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy- pyrido[3,4-J]pyrimidin-6-yl]-l-oxo-3,6-dihydro- 2H -thiopyran-l-ylidene]cyanamide;

(R) -N-(l-(3-(difluoromethyl)-2-fluorophenyl)ethyl)-8-methoxy-6-(4- methoxypiperidin-4-yl)pyrido[3,4-d]pyrimidin-4-amine; and

1-[4-[4-[[ (1R)-l-[3-(difluoromethyl)-2-fluoro-phenyl]ethyl]amino]-8-methoxy- pyrido[3,4-J]pyrimidin-6-yl]-4-methoxy-l-piperidyl]ethanone.

15. A pharmaceutical composition comprising a compound of any one of claims 1-13, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof, and a pharmaceutically acceptable carrier.

16. A method of inhibiting S0S1 in a subject, comprising administering to the subject: a compound of any one of claims 1-14, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof; or a pharmaceutical composition of claim 15, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof.

17. A method of inhibiting the interaction of S0S1 and a RAS-family protein in a cell or inhibiting the interaction of S0S1 and RAC1 in a cell, comprising administering to the cell: a compound of any one of claims 1-14, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof; or a pharmaceutical composition of claim 15, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof.

18. A method of treating or preventing a disease, wherein treating or preventing the disease is characterized by inhibition of the interaction of S0S1 and a RAS-family protein or by inhibition of the interaction of S0S1 and RAC1, the method comprising administering to a subject in need thereof an effective amount of: a compound of any of claims 1-14, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof; or a pharmaceutical composition of claim 15, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof.

19. A method of treating or preventing cancer in a subject in need thereof, comprising administering to the subject an effective amount of: a compound of any of claims 1-14, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof; or a pharmaceutical composition of claim 15, or a pharmaceutically acceptable salt, solvate, isomer, stereoisomer, prodrug, or tautomer thereof.

20. The method of claim 18 or claim 19, wherein the disease or cancer is selected from the group consisting of pancreatic cancer, lung cancer, colorectal cancer, hematological cancer, cholangiocarcinoma, multiple myeloma, melanoma, uterine cancer, endometrial cancer, thyroid cancer, acute myeloid leukemia, bladder cancer, urothelial cancer, gastric cancer, cervical cancer, head and neck squamous cell carcinoma, diffuse large B cell lymphoma, esophageal cancer, chronic lymphocytic leukemia, hepatocellular cancer, breast cancer, ovarian cancer, prostate cancer, glioblastoma, renal cancer and sarcomas.

21. The method of claim 19 or claim 20, wherein the cancer comprises a Ras^MUT or an NF1^LOF mutation.

22. The method of claim 18, wherein the disease is a RASopathy.

23. The method of claim 22, wherein the RASopathy is selected from the group consisting of Neurofibromatosis type 1 (NF1), Noonan Syndrome (NS), Noonan Syndrome with Multiple Lentigines (NSML), Capillary Malformation- Arteriovenous Malformation Syndrome (CM-AVM), Costello Syndrome (CS), Cardio-Facio-Cutaneous Syndrome (CFC), Legius Syndrome, and Hereditary gingival fibromatosis.