Composition for inducing RAS GTP hydrolysis and use thereof

Abstract

This disclosure relates to therapies for treating cancer. In particular, this disclosure relates to a method for treating cancer in a subject requiring treatment for cancer, comprising administering to the subject a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound (which may optionally be administered in combination with additional therapeutic agents (e.g., RAS(OFF) inhibitors, RTK inhibitors, SHP2 inhibitors, or SOS1 inhibitors)), and also relates to a pharmaceutical composition comprising a therapeutically effective amount of the inhibitor(s), a kit comprising the composition, and a method of using the same.

Description

[Background technology]

[0001] Cancer remains one of the most deadly threats to human health. In the United States, cancer affects approximately 1.3 million new cases each year, is the second leading cause of death after heart disease, and accounts for about one in four deaths.

[0002] RAS (KRAS, NRAS, HRAS) proteins regulate cell proliferation and other cellular functions by converting between a guanosine triphosphate (GTP)-bound "on" state ("RAS(ON)") and a guanosine diphosphate (GDP)-bound "off" state ("RAS(OFF)"). The active state of RAS is bound to GTP, which is hydrolyzed to the GDP-bound inactive state. RAS proteins have a slow intrinsic hydrolysis rate (Non-Patent Literature 1), which is enhanced in the presence of RAS GTPase-activating protein (GAP). GDP-bound RAS can be converted to the active state via a slow exchange from GDP to GTP, enhanced by guanine nucleotide exchange factors (GEFs). One way in which oncogenic mutations in RAS increase the amount of GTP-bound RAS protein is by reducing the intrinsic hydrolysis rate and decreasing sensitivity to GAP-mediated hydrolysis enhancement. Based on these observations, RAS variants have historically been thought to be "constitutively" activated in cancer. The amount of active RAS can also increase via the activation of intracellular GEFs, which can be due to either mutations in upstream proteins (e.g., mutations in receptor tyrosine kinases) or non-mutagenic mechanisms (e.g., reactivation of adaptive pathways). In either case, elevated GTP-bound RAS levels lead to excessive cell proliferation.

[0003] KRAS G12CCovalent inhibitors of the "off" form of KRAS have shown promising antitumor activity in cancer patients with oncogenic mutations in KRAS. However, therapeutic inhibition of the RAS pathway may often prove ineffective in the long run, even if initially effective, potentially leading to hyperactivation of RAS pathway signaling or resistance to RAS(OFF) inhibitors through numerous mechanisms, including reactivation of RAS pathway signaling by disabling naturally operating negative feedback mechanisms in these pathways. As a result, cancer cells that were initially sensitive to such inhibitors may become resistant. Most KRAS variants are susceptible to inhibitors that preferentially target their inactive state, suggesting that KRAS variants retain the ability to hydrolyze GTP within cancer cells. These findings spur the exploration of pharmacological interventions that enhance GTP hydrolysis by KRAS variants.

[0004] Therefore, in this field, there is a need for compositions and methods that induce RAS GTP hydrolysis. Furthermore, there is a need for compositions and methods that enhance the sensitivity of cancer cells to RAS(OFF) inhibitors. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Westcover et al., Mol Cancer Res (2015) 13(9):1325-1335 [Overview of the project]

[0006] This disclosure provides compositions and uses thereof for treating RAS-related diseases or disorders (e.g., cancer) comprising RAS(ON)GTP hydrolysis-promoting compounds. For example, this disclosure provides combination therapies useful for treating cancer, comprising a combination of a RAS(ON)GTP hydrolysis-promoting compound and an additional therapeutic agent (e.g., a RAS(OFF) inhibitor or a RAS degrading agent that targets the RAS(OFF) state ("RAS(OFF) degrading agent"). In any embodiment of this specification using a RAS(OFF) inhibitor, a RAS(OFF) degrading agent may be used instead. In some embodiments, the combination comprises two or more therapeutic agents in addition to the RAS(ON)GTP hydrolysis-promoting compound (e.g., a RAS(OFF) inhibitor and a SHP2 inhibitor). In one embodiment, this disclosure is based at least in part on the observation that contacting cancer cells with a RAS(ON)GTP hydrolysis-promoting compound and a RAS(OFF) inhibitor results in a synergistic effect in reducing the viability of cancer cells.

[0007] In other embodiments, the Disclosure provides a method for treating cancer in a subject requiring cancer treatment, the method comprising administering to the subject a therapeutically effective amount of a RAS(ON)GTP hydrolytic compound and a RAS(OFF) inhibitor, wherein the cancer does not contain a RAS mutation at position 61. In some embodiments, the cancer contains a RAS mutation (for example, the RAS mutation is located at position 12 or 13). In some embodiments, the cancer is pancreatic cancer, colorectal cancer, non-small cell lung cancer, gastric cancer, esophageal cancer, ovarian cancer, or uterine cancer. In some embodiments, the cancer is RAS amplification (RAS AMP ) is characterized by. In some embodiments, RAS AMPは It is either wild-type RAS or mutant RAS.

[0008] In each of the above embodiments, the binding of the RAS(ON)GTP hydrolysis-promoting compound to RAS GTP (RAS(ON)) alters the position of glutamine 61 of RAS(ON) toward the gamma phosphate of GTP bound to the RAS(ON) protein, compared to its position in the absence of the RAS(ON)GTP hydrolysis-promoting compound, thereby increasing the GTP hydrolysis rate compared to the hydrolysis rate of RAS(ON) in the absence of the RAS(ON)GTP hydrolysis-promoting compound.

[0009] In some embodiments, the RAS(ON)GTP hydrolysis-promoting compound is a compound of formula Ia or Ib, or a pharmaceutically acceptable salt thereof. In exemplary embodiments, the RAS(ON)GTP hydrolysis-promoting compound is a compound listed in Table 1 or a pharmaceutically acceptable salt thereof.

[0010] In some embodiments, the KRAS(OFF) inhibitor is a KRAS(OFF) inhibitor. In some embodiments, the KRAS(OFF) inhibitor is a KRAS G12C It is an (OFF) inhibitor. In some embodiments, KRAS G12C (OFF) inhibitors are selected from the group consisting of AMG510 (sotrasib), MRTX849 (adagrasib), MRTX1257, GDC-6036 (divalasib), JDQ443 (opnurasib), ERAS-3490, LY3537982 (olomorasib), BI1823911, BPI-421286, JAB-3312, JAB-21000, JAB-21822 (glesilasib), D-1553, D3S-001, HBI-2438, HS-10370, MK-1084, YL-15293, BBO-8520 (ON / OFF inhibitor), FMC-376 (ON / OFF inhibitor), GEC255, and GFH925 (IBI351). In some embodiments, the KRAS(OFF) inhibitor is KRAS G12D It is an (OFF) inhibitor. In some embodiments, KRAS G12D(OFF) inhibitors are selected from the group consisting of MRTX1133, MRTX282, JAB-22000, ERAS-4, ERAS-5024, HRS-4642, BI-2852, ASP3082, TH-Z827, TH-7835, QTX-3046, GFH375 (VS-7375), INCB161734, and KD-8.

[0011] In some embodiments, the KRAS(OFF) inhibitor is a KRAS G12V (OFF) inhibitor. In certain embodiments, the KRAS G12V (OFF) inhibitor is JAB-23000.

[0012] In some embodiments, the KRAS(OFF) inhibitor is a pan-RAS(OFF) inhibitor. In certain embodiments, the pan-RAS(OFF) inhibitor is JAB-23400, JAB-23425, BI-2493, BI-2865, QTX-3034 (preferably G12D), QTX3544 (preferably G12V), ZG2001, BBO-a, BBO-B, or pan KRas-IN-1.

[0013] In some embodiments, the RAS(ON) GTP hydrolysis promoting compound and the RAS(OFF) inhibitor are administered on the same day. In some embodiments, the RAS(ON) GTP hydrolysis promoting compound and the RAS(OFF) inhibitor are administered simultaneously or sequentially. In some embodiments, the RAS(ON) GTP hydrolysis promoting compound and the RAS(OFF) inhibitor are administered on different days.

[0014] In each of the preceding embodiments, the method may further include administering additional anticancer therapy. In some embodiments, the additional anticancer therapy is an EGFR inhibitor, SHP2 inhibitor, SOS1 inhibitor, Raf inhibitor, MEK inhibitor, ERK inhibitor, PI3K inhibitor, PTEN inhibitor, AKT inhibitor, mTORC1 inhibitor, BRAF inhibitor, immune checkpoint inhibitor, CDK4 / 6 inhibitor, HER2 inhibitor, or a combination thereof. In some embodiments, the immune checkpoint inhibitor is a PD-L1 inhibitor or a PD-1 inhibitor. In some embodiments, the additional therapy is an SHP2 inhibitor or a SOS1 inhibitor. In each of the preceding embodiments, the SOS1 inhibitor is RMC-5845, RMC-4948, RMC-0331, BI-1701963, BI-3406, SDR5, MRTX0902, BAY-293, or any combination thereof. In each of the preceding embodiments, the SHP2 inhibitor is SHP099, TNO155, RMC-4550, RMC-4630, JAB-3068, JAB-3312, RLY-1971, ERAS-601, SH3809, PF-07284892, BBP-398, or any combination thereof.

[0015] In some embodiments, the cancer in question progresses under the administration of a RAS(OFF) inhibitor (e.g., the subject is administered a RAS(OFF) inhibitor in the absence of a RAS(ON)GTP hydrolytic compound). In some embodiments, the subject is treated with a RAS(OFF) inhibitor (e.g., the subject has been previously treated with a RAS(OFF) inhibitor, for example, before administration of a RAS(ON)GTP hydrolytic compound). In some embodiments, the subject has acquired resistance to a RAS(OFF) inhibitor (e.g., the subject has acquired a mutation that confers resistance to a RAS(OFF) inhibitor, for example, before administration of a RAS(ON)GTP hydrolytic compound).

[0016] In other embodiments, the Disclosure provides a method for treating RAS protein-related disorders in subjects requiring treatment for such disorders, the method generally comprising administering a therapeutically effective amount of a RAS(ON)GTP hydrolytic compound to the subject.

[0017] In further embodiments, the disclosure provides a method for treating RASopathy in subjects requiring treatment of RASopathy, the method generally comprising administering a therapeutically effective amount of a RAS(ON)GTP hydrolytic compound to the subject. In some embodiments, the RASopathy is cardiac-facial-cutaneous syndrome, Costello syndrome, Regius syndrome, neurofibromatosis type 1, Noonan syndrome, or capillary malformation-arteriovenous malformation syndrome. In some embodiments, binding of the RAS(ON)GTP hydrolytic compound to RAS(ON) alters the position of glutamine 61 of RAS(ON) toward the gamma phosphate of GTP bound to the RAS(ON) protein, compared to its position in the absence of the GTP hydrolytic compound, thereby increasing the rate of GTP hydrolysis compared to the rate of hydrolysis of RAS(ON) in the absence of the RAS(ON)GTP hydrolytic compound. In some embodiments, the method may further include additional RASopathy therapies. In some embodiments, additional RASopathy therapies include EGFR inhibitors, SHP2 inhibitors, SOS1 inhibitors, Raf inhibitors, MEK inhibitors, ERK inhibitors, PI3K inhibitors, PTEN inhibitors, AKT inhibitors, mTORC1 inhibitors, BRAF inhibitors, CDK4 / 6 inhibitors, HER2 inhibitors, or combinations thereof.

[0018] In further embodiments, the disclosure provides a method for inhibiting RAS activity in cells, the method generally comprising contacting cells in which inhibition of RAS activity is desired with an effective amount of a RAS(ON)GTP hydrolytic compound. In some embodiments, the RAS(ON)GTP hydrolytic compound is provided in combination with a RAS(OFF) inhibitor, the RAS(ON)GTP hydrolytic compound synergistically increases sensitivity to the RAS(OFF) inhibitor.

[0019] In other embodiments, the Disclosure provides a method for increasing the sensitivity of cells to a RAS(OFF) inhibitor, the method generally comprising contacting cells for which increased sensitivity to a RAS(OFF) inhibitor is desired with an effective amount of a RAS(ON)GTP hydrolytic compound, wherein the RAS(ON)GTP hydrolytic compound synergistically increases the sensitivity of cells to a RAS(OFF) inhibitor.

[0020] In other embodiments, this disclosure relates to intracellular KRAS G12C KRAS at the cysteine ​​residue at position 12 G12C This provides a method for increasing the crosslinking rate of (OFF) inhibitors, comprising contacting cells with an effective amount of a RAS(ON)GTP hydrolysis-promoting compound.

[0021] In other embodiments, the disclosure provides a pharmaceutical composition comprising a therapeutically effective amount of a RAS(ON)GTP hydrolytic compound. In some embodiments, the pharmaceutical composition comprises a RAS(ON)GTP hydrolytic compound and a RAS(OFF) inhibitor.

[0022] In other embodiments, the Disclosure provides a kit comprising a) a RAS(ON)GTP hydrolysis-promoting compound and b) a RAS(OFF) inhibitor. In some embodiments, the kit further includes a package insert containing instructions for the administration of the pharmaceutical composition(s).

[0023] Any limitations discussed in relation to one embodiment of this disclosure are specifically intended to be applicable to any other embodiment of this disclosure. Furthermore, any compound or composition of this disclosure may be used in any manner of this disclosure, and any compound or composition of this disclosure may be produced or utilized by any manner of this disclosure. [Brief explanation of the drawing]

[0024] [Figure 1]The graphs illustrate the characterization of RAS GTP hydrolysis activation in various RAS mutants using a representative moderate-level RAS(ON)GTP hydrolysis-promoting compound (compound E). Except for those with mutations in the Q61 residue necessary for catalytic hydrolysis activity, all RAS mutants exhibit increased hydrolysis in the presence of the RAS(ON)GTP hydrolysis-promoting compound. [Figure 2] This graph shows the characterization of RAS-GTP hydrolysis activation by various compounds. Compound F represents a class that does not activate GTP hydrolysis by RAS, while the others show varying degrees of RAS-GTP hydrolysis activation. [Figure 3A] This shows the optimal fit of the phospho-(Thr202 / Tyr204;Thr185 / Tyr187)-ERK1 / 2 concentration-response curve of compound C in the KRASG12D mutant cell line AsPC-1, in the presence or absence of 1 μM RMC-4550. [Figure 3B] This shows the optimal fit of the phospho-(Thr202 / Tyr204;Thr185 / Tyr187)-ERK1 / 2 concentration-response curve of compound F in the KRASG12D mutant cell line AsPC-1, in the presence or absence of 1 μM RMC-4550. [Figure 3C] This shows the optimal fit of the cell viability concentration-response curves for compound C in the KRASG12D mutant cell line AsPC-1, in the presence or absence of 1 μM RMC-4550. [Figure 3D] This shows the optimal fit of the cell viability concentration-response curves for compound F in the KRASG12D mutant cell line AsPC-1, in the presence or absence of 1 μM RMC-4550. [Figure 4A] This shows the optimal fit of the cell viability concentration-response curves for compound A and MRTX1133 in the KRASG12D mutant cell line AsPC-1. [Figure 4B] This paper demonstrates the HSA synergy model of compound A and MRTX1133 in the KRASG12D mutant cell line AsPC-1. [Figure 4C]This bar graph shows key points regarding synergistic drug combinations between compound A and MRTX1133 in the KRASG12D mutant cell line AsPC-1. [Figure 4D] This shows the optimal fit of the cell viability concentration-response curves for compound A and MRTX-282 in the KRASG12D mutant cell line AsPC-1. [Figure 4E] This paper demonstrates the HSA synergy model of compound A and MRTX-282 in the KRASG12D mutant cell line AsPC-1. [Figure 4F] This bar graph shows key points regarding the synergistic drug combination of compound A and MRTX-282 in the KRASG12D mutant cell line AsPC-1. [Figure 4G] This shows the optimal fit of the concentration-response curves for compound D and MRTX1133 in the KRASG12D mutant cell line AsPC-1. [Figure 4H] This paper demonstrates the HSA synergy model of compound D and MRTX1133 in the KRASG12D mutant cell line AsPC-1. [Figure 4I] This bar graph shows key points regarding the synergistic drug combination of compound D and MRTX1133 in the KRASG12D mutant cell line AsPC-1. [Figure 5A] This shows the optimal fit of the cell viability concentration-response curves for compound A and MRTX1133 in the KRASG12D mutant cell line HPAC. [Figure 5B] This paper demonstrates the HSA synergy model of compound A and MRTX1133 in the KRASG12D mutant cell line HPAC. [Figure 5C] This bar graph shows key points regarding the synergistic drug combination of compound A and MRTX1133 in the KRASG12D mutant cell line HPAC. [Figure 5D] This shows the optimal fit of the cell viability concentration-response curves for compound A and MRTX-282 in the KRASG12D mutant cell line HPAC. [Figure 5E]This paper demonstrates the HSA synergy model of compound A and MRTX-282 in the KRASG12D mutant cell line HPAC. [Figure 5F] This bar graph shows key points regarding the synergistic drug combination of compound A and MRTX-282 in the KRASG12D mutant cell line HPAC. [Figure 6A] This shows the optimal fit of the cell viability concentration-response curves for compound A and MRTX1133 in the KRASG12D mutant cell line Gp2D. [Figure 6B] This paper demonstrates the HSA synergy model of compound A and MRTX1133 in the KRASG12D mutant cell line Gp2D. [Figure 6C] This bar graph shows key points regarding the synergistic drug combination of compound A and MRTX1133 in the KRASG12D mutant cell line Gp2D. [Figure 6D] This shows the optimal fit of the cell viability concentration-response curves for compound A and MRTX-282 in the KRASG12D mutant cell line Gp2D. [Figure 6E] This paper demonstrates the HSA synergy model of compound A and MRTX-282 in the KRASG12D mutant cell line Gp2D. [Figure 6F] This bar graph shows key points regarding the synergistic drug combination of compound A and MRTX-282 in the KRASG12D mutant cell line Gp2D. [Figure 7A] This shows the optimal fit of the cell viability concentration-response curves for compounds D and MRTX-282 in the RAS wild-type cell line HaCat. [Figure 7B] This paper demonstrates the HSA synergy model of compound D and MRTX-282 in the RAS wild-type cell line HaCat. [Figure 7C] This bar graph shows representative points regarding the non-synergistic drug combination of compound D and MRTX-282 in the RAS wild-type cell line HaCat. [Figure 7D] This shows the optimal fit of the cell viability concentration-response curves for compounds D and MRTX-282 in the RAS wild-type cell line HaCat. [Figure 7E] This paper demonstrates the HSA synergy model of compound D and MRTX-282 in the RAS wild-type cell line HaCat. [Figure 7F] This bar graph shows representative points regarding the non-synergistic drug combination of compound D and MRTX-282 in the RAS wild-type cell line HaCat. [Figure 8A] This shows the optimal fit of the cell viability concentration-response curves for compounds A and AMG510 in the KRASG12C mutant cell line MiaPaCa2. [Figure 8B] This paper demonstrates the HSA synergy model of compounds A and AMG510 in the KRASG12C mutant cell line MiaPaCa2. [Figure 8C] This bar graph shows key points regarding the synergistic drug combination of compound A and AMG510 in the KRASG12C mutant cell line MiaPaCa2. [Figure 8D] This shows the optimal fit of the cell viability concentration-response curves for compound A and MRTX849 in the KRASG12C mutant cell line MiaPaCa2. [Figure 8E] This paper demonstrates the HSA synergy model of compound A and MRTX849 in the KRASG12C mutant cell line MiaPaCa2. [Figure 8F] This bar graph shows key points regarding the synergistic drug combination of compound A and MRTX849 in the KRASG12C mutant cell line MiaPaCa2. [Figure 9A] This is an immunoblot of MiaPaCa2 cells showing whole RAS isolated into wild-type HRAS and NRAS, free KRASG12C, and KRASG12C covalently modified (crosslinked) by sotrasib. [Figure 9B] This bar graph shows the ratio of covalent modification of KRASG12C in MiaPaCa2 cells, either with sotrasib alone or in combination with compound A. [Figure 10A] This shows the optimal fit of the intracellular RAS-RAF complex reporter assay concentration-response curves for compound E in different KRAS mutant proteins. [Figure 10B]This shows the optimal fit of the intracellular RAS-RAF complex reporter assay concentration-response curves for pan KRAS-IN-1 in different KRAS mutant proteins. [Figure 10C] This shows the optimal fit of the concentration-response curves for the intracellular KRASG12V-RAF complex reporter assay of pan KRAS-IN-1 in the presence or absence of 1 μM compound D. [Figure 11A] This shows the optimal fit of the cell viability concentration-response curves for compounds D and pan KRAS-IN-1 in the KRASG12D mutant cell line AsPC-1. [Figure 11B] This paper demonstrates the HSA synergy model of compound D and pan KRAS-IN-1 in the KRASG12D mutant cell line AsPC-1. [Figure 11C] This bar graph shows key points regarding the synergistic drug combination of compound D and pan KRAS-IN-1 in the KRASG12D mutant cell line AsPC-1. [Figure 11D] This shows the optimal fit of the cell viability concentration-response curves for compound C and pan KRAS-IN-1 in the KRASG12D mutant cell line AsPC-1. [Figure 11E] This paper demonstrates the HSA synergy model of compound C and pan KRAS-IN-1 in the KRASG12D mutant cell line AsPC-1. [Figure 11F] This bar graph shows key points regarding the synergistic drug combination of compound C and pan KRAS-IN-1 in the KRASG12D mutant cell line AsPC-1. [Figure 12A] This shows the optimal fit of the cell viability concentration-response curves for compounds D and pan KRAS-IN-1 in the KRASG12D mutant cell line Capan-2. [Figure 12B] This paper demonstrates the HSA synergy model of compound D and pan KRAS-IN-1 in the KRASG12D mutant cell line Capan-2. [Figure 12C]This bar graph shows key points regarding the synergistic drug combination of compound D and pan KRAS-IN-1 in the KRASG12D mutant cell line Capan-2. [Figure 12D] This shows the optimal fit of the cell viability concentration-response curves for compounds C and pan KRAS-IN-1 in the KRASG12D mutant cell line Capan-2. [Figure 12E] This paper demonstrates the HSA synergy model of compound C and pan KRAS-IN-1 in the KRASG12D mutant cell line Capan-2. [Figure 12F] This bar graph shows key points regarding the synergistic drug combination of compound C and pan KRAS-IN-1 in the KRASG12D mutant cell line Capan-2. [Figure 13A] This shows the optimal fit of the cell viability concentration-response curves for compound D and pan KRAS-IN-1 in the KRASG12D mutant cell line NCI-H358. [Figure 13B] This paper demonstrates the HSA synergy model of compound D and pan KRAS-IN-1 in the KRASG12D mutant cell line NCI-H358. [Figure 13C] This bar graph shows key points regarding the synergistic drug combination of compound D and pan KRAS-IN-1 in the KRASG12D mutant cell line NCI-H358. [Figure 13D] This shows the optimal fit of the cell viability concentration-response curves for compound C and pan KRAS-IN-1 in the KRASG12D mutant cell line NCI-H358. [Figure 13E] This paper demonstrates the HSA synergy model of compound C and pan KRAS-IN-1 in the KRASG12D mutant cell line NCI-H358. [Figure 13F] This bar graph shows key points regarding the synergistic drug combination of compound C and pan KRAS-IN-1 in the KRASG12D mutant cell line NCI-H358. [Figure 14A]This shows the optimal fit of the cell viability concentration-response curves for compound D and pan KRAS-IN-1 in the KRASG12D mutant cell line PSN-1. [Figure 14B] This paper demonstrates the HSA synergy model of compound D and pan KRAS-IN-1 in the KRASG12D mutant cell line PSN-1. [Figure 14C] This bar graph shows key points regarding the synergistic drug combination of compound D and pan KRAS-IN-1 in the KRASG12D mutant cell line PSN-1. [Figure 14D] This shows the optimal fit of the cell viability concentration-response curves for compound C and pan KRAS-IN-1 in the KRASG12D mutant cell line PSN-1. [Figure 14E] This paper demonstrates the HSA synergy model of compound C and pan KRAS-IN-1 in the KRASG12D mutant cell line PSN-1. [Figure 14F] This bar graph shows key points regarding the synergistic drug combination of compound C and pan KRAS-IN-1 in the KRASG12D mutant cell line PSN-1. [Figure 15] This demonstrates that the orientation of the KRAS residue Q61 differs between exemplary non-RAS(ON)GTP hydrolytic compounds containing phenol at position A of formula Ia or formula Ib type compounds and exemplary RAS(ON)GTP hydrolytic compounds containing thiazole at the equivalent position. [Figure 16] A schematic diagram illustrating the mechanistic basis for the synergistic benefits of combining a RAS(ON)GTP hydrolysis-promoting compound and a RAS nucleotide exchange inhibitor, which accelerate GTP hydrolysis, is shown. [Figure 17]This schematic diagram illustrates the mechanistic basis for the mechanistic advantages of synergistic combinations between RAS(ON)GTP hydrolysis-promoting compounds, which accelerate GTP hydrolysis, and RAS inhibitors, which bind with high affinity to the RAS(OFF) [GDP-bound] state. Here, the RAS(ON)GTP hydrolysis-promoting compound is shown complexed with cyclophyllin A (CypA), which then binds to RAS(ON) to form a tripocomplex. This complex then catalyzes the hydrolysis of GTP, converting RAS(ON) to RAS(OFF). The resulting RAS(OFF) is then bound by the RAS(OFF) inhibitor. [Modes for carrying out the invention]

[0025] This disclosure generally relates to compositions and methods for modulating RAS activity for the treatment of cancer. In particular, this disclosure provides therapeutic methods for cancers having RAS mutations. In each embodiment, the cancer does not have a mutation at residue Q61. This disclosure provides a method for treating cancer in a subject requiring cancer treatment, the method comprising administering to the subject a therapeutically effective amount of a RAS(ON)GTP hydrolytic compound, or a pharmaceutically acceptable salt or pharmaceutical composition thereof, in combination with an optional RAS(OFF) inhibitor, or a pharmaceutically acceptable salt or pharmaceutical composition thereof. In any embodiment of this specification using an RAS(OFF) inhibitor, a RAS(OFF) degrading agent may be used instead. This disclosure also provides a method comprising a RAS(ON)GTP hydrolytic compound and additional therapeutic agents (e.g., SOS1 inhibitors, SHP2 inhibitors, RTK inhibitors, and / or additional RAS inhibitors). This disclosure also provides a pharmaceutical composition comprising a therapeutically effective amount of an inhibitor, a kit comprising the composition, and a method for using the same.

[0026] Oncogenic RAS mutations that increase the proportion of RAS protein in the GTP-bound state limit the amount of GDP-bound RAS to which RAS(OFF) inhibitors can bind. While we do not wish to be bound by theory, we hypothesize that the compounds disclosed herein enhance the potency of RAS(OFF) inhibitors by increasing the rate of GTP hydrolysis by oncogenic RAS and / or wild-type RAS, thereby increasing the level of GDP-bound RAS. The RAS(ON)GTP hydrolysis-promoting compounds disclosed herein are due to the mutant RAS isoforms having an inherent GAP deficiency, resulting in RAS WT In comparison, RAS G12X It exhibits higher selectivity for RAS(ON)GTP hydrolysis-promoting compounds disclosed herein, because target inhibition is by a catalytic mechanism rather than a stoichiometric mechanism, RAS AMP (For example, mutant RAS AMP It is useful in the context of ). Furthermore, the unique profiles of the RAS(ON)GTP hydrolytic compounds disclosed herein are useful for improving tolerability, including in the context of combination therapy, particularly intra-route combinations.

[0027] Overall method In the practice of this disclosure, unless otherwise specified, prior arts in cell culture, molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry, and immunology are used, and these are within the scope of the art. Such techniques are described in Molecular Cloning: A Laboratory Manual, third edition (Sambrook et al., 2001); Cold Spring Harbor Press; Oligonucleotide Synthesis (P. Herdewijn, ed., 2004); Animal Cell Culture (RIFreshney), ed., 1987); CCBlackwell, eds.); Gene transfer Vectors for Mammalian Cells (JMMiller & MPCalos, eds., 1987); Current Protocols in Molecular Biology (FMAusubel et al., eds., 1987); PCR: The Polymerase Chain Reaction, (Mullis et al., eds., 1994); Current Protocols in Immunology (JEColigan et al. al., eds., 1991);Short Protocols in Molecular Biology(Wiley and Sons, 1999); Manual of Clinical Laboratory Immunology (B. Detrick, NRRose, and JDFolds eds., 2006); Immunochemical Protocols (J. Pound, ed., 2003); Lab Manual in Biochemistry: Immunology and Biotechnology (A. Nigam and A. Ayyagari, eds.This is fully explained in the following literature: 2007); Immunology Methods Manual: The Comprehensive Sourcebook of Techniques (Ivan Lefkovits, ed., 1996); Using Antibodies: A Laboratory Manual (E. Harlow and D. Lane, eds., 1988); and other publications.

[0028] definition In this application, unless otherwise clearly indicated by the context, (i) the term “one (a)” means “one or more”; (ii) the term “or” is used to mean “and / or” unless it is explicitly indicated that it means only alternative expressions or that such alternative expressions are mutually exclusive, however this disclosure supports the definitions that refer only to alternative expressions and to “and / or”; (iii) the terms “comprising” and “including” are understood to encompass itemized components or processes, whether presented by themselves or together with one or more additional components or processes; and (iv) where a scope is indicated, it includes endpoints.

[0029] As used herein, the term “approximately” is used to indicate that a value includes the standard deviation of the error of the device or method used to determine the value. In certain embodiments, unless otherwise specified or otherwise evident from the context (for example, if such a number may exceed 100% of a possible value), the term “approximately” 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%, and 1% or less in either direction (above or below) of the stated value.

[0030] As used herein, the term “adjacent” in the context of describing adjacent atoms refers to divalent atoms directly bonded by a covalent bond.

[0031] Throughout this specification, unless the context requires otherwise, the words “comprise,” “comprises,” and “comprising” mean to include the steps or elements, or groups of steps or elements, described, but not to exclude any other steps or elements, or groups of steps or elements. “Consisting of” means to include, and be limited to, everything that follows the phrase “consisting of.” Thus, the phrase “consisting of” indicates that the listed elements are necessary or essential, and other elements may or may not be present. “Consisting essentially of” means to include any elements listed after this phrase, and, regarding other elements, to be limited to those that do not interfere with or contribute to the activity or effect specified in the disclosure of the listed elements. Thus, the phrase “consisting essentially of” indicates that the listed elements are necessary or essential, but other elements are optional, and may or may not be present, depending on whether they substantially affect the activity or effect of the listed elements.

[0032] As used herein, “compounds of the disclosure” and similar terms mean, whether expressly stated or not, the compounds of formulas Ia and lb and their subformulas, as well as the compounds of Tables 1 and 2, in addition to their salts (e.g., pharmaceutically acceptable salts), solvates, hydrates, stereoisomers (including atropisomers), and tautomers, as described herein, and the RAS(ON)GTP hydrolysis-promoting compounds.

[0033] Those skilled in the art will understand that certain compounds described herein may exist in one or more different isomeric forms (e.g., stereoisomers, geometric isomers, atropisomers, tautomers) or isotopic forms (e.g., one or more atoms, such as deuterium-substituted hydrogen, are substituted with isotopes of different atoms). Unless otherwise specified or made clear from the context, the described structures can be understood to represent any such isomeric or isotopic forms, individually or in combination.

[0034] The compounds described herein may be asymmetric (e.g., having one or more stereocenters). Unless otherwise indicated, all stereoisomers, including enantiomers and diastereomers, are intended. Compounds of this disclosure containing asymmetrically substituted carbon atoms may be isolated in optically active forms or in racemic forms. Methods for preparing optically active forms from optically active starting materials are known in the art, for example, by resolution of racemic mixtures or stereoselective synthesis. Many geometric isomers, such as olefins and C=N double bonds, may also exist among the compounds described herein, and all such stable isomers are intended in this disclosure. Cis and trans geometric isomers of the compounds of this disclosure are described and may be isolated as mixtures of isomers or as separated isomers.

[0035] In some embodiments, one or more compounds described herein may exist in different tautomerized forms. As will be apparent from the context, unless explicitly excluded, references to such compounds encompass all such tautomerized forms. In some embodiments, the tautomerized form arises from the exchange of a single bond with an adjacent double bond and the accompanying transfer of a proton. In certain embodiments, the tautomerized form may be a prototropic tautomer, which is a protonated state of an isomer having the same empirical formula and total charge as the reference form. Examples of moieties having a prototropic tautomerized form include ketone-enol pairs, amide-imoid acid pairs, lactam-lactim pairs, amide-imoid acid pairs, enamine-imine pairs, and cyclic forms in which a proton can occupy two or more positions in a heterocyclic system, such as 1H- and 3H-imidazoles, 1H-, 2H-, and 4H-1,2,4-triazoles, 1H- and 2H-isoindoles, and 1H- and 2H-pyrazoles. In some embodiments, the tautomer morphs are in equilibrium or can be sterically fixed to one morph by appropriate substitution. In certain embodiments, the tautomer morphs arise from acetal interconversion.

[0036] Unless otherwise specified, the structures shown herein also mean that they contain different compounds only in the presence of one or more isotopic enriched atoms. Exemplary isotopes that can be incorporated into the compounds of this disclosure are: 2 H, 3 H, 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 32 P, 33 P, 35 S, 18 F, 36 Cl, 123 I, and 125 It contains isotopes such as hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, and iodine, such as I. Isotope-labeled compounds (e.g.,3 H and 14 Those labeled with 1C may be useful in tissue distribution assays of compounds or substrates. Tritium labeling (i.e., 3 H), and carbon-14 (i.e., 14 C) Isotopes can be useful due to their ease of preparation and detection. Furthermore, heavier isotopes, such as deuterium (i.e., 2 Substitution with H, etc., can lead to increased metabolic stability, resulting in certain therapeutic benefits (e.g., longer in vivo half-life or reduced dosage). In some embodiments, one or more hydrogen atoms are 2 H or 3 Replaced by H, or one or more carbon atoms 13 C or 14 It is replaced by carbon-rich carbon. 15 O, 13 N, 11 C, and 18 Positron-emitting isotopes such as fluorine (F) are useful in positron emission tomography (PET) studies to test substrate receptor occupancy. The preparation of isotopically labeled compounds is known to those skilled in the art. For example, isotopically labeled compounds can be prepared by replacing non-isotopic labeling reagents with isotopic labeling reagents, generally following procedures similar to those disclosed for the compounds of this disclosure as described herein.

[0037] Non-limiting examples of a compound in this disclosure that may include one or more deuterium substitutions (where any position "R" may be deuterium (D)) are: [ka] There are additional examples. [ka] These parts, similar to R 1 One example is the deuteration of the mold portion, where R 1The definition of is found herein. Deuteration of a portion of substituent W in the compounds of this disclosure is also contemplated, where W is defined herein (see, for example, formula Ib and its subformulas, and specific examples of W described herein). Furthermore, deuteration of any available position in any A portion of the compounds of the formulas described herein is also contemplated. Moreover, deuterium substitution may also occur at the linker position of the compounds of this disclosure.

[0038] In further embodiments, silylation substitutions in the linker, as follows, are also considered: [ka]

[0039] As is known from the prior art, many chemical substances can exist in various different solid forms, such as amorphous or crystalline forms (e.g., polymorphs, hydrates, solvates). In some embodiments, the compounds of this disclosure can be used in any such form, including any solid form. In some embodiments, the compounds described or explained herein can be provided or used in hydrate or solvate form.

[0040] In various parts of this specification, substituents of the compounds of this disclosure are disclosed in groups or ranges. This disclosure is specifically intended to include each individual partial combination of members of such groups and ranges. For example, the term "C1-C6 alkyl" is specifically intended to disclose methyl, ethyl, C3 alkyl, C4 alkyl, C5 alkyl, and C6 alkyl individually. Furthermore, where a compound includes multiple positions in which substituents are disclosed in groups or ranges, this disclosure is intended to include individual compounds and groups of compounds (e.g., classes and subclasses) including each and all individual member subcombinations at each position, unless otherwise specified.

[0041] The term “optionally substituted X” (e.g., “optionally substituted alkyl”) is intended to be equivalent to “X which is optionally substituted” (e.g., “alkyl which is optionally substituted”). It is not intended to mean that feature “X” (e.g., alkyl) itself is optional. As described herein, a particular compound of interest may contain one or more “optionally substituted” moieties. Generally, the term “substituted” means that one or more hydrogens of a given moiety are replaced with a suitable substituent, e.g., one of the substituents or groups described herein, whether preceded by the term “optionally”. Unless otherwise indicated, an “optionally substituted” group may have a suitable substituent at each of its substitutable positions, and if two or more positions of any given structure can be replaced with two or more substituents selected from a particular group, the substituents are either the same at all positions or different at all positions. For example, in the term "optionally substituted C1-C6 alkyl-C2-C9 heteroaryl," the alkyl moiety, the heteroaryl moiety, or both may be optionally substituted. The substituent combinations envisioned in this disclosure preferably result in the formation of stable or chemically suitable compounds. As used herein, the term "stable" refers to a compound that remains substantially unchanged when subjected to conditions anticipating the production, detection, and, in certain embodiments, their recovery, purification, and use for one or more purposes disclosed herein.

[0042] Suitable monovalent substituents on the substitutable carbon atoms of the "optionally substituted" group are, independently, deuterium; halogen; -(CH2)0-4R°; -(CH2)0-4OR°; -O(CH2)0-4Ro; -O-(CH2)0-4C(O)OR°; -(CH2)0-4CH(OR°)2; -(CH2)0-4SR°; -(CH2)0-4Ph[may be substituted with R°]; -(CH2)0-4O(CH2)0-1Ph[may be substituted with R°]; -CH=CHPh[may be substituted with R°]; -(CH2)0-4O(CH2)0-1-pyridyl[may be substituted with R°]; 4-8 membered saturated or unsaturated heterocycloalkyl (e.g., pyridyl); 3 ~8-membered saturated or unsaturated cycloalkyl (e.g., cyclopropyl, cyclobutyl, or cyclopentyl);-NO2;-CN;-N3;-(CH2)O-4N(R°)2;-(CH2)O-4N(R°)C(O)R°;-N(R°)C(S)R°;-(CH2)O-4N(R°)C(O)NR°2;-N(R°)C( S)NR°2;-(CH2)0-4N(R°)C(O)OR°;-N(R°)N(R°)C(O)R°;-N(R°)N(R°)C(O)NR°2;-N(R° )N(R°)C(O)OR°;-(CH2)0-4C(O)R°;-C(S)R°;-(CH2)0-4C(O)OR°;-(CH2)0-4-C(O)-N(R o )2;-(CH2)O-4-C(O)-N(R o )-S(O)2-R o;-C(NCN)NR°2;-(CH2)0-4C(O)SR°;-(CH2)0-4C(O)OSiR°3;-(CH2)0-4OC(O)R°;-OC(O)(CH2)0-4SR°;-SC(S)SR°;-(CH2)0- 4SC(O)R°;-(CH2)0-4C(O)NR°2;-C(S)NR°2;-C(S)SR°;-(CH2)0-4OC(O)NR°2;-C(O)N(OR°)R°;-C(O)C(O)R°;-C(O)CH2C(O)R °;-C(NOR°)R°;-(CH2)0-4SSR°;-(CH2)0-4S(O)2R°;-(CH2)0-4S(O)2OR°;-(CH2)0-4OS(O)2R°;-S(O)2NR°2;-(CH2)0-4S(O) R°;-N(R°)S(O)2NR°2;-N(R°)S(O)2R°;-N(OR°)R°;-C(NOR°)NR°2;-C(NH)NR°2;-P(O)2R°;-P(O)R°2;-P(O)(OR°)2;-OP(O)R °2;-OP(O)(OR°)2;-OP(O)(OR°)R°, -SiR°3;-(C1-4 linear or branched alkylene)ON(R°)2; or -(C1-4 linear or branched alkylene)C(O)ON(R°)2, where each R° may be substituted as defined below, independently of hydrogen, -C1-6 aliphatic, -CH2Ph, -O(CH2)0-1Ph, -CH2-(5-6 membered heteroaryl ring), or nitrogen, oxygen or sulfur. These may be 3-6 member saturated, partially unsaturated, or aryl rings having 0-4 heteroatoms independently selected, or, notwithstanding the above definition, two independently existing R° may, together with the intervening atom(s), form a 3-12 member saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, and these may be substituted as defined below.

[0043] A suitable monovalent substituent on R° (or a ring formed by combining two independently existing R° atoms with the intervening atom) is, independently, a halogen, -(CH2)0-2R ● ,-(HaroR ●), -(CH2)O-2OH, -(CH2)O-2OR ● ,-(CH2)0-2CH(OR ● )2;-O(HaroR ● ), -CN, -N3, -(CH2)0-2C(O)R ● , -(CH2)0-2C(O)OH, -(CH2)0-2C(O)OR ● ,-(CH2)0-2SR ● , -(CH2)0-2SH, -(CH2)0-2NH2, -(CH2)0-2NHR ● ,-(CH2)0-2NR ● 2, -NO2, -SiR ● 3. -OSiR ● 3. -C(O)SR ● 、 -(C1-4 straight-chain or branched-chain alkylene)C(O)OR ● , or -SSR ● It may also be the case that, in the formula, each R ● It is either unsubstituted or, if preceded by "halo", substituted by only one or more halogens, and independently selected from C1-4 aliphatic, -CH2Ph, -O(CH2)0-1Ph, or a 5-6 member saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Preferred divalent substituents on the saturated carbon atom of R° include =O and =S.

[0044] Suitable divalent substituents on the saturated carbon atoms of the "optionally substituted" groups include: =O, =S, =NNR * 2. =NNHC(O)R * ,=NNHC(O)OR * ,=NNHS(O)2R * ,=NR * 、=NOR * , -O(C(R * 2))2-3O-, or -S(C(R * 2))2-3S-(In the formula, each R exists independently. *is selected from hydrogen, a C1-6 aliphatic group that may be substituted as defined below, or an unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl ring having from 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur). Suitable divalent substituents bonded to an adjacent substitutable carbon of an "optionally substituted" group include -O(CR * 2)2-3O-, wherein each independently occurring R * is selected from hydrogen, a C1-6 aliphatic group that may be substituted as defined below, or an unsubstituted 5- to 6-membered saturated, partially unsaturated, or aryl ring having from 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur).

[0045] R * Suitable substituents on the aliphatic group of are halogen, -R ● , -(haloR ● ), -OH, -OR ● , -O(haloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2, or -NO2, wherein each R ● is unsubstituted or, when "halo" precedes, is substituted only by one or more halogens and is independently a C1-4 aliphatic group, -CH2Ph, -O(CH2)0-1Ph, or a 5- to 6-membered saturated, partially unsaturated, or aryl ring having from 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur).

[0046] Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include -R † , -NR † 2, -C(O)R † , -C(O)OR † , -C(O)C(O)R[[ID=;39]] † , -C(O)CH2C(O)R † 2, -S(O)2R † , -S(O)2NR † 2, -C(S)NR †2. -C(NH)NR † 2. or -N(R † )S(O)2R † are exemplified; wherein each R † is independently hydrogen, a C1-6 aliphatic group which may be substituted as defined below, an unsubstituted -OPh, or an unsubstituted 3-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, regardless of the above definition, two independently present R † together with the atom(s) intervening therebetween form an unsubstituted 3-12 membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0047] R † suitable substituents on the aliphatic group of are independently halogen, -R ● , -(haloR ● ), -OH, -OR ● , -O(haloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2, or -NO2, wherein each R ● is unsubstituted or, when "halo" precedes, is substituted only by one or more halogens and is independently a C1-4 aliphatic group, -CH2Ph, -O(CH2)0-1Ph, or a 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on the saturated carbon atom of R † include =O and =S.

[0048] Those skilled in the art reading this disclosure will understand that certain compounds described herein may be provided or utilized in any of a variety of forms, such as salt forms, protected forms, prodrug forms, ester forms, isomer forms (e.g., optical or structural isomers), isotopic forms, etc. In some embodiments, reference to a particular compound may relate to a particular form of the compound. In some embodiments, reference to a particular compound may relate to the compound in any form. In some embodiments, for example, a preparation of a single stereoisomer of a compound may be considered a different form of the compound from a racemic mixture of the compound; a particular salt of a compound may be considered a different form from other salt forms of the compound; a preparation containing a structural isomer of the double bond ((Z) or (E)) may be considered a different form from one containing other structural isomers of the double bond ((E) or (Z)); and a preparation in which one or more atoms are isotopes different from those present in the reference preparation may be considered a different form.

[0049] As used herein, the term “administration” means the administration of a composition (e.g., a compound, or a preparation containing a compound as described herein) to a subject or system. Administration also includes administering to a subject a prodrug derivative or analog of a compound, or a pharmaceutically acceptable salt of a compound or composition, which can form an equivalent amount of the active compound in the subject’s body. Administration to animal subjects (e.g., humans) may be by any suitable route. For example, in some embodiments, administration may be bronchial (including bronchial infusion), cheek, enteral, interdermal, intra-arterial, intradermal, intragastric, intramedullary, intramuscular, nasal, intraperitoneal, intrathecal, intravenous, intraventricular, mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, intratracheal (including intratracheal infusion), percutaneous, vaginal, or intravitreous.

[0050] As used herein, the term "acetyl" refers to the group -C(O)CH3.

[0051] As used herein, the term "alkoxy" means -O-C1-C 20An alkyl group is an alkoxy group that is bonded to the rest of the compound via an oxygen atom.

[0052] As used herein, the term “alkyl” refers to a saturated, linear or branched, monovalent hydrocarbon group containing 1 to 20 (e.g., 1 to 10, or 1 to 6) carbon atoms. In some embodiments, the alkyl group is unbranched (i.e., linear), and in some embodiments, the alkyl group is branched. Alkyl groups are exemplified by, but are not limited to, methyl, ethyl, n-propyl and isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl, and neopentyl.

[0053] As used herein, the term "alkylene" refers to a saturated divalent hydrocarbon group derived from a saturated hydrocarbon, either linear or branched, by removing two hydrogen atoms, exemplified by methylene, ethylene, isopropylene, and the like. x -C y The term "alkylene" refers to an alkylene group having x to y carbon atoms. Exemplary values ​​for x are 1, 2, 3, 4, 5, and 6, and exemplary values ​​for y are 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, or 20 (e.g., C1-C6, C1-C 10 , C2-C 20 , C2-C6, C2-C 10 or C2-C 20 Alkylene). In some embodiments, the alkylene may be further substituted with one, two, three, or four substituents as defined herein.

[0054] As used herein, the term “alkenyl” refers to a monovalent, linear or branched group of 2 to 20 carbon atoms (e.g., 2 to 6 or 2 to 10 carbon atoms) containing one or more carbon-carbon double bonds, unless otherwise specified, and is exemplified by ethenyl, 1-propenyl, 2-propenyl, 2-methyl-1-propenyl, 1-butenyl, and 2-butenyl. Alkenyls include both cis and trans isomers. As used herein, the term “alkenylene” refers to a divalent, linear or branched group of 2 to 20 carbon atoms (e.g., 2 to 6 or 2 to 10 carbon atoms) containing one or more carbon-carbon double bonds, unless otherwise specified.

[0055] As used herein, the term "alkynyl" refers to a monovalent linear or branched group consisting of 2 to 20 carbon atoms (e.g., 2 to 4, 2 to 6, or 2 to 10 carbon atoms) containing a carbon-carbon triple bond, exemplified by ethynyl and 1-propynyl.

[0056] As used herein, the term "amino" means -N(R † )2, for example, represents -NH2 and -N(CH3)2.

[0057] As used herein, the term "aminoalkyl" refers to an alkyl moiety in which one or more carbon atoms are substituted with one or more amino moieties.

[0058] As used herein, the term “amino acid” refers to a molecule having a side chain, an amino group, and an acidic group (e.g., -CO2H or -SO3H), and an amino acid is bonded to a parent molecule by a side chain, an amino group, or an acidic group (e.g., a side chain). As used herein, the term “amino acid” in its broadest sense refers to any compound or substance that can be incorporated into a polypeptide chain, for example, by the formation of one or more peptide bonds. In some embodiments, an amino acid has the general structure H2N-C(H)(R)-COOH. In some embodiments, an amino acid is a naturally occurring amino acid. In some embodiments, an amino acid is a synthetic amino acid, in some embodiments, an amino acid is a D-amino acid, and in some embodiments, an amino acid is an L-amino acid. “Standard amino acid” refers to any of the 20 standard L-amino acids commonly found in naturally occurring peptides. Examples of amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, optionally substituted hydroxylnorvaline, isoleucine, leucine, lysine, methionine, norvaline, ornithine, phenylalanine, proline, pyrrolicine, selenocysteine, serine, taurine, threonine, tryptophan, tyrosine, and valine.

[0059] As used herein, “amino acid substitution” refers to the substitution of a wild-type amino acid in a protein with a non-wild-type amino acid. Amino acid substitutions can be caused by gene mutations and can alter one or more properties of a protein (for example, altered binding affinity or specificity, altered enzyme activity, altered structure, or altered function).

[0060] As used herein, the term “aryl” refers to a monovalent monocyclic, bicyclic, or polycyclic ring system formed by carbon atoms, wherein the ring bonded to the pendant group is aromatic. Examples of aryl groups include phenyl, naphthyl, phenantrenyl, and anthracenyl. The aryl ring may be bonded to its pendant group by any heteroatom or carbocyclic atom that results in a stable structure, and unless otherwise specified, any of the ring atoms may be optionally substituted.

[0061] As used herein, the term "C0" represents a bond. For example, part of the term -N(C(O)-(C0-C5alkylene-H)- includes -N(C(O)-(C0alkylene-H)-, which is also represented by -N(C(O)-H)-.

[0062] As used herein, the terms "carbocyclic" and "carbocyclyl" refer to C3- substituted with a monovalent of any choice. 12 This refers to monocyclic, bicyclic, or tricyclic structures, which may be bridged, condensed, or spirocyclic, where all rings are formed of carbon atoms and at least one ring is non-aromatic. Examples of carbocyclic structures include cycloalkyl, cycloalkenyl, and cycloalkynyl groups. Examples of carbocyclyl groups include cyclohexyl, cyclohexenyl, cyclooctinyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indenyl, indanyl, and dekalinyl. A carbocyclic ring can be bonded to its pendant group at any ring atom that results in a stable structure, and unless otherwise specified, any of the ring atoms may be optionally substituted.

[0063] As used herein, the term "carbonyl" refers to a C(O) group, which can also be represented as C=O.

[0064] As used herein, the term "carboxyl" means -CO2H, (C=O)(OH), COOH, or C(O)OH, or the corresponding aprotonated group.

[0065] The term “combination therapy” refers to a treatment method comprising administering to a subject, as part of a treatment regimen, at least two therapeutic agents, optionally as one or more pharmaceutical compositions. For example, combination therapy may include the administration of a single pharmaceutical composition comprising at least two therapeutic agents and one or more pharmaceutically acceptable carriers, excipients, diluents, or surfactants. Combination therapy may include the administration of two or more pharmaceutical compositions, each composition comprising one or more therapeutic agents and one or more pharmaceutically acceptable carriers, excipients, diluents, or surfactants. In various embodiments, at least one of the therapeutic agents is a RAS(ON)GTP hydrolysis promoting compound (e.g., any one or more such KRAS(ON)GTP hydrolysis promoting compounds disclosed herein or known in the art). In various embodiments, at least one of the therapeutic agents is a KRAS(OFF) inhibitor (e.g., any one or more KRAS(OFF) inhibitors disclosed herein or known in the art). In some embodiments, at least one of the therapeutic agents is a KRAS G12C (OFF) inhibitors (e.g., any one or more KRASGs disclosed herein or known in the art) 12C (OFF) inhibitors. In some embodiments, at least one of the therapeutic agents is a KRAS G12D (OFF) inhibitors (e.g., any one or more KRAS disclosed herein or known in the art) G12D It is an inhibitor. In some embodiments, at least one of the therapeutic agents is KRAS G12V (OFF) inhibitors (e.g., any one or more KRAS disclosed herein or known in the art) G12VThe inhibitor is a pan-RAS(OFF) inhibitor. In some embodiments, at least one of the therapeutic agents is a pan-RAS(OFF) inhibitor (e.g., any one or more pan-RAS(OFF) inhibitors disclosed herein or known in the art). Two or more agents may be optionally administered simultaneously (as a single or separate composition) or sequentially (as separate compositions). The therapeutic agents may be administered in an effective dose. In some embodiments, the effective dose of one or more therapeutic agents may be less when used in combination therapy than the therapeutic dose of the same therapeutic agent when used as a monotherapy, for example, due to the addition or synergistic effect of combining two or more therapeutic agents.

[0066] As used herein, the term "cyano" refers to the -CN group.

[0067] As used herein, the term "cycloalkyl" refers to a monovalent saturated cyclic hydrocarbon group, which, unless otherwise specified, may be crosslinked, condensed, or a spirocyclic group having 3 to 8 ring carbons, exemplified by cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cycloheptyl.

[0068] As used herein, the term "cycloalkenyl" refers to a monovalent non-aromatic saturated cyclic hydrocarbon group, which, unless otherwise specified, may be bridged, condensed, or a spirocyclic group having 3 to 8 ring carbons and containing one or more carbon-carbon double bonds.

[0069] As used herein, the term “diastereomer” means stereoisomers that are not mirror images of each other and cannot be superimposed on each other.

[0070] As used herein, the term “dosage form” refers to a physically distinct unit of a compound (e.g., a compound of this disclosure) for administration to a subject. Each unit contains a predetermined amount of the compound. In some embodiments, such an amount is a unit dose (or its entire fraction) appropriate for administration according to an administration regimen (i.e., using a therapeutic administration regimen) that has been measured to correlate with a desired or beneficial outcome when administered to a suitable population. Those skilled in the art will understand that the total amount of a therapeutic composition or compound administered to a particular subject may be determined by one or more attending physicians and may involve administration of multiple dosage forms.

[0071] As used herein, the term “dosage regimen” refers to a set of unit doses (usually two or more) administered individually to a subject, typically separated by a period of time. In some embodiments, a given therapeutic compound (e.g., a compound of this disclosure) has a recommended dosage regimen, which may consist of one or more doses. In some embodiments, the dosage regimen comprises multiple doses, each separated from the others by a period of time equal in length, and in some embodiments, the dosage regimen comprises multiple doses and at least two different periods separating the individual doses. In some embodiments, all doses within the dosage regimen are identical unit doses. In some embodiments, different doses within the dosage regimen are different amounts. In some embodiments, the dosage regimen comprises a first dose at a first dose, followed by one or more further doses at a second dose different from the first dose. In some embodiments, the dosage regimen comprises a first dose at a first dose, followed by one or more further doses at a second dose identical to the first dose. In some embodiments, the administration regimen, when administered across relevant populations, correlates with a desired or beneficial outcome (i.e., it is a therapeutic administration regimen).

[0072] The term “disability” is used in this disclosure to mean, and is interchangeable with, the terms “disease,” “condition,” or “illness,” unless otherwise indicated.

[0073] As used herein, the term “enantiomer” means each individual optically active form of the compound of the Disclosure having an optical purity or enantiomeric excess (measured by methods standard in the art) of at least 80% (i.e., at least 90% of one enantiomer and up to 10% of the other enantiomer), preferably at least 90%, and more preferably at least 98%.

[0074] The term "guanidyl" refers to the structure: [ka] (wherein each R independently refers to a group having any chemically suitable substituent as described herein.)

[0075] As used herein, the term "guanidinoalkylalkyl" refers to an alkyl moiety in which one or more carbon atoms are substituted with one or more guanidyl moieties.

[0076] As used herein, the term "haloacetyl" refers to an acetyl group in which at least one hydrogen atom is substituted with a halogen.

[0077] As used herein, the term "haloalkyl" refers to an alkyl moiety in which one or more carbon atoms are substituted with one or more identical or different halogen moieties.

[0078] As used herein, the term "halogen" refers to a halogen selected from bromine, chlorine, iodine, or fluorine.

[0079] As used herein, the term “heteroalkyl” refers to an “alkyl” group (as defined herein) in which at least one carbon atom is replaced by a heteroatom (e.g., an O, N, or S atom). The heteroatom may appear in the middle or at the ends of the group.

[0080] As used herein, the term “heteroaryl” refers to a monovalent, monocyclic or polycyclic cyclic structure containing at least one complete aromatic ring. That is, these contain 4n+2 π electrons within the monocyclic or polycyclic ring system and at least one ring heteroatom selected from N, O, or S within the aromatic ring. Exemplary unsubstituted heteroaryl groups have 1 to 12 (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9) carbon atoms. The term “heteroaryl” includes bicyclic, tricyclic, and tetracyclic groups in which any of the above heteroaromatic rings are fused to one or more aryl or carbocyclic rings, such as a phenyl ring or a cyclohexane ring. Examples of heteroaryl groups include, but are not limited to, pyridyl, pyrazolyl, benzoxazolyl, benzimidazolyl, benzothiazolyl, imidazolyl, thiazolyl, quinolinyl, tetrahydroquinolinyl, and 4-azaindylol. The heteroaryl ring can be bonded to its pendant group at any ring atom that provides a stable structure, and unless otherwise specified, any of the ring atoms may be optionally substituted. In some embodiments, the heteroaryl is substituted with 1, 2, 3, or 4 substituents.

[0081] As used herein, the term “heterocycloalkyl” refers to a monovalent, monocyclic, bicyclic, or polycyclic ring system, which may be bridged, condensed, or spirocyclic, in which at least one ring is non-aromatic, and the non-aromatic ring contains 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Five-membered rings have 0 to 2 double bonds, and six- and seven-membered rings have 0 to 3 double bonds. Exemplary unsubstituted heterocycloalkyl groups have 1 to 12 (e.g., 1 to 11, 1 to 10, 1 to 9, 2 to 12, 2 to 11, 2 to 10, or 2 to 9) carbon atoms. The term “heterocycloalkyl” also refers to a heterocyclic compound having a bridged polycyclic structure in which one or more carbons or heteroatoms bridge two non-adjacent members of a monocyclic ring, e.g., a quinuclidinyl group. The term "heterocycloalkyl" includes bicyclic, tricyclic, and tetracyclic groups in which any of the above heterocyclic rings is fused to one or more aromatic, carbocyclic, heteroaromatic, or heterocyclic rings, such as an aryl ring, cyclohexane ring, cyclohexene ring, cyclopentane ring, cyclopentene ring, pyridine ring, or pyrrolidine ring. Examples of heterocycloalkyl groups are pyrrolidinyl, piperidinyl, 1,2,3,4-tetrahydroquinolinyl, decahydroquinolinyl, dihydropyrrolopyridine, and decahydronaphthilidinyl. Heterocycloalkyl rings can be bonded to their pendant group at any ring atom that results in a stable structure, and unless otherwise specified, any of the ring atoms may be optionally substituted.

[0082] As used herein, the term "hydroxy" refers to the -OH group.

[0083] As used herein, the term "hydroxyalkyl" refers to an alkyl moiety in which one or more carbon atoms are substituted with one or more -OH moieties.

[0084] As used herein, the term “isomer” means any tautomer, stereoisomer, atropisomer, enantiomer, or diastereomer of any of the compounds of this disclosure. The compounds of this disclosure may have one or more chiral centers or double bonds and are therefore recognized to exist as stereoisomers, e.g., double bond isomers (i.e., geometric E / Z isomers), or diastereomers (i.e., enantiomers (i.e., (+) or (-), or cis / trans isomers)). According to this disclosure, the chemical structures illustrated herein, i.e., the compounds of this disclosure, are recognized to exist as all corresponding stereoisomers, i.e., stereoisomerically pure forms (i.e., geometrically pure, enantiomerically pure, or diastereoisomerically pure forms) and enantiomers. This includes both thiomeric and stereoisomer mixtures, such as racemic compounds. Mixtures of enantiomers and stereoisomers of the compounds disclosed herein can typically be separated into their constituent enantiomers or stereoisomers by known methods, such as chiral phase gas chromatography, chiral phase high-performance liquid chromatography, crystallization of the compound as a chiral salt complex, or crystallization of the compound in a chiral solvent. Enantiomers and stereoisomers can also be obtained from stereoisomerically or enantiomerically pure intermediates, reagents, and catalysts by known asymmetric synthesis methods.

[0085] As used herein, the term “inhibitor” refers to a compound that prevents a biomolecule (e.g., protein, nucleic acid) from completing or initiating a reaction. Inhibitors can inhibit a reaction by competitive, non-competitive, or non-competitive means, for example. With respect to its binding mechanism, an inhibitor can be an irreversible or reversible inhibitor. Exemplary inhibitors include, but are not limited to, nucleic acids, DNA, RNA, shRNA, siRNA, proteins, protein mimics, peptides, peptide mimics, antibodies, small molecules, chemicals, enzymes, receptors, or analogs that mimic the binding sites of other proteins. In some embodiments, the inhibitor is a small molecule, e.g., a low molecular weight organic compound, e.g., an organic compound having a molecular weight (MW) of less than 1200 Daltons (Da). In some embodiments, the MW is less than 1100 Da. In some embodiments, the MW is less than 1000 Da. In some embodiments, the MW is less than 900 Da. In some embodiments, the MW is less than 800 Da. In some embodiments, the MW is less than 700 Da. In some embodiments, the MW is less than 600 Da. In some embodiments, the MW range of the small molecule is 600 Da to 700 Da (including both ends). In some embodiments, the MW range of the small molecule is 600 Da to 800 Da (including both ends). Examples of small molecule inhibitors include cyclic and acyclic compounds. Examples of small molecule inhibitors include natural products, as well as their derivatives and analogs. Small molecule inhibitors may include, for example, covalent crosslinking groups capable of forming covalent crosslinks with the amino acid side chains of the target protein.

[0086] As used herein, the term “linker” refers to a divalent organic moiety that connects a first moiety (e.g., a macrocyclic moiety) to a second moiety (e.g., a crosslinking group). In some embodiments, the linker results in a compound capable of achieving an IC50 of 2 μM or less in the Ras-RAF disruption assay protocol provided herein.

[0087] The purpose of this biochemical assay is to measure the ability of the test compound to facilitate the formation of a ternary complex between the nucleotide load Ras isoform and cyclophyllin A, and the resulting ternary complex is BRAF. RBD It disrupts binding to constructs and inhibits Ras signaling via RAF effectors.

[0088] In an assay buffer containing 25 mM HEPES (pH 7.3), 0.002% Tween 20, 0.1% BSA, 100 mM NaCl, and 5 mM MgCl2, untagged cyclophyllin A, His6-K-RasGMPPNP (or other Ras variants), and GST-BRAF were tested. RBD These compounds were combined in a 384-well assay plate at final concentrations of 25 μM, 12.5 nM, and 50 nM, respectively. The compounds were present in the plate wells as a 10-point 3-fold dilution series, starting at a final concentration of 30 μM. After incubation at 25°C for 3 hours, the mixture of anti-His Eu-W1024 and anti-GST allophycocyanin was added to the assay sample wells at final concentrations of 10 nM and 50 nM, respectively, and the reaction was incubated for a further 1.5 hours. The TR-FRET signal was read using a microplate reader (excitation 320 nm, fluorescence 665 / 615 nm). Compounds that promote the disruption of the Ras:RAF complex were identified as those that induced a decrease in the TR-FRET ratio compared to the DMSO control well.

[0089] In some embodiments, the linker contains 20 or fewer linear atoms. In some embodiments, the linker contains 15 or fewer linear atoms. In some embodiments, the linker contains 10 or fewer linear atoms. In some embodiments, the linker has a molecular weight of less than 500 g / mol. In some embodiments, the linker has a molecular weight of less than 400 g / mol. In some embodiments, the linker has a molecular weight of less than 300 g / mol. In some embodiments, the linker has a molecular weight of less than 200 g / mol. In some embodiments, the linker has a molecular weight of less than 100 g / mol. In some embodiments, the linker has a molecular weight of less than 50 g / mol.

[0090] As used herein, the term “mutation” refers to any modification of a nucleic acid or polypeptide that results in a change to the nucleic acid or polypeptide. The term “mutation” can include, for example, point mutations, deletions or insertions of one or more residues within a polynucleotide, changes occurring within the protein-coding region of a gene, in addition to changes in regions outside the protein-coding region, such as, but not limited to, modifications in regulatory or promoter sequences, in addition to amplification, or chromosomal disruption or translocation. In certain embodiments, a mutation results in an amino acid substitution in the encoded protein.

[0091] "Patient" or "subject" is a mammal, such as a human, mouse, rat, guinea pig, dog, cat, horse, cattle, pig, or a non-human primate, such as a monkey, chimpanzee, baboon, or rhesus macaque.

[0092] The terms “prevent” or “prevention” in relation to a subject refer to preventing the subject from contracting a disease or disorder. Prevention includes prophylactic treatment. For example, prevention may include administering a compound disclosed herein to a subject before the subject contracts a disease, and this administration prevents the subject from contracting the disease.

[0093] As used herein, the term “pharmaceutical composition” means a compound such as the compounds of this disclosure, or a pharmaceutically acceptable salt thereof, formulated with pharmaceutically acceptable excipients.

[0094] As used herein, “pharmaceutically acceptable excipient” means any inert component that is non-toxic and non-inflammatory in the subject matter (e.g., a vehicle capable of suspending or dissolving an active compound). Typical excipients include, for example, antifouling agents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (colorants), emollients, emulsifiers, fillers (diluents), film-forming agents or coatings, flavoring agents, fragrances, flow enhancers, lubricants, preservatives, printing inks, adsorbents, suspending agents or dispersants, sweeteners, or hydration water. Examples of excipients include, but are not limited to, optionally substituted butylated hydroxytoluene (BHT), calcium carbonate, dibasic calcium phosphate, calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, optionally substituted hydroxypropylcellulose, optionally substituted hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolic acid, sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol. Those skilled in the art are familiar with a wide variety of agents and materials useful as excipients.See, for example, 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, the composition comprises at least two different pharmaceutically acceptable excipients.

[0095] As used herein, the term “pharmaceutically acceptable salt” refers to salts of the compounds described herein that are suitable for use in contact with human and other animal tissues without causing excessive toxicity, irritation, allergic reactions, etc., within the normal range of reasonable medical judgment, and that are balanced by a reasonable benefit / risk ratio. Pharmacopoeia-acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (Eds. PHStahl and CGWermuth), Wiley-VCH, 2008. Salts can be prepared in situ during the final isolation and purification of the compounds described herein, or separately by reacting a free base with a suitable organic acid.

[0096] The terms "RAS inhibitor" and "inhibitor of RAS" are used interchangeably and refer to any inhibitor that targets the RAS protein, i.e., selectively binds to or selectively inhibits the RAS protein.

[0097] As used herein, the term “RAS(OFF) inhibitor” refers to an inhibitor that targets, i.e., selectively binds to or selectively inhibits the GDP-bound inactive state of RAS (e.g., more selectively than GTP-bound active RAS). Inhibition of the GDP-bound inactive state of RAS can be achieved, for example, by inhibiting the exchange of GDP with GTP, thereby sequestering the inactive state and inhibiting the adaptation of RAS to its active conformation. In certain embodiments, RAS(OFF) inhibitors can also bind to or inhibit the GTP-bound active state of RAS (e.g., with lower affinity or inhibition constant than GDP-bound inactive RAS). RAS(OFF) inhibitors may be mutaselective, such as being selective to G12C, G12D, or G12V variants. RAS(OFF) inhibitors may be selective to multiple variants, or to one or more variants and the wild type (in either case, “pan-RAS(OFF)” inhibitors). Methods for measuring RAS(OFF) inhibition are known in the art.

[0098] As used herein, the term “RAS(ON) inhibitor” refers to a non-covalent triple complex formation inhibitor that targets, i.e., selectively binds to, or selectively inhibits (e.g., more selectively than GDP-bound inactive RAS) the GTP-bound activated RAS. Examples of inhibition of the GTP-bound activated RAS include, for example, the inhibition of oncogenic signaling from the GTP-bound activated RAS. In some embodiments, a RAS(ON) inhibitor is an inhibitor that selectively binds to and inhibits the GTP-bound activated RAS. In certain embodiments, a RAS(ON) inhibitor may also bind to, or inhibit, the GDP-bound inactive RAS (e.g., with lower affinity or inhibition constant than GTP-bound activated RAS). RAS(ON) inhibitors are non-covalent conjugates of the GTP-bound form of RAS, and those skilled in the art are familiar with methods for determining whether crosslinking has occurred. In some embodiments, the RAS(ON) inhibitor does not contain crosslinking groups such as those found in the Art (e.g., WO2020 / 132597, WO2021 / 091982, WO2021 / 091967, WO2022 / 235864, WO2022 / 235870, WO2023 / 060253, PCT / US2023 / 037057, and WO2023 / 133543). In some embodiments, the RAS(ON) inhibitor has a molecular weight of 800 to 1200 Da (including both ends). References to the term RAS(ON) inhibitors include, but are not limited to, any one or more RAS(ON) inhibitors selected from those disclosed in WO2021 / 091956, WO2022 / 060836, U.S. Provisional Application No. 63 / 351,146, or WO2023 / 240263, each of which is invoked by reference, either as a whole or in combination, as any RAS(ON) inhibitor. In some embodiments, compounds from WO2021 / 091956, WO2022 / 060836, and WO2023 / 240263 that contain phenol at the A position are excluded. Methods for determining RAS(ON) inhibition are known in the art.For example, see WO2021 / 091956, WO2022 / 060836, and WO2023 / 240263.

[0099] As used herein, "RAS(ON)GTP hydrolysis-promoting compound" refers to a compound that forms a triple complex (i.e., CYPA-RAS(ON)GTP hydrolysis-promoting compound-RAS(ON) isoform), and RAS(ON) mutant isoforms in the absence of the compound (RAS MUT ) and / or RAS wild-type isoform (RAS WT It exhibits a higher RAS(ON)GTP hydrolysis rate than the intrinsic hydrolysis rate of KRAS. In some embodiments, the RAS(ON)GTP hydrolysis-promoting compound exhibits a hydrolysis rate more than 14 times the intrinsic hydrolysis rate ("strong hydrolysant"). In some embodiments, the RAS(ON)GTP hydrolysis-promoting compound exhibits a hydrolysis rate more than 5 to 14 times the intrinsic hydrolysis rate ("moderate hydrolysant"). In some embodiments, the RAS(ON)GTP hydrolysis-promoting compound exhibits a hydrolysis rate greater than 1 times the intrinsic hydrolysis rate but less than 5 times the intrinsic hydrolysis rate ("weak hydrolysant"). In some embodiments, KRAS G12Vは These isoforms are used to determine whether a hydrolyzing agent is strong, moderate, or weak. Methods for measuring hydrolysis are known in the art, such as those described herein. In some embodiments, the RAS(ON)GTP hydrolytic compounds are RAS(ON) inhibitors. All RAS(ON)GTP hydrolytic compounds retain catalytic water near the γ-phosphorus atom of GTP (distance < 5 angstroms) and position the delta carbon of Q61 of RAS(ON) within 8 angstroms of the γ-phosphorus atom: these parameters can be determined by those skilled in the art. Further descriptions of RAS(ON)GTP hydrolytic compounds are provided herein.

[0100] As used herein, “RAS(OFF) decomposer” is a RAS decomposer that targets the OFF state. Such decomposers are known in the art. Non-limiting examples of RAS(OFF) decomposers can be found in one or more of the following applications: WO2024055112, WO2024054625, WO2024050742, WO2024044334, WO2024040080, WO2024034657, WO2024034593, WO2024034591, WO2024034123, WO2024029613, WO2024020159, WO 2024019103, WO2024017392, WO2023185864, WO2023171781, WO2023141570, WO2023138524, WO2023130012, WO2023116934, WO2023099620, WO2023081476, WO2023077441, and CN115785199, each of which is incorporated herein by reference in whole.

[0101] The terms “RAS pathway” and “RAS / MAPK pathway” are used interchangeably herein and refer to a signal transduction cascade downstream of various cell surface growth factor receptors, where the activation of RAS (and its diverse isoforms and allotypes) is a central event driving various cellular effector events that determine cell proliferation, activation, differentiation, mobility, and other functional properties. SHP2 carries a positive signal from the growth factor receptor to the RAS activation / deactivation cycle, which is regulated by guanine nucleotide exchange factors (GEFs such as SOS1) that load GTP onto RAS to produce functionally active, GTP-bound RAS, and GTPase-activating proteins (GAPs such as NF1) that facilitate signal termination by converting GTP to GDP. The GTP-bound RAS produced by this cycle carries essential positive signals to a series of serine / threonine kinases, including RAFs and MAP kinases, from which further signals extend to various cellular effector functions.

[0102] As used herein, the term “stereoisomer” means all possible different isomeric and structural forms that a compound may have (e.g., any compound of any formula described herein), in particular all possible stereochemical and structural isomeric forms of the basic molecular structure, including atropisomers, all diastereomers, enantiomers, or conformational isomers. Some of the compounds in this disclosure may exist in different tautomers, all of which are included in the scope of this disclosure.

[0103] As used herein, the term "sulfonyl" refers to the -S(O)2- group.

[0104] A “therapeutic agent” is any substance, such as a compound or composition, that can treat a disease or disorder. In some embodiments, therapeutic agents useful in combination with the present disclosure include RAS inhibitors and cancer chemotherapy. Many such therapeutic agents are known in the art and are disclosed herein.

[0105] The term “therapeutic dose” means a quantity sufficient to treat a disease, disorder, or condition when administered to a population suffering from or suspected of having a disease, disorder, or condition, according to a therapeutic dosing regimen. In some embodiments, a therapeutic dose is a quantity that reduces the incidence or severity of one or more symptoms of a disease, disorder, or condition, or delays their onset. Those skilled in the art will understand that the term “therapeutic dose” does not actually require that successful treatment be achieved in a particular individual. Rather, a therapeutic dose may be a quantity that, when administered to subjects requiring such treatment, elicits a specific desired pharmacological response in a significant number of subjects. It is specifically understood that a particular subject may actually be “refractory” to the “therapeutic dose.” In some embodiments, a reference to a therapeutic dose may refer to a quantity measured in one or more specific tissues (e.g., tissues affected by the disease, disorder, or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine). Those skilled in the art will understand that in some embodiments, the therapeutically effective dose may be formulated or administered as a single dose. In some embodiments, the therapeutically effective dose may be formulated or administered in multiple doses, for example, as part of an administration regimen.

[0106] As used herein, the term "thiocarbonyl" refers to the -C(S)- group.

[0107] The term “treatment” (and additionally, “to treat” or “to treat”) in its broadest sense refers to any administration of a substance (e.g., a compound of the present disclosure) that partially or completely alleviates, improves, reduces, inhibits, delays the onset, reduces the severity, or decreases the 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 subjects who show no signs of the disease, disorder, or condition in question, or to subjects who show only the initial signs of the disease, disorder, or condition in question. Alternatively, or in addition, in some embodiments, treatment may be administered to subjects who show one or more established signs of the disease, disorder, or condition in question. In some embodiments, treatment may be administered to subjects diagnosed with suffering from the disease, disorder, or condition in question. In some embodiments, treatment may be administered to subjects known to have one or more susceptibility factors that are statistically correlated with an increased risk of progression of the disease, disorder, or condition in question.

[0108] The term "tricomplex" refers to a mechanism of action involving the formation of a high-affinity tricomplex between a synthetic ligand (e.g., a RAS(ON)GTP hydrolysis promoter) and two intracellular proteins that do not interact under normal physiological conditions (RAS, the target protein of interest, and cyclophyllin A, a cytosolic chaperone protein widely expressed in cells). Such tricomplexes are known in the art. See, for example, WO2020 / 132597, WO2021 / 091956, WO2021 / 091967, WO2021 / 091982, WO2022 / 060836, WO2022 / 235864, WO2022 / 235 / 870, WO2023 / 060253, WO2023 / 133543, and WO2023 / 240263.

[0109] The term "wild-type" refers to an entity with a structure or activity that is found in a "normal" state or context in nature (as opposed to mutants, diseases, modified organisms, etc.). Those skilled in the art will understand that wild-type genes and polypeptides often exist in multiple different forms (e.g., alleles).

[0110] I. Composition This specification provides compounds that promote the hydrolysis of RAS(ON) GTP and their uses. Also provided are pharmaceutical compositions comprising one or more such compounds, or pharmaceutically acceptable salts thereof, and pharmaceutically acceptable excipients. RAS(ON)GTP hydrolytic compounds may be used in methods of modulating RAS (e.g., intrasubjective or intracellular) and methods of treating cancer, as described herein. This disclosure, in particular, provides compositions, methods, and kits for treating or preventing diseases or disorders (e.g., cancer) by RAS(ON)GTP hydrolytic compounds in combination with RAS(OFF) inhibitors.

[0111] RAS proteins (KRAS, HRAS, and NRAS) play essential roles in various human cancers and are therefore suitable targets for anticancer therapy. In fact, mutations in RAS proteins account for approximately 30% of all human cancers in the United States, many of which are lethal. Dysregulation of RAS proteins due to mutation activation, overexpression, or upstream activation is common in human tumors, and mutation activation in RAS is frequently found in human cancers. RAS switches between a GDP-bound "off" state and a GTP-bound "on" state. This state switching is facilitated by the interaction of guanine nucleotide exchange factor (GEF) proteins (e.g., SOS1) that load GTP into RAS and GTPase-activating protein (GAP) proteins (e.g., NF1) that hydrolyze GTP, thereby inactivating RAS. In addition, the SH2 domain-containing protein tyrosine phosphatase 2 (SHP2) associates with receptor signaling machinery and, after becoming active during RTK activation, promotes RAS activation. Mutations in the RAS protein can lead to constitutively active signaling pathways that lock the protein in an "on" state, resulting in uncontrolled cell proliferation. For example, activation of a mutation at codon 12 in the RAS protein inhibits both the GAP-dependent and intrinsic hydrolysis rates of GTP, significantly distorting the population of RAS mutant proteins into an "on" (GTP-bound) state (RAS(ON)), leading to oncogenic MAPK signaling. In particular, RAS exhibits picomolar affinity for GTP, allowing it to be activated even in the presence of low concentrations of this nucleotide. Mutations at codon 13 (e.g., G13D) and codon 61 (e.g., Q61K) of RAS also contribute to oncogenic activity in some cancers.

[0112] a) RAS(ON)GTP hydrolysis promoting compound The compositions of the present disclosure may include one or more RAS(ON) GTP hydrolysis promoting compounds. The RAS(ON) GTP hydrolysis promoting compounds of the present disclosure form a high-affinity ternary complex with two intracellular proteins that do not interact under normal physiological conditions, namely RAS and cyclophilin A (CypA), a cytosolic chaperone that is widely expressed in cells. Further, the present disclosure provides a non-covalent binding of RAS by a RAS(ON) GTP hydrolysis promoting compound that promotes a catalytically competent orientation, in which orientation the glutamine 61 (Q61) side chain coordinates a catalytic water and promotes a nucleophilic attack on the γ-phosphate GTP bound to the RAS protein. See Figure 15.

[0113] Therefore, as used herein, the structure of Formula Ia:

Chemical formula

[0114] In some embodiments, the present disclosure relates to compounds of structural formula Ib that do not crosslink with the RAS protein: [ka] or a pharmaceutically acceptable salt thereof, where the dotted line represents 0, 1, 2, 3, or 4 non-adjacent double bonds; A is a C2-C4 alkylene that has been optionally substituted, a C1-C4 heteroalkylene that has been optionally substituted, or a C2-C4 alkenylene that has been optionally substituted; B does not exist, or -NH-, -N(CH3)-, -O-, -CH(R 9 )-, or >C=CR 9 R 9’ [In the formula, carbon is -N(R 11 ) bonded to the carbonyl carbon of C(O)-, optionally substituted 3- to 6-membered cycloalkylenes, optionally substituted 3- to 6-membered heterocycloalkylenes, optionally substituted 6-membered arylenes, or 5- to 6-membered heteroarylenes; G is a C1-C4 alkylene that has been optionally substituted, a C1-C4 alkenylene that has been optionally substituted, a C1-C4 heteroalkylene that has been optionally substituted, and -C(O)O-CH(R 6 )-[In the formula, the second C is -C(R 7 R 8 )-bound, -C(O)NH-CH(R 6 )-[In the formula, the second C is -C(R 7 R 8 )-bound, optionally substituted C1-C4 heteroalkylene, or 3- to 8-membered heteroarylene; L either does not exist or is a linker; W is hydrogen, cyano, optionally substituted amino, optionally substituted amide, optionally substituted C1-C4 alkoxy, optionally substituted C1-C4 hydroxyalkyl, optionally substituted C1-C4 aminoalkyl, optionally substituted C1-C4 haloalkyl, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 guanidinoalkyl, optionally substituted C0-C4 alkyl, optionally substituted 3-11 member heterocycloalkyl, optionally substituted 3-10 member cycloalkyl, optionally substituted 6-10 member aryl, or optionally substituted 3-10 member heteroaryl, where W is not crosslinked to RAS; Z is -C(O)- or -S(O)2-; X 1 This is a C1-C2 alkylene, NR, O, or S(O) that has been optionally substituted. n and; X 2 is either O or NH; X 3 is N or CH; n is 0, 1, or 2; R is hydrogen, cyano, optionally substituted C1-C4 alkyl, optionally substituted C2-C4 alkenyl, optionally substituted C2-C4 alkynyl, C(O)R', C(O)OR', C(O)N(R')2, S(O)R', S(O)2R', or S(O)2N(R')2; Each R ’ These are independently H or optionally substituted C1-C4 alkyl groups; Y 1 is C, CH, or N; Y 2 , Y 3 , Y 4 , and Y 7 Independently, C or N; Y 5 is CH, CH2, or N; Y 6 is C(O), CH, CH2, or N; R 1This is a cyano, an optionally substituted C1-C6 alkyl, an optionally substituted C1-C6 heteroalkyl, an optionally substituted 3-6 member cycloalkyl, an optionally substituted 3-6 member cycloalkenyl, an optionally substituted 3-6 member heterocycloalkyl, an optionally substituted 6-10 member aryl, or an optionally substituted 5-10 member heteroaryl, or R 1 and R 2 These combine with the atoms they bond to to form optionally substituted 3- to 14-membered heterocycloalkyl groups; R 2 It is either absent, or hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, or optionally substituted 5 or 6-membered heteroaryl; R 3 It does not exist, or R 2 and R 3 These combine with the atoms they bond to to form optionally substituted 3- to 8-membered cycloalkyl groups or optionally substituted 3- to 14-membered heterocycloalkyl groups; R 4 It is either absent, or a methyl atom optionally substituted with hydrogen, halogen, cyano, or 1 to 3 halogens; R 5 These are C1-C4 alkyl, cyano, hydroxy, or C1-C4 alkoxy, cyclopropyl, or cyclobutyl atoms optionally substituted with hydrogen or halogens; R 6 R is hydrogen or methyl; 7 is hydrogen, halogen, or optionally substituted C1-C3 alkyl, or R 6 and R 7 These combine with the carbon atoms to which they are bonded to form optionally substituted 3- to 6-membered cycloalkyl groups or optionally substituted 3- to 7-membered heterocycloalkyl groups; R 8 is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R 7 and R 8 combine with the carbon atom to which they are attached to form C=CR 7’ R 8’ ; C=N(OH), C=N(O-C1-C3 alkyl), C=O, C=S, C=NH, optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; R 7a and R 8a are independently hydrogen, halo, optionally substituted C1-C3 alkyl, or combine with the carbon to which they are attached to form carbonyl; R 7’ is hydrogen, halogen, or optionally substituted C1-C3 alkyl; R 8’ is hydrogen, halogen, hydroxy, cyano, optionally substituted C1-C3 alkoxy, optionally substituted C1-C3 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 8-membered cycloalkyl, optionally substituted 3- to 14-membered heterocycloalkyl, optionally substituted 5- to 10-membered heteroaryl, or optionally substituted 6- to 10-membered aryl, or R 7’ and R 8’ combine with the carbon atom to which they are attached to form optionally substituted 3- to 6-membered cycloalkyl, or optionally substituted 3- to 7-membered heterocycloalkyl; R 9is a hydrogen atom, a fluorine atom, an optionally substituted C1-C6 alkyl group, an optionally substituted C1-C6 heteroalkyl group, an optionally substituted 3- to 6-membered cycloalkyl group, or an optionally substituted 3- to 7-membered heterocycloalkyl group; R 9 And L combine with the atoms they bond to to form optionally substituted 3- to 14-membered heterocycloalkyl groups; R 9’ is a C1-C6 alkyl group substituted with hydrogen or optionally; R 10 These are hydrogen, halo, hydroxyl, C1-C3 alkoxy, or C1-C3 alkyl; R 10a is hydrogen or halo; R 11 is hydrogen or a C1-C3 alkyl group; and R 16 It is either hydrogen or a C1-C3 alkyl group.

[0115] In some embodiments, the present disclosure relates to compounds of formula Ia. [ka] Provided, in the formula, R 1 This is a cyano, halogen (e.g., fluoro), optionally substituted C1-C6 alkyl (e.g., C1-C6 haloalkyl or C1-C6 fluoroalkyl), optionally substituted C1-C6 heteroalkyl, optionally substituted 3-6 member cycloalkyl, optionally substituted 3-6 member cycloalkenyl, optionally substituted 3-6 member heterocycloalkyl, optionally substituted 6-10 member aryl, or optionally substituted 5-10 member heteroaryl, or R 1 and R 2 These combine with the atoms they bond to to form optionally substituted 3- to 14-membered heterocycloalkyl groups; R 2It is either absent, or it is hydrogen, halogen (e.g., fluoro), optionally substituted C1-C6 alkyl, C1-C6 haloalkyl (e.g., C1-C6 fluoroalkyl), optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, or optionally substituted 5 or 6-membered heteroaryl; R 3 It does not exist, or R 2 and R 3 These combine with the atoms they bond to to form optionally substituted 3- to 8-membered cycloalkyl groups or optionally substituted 3- to 14-membered heterocycloalkyl groups; R 4 It is either absent, or methyl is optionally substituted with hydrogen, halogen (e.g., fluoro), cyano, or 1 to 3 halogens; R 5 These are C1-C4 alkyl, cyano, hydroxy, or C1-C4 alkoxy, cyclopropyl, or cyclobutyl atoms optionally substituted with hydrogen, halogen (e.g., fluoro), or halogen (e.g., fluoro); Y 5 is CH, CH2, CF, CHF, CF2, or N; Y 6 is C(O), CH, CF, CH2, CF2, or N; The remaining variables are as defined above.

[0116] In some embodiments, the present disclosure relates to compounds of formula Ib. [ka] Provided, in the formula, R 1This is a cyano, halogen (e.g., fluoro), optionally substituted C1-C6 alkyl (e.g., C1-C6 haloalkyl or C1-C6 fluoroalkyl), optionally substituted C1-C6 heteroalkyl, optionally substituted 3-6 member cycloalkyl, optionally substituted 3-6 member cycloalkenyl, optionally substituted 3-6 member heterocycloalkyl, optionally substituted 6-10 member aryl, or optionally substituted 5-10 member heteroaryl, or R 1 and R 2 These combine with the atoms they bond to to form optionally substituted 3- to 14-membered heterocycloalkyl groups; R 2 It is either absent, or it is hydrogen, halogen (e.g., fluoro), optionally substituted C1-C6 alkyl, C1-C6 haloalkyl (e.g., C1-C6 fluoroalkyl), optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 7-membered heterocycloalkyl, optionally substituted 6-membered aryl, or optionally substituted 5 or 6-membered heteroaryl; R 3 It does not exist, or R 2 and R 3 These combine with the atoms they bond to to form optionally substituted 3- to 8-membered cycloalkyl groups or optionally substituted 3- to 14-membered heterocycloalkyl groups; R 4 It is either absent, or methyl is optionally substituted with hydrogen, halogen (e.g., fluoro), cyano, or 1 to 3 halogens; R 5 These are C1-C4 alkyl, cyano, hydroxy, or C1-C4 alkoxy, cyclopropyl, or cyclobutyl atoms optionally substituted with hydrogen, halogen (e.g., fluoro), or halogen (e.g., fluoro); Y 5 is CH, CH2, CF, CHF, CF2, or N; Y6 is C(O), CH, CF, CH2, CF2, or N; The remaining variables are as defined above.

[0117] In some embodiments, A is one of the following: [ka]

[0118] In some embodiments, A is one of the following: [ka]

[0119] In some embodiments, R 1 teeth, [ka] That is the case.

[0120] In some embodiments, R 1 teeth, [ka] That is the case.

[0121] In some embodiments, R 1 teeth, [ka] And, In the formula, Z 1 is N or CH; m is either 1 or 2; R 18 , R 19 , R 20 , and R 21Each of these is independently selected from hydrogen, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 heteroalkyl, optionally substituted 3- to 6-membered cycloalkyl, optionally substituted 3- to 6-membered cycloalkenyl, optionally substituted 3- to 6-membered heterocycloalkyl, optionally substituted 6- to 10-membered aryl, or optionally substituted 5- to 10-membered heteroaryl; or R 18 and R 20 They combine with the atoms they bond to to form optionally substituted 3- to 8-membered cycloalkyl groups or optionally substituted 3- to 8-membered heterocycloalkyl groups; or R 20 and R 21 They combine with the atoms they bond to to form optionally substituted 3- to 8-membered heterocycloalkyl groups; or R 19 and R 20 These combine with the atoms they bond to to form optionally substituted 4- to 8-membered heterocycloalkyl groups.

[0122] In some embodiments, R 1 teeth, [ka] That is the case.

[0123] In some embodiments, R 1 teeth, [ka] That is the case.

[0124] In some embodiments, R 18 It is methyl.

[0125] In some embodiments, R 1 teeth, [ka] That is the case.

[0126] In some embodiments, B is -CHR 9 - In some embodiments, R 9 B is an optionally substituted C1-C6 alkyl or an optionally substituted 3- to 6-membered cycloalkyl. In some embodiments, B is an optionally substituted 6-membered arylene. In some embodiments, B is absent.

[0127] In some embodiments, the linker has a structure having the structure of formula II: A 1 -( B 1 ) f -(C 1 ) g -( B 2 ) h -(D 1 )-(B 3 ) i -(C 2 ) j -( B 4 ) k -A 2 Formula II In the formula, A 1 This is the bond between the linker and B; A 2 This is the bond between W and the linker; B 1 B 2 B 3 , and B 4 These are, independently, a C1-C2 alkylene substituted by choice, a C1-C3 heteroalkylene substituted by choice, O, S, and NR. N Selected from; R N This includes hydrogen, optionally substituted C1-4 alkyl, optionally substituted C1-C3 cycloalkyl, and optionally substituted C 2- C4 alkenyl, optionally substituted C2-C4 alkynyl, optionally substituted 3-14 member heterocycloalkyl, optionally substituted 6-10 member aryl, or optionally substituted C1-C7 heteroalkyl; C1 and C 2 Each is independently selected from carbonyl, thiocarbonyl, sulfonyl, or phosphoryl; f, g, h, i, j, and k are each independently 0 or 1; D 1 C1-C, which were replaced by arbitrary selection. 10 Alkylene, optionally substituted C2-C 10 Alkenylene, optionally substituted C2-C 10 Alkynylene, optionally substituted 3-14 member heterocycloalkylene, optionally substituted 5-10 member heteroarylene, optionally substituted 3-8 member cycloalkylene, optionally substituted 6-10 member arylene, optionally substituted C2-C 10 Polyethylene glycolen, or optionally substituted C1-C 10 Heteroalkylene, or A 1 -( B 1 ) f -(C 1 ) g -( B 2 ) h -to-(B 3 ) i -(C 2 ) j -( B 4 ) k -A 2 It is a chemical bond that connects two things together.

[0128] In some embodiments, the linker is acyclic. In some embodiments, the linker has the structure of formula IIa: [ka] In the formula, X a It either does not exist or is N; R 14 It is either absent, hydrogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C3 cycloalkyl; L 2It does not exist, or it is a C1-C4 alkylene that is optionally substituted with -C(O)-, -SO2-, or an optionally substituted C1-C4 heteroalkylene. In the formula, X a , R 14 , or L 2 At least one of these exists.

[0129] In some embodiments, the linker is a cyclic group or contains a cyclic group. In some embodiments, the linker has the structure of formula IIb: [ka] In the formula, o is either 0 or 1; X b is C(O) or SO2; R 15 is a C1-C6 alkyl group substituted with hydrogen or optionally; Cy is an optionally substituted 3- to 8-membered cycloalkylene, an optionally substituted 3- to 8-membered heterocycloalkylene, an optionally substituted 6- to 10-membered arylene, or an optionally substituted 5- to 10-membered heteroarylene; L 3 It does not exist, or it is -C(O)-, -SO2-, optionally substituted C1-C4 alkylene, or optionally substituted C1-C4 heteroalkylene.

[0130] In some embodiments, the linker is absent.

[0131] In some embodiments, W is hydrogen. In some embodiments, W is optionally substituted cyclopropyl, optionally substituted cyclobutyl, optionally substituted cyclopentyl, optionally substituted cyclohexyl, optionally substituted piperidine, optionally substituted piperazine, optionally substituted pyridine, or optionally substituted phenyl. In some embodiments, W is optionally substituted amino. In some embodiments, W is optionally substituted amide. In some embodiments, W is optionally substituted C1-C4 alkoxy. In some embodiments, W is optionally substituted C1-C4 alkyl. In some embodiments, W is optionally substituted C1-C4 hydroxyalkyl. In some embodiments, W is optionally substituted C1-C4 aminoalkyl. In some embodiments, W is optionally substituted C1-C4 haloalkyl. In some embodiments, W is optionally substituted C1-C4 guanidinoalkyl. In some embodiments, W is a C0-C4 alkyl, or an optionally substituted 3- to 11-membered heterocycloalkyl. In some embodiments, W is an optionally substituted 3- to 10-membered cycloalkyl. In some embodiments, W is an optionally substituted 3- to 10-membered heteroaryl. In some embodiments, W is an optionally substituted 6- to 10-membered aryl.

[0132] In some embodiments, a potent hydrolyzing agent is intended, where the potent hydrolyzing agent is a compound comprising one of the following cores: [ka] And in the formula, [ka] It is one of the following: [ka] A compound, or a pharmaceutically acceptable salt thereof. In some embodiments, [ka] The following: [ka] It is one of them.

[0133] In some embodiments, a moderate hydrolyzing agent is intended, where the moderate hydrolyzing agent is a compound comprising one of the following cores: [ka] And in the formula, [ka] The following is true: [ka] A compound, or a pharmaceutically acceptable salt thereof. Examples of such compounds can be found, for example, in WO2021 / 091956, WO2022 / 060836, and WO2023 / 240263, each of which is incorporated herein by reference in whole.

[0134] In some embodiments, a weak hydrolysant is intended, where the weak hydrolysant is a compound comprising one of the following cores: [ka] And in the formula, [ka] It is one of the following: [ka] A compound, or a pharmaceutically acceptable salt thereof. Examples of such compounds can be found, for example, in WO2021 / 091956, WO2022 / 060836, and WO2023 / 240263, each of which is invoked in whole by reference.

[0135] This specification further describes compounds having the structure of formula Ic, Id, or Ie: [ka] or provide a pharmaceutically acceptable salt thereof, here: In formula Ic: R w It is methylcyclopropyl; R y These are CH3, CH2F, CHF, or CHF3; R z However, it is hydrogen or N-methylpiperazinyl; R 10 H is; A 1 is -CH2-, -O-, or -NCH3; In formula Id: R w These are methylcyclopropyl or dimethylcyclopropyl; R y It is CH3; R z However, it is hydrogen or N-methylpiperazinyl; R 10 H is; A 1 is -CH2-, -O-, or -NCH3; or In formula Ie: Either an E or Z n double bond exists; R w These are methylcyclopropyl or dimethylcyclopropyl; Ry It is CH3; R z is hydrogen or N-methylpiperazinyl; R 10 H is; A 1 It is -CH2-.

[0136] In some embodiments, a strong hydrolyzing agent is used in the methods disclosed herein. In some embodiments, a moderate hydrolyzing agent is not used in the methods of the present invention. In some embodiments, a weak hydrolyzing agent is not used in the methods of the present invention.

[0137] In some embodiments, the compounds of the disclosure are selected from Table 1, or their pharmaceutically acceptable salts or stereoisomers. [Table 1]

[0138] In some embodiments, the RAS(ON)GTP hydrolysis-promoting compounds of the present disclosure are selected from Table 2, or from their pharmaceutically acceptable salts or stereoisomers. In some embodiments, the compounds of the present disclosure are selected from Table 2, or from their pharmaceutically acceptable salts or atropisomers. [Table 2-1] [Table 2-2]

[0139] The RAS(ON)GTP hydrolysis-promoting compounds described herein may be prepared from commercially available starting materials or synthesized using known organic, inorganic, or enzymatic processes.

[0140] The compounds of the present invention can be prepared by methods known to those skilled in the art, in combination with known synthetic organic chemistry techniques, such as those disclosed in WO2021 / 091956, WO2022 / 060836 and WO2023 / 240263, each of which is incorporated herein by reference. For example, the compounds of the present invention can be synthesized by methods shown in the following scheme, in combination with synthetic methods known in the field of organic synthesis, or variations thereof as understood by those skilled in the art. These methods include, but are not limited to, those described in the following scheme. Scheme 1. General synthesis of macrocyclic esters [ka]

[0141] A common synthesis of alternatives to macrocyclic esters is outlined in Scheme 1. A well-substituted indolylboronic acid ester (1) can be prepared in four steps, starting with a protected 3-(5-bromo-2-iodo-1H-indole-3-yl)-2,2-dimethylpropan-1-ol and a well-substituted boronic acid, and involving palladium-mediated coupling, alkylation, deprotection, and palladium-mediated borylation reactions.

[0142] (S)-2-amino-3-(4-bromothiazole-2-yl)propanoic acid (2) can be coupled with methyl(S)-hexahydropyridazine-3-carboxylate to prepare methyl-amino-3-(4-bromothiazole-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (3).

[0143] In the presence of a Pd catalyst, coupling of methyl-amino-3-(4-bromothiazole-2-yl)propanoyl)hexahydropyridazine-3-carboxylate (3) with a appropriately substituted indolylboronic acid ester (1), followed by hydrolysis and macrolactonization steps, yields a appropriately protected macrocyclic intermediate (5), from which the final macrocyclic ester can be prepared. Deprotection and coupling with an appropriately substituted carboxylic acid (or other coupling partner) can yield the macrocyclic product. Additional deprotection or functionalization steps may be required to prepare the final compound 6.

[0144] Furthermore, with respect to Scheme 1, thiazole can be replaced with an alternative, optionally substituted 5-6 member heteroarylene, an optionally substituted 3-6 member cycloalkylene, an optionally substituted 3-6 member heterocycloalkylene (e.g., morpholino), or an optionally substituted 6 member arylene (e.g., phenyl). Scheme 2. General synthesis of alternative macrocyclic esters [ka]

[0145] Alternatively, macrocyclic esters can be prepared as described in Scheme 2. A well-substituted and protected indolylboronic acid ester (7) can be coupled with (S)-2-amino-3-(4-bromothiazole-2-yl)propanoic acid in the presence of a Pd catalyst, followed by iodination, deprotection, and ester hydrolysis. After coupling with methyl(S)-hexahydropyridazine-3-carboxylate, hydrolysis and macrolactonization can be followed to obtain an iodine intermediate (11). Subsequent palladium-mediated borylation and coupling with a well-substituted iodoaryl or iodoheteroaryl intermediate in the presence of a Pd catalyst can yield a well-protected macrocyclic intermediate. Alkylation, deprotection, and coupling with a well-substituted carboxylic acid (or other coupling partner) can yield the macrocyclic product. Additional deprotection or functionalization steps may be required to prepare the final compound 6.

[0146] Furthermore, with respect to Scheme 2, thiazole can be replaced with an alternative, optionally substituted 5- to 6-membered heteroarylene, an optionally substituted 3- to 6-membered cycloalkylene, an optionally substituted 3- to 6-membered heterocycloalkylene (e.g., morpholino), or an optionally substituted 6-membered arylene (e.g., phenyl).

[0147] The compounds listed in Table 1 of this specification were prepared using the methods disclosed herein, or using the methods described herein in combination with the knowledge of those skilled in the art.

[0148] As described herein, RAS(ON)GTP hydrolysis-promoting compounds increase the hydrolysis rate of RAS GTP (guanosine triphosphate) compared to the hydrolysis rate of RAS GTP (guanosine triphosphate) in the absence of the compound. GTP hydrolysis refers to the process in which GTP molecules are cleaved into GDP (guanosine diphosphate) and inorganic phosphate (Pi) in the presence of water. In various embodiments, the GTP hydrolysis rate increases by about 5 to 100% (e.g., at least about or about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or about 100% (including all values ​​and ranges between these values)), 10 to 95%, 15 to 90%, 20 to 85%, 25 to 75%, 30 to 70%, 35 to 65%, 40 to 60%, 45 to 55%, or 50% compared to the GTP hydrolysis rate in the absence of the compound. In various embodiments, the GTP hydrolysis rate increases by approximately 2 to 100 times (e.g., at least approximately or approximately 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 times (including all values ​​and the range between these values)) compared to the GTP hydrolysis rate in the absence of the inhibitor.

[0149] Assays for measuring the rate of GTP hydrolysis are known in the art and are intended herein. For example, the analysis of GTP hydrolysis rates can be performed in cell culture systems or cell-free systems. Thin-layer chromatography (TLC) is a technique used to separate and visualize different molecules, including nucleotides such as GTP and GDP. In a typical GTP hydrolysis assay, a reaction mixture containing GTP, RAS, and with or without the RAS(ON)GTP hydrolysis-promoting compound of this disclosure is incubated for a sufficient amount of time under appropriate conditions. The reaction is then stopped, and the reaction products (GDP and Pi) can be separated and quantified by TLC. A colorimetric assay can be used that relies on the detection of a colored product produced by the reaction between Pi and a specific reagent (such as molybdic acid or malachite green). In this type of assay, the GTP hydrolysis reaction is carried out in the presence of a colorimetric reagent, and the absorbance of the colored product is measured over time. Fluorescence-based assays, including fluorescent nucleotides such as mantle GTP that fluoresce during hydrolysis, can also be used. A reaction mixture containing a fluorescent nucleotide and RAS (with or without a RAS(ON)GTP hydrolysis-promoting compound) is prepared, and the decrease in fluorescence over time is observed using a fluorescence spectrophotometer. The radioactive assay may also use radiolabeled GTP ([γ-32P]GTP or [α-32P]GTP), which enables highly sensitive detection of the reaction products (GDP and Pi) by scintillation counting. Additional methods are described herein, for example, in the following examples.

[0150] In some embodiments, hydrolysis is increased in the presence of a RAS(ON)GTP hydrolysis-promoting compound for RAS proteins containing the G12C amino acid substitution, compared to wild-type RAS or other RAS variants. In some embodiments, GTP hydrolysis is increased for RAS containing the G12D amino acid substitution, compared to wild-type RAS or other KRAS variants. In some embodiments, GTP hydrolysis is increased for RAS containing the G12V amino acid substitution, compared to wild-type RAS or other RAS variants. In some embodiments, GTP hydrolysis is increased for RAS containing the G13D amino acid substitution, compared to wild-type RAS or other RAS variants. In each of the above embodiments, the RAS does not have a mutation at residue 61 compared to wild-type RAS.

[0151] b) RAS(OFF) inhibitors and RAS(OFF) degrading agents The compositions described herein may comprise one or more RAS(OFF) inhibitors. Numerous mutaselective and pan-RAS inhibitors are disclosed. RAS(OFF) inhibitors may be administered or formulated in combination with the RAS(ON)GTP hydrolytic compounds described herein. RAS(OFF) inhibitors are designed to inhibit RAS activity by targeting different regions of the inactive RAS protein and blocking its activation and downstream signaling.

[0152] In some embodiments, the RAS(OFF) inhibitor is a KRAS(OFF) inhibitor having a molecular weight of less than 700 Da. The term "KRAS(OFF) inhibitor" refers to any RAS(OFF) inhibitor that binds to KRAS in its GDP-bound "off" position. In some embodiments, the KRAS(OFF) inhibitor is a KRAS G12C It is specific to the mutation. KRAS G12C (OFF) inhibitors are KRAS G12C Using covalent groups that allow for selective targeting of mutant proteins, many such inhibitors contain a pyrimidine core. All KRASG12C(OFF) inhibitors contain a pyrimidine core. G12CKRAS targets identical cysteine ​​residues within mutant proteins, inducing a structural change that fixes the protein in an inactive state. G12C Examples of (OFF) inhibitors include AMG510 (sotrasib), MRTX849 (adaglasib), MRTX1257, GDC-6036 (divalasib), JDQ443 (opnurasib), ERAS-3490, LY3537982 (olomorasib), BI1823911, BPI-421286, JAB-3312, JAB-21000, JAB-21822 (glesilasib), D-1553, D3S-001, HBI-2438, HS-10370, MK-1084, YL-15293, BBO-8520 (ON / OFF inhibitor), FMC-376 (ON / OFF inhibitor), GEC255, and GFH925 (IBI351). In some embodiments, the KRAS(OFF) inhibitor is selected from AMG510 and MRTX849. In some embodiments, the KRAS(OFF) inhibitor is AMG510. In some embodiments, the KRAS(OFF) inhibitor is selected from BPI-421286, JNJ-74699157 (ARS-3248), LY3537982, MRTX1257, ARS853, ARS1620, and GDC-6036. [ka]

[0153] In some embodiments, the KRAS(OFF) inhibitor is KRAS G12D It is specific to the mutation. Many KRAS G12D (OFF) inhibitors were developed using RAS(OFF)G12C inhibitors as a starting point, and therefore share the G12C inhibitor skeleton when combined with other chemical components such as piperazine compounds. G12DNon-limiting examples of (OFF) inhibitors include MRTX1133, MRTX282, JAB-22000, ERAS-4, ERAS-5024, HRS-4642, BI-2852, ASP3082, TH-Z827, TH-Z835, QTX-3046, GFH375 (VS-7375), INCB161734, and KD-8. In some embodiments, the KRAS(OFF) inhibitor is MRTX1133.

[0154] References to "MRTX1133," "TH-Z827," "TH-Z835," and "KD-8" in this specification refer to the following compounds: [ka] It means...

[0155] In some embodiments, the small molecule RAS(OFF) inhibitor is KRAS G12V It is specific to the mutation. In some embodiments, the small molecule RAS(OFF) inhibitor is KRAS G13DThey are specific to the mutation. In some embodiments, small molecule RAS(OFF) inhibitors are specific to pan-RAS(OFF) inhibitors. In some embodiments, references to the term RAS(OFF) inhibitor include any such RAS(OFF) inhibitor disclosed in any one of the following patent applications: WO2024056063, WO2024055112, WO2024054926, WO2024054647, WO2024054625, WO2024051763, WO2024051721, WO2024050742, WO2024050640, WO2024046406, WO2024046370, WO2024 045066, WO2024044667, WO2024044649, WO2024044334, WO2024041621, WO2024041606, WO2024041589, WO2024041573, WO2024040131, WO202 4040109, WO2024040080, WO2024036270, WO2024034657, WO2024034593, WO2024034591, WO2024034123, WO2024032747, WO2024032704, WO20 24032703, WO2024032702, WO2024031088, WO2024030647, WO2024030633, WO2024029613, WO2024022507, WO2024022444, WO2024020159, WO2 024019103, WO2024017859, WO2024017392, WO2024015731, WO2024015262, WO2024012456, WO2024009191, WO2024008179, WO2024008178, WO 2024008068, WO2024006445, WO2024006424, WO2024002373, WO2023287896, WO2023287730, WO2023284881, WO2023284730, WO2023284537, W O2023283933, WO2023283213, WO2023280280, WO2023280136, WO2023280026, WO2023278600, WO2023274383, WO2023327324, WO2023246914,WO2023246903, WO2023246777, WO2023244713, WO2023244615, WO2023244604, WO2023244600, WO2023244599, WO2023230190, WO2023226630, WO202322 5302, WO2023225252, WO2023220421, WO2023219941, WO2023217148, WO2023215802, WO2023215801, WO2023213269, WO2023212548, WO2023208005, WO2 023205719, WO2023199180, WO2023198191, WO2023197984, WO2023190748, WO2023185864, WO2023183755, WO2023183585, WO2023179703, WO202317962 9, WO2023173017, WO2023173016, WO2023173014, WO2023172737, WO2023171781, WO2023159087, WO2023159086, WO2023154766, WO2023152255, WO2023 151674、WO2023151621、WO2023150394、WO2023150284、WO2023143623、WO2 023143605、WO2023143352、WO2023143352、WO2023143312、WO2023141570、W O2023141300, WO2023138662, WO2023138601, WO2023138589, WO2023138524, WO2023133183, WO2023133181, WO2023130012, WO2023125989, WO2023125 627, WO2023122662, WO2023122154, WO2023120742, WO2023119677, WO2023117681, WO2023116934, WO2023116895, WO2023114733, WO2023105491, WO20 23104018, WO2023103906, WO2023103523, WO2023101928, WO2023099624, WO2023099624, WO2023099620, WO2023099612, WO2023099608, WO2023099592WO2023098832, WO2023098425, WO2023097227, WO2023081840, WO2023081476, WO2023078424, WO2023077441, WO2023072297, WO2023072188, WO20230 66371, WO2023064857, WO2023061463, WO2023061294, WO2023057985, WO2023056951, WO2023056421, WO2023051586, WO2023049697, WO2023046135, W O2023045960, WO2023041059, WO2023041059, WO2023040989, WO2023040513, WO2023039240, WO2023039020, WO2023036282, WO2023034290, WO202303 0517, WO2023030495, WO2023030385, WO2023025116, WO2023020523, WO2023020521, WO2023020519, WO2023020518, WO2023020347, WO2023018812, WO2 023018810, WO2023018809, WO2023018699, WO2023014979, WO2023014006, WO2023004102, WO2023003417, WO2023001141, WO2023001123, WO20222716 58, WO2022269508, WO2022266167, WO2022266069, WO2022266015, WO2022265974, WO2022261154, WO2022261154, WO2022251576, WO2022251296, WO202 2237815, WO2022232332, WO2022232331, WO2022232320, WO2022232318, WO2022223037, WO2022221739, WO2022221528, WO2022221386, WO2022216762 (e.g., compound 44 or compound 66a), WO2022192794, WO2022192790, WO2022188729, WO2022187411, WO2022184178, WO2022173870, WO2022173678, WO2022135346,WO2022133731、WO2022133038、WO2022133345、WO2022132200、WO2022119748、WO2022109485、WO2022109487、WO2022066805、WO2022002102、WO2022002018、WO2021259331、WO2021257828、WO2021252339、WO2021248095、WO2021248090、WO2021248083、WO2021248082、WO2021248079、WO2021248055、WO2021245051、WO2021244603、WO2021239058、WO2021231526、WO2021228161、WO2021219090、WO2021219090、WO2021219072、WO2021218939、WO2021217019、WO2021216770、WO2021215545、WO2021215544、WO2021211864、WO2021190467、WO2021185233、WO2021180181、WO2021175199、2021173923、WO2021169990、WO2021169963、WO2021168193、WO2021158071、WO2021155716、WO2021152149、WO2021150613、WO2021147967、WO2021147965、WO2021143693、WO2021142252、WO2021141628、WO2021139748、WO2021139678、WO2021129824、WO2021129820、WO2021127404、WO2021126816、WO2021126799、WO2021124222、WO2021121371、WO2021121367、WO2021121330、WO2021113595、WO2021107160、WO2021106231、WO2021088458、WO2021086833、WO2021085653、WO2021081212、WO2021058018、WO2021057832、WO2021055728、WO2021031952、WO2021027911、WO2021023247、WO2020259513、WO2020259432、WO2020234103、WO2020233592、WO2020216190、WO2020178282、WO2020146613、WO2020118066、WO2020113071、WO2020106647、WO2020102730、WO2020101736、WO2020097537、WO2020086739、WO2020081282、WO2020050890、WO2020047192、WO2020035031、WO2020028706、WO2019241157、WO2019232419、WO2019217691、WO2019217307、WO2019215203、WO2019213526、WO2019213516、WO2019155399、WO2019150305、WO2019110751、WO2019099524、WO2019051291、WO2018218070、WO2018218071、WO2018218069、WO2018217651、WO2018206539、WO2018143315、WO2018140600、WO2018140599、WO2018140598、WO2018140514、WO2018140513、WO2018140512、WO2018119183、WO2018112420、WO2018068017、WO2018064510、WO2017201161、WO2017172979、WO2017100546、WO2017087528、WO2017058807、WO2017058805、WO2017058728、WO2017058902、WO2017058792、WO2017058768、WO2017058915、WO2017015562、WO2016168540、WO2016164675、WO2016049568、WO2016049524、WO2015054572、WO2014152588、WO2014143659、WO2013155223、CN117683051、CN117645627、CN117624194、CN117624190、CN117586280、CN117486901、CN117466917、CN117462688、CN117362315、CN117327102、CN117327094、CN117327074、CN117285590、CN117263959、CN117247382、CN117186095、CN117164605、CN、 116969977、CN116925075、CN116891489、CN116731045、CN116731044、CN116554208、CN116514846、CN116478184、CN116478141、CN116410145、CN116375742、CN116354988、CN116332948、CN116332938、CN116327956、CN116262759、CN116217592、CN116199703、CN116162099、CN116143806、CN116143805、CN116120315、CN116102559、CN115960105、CN115894520、CN115872979、CN115850267、CN115785199、CN115785124、CN115785124、CN115724842、CN115716840、CN115703775、CN115611923、CN115611898、CN115583937、CN115572278、CN115557949、CN115521312、CN115504976、CN115490709、CN115466272、CN115433183、CN115433179、CN115403575、CN115385938、CN115385937、CN115385912、CN115381786、CN115368383、CN115368382、CN115368381、CN115353506、CN115322158、CN115304623、CN115304602、CN115197245、CN115181106、CN114989195、CN114989166、CN114989147、CN114920741、CN114920739、CN114907387、CN114874234、CN114874201、CN114716436、CN114716435、CN114685532、CN114685460、CN114591319、CN114539293、CN114539286、CN114539246、CN114437107、CN114437084、CN114409653、CN114380827、CN114195804、CN114195788、CN114057776、CN114057744、CN114057743、CN113999226, CN113980032, CN113980014, CN113929676, CN113754653, CN113683616, CN113563323, CN113527299, CN1 13527294, CN113527293, CN113493440, CN113429405, CN113248521, CN113087700, CN113024544, CN113004269, CN11292 0183, CN112778284, CN112390818, CN112390788, CN112300196, CN112300194, CN112300173, CN112225734, CN112142735, CN112110918, CN112094269, CN112047937, and CN109574871, each of these, including the compound structures disclosed therein, are incorporated herein by reference in their entirety.

[0156] In some embodiments, KRAS(OFF) inhibitors are peptide-based inhibitors. For example, peptide-based RAS(OFF) inhibitors have been developed that target specific regions of the RAS protein, such as the switch II region or the RAS effector interface. Non-limiting examples include K-Ras binding peptide (KRpep-2d), Ras inhibitory peptide (RasIn), and LUNA18 (NCT05012618). Peptide-based RAS(OFF) inhibitors are a class of compounds that target the RAS protein by interfering with its interaction with downstream effectors or other signaling proteins. These inhibitors are typically designed to mimic the binding motif of RAS interacting proteins or other RAS effectors, such as RAF or PI3K. By binding to RAS at the same site as these effectors, peptide-based inhibitors can effectively compete with these proteins and prevent activation of downstream signaling pathways.

[0157] Peptide-based RAS(OFF) inhibitors can be further classified into two main categories: those targeting the RAS-effector interface and those targeting other regions of the RAS protein. Peptide-based inhibitors targeting the RAS-effector interface are designed to bind to the switch region of RAS, which is important in interactions with downstream effectors such as RAF or PI3K. These inhibitors typically contain amino acid residues similar to those found in the binding motif of RAS-interacting proteins or effectors, and are often designed to form hydrogen bonds or other interactions with key residues on the RAS surface.

[0158] Peptide-based RAS(OFF) inhibitors that target other regions of the RAS protein are typically designed to interfere with other interactions that are important for RAS activation or signaling. For example, some peptide-based inhibitors are designed to bind to the hypervariable region of the RAS, which is thought to play a role in protein membrane localization and fixation. By binding to this region, peptide-based inhibitors may interfere with the proper localization of the RAS to the plasma membrane, which is necessary for activation and signaling.

[0159] Several common motifs have been identified as important for the binding of RAS-interacting proteins and effectors and are often used in the design of peptide-based inhibitors. An example is the RAF-binding domain (RBD), found in many RAS-interacting proteins and crucial for the interaction between RAS and downstream effectors such as RAFs. The RBD contains a conserved amino acid sequence (Arg-Xaa-Arg) important for binding to RAS, and this motif has been incorporated into several peptide-based inhibitors designed to disrupt RAS-RAF interactions. Another example is the RAS-binding domain (RBD) of PI3K, which is important for the interaction between RAS and its downstream effector. The PI3K RBD contains several conserved amino acid residues (such as Arg-Arg-Trp) important for binding to RAS, and these motifs have been used in the design of peptide-based inhibitors targeting RAS-PI3K interactions. Other common motifs used in peptide-based RAS(OFF) inhibitors include sequences that mimic the structure of the Ras-binding domain (RBD) of other RAS-interacting proteins such as RalGDS and SOS, as well as the switch region of RAS itself. These motifs are typically used to optimize the binding affinity and selectivity of the inhibitor for a desired target protein or interaction.

[0160] In some embodiments, RAS(OFF) inhibitors are antibodies or antigenic binding peptides specific to RAS(OFF). For example, antibodies have been developed that bind to specific regions of the RAS protein, such as the switch II region or the RAS effector interface. For instance, several antibodies have been developed that target the switch region of the RAS protein, which is crucial for the activation of these proteins and their interaction with downstream effectors. By binding to the switch region, these antibodies can block the conformational changes necessary for RAS activation and downstream signaling. Other approaches involve the use of antibodies that target RAS-interacting proteins or downstream effectors, such as RAF or PI3K. By binding to the target protein, these antibodies can disrupt the RAS-dependent signaling pathway, potentially inhibiting the proliferation and survival of cancer cells. Furthermore, several antibodies have been developed that can induce the internal translocation and degradation of RAS proteins, leading to their depletion and inhibition of downstream signaling. For example, several antibodies have been developed that recognize the unique structures of mutant RAS proteins and target them for degradation via the ubiquitin-proteasome pathway. Non-specific examples of KRAS(OFF)-specific inhibitory antibodies include anti-p21ser and K27(DARPin) (see, for example, Khan et al, Biochim Biophys Acta Mol Cell Res. 2020 Feb;1867(2):118570).

[0161] In any embodiment of this specification using an RAS(OFF) inhibitor, RAS(OFF) degrading agents targeting the OFF state of the RAS may be used as an alternative. These degrading agents are known in the art. RAS degrading agents may be found, for example, in one or more of the following applications: WO2024055112, WO2024054625, WO2024050742, WO2024044334, WO2024040080, WO2024034657, WO2024034593, WO2024034591, WO2024034123, WO2024029613, WO2024020159, WO2024019103, WO2024017392, WO2023185864, WO2023171781, WO2023141570, WO2023138524, WO2023130012, WO2023116934, WO2023099620, WO2023081476, WO2023077441, and CN115785199, each of which includes the compound structure disclosed in the said document, is incorporated herein by reference. The RAS(OFF) inhibitory structures disclosed herein provide means for inhibiting RAS(OFF).

[0162] c) RTK inhibitors The compositions and methods described herein may, in combination with one or more receptor tyrosine kinase inhibitors, include RAS(ON)GTP hydrolysis-promoting compounds. Receptor tyrosine kinase (RTK) inhibitors are a type of molecule (e.g., small molecules, antibodies, and nucleic acids) that binds to receptor tyrosine kinases or their ligands and blocks their activity. RTKs are proteins found on the surface of cells and play important roles in cell signaling and proliferation, and have been developed as therapeutic agents for various diseases, including cancer, diabetes, and autoimmune disorders.

[0163] i) EGFR inhibitors In some embodiments, the compositions and methods described herein may comprise one or more EGFR inhibitors. The EGFR inhibitors may be administered or formulated in combination with the RAS(ON)GTP hydrolysis-promoting compounds described herein and / or any additional therapeutic agents described herein. EGFR inhibitors include, but are not limited to, small molecule antagonists, antibody inhibitors, or specific antisense nucleotides or siRNAs. Useful antibody inhibitors of EGFR include cetuximab (Erbitux®), panitumumab (Vectibix®), zaltumumab, nimotuzumab, and matuzumab. Further antibody-based EGFR inhibitors include any anti-EGFR antibody or antibody fragment capable of partially or completely blocking 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 may be a monoclonal antibody Mab E7.6.3 (Yang, 1999 (see above)), or Mab C225 (ATCC accession number HB-8508), or an antibody or antibody fragment having binding specificity thereto.

[0164] Examples of small molecule antagonists of EGFR include gefitinib (Iressa®), razertinib, erlotinib (Tarceva®), and lapatinib (TykerB®). See, for example, 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®). In some embodiments, the EGFR inhibitor is one or more of cetuximab, gefitinib (Iressa), erlotinib (Tarceva), and afatinib (Gilotrif). 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. The EGFR inhibitor may be ERAS-801. In some embodiments, the EGFR inhibitor is an ERBB inhibitor. In humans, the ERBB family includes HER1 (EGFR, ERBB1), HER2 (NEU, ERBB2), HER3 (ERBB3), and HER (ERBB4). In some embodiments, references to the term EGFR inhibitor include any such EGFR inhibitor disclosed in any one of the following patent applications: WO2023041071, WO2023049312, WO2023020600, WO2023284747, WO2022206797, WO2022258977, WO2022033416, WO2022033410, WO2022105908, WO2022100641, WO2022014639, WO2022007841, WO2021018009,WO2021057882, WO2021252661, WO2021018003, WO2021073498, WO2021238827, WO2020254547, WO2020216371, WO2020147838, WO202 0207483, WO2020254572, WO2020001350, WO2021001351, WO2019164948, WO2019218958, WO2019046775, WO2019015655, WO20181217 58, WO2018218963, WO2017220007, WO2017205459, WO2017161937, WO2016192609, WO199633980, WO199630347, WO199730034, WO199 730044, WO199738994, WO199749688, WO199802434, WO199738983, WO199519774, WO199519970, WO199713771, WO199802437, WO1998 02438, WO199732881, WO199833798, WO199732880, WO199732880, WO199702266, WO199727199, WO199807726, WO1997 / 34895, WO1996 31510, WO199814449, WO199814450, WO199814451, WO199509847, WO199719065, WO199817662, WO199935146, WO199935132, WO19990 7701, WO199220642, DE19629652, EP682027, EP837063, EP0787772, EP0520722, EP0566226, CN115960018, CN110283162, CN114044774, CN111973601, CN111973602, and CN113896744, each of these, including the compound structures disclosed in the said documents, are specifically incorporated herein by reference, and the whole thereof is incorporated herein by reference.

[0165] ii) HER2 inhibitors In some embodiments, the compositions and methods described herein may comprise one or more HER2 inhibitors. The HER2 inhibitors may be administered or formulated in combination with the RAS(ON)GTP hydrolysis-promoting compounds described herein and / or any additional therapeutic agents described herein. In some embodiments, the HER2 inhibitors are one or more of tucatinib, rastuzumab (Herceptin), pertuzumab (Perjeta), lapatinib (Tykerb), ad-trastuzumab ethansine (Kadcyla), and neratinib (Nerlynx). Non-exclusive examples of HER2 inhibitors include monoclonal antibodies, e.g., trastuzumab (Herceptin®) and pertuzumab (Perjeta®); small molecule tyrosine kinase inhibitors, e.g., gefitinib (Iressa®), erlotinib (Tarceva®), pyritinib, CP-654577, CP-724714, canertinib (CI1033), HKI-272, lapatinib (GW-572016; Tykerb®), PKI-166, AEE788, BMS-599626, HKI-357, BIBW2992, ARRY-334543, and JNJ-26483327. In some embodiments, references to the term HER2 inhibitor include any such HER2 inhibitor disclosed in any one of the following patent applications: WO2021156178, WO2021156180, WO2021213800, WO2021088987, WO2013561183, and WO2013056108, each of which includes the compound structure disclosed in the said document, which is specifically incorporated herein by reference.

[0166] iii) MET inhibitors In some embodiments, the compositions and methods described herein may comprise one or more MET inhibitors. The MET inhibitors may be administered or formulated in combination with the RAS(ON)GTP hydrolysis-promoting compounds described herein and / or any additional therapeutic agents described herein. In some embodiments, the MET inhibitors are one or more of crizotinib (Xalkori), cabozantinib (Cometriq, Cabometyx), capmatinib (Tabrecta), tepotinib (Tepmetko), savolitinib (Volitinib), onartuzumab (MetMab), foretinib (GSK1363089), MGCD-265 (Amuvatinib), SU11274, and SU5416. In some embodiments, references to the term MET inhibitor include any such MET inhibitor disclosed in any one of the following patent applications: WO2022226168, WO2021222045, WO2020047184, WO2020015744, WO2020244654, WO2020156453, WO2019206268, WO2018077227, WO2017012539, WO2016015653, WO2016012963, WO20120156 77, WO2011162835, WO2010089507, WO2009091374, WO2009056692, WO2008051547, WO2007130468, US2012237524, CN103497177, CN107311983, CN107382968, CN110218191, and TW201331206, each of these, including the compound structures disclosed in the said documents, are specifically incorporated herein by reference, and the whole thereof is incorporated herein by reference.

[0167] iv) AXL inhibitors In some embodiments, the compositions and methods described herein may comprise one or more AXL inhibitors. The AXL inhibitors may be administered or formulated in combination with the RAS(ON)GTP hydrolysis-promoting compounds described herein and / or any additional therapeutic agents described herein. AXL is a receptor tyrosine kinase belonging to the TAM family of receptors, which also includes TYRO3 and MERTK. In some embodiments, the AXL inhibitor is one or more of vemcentib, BGB324, R428, SGI-7079, TP-0903, BMS-777607, UNC2025, and TP-0903. In some embodiments, references to the term AXL inhibitor include any such AXL inhibitor disclosed in any one of the following patent applications: WO2023045816, WO2022237843, WO2022246179, WO2021012717, WO2021088787, WO2021067772, WO2021239133, WO2021204713, WO2020238802, WO201 9039525, WO2019101178, WO2019074116, WO2017146236, WO2016097918, WO2015012298, WO2010005876, WO2010083465, CN115073367, and JP2022171109, each of these, including the compound structures disclosed in the said documents, are specifically incorporated herein by reference, and the whole thereof is incorporated herein by reference.

[0168] V) IGFR inhibitors In some embodiments, the compositions and methods described herein may comprise one or more insulin-like growth factor receptor 1 (IGF-1R) inhibitors. The IGFR inhibitor may be administered or formulated in combination with the RAS(ON)GTP hydrolytic compounds described herein and / or any additional therapeutic agents described herein. IGFR inhibitors have been developed to target the IGFR receptor, which plays a critical role in cancer progression and metastasis. In some embodiments, the IGFR inhibitor is one or more of lincitinib, AXL1717, OSI-906 (lincitinib), BMS-754807, BI836845, AZ12253801, PQIP (pyrrolo[1,2-a]quinoxaline), and NVP-AEW541. In some embodiments, references to the term IGFR inhibitor include any such IGFR inhibitor disclosed in any one of the following patent applications: WO2022115946, WO2022217923, WO2021203861, WO2021246413, WO2020116398, WO2019046600, WO2018195250, WO201822152 1, WO2018204872, WO2017072196, WO2016173682, WO2015162291, WO2015162292, WO2010066868, WO2006069202, and CN112125916, each of these, including the compound structures disclosed in the said documents, are specifically incorporated herein by reference, and the whole thereof is incorporated herein by reference.

[0169] v) RET inhibitors In some embodiments, the compositions and methods described herein may include one or more transfection-conversion (RET) inhibitors. The RET inhibitors may be administered or formulated in combination with the RAS(ON)GTP hydrolysis-promoting compounds described herein and / or any additional therapeutic agents described herein. RETs play important roles in various cellular processes, including cell proliferation, differentiation, survival, and migration. RETs are activated by binding to their ligands, such as ligands of the glial cell line-derived neurotrophic factor (GDNF) family, which leads to the activation of downstream signaling pathways that facilitate these cellular processes. In some embodiments, the RET inhibitors are one or more of pralcetinib, serpercatinib (LOXO-292), BLU-667, RXDX-105, TPX-0046, GSK3179106, molydastat (BAY85-3934), and RPI-1 (retrofin). In some embodiments, references to the term RET inhibitor include any such RET inhibitor disclosed in any one of the following patent applications: WO2021211380, WO2021057963, WO2021043209, WO2021222017, WO2020035065, WO2020114487, WO2020200314, WO2020200316, WO2020114494, WO2018071447, WO2018213329, WO2017079140, WO2014050781, CN113943285, CN113683610, CN113683611, CN113620944, CN113620945, CN113527291, CN113527292, CN113527290, CN113135896, CN111057075, CN111233899, and CN111362923, each of these, including the compound structure disclosed in the said document, is specifically incorporated herein by reference, and the whole thereof is incorporated herein by reference.

[0170] vi) ROS1 inhibitors In some embodiments, the compositions and methods described herein may comprise one or more c-ros oncogene 1 (ROS1) inhibitors. The ROS1 inhibitor may be administered or formulated in combination with the RAS(ON)GTP hydrolysis-promoting compounds described herein and / or any additional therapeutic agents described herein. ROS1 is a receptor tyrosine kinase belonging to the insulin receptor family and plays a role in various cellular processes, including cell proliferation, differentiation, survival, and migration. In some embodiments, the ROS1 inhibitor is one or more of taretrectinib, DS-6051b, TPX-0131, GZD824, and PF-06463922. In some embodiments, references to the term ROS1 inhibitor include any such ROS1 inhibitor disclosed in any one of the following patent applications: WO2021098703, WO2020024825, and US2017079972, each of which includes the compound structure disclosed in said document, which is specifically incorporated herein by reference.

[0171] vii) PDGFR inhibitors In some embodiments, the compositions and methods described herein may comprise one or more platelet-derived growth factor receptor (PDGFR) inhibitors. The PDGFR inhibitors may be administered or formulated in combination with the RAS(ON)GTP hydrolytic compounds described herein and / or any additional therapeutic agents described herein. PDGFR is a family of receptor tyrosine kinases consisting of two members, PDGFRα and PDGFRβ. They are activated by binding to their ligands, such as platelet-derived growth factor (PDGF), which leads to the activation of downstream signaling pathways that promote cell proliferation, growth, and survival. In some embodiments, the PDGFR inhibitor is one or more of CP-673451, imatinib, nintedanib (Ofev), sunitinib (Stent), pazopanib (Botrian), regorafenib (Stivarga), and dasatinib (Sprycel).

[0172] viii) FGFR inhibitors In some embodiments, the compositions and methods described herein may comprise one or more fibroblast growth factor receptor (FGFR) inhibitors. The FGFR inhibitors may be administered or formulated in combination with the RAS(ON)GTP hydrolytic compounds described herein and / or any additional therapeutic agents described herein. FGFR is a family of receptor tyrosine kinases consisting of four members: FGFR1-4. FGFRs are activated by binding to their ligand, fibroblast growth factor (FGF), which leads to the activation of downstream signaling pathways that promote cell proliferation, differentiation, and survival. In some embodiments, the FGFR inhibitor is an inhibitor of FGFR2. In some embodiments, the EGFR inhibitor is an inhibitor of FGFR4. In some embodiments, the FGFR inhibitor is one or more of futivatinib (TAK-659), erdafitinib (Barvasa), infiglatinib (Truseltiq), Debio1347, and logaratinib (BAY1163877).In some embodiments, references to the term FGFR inhibitor include any such FGFR inhibitor disclosed in any one of the following patent applications: WO2022033472, WO2022152274, WO2022166469, WO2022206939, WO2021037219, WO2021089005, WO2021113462, WO2020185532, WO2019213544, WO2020164603, WO2019154364, WO2019034076, WO2019213506, WO2019223766, WO2018028438, WO201815 3373, WO2018121650, WO2018010514, WO2017028816, WO2017118438, WO2016134320, WO2015008844, WO2014172644, WO2014007951, WO2013179033, WO2013087578, WO2012047699, CN105906630, CN115869315, CN115141176, CN115043832 and CN115028634, each of these, including the compound structures disclosed in the said documents, are specifically incorporated herein by reference, and the whole of them is incorporated herein by reference.

[0173] ix) VEGF inhibitors In some embodiments, the compositions and methods described herein may comprise one or more vascular endothelial growth factor (VEGF) signaling inhibitors. VEGF signaling inhibitors are a class of drugs that target signaling pathways mediated by VEGF and its receptors. VEGF plays a crucial role in angiogenesis, the process of forming new blood vessels from existing ones, and is overexpressed in many types of cancer, making it an attractive target for cancer treatment. VEGF inhibitors may be administered or formulated in combination with the RAS(ON)GTP hydrolytic compounds described herein, and / or any additional therapeutic agents described herein. In some embodiments, the VEGF inhibitor is an antibody or antigen-binding domain that specifically binds to VEGF (e.g., bevacizumab), or a soluble VEGF receptor or its ligand-binding domain, e.g., VEGF-TRAP®, and an anti-VEGF receptor agent (e.g., an antibody or antigen-binding domain that specifically binds to it). In some embodiments, the VEGF inhibitor is one or more of bevacizumab, aflibercept, ramucirumab, sorafenib, sunitinib, and pazopanib.

[0174] d) SHP inhibitors In some embodiments, the compositions and methods described herein may comprise one or more SHP inhibitors. The SHP inhibitor may be administered or formulated in combination with the RAS(ON)GTP hydrolysis-promoting compounds described herein and / or any additional therapeutic agents described herein. In some embodiments, the SHP inhibitor is an inhibitor of SHP1. In some embodiments, the SHP inhibitor is an inhibitor of SHP2. In some embodiments, the SHP1 inhibitor is SB6299 aka DA-4511. In some embodiments, the SHP2 inhibitor is one or more of SHP099, TNO155, RMC-4550, RMC-4630, JAB-3068, JAB-3312, RLY-1971, ERAS-601, SH3809, PF-07284892, or BBP-398. In some embodiments, references to the term SHP2 inhibitor include any such SHP2 inhibitor disclosed in any one of the following patent applications: WO2023282702, WO2023280283, WO2023280237, WO2023018155, WO2023011513, WO2022271966, W O2022271964, WO2022271911, WO2022259157, WO2022242767, WO2022241975, WO2022237676, W O2022237367, WO2022237178, WO2022235822, WO20222084008, WO2022135568, WO2022063190, WO2022043865, WO2022042331, WO2022033430, WO2022017444, WO2022007869, WO2021259077, WO2021249449, WO2021249057, WO2021244659, WO2021218755, WO2021176072, WO2021171261, WO2021149817, WO2021148010, WO2021147879, WO2021143823, WO2021143701, WO2021143680, WO2021281752, WO2021121397, WO2021119525, WO2021115286, WO2021110796, WO2021088945,WO2021073439、WO2021061706、WO2021061515、WO2021043077、WO2021033153、WO2021028362、WO2021033153、WO2021028362、WO2021018287、WO2020259679、WO2020249079、WO2020210384、WO2020201991、WO2020181283、WO2020177653、WO2020165734、WO2020165733、WO2020165732、WO2020156243、WO2020156242、WO2020108590、WO2020104635、WO2020094104、WO2020094018、WO2020081848、WO2020073949、WO2020073945、WO2020072656、WO2020065453、WO2020065452、WO2020063760、WO2020061103、WO2020061101、WO2020033828、WO2020033286、WO2020022323、WO2019233810、WO2019213318、WO2019183367、WO2019183364、WO2019182960、WO2019167000、WO2019165073、WO2019158019、WO2019152454、WO2019051469、WO2019051084、WO2018218133、WO2018172984、WO2018160731、WO2018136265、WO2018136264、WO2018130928、WO2018129402、WO2018081091、WO2018057884、WO2018013597、WO2017216706、WO2017211303、WO2017210134、WO2017156397、WO2017100279、WO2017079723、WO2017078499、WO2016203406、WO2016203405、WO2016203404、WO2016196591、WO2016191328、WO2015107495、WO2015107494、WO2015107493、WO2014176488、WO2014113584、CN115677661、CN115677660、CN115611869、CN115521305, CN115490697, CN115466273, CN115394612, CN115304613, CN11530461 2, CN115300513, CN115197225, CN114957162, CN114920759, CN114716448, CN1146718 79, CN114539223, CN114524772, CN114213417, CN114195799, CN114163457, CN11389 6710, CN113248521, CN113248449, CN113135924, CN113024508, CN112920131, CN1128 23796, CN112409334, CN112402385, CN112174935, 111848599, CN111704611, CN111393459, CN111265529, CN110143949, CN108113848, US11179397, US11044675, US11034705, US11033547, US11001561, US10988466, US10954243, US10934302, or US10858359, each of these, including the compound structure disclosed in the said document, is specifically incorporated herein by reference.

[0175] e) SOS1 inhibitors In some embodiments, the compositions and methods described herein may comprise one or more SOS1 inhibitors. The SOS1 inhibitors may be administered or formulated in combination with the RAS(ON)GTP hydrolysis-promoting compounds described herein and / or any additional therapeutic agents described herein. In some embodiments, the SOS1 inhibitors are one or more of RMC-5845, RMC-4948, RMC-0331, BI-1701963, BI-3406, SDR5, MRTX-0902, and BAY-293. In some embodiments, references to the term SOS1 inhibitor include any such SOS1 inhibitor disclosed in any one of the following patent applications: WO2023029833, WO2023041049, WO2023022497, WO2022184116, WO2022170952, WO2022170917, WO2022171184, WO2022170802, WO202216 1461, WO2022121813, WO2022028506, WO2022139304, WO2021228028, WO2019122129, CN115215847, CN115028644, CN114685488, CN111393519, each of these, including the compound structures disclosed in the said documents, are specifically incorporated herein by reference, and the whole thereof is incorporated herein by reference.

[0176] f) Pharmaceutical compositions This disclosure provides a pharmaceutical composition comprising a RAS(ON)GTP hydrolysate compound in combination with one or more RAS(OFF) inhibitors, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0177] In some embodiments, the compound is present in the pharmaceutical composition in a unit dose appropriate for administration in a treatment regimen that exhibits a statistically significant probability of achieving a predetermined therapeutic effect when administered to the relevant population. In some embodiments, the pharmaceutical composition may be specifically formulated for administration in solid or liquid form, including those suitable for oral administration, e.g., oral tablets (aqueous or nonaqueous solutions or suspensions), tablets, e.g., buccal, sublingual, and those targeted for intracellular absorption, boluses, powders, granules, and pastes for application to the tongue; parenteral administration, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection, e.g., as sterile solutions or suspensions, or as sustained-release formulations; topical application, e.g., as creams, ointments, or controlled-release patches or sprays applied to the skin, lungs, or oral cavity; vaginal or rectal administration, e.g., as pessaries, creams, or foams; sublingual administration; intraocular administration; transdermal administration; or administration via nasal, intrapulmonary, and other mucosal surfaces.

[0178] The compounds described herein, whether expressly stated or not, may be provided or used in salt form, for example, pharmaceutically acceptable salt form, unless expressly stated otherwise.

[0179] The compounds of this disclosure may have ionic groups so that they can be prepared as pharmaceutically acceptable salts. These salts may be acid addition salts with inorganic or organic acids, or, in the case of the acidic forms of the compounds of this disclosure, the salts may be prepared from inorganic or organic bases. In some embodiments, the compounds are prepared or used as pharmaceutically acceptable salts prepared as addition products of pharmaceutically acceptable acids or bases. Suitable pharmaceutically acceptable acids and bases are well known in the art for forming acid addition salts, such as hydrochloric acid, sulfuric acid, hydrobromic acid, acetic acid, lactic acid, citric acid, or tartaric acid, and for forming basic salts, such as potassium hydroxide, sodium hydroxide, ammonium hydroxide, caffeine, and various amines. Methods for preparing suitable salts are well established in the art.

[0180] Typical acid addition salts include acetate, adipine, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphor sulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, and 2-(optionally substituted) hydroxyethane. Examples include sulfonates, lactobionates, lactates, laurates, lauryl sulfates, malates, maleates, malons, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectinates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propions, stearates, succinates, sulfates, tartrates, thiocyanates, toluenesulfonates, undecanoic acid, and valerates. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, as well as non-toxic ammonium, quaternary ammonium, and amine cations (including, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine).

[0181] For use as a treatment for a subject, the compounds of this disclosure, or pharmaceutically acceptable salts thereof, can be formulated as pharmaceutical or veterinary compositions. Depending on the subject to be treated, the method of administration, and the type of treatment desired, e.g., prevention, prophylaxis, or therapeutic treatment, the compounds, or pharmaceutically acceptable salts thereof, are formulated in a manner consistent with these parameters. A summary of such techniques can be found in Remington: The Science and Practice of Pharmacy, 21st Edition, Lippincott Williams & Wilkins, (2005); and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and JCBoylan, 1988-1999, Marcel Dekker, New York, each of which is incorporated herein by reference in whole.

[0182] Each composition can be prepared according to conventional mixing, granulation, or coating methods, and the pharmaceutical compositions may contain about 0.1% to about 99%, about 5% to about 90%, or about 1% to about 20% (by weight or by volume) of the compounds of the Disclosure or their pharmaceutically acceptable salts. In some embodiments, the compounds described herein or their pharmaceutically acceptable salts may be present in total at an amount of 1.95% of the total weight of a composition such as a pharmaceutical composition.

[0183] The composition can be provided in dosage forms suitable for intra-articular, oral, parenteral (e.g., intravenous, intramuscular), rectal, cutaneous, subcutaneous, topical, transdermal, sublingual, transnasal, intravaginal, intrabladderal, intraurethral, ​​intrathecal, epidural, transaural, or intraocular administration, or for injection, inhalation, or direct contact with the nasal, genitourinary, reproductive, or oral mucosa. Accordingly, the pharmaceutical composition may be in the form of, for example, tablets, capsules, pills, powders, granules, suspensions, emulsions, solutions, gels containing hydrogels, pastes, ointments, creams, plasters, oral medications, infiltration delivery devices, suppositories, enemas, injections, implants, sprays, preparations suitable for iontophoresis delivery, or aerosols. The composition can be formulated according to conventional pharmaceutical regulations.

[0184] The formulations can be prepared in a manner suitable for systemic administration or local or local administration. Systemic formulations may be designed for injection (e.g., intramuscular, intravenous, or subcutaneous injection), or they may be prepared for transdermal, transmucosal, or oral administration. The formulations generally contain diluents, and optionally adjuvants, buffers, preservatives, etc. The compound, or a pharmaceutically acceptable salt thereof, may also be administered in a liposome composition or as a microemulsion.

[0185] For injection, the formulation may be prepared in conventional forms, such as a solution or suspension, or as a solid suitable for solution or suspension in liquid before injection, or as an emulsion. Suitable excipients include, for example, water, saline, dextrose, and glycerol. Such compositions may also contain certain amounts of non-toxic auxiliary substances (e.g., wetting agents or emulsifiers), pH buffers, such as sodium acetate and sorbitan monolaurate.

[0186] Various sustained-release systems for drugs have also been devised. See, for example, U.S. Patent No. 5,624,677.

[0187] Systemic administration may also include relatively non-invasive methods, such as suppositories, transdermal patches, transmucosal delivery, and intranasal administration. Oral administration is also suitable for the compounds of this disclosure or pharmaceutically acceptable salts thereof. Preferred forms include syrups, capsules, and tablets, as understood in the art.

[0188] Each compound described herein, or a pharmaceutically acceptable salt thereof, can be formulated in various ways known in the art. For example, the first and second agents of a combination therapy can be formulated together or individually. Other modalities of combination therapy are described herein.

[0189] Individually formulated or separately manufactured preparations can be packaged together as a kit. Non-limiting examples include, but are not limited to, kits containing two pills, pills and powder, suppositories and liquids in vials, or two topical creams. A kit may include optional components that facilitate the administration of a unit dose to a target, such as vials for reconstituting powder forms, syringes for injection, customized intravenous delivery systems, or inhalers. In addition, a unit dose kit may include instructions for preparation or administration of the composition. A kit may be manufactured as a single-use unit dose for a particular target (where the efficacy of individual compounds or their pharmaceutically acceptable salts changes at a constant concentration or as treatment progresses), as a multi-use kit for a specific target, or as a multi-use kit suitable for administration to multiple targets ("bulk packaging"). The components of the kit can be assembled into cartons, blister packs, bottles, tubes, etc.

[0190] Preparations for oral use include tablets containing the active ingredient(s) in a mixture with pharmaceutically acceptable, non-toxic excipients. These excipients may include, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugars, mannitol, microcrystalline cellulose, starch containing potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate); granulators and disintegrants (e.g., cellulose derivatives containing microcrystalline cellulose, starch containing potato starch, croscarmellose sodium, alginate, or arginine); binders (e.g., sucrose, glucose, sorbitol, acacia, arginine, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, aluminum magnesium silicate, sodium carboxymethylcellulose, methylcellulose, optionally substituted hydroxypropylmethylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethylene glycol); as well as smoothers, lubricants, and anti-adhesives (e.g., magnesium stearate, zinc stearate, stearic acid, silica, hydrogenated vegetable oil, or talc). Other pharmaceutically acceptable excipients may include colorants, flavoring agents, plasticizers, humectants, buffering agents, and the like.

[0191] Two or more compounds can be mixed or dispensed in a tablet, capsule, or other vehicle. In one example, the first compound is contained inside the tablet, the second compound is on the outside, and a substantial portion of the second compound is released before the first compound.

[0192] Formulations for oral use may be provided as chewable tablets, or as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate, or kaolin), or as soft gelatin capsules in which the active ingredient is mixed with water or an oily medium (e.g., peanut oil, liquid paraffin, or olive oil). Powders, granules, and pellets can be prepared using the above-mentioned components below tablets and capsules by conventional methods, for example, using a mixer, fluidized bed apparatus, or spray dryer.

[0193] Dissolution or diffusion-controlled release can be achieved by appropriate coatings for tablet, capsule, pellet, or granule formation of the compound, or by incorporating the compound, or a pharmaceutically acceptable salt thereof, into a suitable matrix. Controlled release coatings may include one or more of the above-mentioned coating materials, or, for example, shellac, beeswax, glycowax, castor wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glycerol palmitostearate, ethylcellulose, acrylic resin, dl-polylactic acid, cellulose acetate / butyrate, polyvinyl chloride, polyvinyl acetate, vinylpyrrolidone, polyethylene, polymethacrylate, methyl methacrylate, 2-(optionally substituted) hydroxyl methacrylate, methacrylate hydrogel, 1,3-butylene glycol, ethylene glycol methacrylate, or polyethylene glycol. In release-controlled matrix formulations, examples of matrix materials include hydrated methylcellulose, carnauba wax and stearyl alcohol, Carbopol 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, or halogenated fluorocarbons.

[0194] Liquid forms in which the compounds of this disclosure, or pharmaceutically acceptable salts thereof, and compositions may be incorporated for oral administration include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and emulsions flavored with edible oils such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0195] In general, when administered to humans, the oral dose of any of the compounds of this disclosure, or any of its pharmaceutically acceptable salts, depends on the properties of the compound and can be quickly determined by those skilled in the art. The dose may be, for example, about 0.001 mg to about 2000 mg / day, about 1 mg to about 1000 mg / day, about 5 mg to about 500 mg / day, about 100 mg to about 1500 mg / day, about 500 mg to about 1500 mg / day, about 500 mg to about 2000 mg / day, or any of these range variables.

[0196] In some embodiments, the pharmaceutical composition may further include additional compounds having antiproliferative (e.g., anticancer) activity. Depending on the method of administration, the compounds, or pharmaceutically acceptable salts thereof, are formulated into a suitable composition that allows for easy delivery. Each compound of the combination therapy, or a pharmaceutically acceptable salt thereof, can be formulated in various ways known in the art. For example, the first and second agents of the combination therapy can be formulated together or individually. Preferably, the first and second agents are formulated together for simultaneous or near-simultaneous administration.

[0197] It will be understood that the compounds and pharmaceutical compositions of this disclosure can be formulated and used in combination therapy, that is, the compounds and pharmaceutical compositions can be formulated or administered simultaneously with, before, or after one or more other desired therapeutic agents or medical procedures. In specific combinations of treatments (therapeutic agents or procedures) using combination regimens, the suitability of the desired therapeutic agent or procedure and the desired therapeutic effect to be achieved will be taken into consideration. Furthermore, it will be understood that the treatments used may achieve the desired effect for the same disorder, or different effects (e.g., control of any adverse effects).

[0198] As described herein, each drug in combination therapy may be administered independently once to four times a day for a period of one to one year, and may also be administered throughout the patient's lifetime. Chronic, long-term administration may be indicated.

[0199] II. Method In one embodiment, the present disclosure relates to a method for treating a disease or disorder characterized by abnormal RAS activity (e.g., cancer or RASopathy). In some embodiments, the disease or disorder is cancer (e.g., cancer having one or more RAS mutations that cause abnormal RAS activity). In prior embodiments, the method generally involves administering to a subject a therapeutically effective amount of a RAS(ON)GTP hydrolytic compound and a RAS(OFF) inhibitor. Suitable RAS(ON)GTP hydrolytic compounds and further therapeutic agents useful in the methods disclosed herein are described in Section I, which is incorporated herein by reference.

[0200] Accordingly, this disclosure provides a method for treating cancer in a subject requiring cancer treatment, the method comprising administering to the subject a therapeutically effective amount of one or more RAS(ON)GTP hydrolysis-promoting compounds described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising one or more RAS(ON)GTP hydrolysis-promoting compounds described herein, or a salt thereof.

[0201] This disclosure also provides a method for treating cancer in subjects requiring treatment, wherein the cancer includes mutations in the RAS. In one embodiment, the addition of a RAS(ON)GTP hydrolysis-promoting compound, or a pharmaceutically acceptable salt or pharmaceutical composition thereof, synergistically increases the activity of a RAS(OFF) inhibitor, or a pharmaceutically acceptable salt or pharmaceutical composition thereof. To determine the synergistic effect of the combination, any method for determining whether two compounds exhibit a synergistic effect (e.g., the method described herein) may be used.

[0202] Several mathematical models have been developed to determine whether two compounds act synergistically, that is, beyond mere additive effects. For example, Loewe Additivity (Loewe (1928) Physiol. 27:47-187), Bliss Independence (Bliss (1939) Ann. Appl. Biol. 26:585-615), Highest Single Agent, ZIP (Yadav et al (2015) Comput Struct Biotech J 13:504-513), and other models (Chou & Talalay (1984) Adv Enzyme Regul 22:27-55.#6382953; and Greco et al. (1995) Pharmacol Rev 47(2):331-85.#7568331) are well-known models in the pharmaceutical industry and can be used to determine whether a synergistic effect has been detected and to calculate a "synergy score" indicating the magnitude of such a synergistic effect. An additional model for determining the synergistic effect of the two compounds can be found in the following examples.

[0203] Generally, mathematical models use data obtained from single drug values ​​to determine the predicted additive effect of a combination and compare it to the observed effect of the combination. If the observed effect is greater than the predicted effect, the combination is considered synergistic. For example, the Bliss independence model compares the observed combination response (Yo) to the predicted combination response (Yp) obtained under the assumption that there is no effect from drug-drug interactions. Typically, if Yo is greater than Yp, the combination effect is considered synergistic.

[0204] In some embodiments, the “synergistic effect” as used herein refers to a combination of a RAS(ON)GTP hydrolytic compound, or a pharmaceutically acceptable salt thereof, and an additional therapeutic agent (e.g., a RAS(OFF) inhibitor) or a pharmaceutically acceptable salt thereof, which produces an effect that results in any beneficial or desired outcome, including, for example, in vitro results and clinical outcomes or evaluation items described herein, which is greater than the sum of the effects observed when the RAS(ON)GTP hydrolytic compound, or a pharmaceutically acceptable salt thereof, and the additional therapeutic agent (e.g., a RAS(OFF) inhibitor) or a pharmaceutically acceptable salt thereof are administered alone. The higher the hydrolytic capacity of the RAS(ON)GTP hydrolytic compound, the greater the synergistic effect observed with the RAS(OFF) inhibitor.

[0205] In some embodiments, the present disclosure provides a method for treating cancer in a subject requiring cancer treatment, the method comprising administering to the subject a therapeutically effective amount of one or more RAS(ON)GTP hydrolytic compounds described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions and receptor tyrosine kinase (RTK) inhibitors.

[0206] In some embodiments, the present disclosure provides a method for treating cancer in a subject requiring cancer treatment, the method comprising administering to the subject a therapeutically effective amount of one or more RAS(ON)GTP hydrolytic compounds described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions and an SHP2 inhibitor.

[0207] In some embodiments, the present disclosure provides a method for treating cancer in a subject requiring cancer treatment, the method comprising administering to the subject a therapeutically effective amount of one or more RAS(ON)GTP hydrolytic compounds described herein, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions and an SOS1 inhibitor.

[0208] RAS(ON)GTP hydrolytic compounds and additional therapeutic agents may be administered simultaneously or sequentially. RAS(ON)GTP hydrolytic compounds and RAS(OFF) inhibitors may be administered as a single formulation or as separate formulations. In some embodiments, the RAS(ON)GTP hydrolytic compound is administered in a first period; an additional therapeutic agent is administered in a second period, where the first and second periods do not overlap and the first period precedes the second; and the additional therapeutic agent and the RAS(ON)GTP hydrolytic compound are administered in a second period, where the first and second periods do not overlap and the first period precedes the second period.

[0209] In some embodiments, the cancer is colorectal cancer, non-small cell lung cancer, small cell lung cancer, pancreatic cancer, appendiceal cancer, melanoma, acute myeloid leukemia, small intestine cancer, ampulla cancer, germ cell carcinoma, cervical cancer, cancer of unknown primary origin, endometrial cancer, esophageal cancer, GI neuroendocrine cancer, ovarian cancer, sex cord-stromal tumor cancer, hepatobiliary cancer, or bladder cancer. In some embodiments, the cancer is appendiceal cancer, endometrial cancer, or melanoma. In some embodiments, the cancer is non-small cell lung cancer. In some embodiments, the cancer is pancreatic cancer.

[0210] In some embodiments, the compounds of the present disclosure, or pharmaceutically acceptable salts thereof, pharmaceutical compositions containing such compounds or salts, and methods provided herein can be used to treat a wide variety of cancers, including astrocytic cell carcinoma, breast cancer, cervical cancer, colorectal cancer, endometrial cancer, esophageal cancer, gastric cancer, head and neck cancer, hepatocellular carcinoma, laryngeal cancer, lung cancer, pharyngeal cancer, ovarian cancer, prostate cancer, as well as thyroid cancer and sarcoma. Other cancers include, for example, heart: sarcomas (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyosarcoma, fibroma, lipoma, and teratoma; lung: bronchogenic carcinoma (squamous cell carcinoma, anaplastic small cell carcinoma, anaplastic large cell carcinoma, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondrotoxic hamartoma, mesothelioma; gastrointestinal tract: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (cancer, lymphoma, leiomyosarcoma), pancreas (tubules) Cancer (insulinoma, glucagonoma, gastrinoma, carcinoid tumor, lipoma), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, chorioadenoma, hamartoma, leiomyoma); urogenital system: e.g., kidney (adenocarcinoma, Wilms' tumor (nephroblastoma), lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testes (Seminoma, teratoma, embryonic carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma); Liver: e.g., liver cancer (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Biliary tract: e.g., gallbladder cancer, duodenal papilla cancer, cholangiocarcinoma; Bone: e.g., osteosarcoma (osteogenic sarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticular cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, Osteochondroma (osteochondrial exostosis), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma, and giant cell tumor; nervous system: e.g., skull (osteoma, hemangioma, granuloma, xanthomas, degenerative osteitis), meninges (meningioma, meningiosarcoma, gliomas), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germ cell tumor (pineal glandoma), glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumor), spinal neurofibroma, neurofibromatosis type 1, meningioma, glioma, sarcoma);Gynecological conditions: For example, the uterus (endometrial cancer, uterine cancer, endometrial cancer), cervix (cervical cancer, precancerous cervical dysplasia), ovaries (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassifiable cancer), granulosa-theca cell tumor, Sertoli-Leydig cell tumor, undifferentiated germ cell tumor, malignant teratoma), vulva (squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, staphylosarcoma (fetal rhabdomyosarcoma)). ), fallopian tubes (carcinoma); hematological system: e.g., blood (myeloid leukemia (acute and chronic), acute lymphoblastic myeloproliferative neoplasm), multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin lymphoma (malignant lymphoma); skin: e.g., malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, dysplastic nevus, lipoma, hemangioma, dermatofibroma, keloid, psoriasis; and adrenal gland: e.g., neuroblastoma.

[0211] In some embodiments, the cancer includes RAS mutations such as the RAS mutations described herein. In some embodiments, the mutations are the following KRAS variants: G12D, G12V, G12C, G13D, G12R, G12A, G12S, A146T, G13C, K117N, A146V, G12F, L19F, Q22K, V14I, A59T, A146P, G13R, G12L, or G13V, and combinations thereof; the following HRAS variants: G13R, G12S, G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, or G12R, and combinations thereof; and the following NRAS variants: G12D, G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T, G60E, A59D, E132K, E49K, T50I, A146V, or A59T, and combinations thereof; or selected from any combination of the above, in some embodiments the cancer contains a KRAS mutation selected from the group consisting of G12C, G12D, G13C, G12V, G13D, G12R, and G12S. In some embodiments the cancer contains the G12C variant of NRAS. In some embodiments, the cancer contains RAS mutations selected from the group consisting of G12C, G13C, G12A, G12D, G13D, G12S, G13S, G12V, and G13V. In some embodiments, the cancer contains at least two RAS mutations selected from the group consisting of G12C, G13C, G12A, G12D, G13D, G12S, G13S, G12V, and G13V.

[0212] In some embodiments, the compounds of the Disclosure bind to or inhibit two or more RAS variants. In some embodiments, the compounds may inhibit both KRAS G12D and KRAS G12V. In some embodiments, the compounds may bind to or inhibit both KRAS G12V and KRAS G12S. In some embodiments, the compounds of the Disclosure bind to or inhibit wild-type RAS in addition to RAS variants. In some embodiments, the compounds of the Disclosure bind to or inhibit one or more additional RAS variants (e.g., K-,H- or N-RAS). amp and KRAS G12D, G12V, G12C, G13D, G12R, G12A, G12S, A146T, G13C, K117N, A146V, G12F, L19F, Q22K, V14I, A59T, A146P, G13R, G12L, or G13V; K-, H- or N-RAS amp and HRAS, G13R, G12S, G12D, G13V, G13D, G12C, K117N, A59T, G12V, G13C, G13S, A18V, D119N, G13N, A146T, A66T, G12A, A146V, G12N, or G12R; or K-, H- or N-RAS amp And in addition to NRAS G12D, G13R, G13D, G12S, G12C, G12V, G12A, G13V, G12R, P185S, G13C, A146T, G60E, A59D, E132K, E49K, T50I, A146V, or A59T), RAS amp To bind to or inhibit.

[0213] In some embodiments, the cancer is non-small cell lung cancer, and the RAS mutations include KRAS mutations such as KRAS G12C, KRAS G12V, or KRAS G12D. In some embodiments, the cancer is colorectal cancer, and the RAS mutations include KRAS mutations such as KRAS G12C, KRAS G12V, or KRAS G12D. In some embodiments, the cancer is pancreatic cancer, and the RAS mutations include NRAS mutations such as NRAS G12D. In some embodiments, the cancer is melanoma.

[0214] In some embodiments, the cancer includes a RAS mutation and an STK11LoF, KEAP1, EPHA5, or NF1 mutation. In some embodiments, the cancer is non-small cell lung cancer and includes a KRAS G12C mutation. In some embodiments, the cancer is non-small cell lung cancer and includes a KRAS G12C mutation and an STK11LoF mutation. In some embodiments, the cancer is non-small cell lung cancer and includes a KRAS G12C mutation and an STK11LoF mutation. In some embodiments, the cancer includes a KRAS G13C RAS ​​mutation and an STK11LoF, KEAP1, EPHA5, or NF1 mutation. In some embodiments, the cancer is non-small cell lung cancer and includes a KRAS G12D mutation. In some embodiments, the cancer is non-small cell lung cancer and includes a KRAS G12V mutation. In some embodiments, the cancer is colorectal cancer and includes a KRAS G12C mutation. In some embodiments, the cancer is pancreatic cancer and includes a KRAS G12D mutation. In some embodiments, the cancer is pancreatic cancer and contains the KRAS G12V mutation. In some embodiments, the cancer is pancreatic cancer and contains the KRAS G12R mutation. In some embodiments, the cancer is endometrial cancer and contains the KRAS G12C mutation. In some embodiments, the cancer is gastric cancer and contains the KRAS G12C mutation.

[0215] Methods for detecting mutations in KRAS, HRAS, or NRAS nucleotide sequences are known to those skilled in the art. These methods include, but are not limited to, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assays, polymerase chain reaction-single-strand higher-order structure polymorphism (PCR-SSCP) assays, real-time PCR assays, PCR sequencing, mutant allele-specific PCR amplification (MASA) assays, direct sequencing, primer extension reactions, electrophoresis, oligonucleotide ligation assays, hybridization assays, TaqMan assays, SNP genotyping assays, high-resolution thawing assays, and microarray analysis. In some embodiments, G12C KRAS, HRAS, or NRAS mutations in a sample are evaluated by real-time PCR. In real-time PCR, a fluorescent probe specific to the KRAS, HRAS, or NRAS G12C mutation is used. If a mutation is present, the probe binds and fluorescence is detected. In some embodiments, KRAS, HRAS, or NRAS G12C mutations are identified using a method that directly sequences a specific region (e.g., exon 2 or exon 3) within the KRAS, HRAS, or NRAS gene. This method identifies all possible mutations within the sequenced region.

[0216] Methods for detecting mutations in KRAS, HRAS, or NRAS proteins are known to those skilled in the art. These methods include, but are not limited to, the detection of KRAS, HRAS, or NRAS variants using a specific binder (e.g., antibody) for the mutant protein, protein electrophoresis and Western blotting, and direct peptide sequencing. Other methods include ctDNA analysis (e.g., Cescon et al., Nature Cancer 1:276-290 (2020)) and the use of highly sensitive diagnostic assays (using the CE-IVD mark) such as TheraScreen PCR; AmoyDx; PNAClamp; RealQuality; EntroGen; LightMix; StripAssay; Hybcell plexA; Devyser; Surveyor; Cobas; and TheraScreen Pyro, which are incorporated herein by reference, for example, Domagala, et al., Pol J Pathol 3:145-164 (2012). For example, see also WO2020 / 106640.

[0217] Various samples can be used in methods for determining whether a tumor or cancer contains G12C or other KRAS, HRAS, or NRAS mutations. In some embodiments, the sample is taken from a subject having a tumor or cancer. In some embodiments, the sample is a fresh tumor / cancer sample. In some embodiments, the sample is a frozen tumor / cancer sample. In some embodiments, the sample is a (CTC) sample. In some embodiments, the sample is processed to become a cell lysate. In some embodiments, the sample is processed to become DNA or RNA.

[0218] Also provided is a method for inhibiting or binding to intracellular RAS proteins, comprising contacting cells with a combination comprising an effective amount of the RAS(ON)GTP hydrolysis-promoting compound of this disclosure, or a pharmaceutically acceptable salt thereof, and optionally an additional therapeutic agent (e.g., a RAS(OFF) inhibitor, an RTK inhibitor, an SHP2 inhibitor, or an SOS1 inhibitor), or a pharmaceutically acceptable salt thereof. The cells may be cancer cells. The cancer cells may be of any type of cancer described herein. The cells may be in vivo or in vitro.

[0219] In some embodiments of any of the methods described herein, prior to treatment by the composition or method of the present invention, the patient has been treated with one or more of the following: chemotherapy, targeted anticancer agents, radiotherapy, and surgery, optionally, the previous treatment was unsuccessful; and / or the patient has undergone surgery, optionally, the surgery was unsuccessful; and / or the patient has been treated with a platinum-based chemotherapeutic agent, optionally, the patient was previously determined not to respond to treatment with the platinum-based chemotherapeutic agent; and / or the patient has been treated with a kinase inhibitor, optionally, the previous treatment with the kinase inhibitor was unsuccessful; and / or the patient has been treated with one or more other therapeutic agents.

[0220] In various embodiments, the Disclosure provides a method for treating cancer in a subject, comprising administering to the subject a RAS(ON)GTP hydrolysis-promoting compound, or a combination thereof, in combination with an additional therapeutic agent as optionally described herein (e.g., a RAS(OFF) inhibitor, an RTK inhibitor, an SHP2 inhibitor, or an SOS1 inhibitor), wherein the subject has one or more tumors that are resistant or unresponsive to the treatment. In various embodiments, the subject has one or more tumors that are resistant or unresponsive to one or more treatments selected from the group consisting of surgery, radiation, chemotherapy, biological agents, small molecule, cell-based therapies, hormone therapy, and immunotherapy. In various embodiments, the treatment is standard treatment, first-line treatment, second-line treatment, or third-line treatment. In various embodiments, the subject has one or more tumors that have progressed during one or more treatments which are standard treatment, first-line treatment, second-line treatment, or third-line treatment.

[0221] First-line therapy is defined as treatment administered to a patient with cancer who has received no prior treatment. Second-line therapy is defined as treatment administered to a patient with cancer who has previously received first-line therapy but has experienced disease progression during first-line treatment. Third-line therapy is defined as treatment administered to a patient with cancer who has previously received first and second-line therapy but has experienced disease progression during second-line treatment. Each specific type of cancer has first-line, second-line, and third-line therapies. First, second, and third-line therapies for each type of cancer are publicly known in the relevant art. Furthermore, FDA-approved drug labels indicate whether a particular drug is approved as a first, second, or third-line therapy.

[0222] Several criteria and definitions published in the literature can be used to determine the effectiveness of one or more treatments for a target tumor affected by cancer. Based on these criteria, a tumor is defined as “responsive,” “stable,” or “progressive” depending on whether it improves, remains the same, or worsens during treatment, respectively.

[0223] Examples of commonly used criteria published in the literature include the Criteria for Evaluation of Solid Tumors (RECIST), the Modified Criteria for Evaluation of Solid Tumors (mRECIST), the PET Criteria for Evaluation of Solid Tumors (PERCIST), the Choi Criteria, the Lugano Criteria, the European Association for the Study of the Liver (EASL) Criteria, the Criteria for Evaluation of Treatment Response to Hepatocellular Carcinoma (RECICL), and the WHO Criteria for Tumor Response.

[0224] In various embodiments, the Disclosure provides a method for treating cancer in a subject, comprising administering to the subject a RAS(ON)GTP hydrolytic compound of the Disclosure, or a pharmaceutically acceptable salt thereof, and optionally an additional therapeutic agent described herein (e.g., a RAS(OFF) inhibitor, an RTK inhibitor, an SHP2 inhibitor, or an SOS1 inhibitor), wherein the subject is intolerant to standard treatment, first-line therapy, second-line therapy, or third-line therapy. In various embodiments, the Disclosure provides a method for treating cancer in a subject, comprising administering to the subject a RAS(ON)GTP hydrolytic compound of the Disclosure, or a pharmaceutically acceptable salt thereof, and optionally an additional therapeutic agent described herein (e.g., a RAS(OFF) inhibitor, an RTK inhibitor, an SHP2 inhibitor, or an SOS1 inhibitor), wherein the subject has experienced tumor recurrence after surgical resection of a primary tumor. In various embodiments, the Disclosure provides a method for treating cancer in a subject, comprising administering to the subject a RAS(ON)GTP hydrolytic compound, or a pharmaceutically acceptable salt thereof, and optionally an additional therapeutic agent as described herein (e.g., a RAS(OFF) inhibitor, an RTK inhibitor, an SHP2 inhibitor, or an SOS1 inhibitor), wherein the subject has a tumor that cannot be surgically resected. In various embodiments, the Disclosure provides a method for treating cancer in a subject, comprising administering to the subject a RAS(ON)GTP hydrolytic compound, or a pharmaceutically acceptable salt thereof, and optionally an additional therapeutic agent as described herein (e.g., a RAS(OFF) inhibitor, an RTK inhibitor, an SHP2 inhibitor, or an SOS1 inhibitor), wherein the subject has no possible treatment options.

[0225] Some therapies used to treat cancer (e.g., chemotherapy) are cytotoxic and carry significant side effects and toxicities associated with poor outcomes and poor response to treatment. Before administering such treatments, clinicians rely on several assessment tools to evaluate the risk that a patient with cancer will experience treatment-related toxicities and adverse events. Based on the results of these assessments, a patient with cancer is considered intolerant to treatment if they are determined to be at high risk of experiencing treatment-related toxicities and adverse events resulting in a poor outcome. Examples of commonly used assessment tools used to determine treatment intolerance include the Karnovsky Performance Status (KPS), the East Coast Cancer Clinical Trials Group Performance Status (ECOG PS), the Timed Get Up and Go (TUG), the Short-Term Physical Ability Battery (SPPB), the Comprehensive Assessment of Geriatric Function (CGA), the Cancer Aging Research Group (CARG) score, and the Chemotherapy Risk Assessment Scale for Elderly Patients (CRASH).

[0226] In some embodiments, the progression of the cancer in question is reduced or prevented. Disease progression of cancer (e.g., the cancers described herein) can be evaluated by one or more of several established methods. Those skilled in the art can monitor the subject by direct observation to evaluate how the symptoms exhibited by the subject have changed (e.g., reduction or disappearance of symptoms) in response to treatment (e.g., the treatment methods disclosed herein). The subject can also be examined by MRI, CT scan, or PET analysis to determine whether the tumor has metastasized or whether the tumor size has changed (e.g., decreased in response to treatment (e.g., the treatment methods described herein)). Optionally, cells can be extracted from the subject by biopsy or procedure, or tumor DNA can be isolated from the subject's blood, and quantitative biochemical analysis can be performed to assess the relative cancer burden and determine the presence or appearance of specific mutations that may be involved in resistance. Based on these analytical results, those skilled in the art can prescribe higher / lower doses or more frequent / less frequent administration in subsequent treatment rounds.

[0227] In various embodiments, the Disclosure provides a method for treating cancer in a subject, comprising administering to the subject a RAS(ON)GTP hydrolysis-promoting compound and optionally an additional therapeutic agent (e.g., a RAS(OFF) inhibitor, an RTK inhibitor, an SHP2 inhibitor, or an SOS1 inhibitor). In various embodiments, the administration reduces tumor size or inhibits tumor growth. In various embodiments, the administration induces tumor cell death, apoptosis, or necrosis.

[0228] The methods described herein are intended to reduce tumor size or tumor volume in a subject, or to reduce metastasis in a subject. In various embodiments, the methods reduce tumor size by 10%, 20%, 30%, or more. In various embodiments, the methods reduce tumor size by about or at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or 100% (or all values ​​and ranges between these values).

[0229] In one embodiment, the present disclosure provides a method for treating a RAS protein-related disorder in a subject, where the pathology of the RAS-related disorder is partially mediated by increased signaling in the RAS / MAPK pathway. In various embodiments, the method generally involves administering a therapeutically effective amount of a RAS(ON)GTP hydrolytic compound to the subject. In some embodiments, the RAS protein-related disorder is RASopathy. RASopathy is a group of hereditary disorders caused by mutations in genes involved in the RAS / MAPK signaling pathway. RASopathy is characterized by a variety of clinical features and may affect multiple organ systems, including the cardiovascular, musculoskeletal, nervous, and cutaneous systems.

[0230] In one embodiment, the present disclosure relates to a method for treating a disease or disorder characterized by abnormal RAS activity (e.g., cancer or RASopathy). In some embodiments, the disease or disorder is cancer (e.g., cancer having one or more RAS mutations that cause abnormal RAS activity). A non-limiting example of non-cancerous RAS-related diseases or disorders is shown in Table 3. In each embodiment, the method generally involves administering a therapeutically effective amount of a RAS(ON)GTP hydrolytic compound to a subject. In some embodiments, the method involves administering the RAS(ON)GTP hydrolytic compound in combination with one or more therapeutic agents. Suitable RAS(ON)GTP hydrolytic compounds and further therapeutic agents are described herein. [Table 3-1] [Table 3-2]

[0231] In some embodiments, the method includes treating RASopathy selected from Noonan syndrome, Costello syndrome, cardiac-facial-cutaneous syndrome, neurofibromatosis type 1, and Regius syndrome. Although each RASopathy has its own unique characteristics, they all share certain similarities, such as facial deformities, cardiac abnormalities, developmental delays, and an increased risk of certain cancers.

[0232] RASopathy is usually diagnosed through a combination of clinical evaluation, genetic testing, and imaging studies. Treatment and management of RASopathy depend on the specific type and severity of the disorder, but may include medication, surgery, and supportive therapies such as physiotherapy and occupational therapy.

[0233] a) Combination therapy The methods of the present disclosure may include the RAS(ON)GTP hydrolysate compound of the present disclosure in combination with additional therapeutic agents (e.g., non-pharmacological treatments or therapeutic agents). When administering one or more doses of the additional therapies (e.g., non-pharmacological treatments or therapeutic agents) alone, the dose may be reduced from the standard dose. For example, the dose may be determined empirically from the combination and order of drugs, or estimated by isobolographic analysis (e.g., Black et al., Neurology 65:S3-S6 (2005)).

[0234] The compounds of this disclosure may be administered before, after, or concurrently with one or more additional therapies. When combined, the dosage of the compounds of this disclosure and the dosage of one or more additional therapies (e.g., non-pharmacological treatments or therapeutic agents) may provide a therapeutic effect (e.g., a synergistic or additive therapeutic effect). The compounds of this disclosure and additional therapies (e.g., anticancer agents) may be administered together (e.g., in a single pharmaceutical composition) or separately, and if administered separately, they may be administered simultaneously or sequentially. Such sequential administrations may be close in time or far apart.

[0235] In some embodiments, additional therapy is the administration of side effect limiting agents (e.g., agents intended to reduce the occurrence or severity of side effects of treatment). For example, in some embodiments, the compounds of the present disclosure may also be used in combination with therapeutic agents for treating nausea. Examples of agents that may be used to treat nausea include dronabinol, granisetron, metoclopramide, ondansetron, and prochlorperazine, or pharmaceutically acceptable salts thereof.

[0236] In some embodiments, one or more additional therapies include non-pharmacological treatment (e.g., surgery or radiotherapy). In some embodiments, one or more additional therapies include therapeutic agents (e.g., compounds or biologics that are anti-angiogenic agents, signaling inhibitors, antiproliferative agents, glycolysis inhibitors, or autophagy inhibitors). In some embodiments, one or more additional therapies include non-pharmacological treatment (e.g., surgery or radiotherapy) and therapeutic agents (e.g., compounds or biologics that are anti-angiogenic agents, signaling inhibitors, antiproliferative agents, glycolysis inhibitors, or autophagy inhibitors). In other embodiments, one or more additional therapies include two therapeutic agents. In yet another embodiment, one or more additional therapies include three therapeutic agents. In some embodiments, one or more additional therapies include four or more therapeutic agents.

[0237] In this section on combination therapies, all references to the listed medications, whether explicitly stated or not, are incorporated by reference.

[0238] i) Nonpharmacological treatment Examples of non-pharmacological 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.

[0239] In some embodiments, the compounds of the present disclosure may be used as adjuvant therapy after surgery. In some embodiments, the compounds of the present disclosure may be used as neoadjuvant therapy before surgery.

[0240] Radiotherapy may be used in subjects (e.g., mammals (e.g., humans)) to inhibit abnormal cell proliferation or to treat hyperproliferative disorders such as cancer. Techniques for administering radiotherapy are known in the art. Radiotherapy may be administered via one or a combination of several methods, including, but not limited to, external beam therapy, internal radiation therapy, implanted radiation, stereotactic radiotherapy, total body radiation therapy, radiotherapy, and permanent or temporary intratissue brachytherapy. As used herein, the term “brachytherapy” refers to radiotherapy delivered by spatially restricted radioactive material inserted into the body at or near a tumor or other site of proliferative tissue disease. The term is intended to include, but is not limited to, exposure to radioisotopes (e.g., At-211, I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, and radioisotopes of Lu). Suitable radioactive sources for use as cell modifiers in this disclosure include both solids and liquids. For non-limiting illustrative purposes, the radioactive sources may be radionuclides (such as I-125, I-131, Yb-169, Ir-192 as solid sources, or I-125 as a solid source), or other radionuclides that emit photons, beta particles, gamma rays, or other therapeutic radiation. The radioactive material may also be any solution of the radionuclide(s), for example, a fluid made from a solution of I-125 or I-131, or the radioactive fluid may be produced using a suitable fluid slurry containing small particles of a solid radionuclide such as Au-198 or Y-90. Furthermore, the radionuclide(s) may be embedded in a gel or radioactive microspheres.

[0241] In some embodiments, the compounds of the Disclosure can make abnormal cells more sensitive to radiotherapy intended to kill or inhibit the proliferation of such cells. Therefore, the Disclosure further relates to a method for sensitizing abnormal cells in mammals to radiotherapy, the method comprising administering to a mammal an amount of the compound of the Disclosure effective in sensitizing the abnormal cells to radiotherapy. The amount of the compound in this method can be determined according to the means for determining an effective amount of such compound described herein. In some embodiments, the compounds of the Disclosure may be used as adjuvant therapy after radiotherapy or as neoadjuvant therapy before radiotherapy.

[0242] In some embodiments, the non-pharmacological treatment is T cell adoptive transfer (ACT) therapy. In some embodiments, the T cells are activated T cells. The T cells 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, CAR-T cells can be generated by introducing a suitable expression vector encoding a CAR into T cells. The source of T cells is obtained from a subject before proliferation and genetic modification of the T cells. T cells can be obtained from multiple sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, umbilical cord blood, thymic tissue, tissue from infection sites, ascites, pleural fluid, splenic tissue, and tumors. In certain embodiments of this disclosure, any number of T cell lines available in the art may be used. In some embodiments, the T cells are autologous T cells. Whether before or after genetic modification of T cells to express a desired protein (e.g., CAR), T cells are, for example, U.S. Patent Nos. 6,352,694, 6,534,055, 6,905,680, 6,692,964, 5,858,358, 6,887,466, 6,905,681, and 7,14 Activation and propagation can be generally carried out using the methods described in Nos. 4,575, 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.

[0243] ii) Therapeutic agents The therapeutic agent may be a compound used to treat cancer or related conditions.

[0244] For example, the therapeutic agent may be a steroid. Therefore, in some embodiments, one or more additional therapies include a steroid. Preferred steroids include 21-acetoxypregnenolone, alclomethasone, algestone, amcinonide, beclomethasone, betamethasone, budesonide, chloroprednisone, clobetasol, crocortol, cloprednol, corticosterone, cortisone, cortibazole, deflazacort, desonide, dexoxymethasone, dexamethasone, diflorasone, diflucortol, difprednate, enoxolone, fluazacort, fluchloronide, flumethasone, flunisolide, fluocinolone acetonide, fluocinonide, flucortin butyl, flucortolone, fluorometholone, fluperolone acetate, flupredniden acetate, fluprednisolone, and flulandre. This may include, but is not limited to, nolid, fluticasone propionate, formocortal, halcinonide, halobetazole propionate, halomethasone, hydrocortisone, loteprednol etavonate, mazipredone, medrisone, meprednisone, methylprednisolone, mometasone furoate, paramethasone, prednicarbate, prednisolone, prednisolone 25-diethylaminoacetate, sodium prednisolone phosphate, prednisone, prednival, prednylidene, rimexolone, thixocortol, triamcinolone, triamcinolone acetonide, triamcinolone benetonide, triamcinolone hexaacetonide, and salts or derivatives thereof.

[0245] Further examples of therapeutic agents that may be used in combination therapy with the compounds of this disclosure include the 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, as well as International Patent Application No. WO0 Issues 1 / 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.

[0246] The therapeutic agent may be a biologic used to treat cancer or related conditions (e.g., cytokines (e.g., interferons or interleukins such as IL-2)). In some embodiments, the biologic is an immunoglobulin-based biologic, such as a monoclonal antibody (e.g., a humanized antibody, a fully human antibody, an Fc fusion protein, or a functional fragment thereof), that acts on a target to stimulate an anti-cancer response or antagonize an antigen important to cancer. Antibody-drug conjugates are also included.

[0247] The therapeutic agent may be a T-cell checkpoint inhibitor. In one embodiment, the checkpoint inhibitor is an inhibitory antibody (e.g., a monospecific antibody, e.g., a monoclonal antibody). The antibody may be, for example, 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 a drug (e.g., an antibody) that interacts with a checkpoint protein. In some embodiments, the checkpoint inhibitor is a drug, such as an antibody, that interacts with a 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 protein). In some embodiments, the checkpoint inhibitor is a PD-1 inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor). In some embodiments, the checkpoint inhibitor is a PD-L1 inhibitor or antagonist (e.g., an inhibitory antibody or small molecule inhibitor). 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., 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, CHK1, CHK2, A2aR, B-7 family ligands, or combinations thereof.In some embodiments, the checkpoint inhibitor is pembrolizumab, nivolumab, PDR001 (NVS), REGN2810 (Sanofi / Regeneron), PD-L1 antibody, e.g., avelumab, durvalumab, atezolizumab, pizilizumab, JNJ-63723283 (JNJ), BGB-A317 (BeiGene & Celgene), or Preusser, M. et al. The checkpoint inhibitors disclosed in al. (2015) Nat. Rev. Neurol. (including, but not limited to, ipilimumab, tremelimumab, nivolumab, pembrolizumab, AMP224, AMP514 / MEDI0680, BMS936559, MEDl4736, MPDL3280A, MSB0010718C, BMS986016, IMP321, lirirumab, IPH2101, 1-7F9, and KW-6002) are disclosed.

[0248] The therapeutic agent may be an anti-TIGIT antibody, such as MBSA43, BMS-986207, MK-7684, COM902, AB154, MTIG7192A, or OMP-313M32 (ethigirimab).

[0249] The therapeutic agent may be a drug that treats cancer or related conditions (e.g., cytotoxic agents, non-peptide small molecules, or other compounds useful in treating cancer or related conditions, collectively referred to as "anticancer agents"). Anticancer agents may be, for example, chemotherapeutic agents or targeted therapy agents.

[0250] Examples of anticancer agents include mitotic inhibitors, intercalating antibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological reaction modifiers, alkylating agents, antimetabolites, folate analogs, pyrimidine analogs, purine analogs and related inhibitors, vinca alkaloids, epipodophyllotoxin, antibiotics, L-asparaginase, topoisomerase inhibitors, interferons, platinum coordination complexes, anthracendione-substituted ureas, methylhydrazine derivatives, corticosteroids, progestins, estrogens, anti-estrogens, androgens, anti-androgens, and gonadotropin-releasing hormone analogs. Further examples of anticancer agents include leucovorin (LV), irenotecan, oxaliplatin, capecitabine, paclitaxel, and doxetaxel. In some embodiments, one or more additional therapies comprise two or more anticancer agents. Two or more anticancer agents can be used in a mixture and administered in combination or separately. Suitable administration regimens for combination anticancer agents are known in the art and are described, for example, in Saltz et al., Proc. Am. Soc. Clin. Oncol. 18:233a (1999) and Douillard et al., Lancet 355 (9209):1041-1047 (2000).

[0251] Other non-exclusive examples of anticancer drugs include: Gleevec® (imatinib mesylate); Kyprolis® (carfilzomib); Velcade® (bortezomib); Casodex (bicalutamide); Iressa® (gefitinib); alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan, and pigosulfan; aziridines such as benzodopa, carbocon, metsuredopa, and uredopa; ethyleneimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolomellamine; acetogenins (especially bratacin and bratacinone); camptothecin (including its synthetic analog topotecan); briostatin; calistatin; CC-1065 (its adzeresin, carzeresin, and b Including iseresin synthetic analogs); cryptophycin (specifically cryptophycin 1 and cryptophycin 8); drastatin; duocalmycin (including synthetic analogs KW-2189 and CB1-TM1); eryuterobin; pancratistatin; sarcodictiin A; spongistatin; nitrogen mustards such as chlorambucil, chlornafadin, colophosphamide, estramustine, ifosfamide, mechloretamine, mechloretamine oxide hydrochloride, melphalan, nobenbitin, fenesterine, prednimustine, trophosphamide, and uracil mustard; nitrosoureas such as camulstine, chlorozotosine, fotemustine, lomustine, nimustine, and ranimustine; antibiotics such as engine antibiotics (e.g., calicheamicin such as calicheamicin gamma II and calicheamicin omega II (e.g., Agnew, Chem. Intl.) See Ed Engl. 33:183-186 (1994); Dyne sewing machines such as Dyne sewing machine A; Bisphosphonates such as chlordronate; Espera sewing machines;Neocardinostatin chromophore and related pigment protein enediin antibiotic chromophore, acrasinomycin, actinomycin, ausuramycin, azaserin, bleomycin, kactinomycin, calicheamicin, carabicin, kaminomycin, carminomycin, cardinophilin, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, adriamycin (doxorubicin), morpholino-doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, deoxy Doxorubicin, epirubicin, esorubicin, idarubicin, marcelomycin, mitomycin, e.g., mitomycin C, mycophenolic acid, nogaramycin, olibomycin, peplomycin, potophyllomycin, puromycin, keramycin, rhodorubicin, streptonigrin, streptozocin, tubercidine, ubenimex, dinostatin, zolbicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogs such as denopterin, pteropterin, and trimethrexate; fludarabine, 6-mercaptoxin Purine analogs such as putopurine, thiamipurine, and thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, and phloxuridine; androgens such as carsterone, dromostanolone propionate, epithiostanol, mepitiostane, and testolactone; anti-adrenal agents such as aminoglutethimide, mitotane, and trilostane; folic acid infusions such as phloric acid; acegraton; aldofamide glycoside; aminolevulinic acid; enyluracil; and Musaclin; Bestlabsil; Bisanthren; Edatrexate; Defofamine; Demecolsin; Diadiquan; Elfomithine; Erliptinium acetate; Epotilon B and other Epotilons; Etoglucid; Gallium nitrate; Hydroxyurea; Lentinan; Mytansinoids such as Ronidynin, Mytansin and Ansamitosin; Mitoguazone; Mitoxantrone; Mopidamol; Nitracline; Pentostatin; Fenamet; Pirarubicin; Rosoxantrone; Podophyllic acid; 2-Ethylhydrazide;Procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, OR); Lazoxane; Rhizoxin; Schizophyllan; Spirogermanium; Tenuazonic acid; Triadicone; 2,2',2''-Trichlorotriethylamine; Trichothecenes such as T-2 toxin, Veraculine A, Loridine A, and Anguidine; Urethane; Vindesine; Dacarbazine; Mannomustine; Mitobronitol; Mitractol; Pipobroman; Gacytosine; Arabinoside ("Ara-C"); Si Clophosphamide; thiotepa; taxoids, e.g., Taxol® (paclitaxel), Abraxane® (chromophore-free, albumin-modified nanoparticle formulation of paclitaxel), and Taxotere® (doxetaxel); chlorambucil; tamoxifen (Nolvadex®); raloxifene; aromatase-inhibiting 4(5)-imidazole; 4-hydroxytamoxifen; trioxyfen; Keoxyfen; LY117018; Onapristone; Toremifene (Fareston®); Flutamide, Nilutamide, Bicalutamide, Leuprolide, Goserelin; Chlorambucil; Gemzar® Gemcitabine; 6-Thiogunine; Mercaptopurine; Platinum analogs such as cisplatin, oxaliplatin, and carboplatin; Vinblastine; Platinum; Etoposide (VP-16); Ifosfamide; Mitoxantrone; Vincristine; Examples include Navelbine® (vinorelbine); novantron; teniposide; edatrexate; daunomycin; aminopterin; ibandronate; irinotecan (e.g., CPT-11); topoisomerase inhibitor RFS2000; difluoromethylornithine (DMFO); retinoids such as retinoic acid; esperamicin; capecitabine (e.g., Xeloda®); and any pharmaceutically acceptable salts of the above.

[0252] Non-exclusive examples of additional anticancer drugs include trastuzumab (Herceptin®), bevacizumab (Avastin®), cetuximab (Erbitux®), rituximab (Rituxan®), Taxol®, Arimidex®, ABVD, Avisin, avagovomab, acridine carboxamide, adecatumumab, 17-N-allylamino-17-demethoxygeldanamycin, and alafara. Zin, Arbocidib, 3-aminopyridine-2-carboxyaldehyde thiosemicarbazone, Amonafide, Anthracendione, Anti-CD22 Antitoxin, Antineoplastic Agents (e.g., cell cycle nonspecific antineoplastic agents, and other antineoplastic agents described herein), Antitumor Herbs, Apadicone, Atiprimod, Azathioprine, Belotecan, Bendamustine, BIBW2992, Bilicodal, Brostalicin, Briostatin, Butionine Sulfoximine, CBV (Chemotherapy) Kalikrin, dichloroacetic acid, discodermolide, erusamitrusine, enocitabine, eribulin, exatecan, exislind, ferginol, forodesine, phosfestrol, ICE chemotherapy regimen, IT-101, imexon, imiquimod, indocarbazole, irofluben, lanikidal, larotaxel, lenalidomide, lucanton, lulutotecan, maphosphamide, mitozolomid, napoxidine, nedaplatin, olaparib, ortata Examples include Xicel, PAC-1, Pawpaw, Pixantrone, proteasome inhibitors, rebeccamycin, reximod, rubitecan, SN-38, salinosporamide A, sapacitabine, Stanford V, swinesonin, talaporfin, talikidal, tegafur-uracil, temodal, tesetaxel, triplatin tetranitrate, tris(2-chloroethyl)amine, troxacitabine, uramustine, bajimezan, vinflunin, ZD6126, and zoskidal.

[0253] Further non-limiting examples of anticancer drugs include natural products, e.g., vinca alkaloids (e.g., vinblastine, vincristine, and vinorelbine), epipodophyllotoxins (e.g., etoposide and teniposide), antibiotics (e.g., dactinomycin (actinomycin D), daunorubicin, and idarubicin), anthracyclines, mitoxantrone, bleomycin, plicamycin (mitramycin), mitomycin, and enzymes (e.g., L-asparagine). L-asparaginase (which does not give cells that do not metabolize and synthesize asparagine of their own), antiplatelet agents, antiproliferative / antimitotic alkylating agents, e.g., nitrogen mustard (e.g., mechloretamine, cyclophosphamide and analogs, melphalan, and chlorambucil), ethyleneimine and methylmelamine (e.g., hexamethylmelamine and thiotepa), CDK inhibitors (e.g., CDK4 / 6 inhibitors, e.g., abemaciclib, ribociclib, palbociclib;Cericiclib (UCN-01, P1446A-05, PD-0332991, Dinacyclib (P27-00, AT-7519, RGB286638, and SCH727965)), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine (BCNU) and its analogs, and streptozocin), trazeneth-dacarbazine (DTIC), antiproliferative / antimitotic antimetabolites (e.g., folate 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-coordinate complexes (e.g., cisplatin and carboplatin) , procarbazine, hydroxyurea, mitotane, aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g., trichostatin, sodium butyrate, apicidan, suberoylanilide hydroamic acid, vorinostat, LBH589, romidepsin, ACY-1215, and panobinostat), mTOR inhibitors (e.g., bisutucertib, temsirolimus, everolimus, sapanacertib) (sapanasertib), ridafololimus, and sirolimus), KSP(Eg5) inhibitors (e.g., Array520), DNA binders (e.g., Zalypsis®), PI3K inhibitors, e.g., PI3K delta inhibitors (e.g., GS-1101 and TGR-1202), PI3K delta and gamma inhibitors (e.g., CAL-130), copanlisib, alpelisib, and idelalisib;Multiple kinase inhibitors (e.g., TG02 and sorafenib), hormones (e.g., estrogen) and hormone agonists, e.g., luteinizing hormone-releasing hormone (LHRH) agonists (e.g., goserelin, leuprolide and triptorelin), BAFF neutralizing antibodies (e.g., LY2127399), IKK inhibitors, p38MAPK inhibitors, anti-IL-6 (e.g., CNT0328), telomerase inhibitors (e.g., GRN163L), aurora kinase inhibitors (e.g., MLN8237), cell surface monoclonal antibodies (e.g., anti-CD38 (HUMAX-CD38), anti-CSl (e.g., elotuzumab), HSP90 inhibitors ( Examples include 17AAG and KOS953), P13K / Akt inhibitors (e.g., perifosine), Akt inhibitors (e.g., GSK-2141795), PKC inhibitors (e.g., Enzastaurin), FTIs (e.g., Zanestra®), 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.

[0254] In some embodiments, the anticancer agent is selected from mechloretamine, camptothecin, ifosfamide, tamoxifen, raloxifene, gemcitabine, Navelbine®, sorafenib, or any analogue or derivative variant of the foregoing.

[0255] In some embodiments, the anticancer agent is an ALK inhibitor. 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 WO05016894.

[0256] In some embodiments, the anticancer agent is an inhibitor of a downstream member of the receptor tyrosine kinase (RTK) / growth factor receptor (e.g., SHP2 inhibitors (e.g., SHP099, TNO155, RMC-4550, RMC-4630, JAB-3068, JAB-3312, RLY-1971, ERAS-601, SH3809, PF-07284892, or BBP-398, or any other SHP2 inhibitor known in the art)), an SOS1 inhibitor (e.g., RMC-5845, BI-1 These include 701963, BI-3406, SDR5, or BAY-293, or any other SHP2 inhibitor known in the art), RAS inhibitors (e.g., BI-2852 or any other RAS inhibitor known in the art), RAS degraders, Raf inhibitors, MEK inhibitors, ERK inhibitors, PI3K inhibitors, PTEN inhibitors, AKT inhibitors, or mTOR inhibitors (e.g., mTORC1 inhibitors or mTORC2 inhibitors). In some embodiments, the anticancer agent is JAB-3312.

[0257] In some embodiments, therapeutic agents that may be combined with the compounds of this disclosure are inhibitors of the MAP kinase (MAPK) pathway (or "MAPK inhibitors"). MAPK inhibitors include, but are not limited to, one or more MAPK inhibitors described in Cancers (Basel) 2015 Sep;7(3):1758-1784. For example, MAPK inhibitors include trametinib, binimetinib, selumetinib, cobimetinib, LErafAON (NeoPharm), ISIS5132; vemurafenib, pimacertib, TAK733, RO4987655 (CH4987655); CI-1040; PD-0325901; CH5126766; MAP855; AZD6244; refametinib (RDEA119 / BAY86-9766); GDC-0973 / XL581; AZD8330 (ARRY-424704 / ARRY-704); RO5126766 (Roche, PLoS One. 2014 Nov 25; 9(11)); and GSK1120212 (or JTP-74057, Clin Cancer). One or more of the following may be selected (as described in Res.2011 Mar 1;17(5):989-1000). The MAPK inhibitor may be PLX8394, LXH254, GDC-5573, or LY3009120.

[0258] In some embodiments, the anticancer agent is an interferant or inhibitor of the RAS-RAF-ERK, PI3K-AKT-TOR, or PI3K-AKT signaling pathway. Examples of PI3K / AKT inhibitors include, but are not limited to, one or more PI3K / AKT inhibitors listed 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.

[0259] In some embodiments, the anticancer agent is a PD-1 or PD-L1 antagonist.

[0260] 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 immunotherapies, such as immune checkpoint inhibitors.

[0261] Examples of MEK inhibitors include, but are not limited to, pimacertib, selumetinib, cobimetinib (Cotellic®), trametinib (Mekinist®), and binimetinib (Mektovi®). In some embodiments, the 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 ΔE51-Q58;ΔF53-Q58;E203K;L177M;C121S;F53L;K57E;Q56P; and K57N.

[0262] Examples of PI3K inhibitors include wartmannin, a 17-hydroxywartmannin analog described in WO06 / 044453; 4-[2-(1H-indazole-4-yl)-6-[[4-(methylsulfonyl)piperazine-1-yl]methyl]thieno[3,2-d]pyrimidine-4-yl]morpholine (also known as pictilisib or GDC-0941, described in WO09 / 036082 and WO09 / 055730); 2-methyl-2-[4-[3-methyl-2-oxo-8-(quinoline-3-yl)-2,3-dihydroimidazo[ 4,5-c]quinoline-1-yl]phenyl]propionitrile (also known as BEZ235 or NVP-BEZ235, described in WO06 / 122806); (S)-1-(4-((2-(2-aminopyrimidine-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidine-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (described in WO08 / 070740); LY294002(2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (Axon Available from Medchem); PI103 hydrochloride (3-[4-(4-morpholinylpyrido-[3',2':4,5]flo[3,2-d]pyrimidine-2-yl]phenol hydrochloride (Available from Axon Medchem); PIK75 (2-methyl-5-nitro-2-[(6-bromoimidazo[1,2-a]pyridine-3-yl)methylene]-1-methylhydrazide-benzenesulfonic acid, monohydrochloride) (Available from Axon Medchem); PIK90 (N-(7,8-dimethoxy-2,3-dihydroimidazo[1,2-c]quinazolin-5-yl)-nicotinamide (Axon Available from Medchem); AS-252424 (5-[1-[5-(4-fluoro-2-hydroxyphenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidined-2,4-dione (available from Axon Medchem); TGX-221 (7-methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrinidine-4-one (available from Axon Medchem); XL-765; and XL-147 are examples, but are not limited to these.Other PI3K inhibitors include demethoxypyridine, perifosine, CAL101, PX-866, BEZ235, SF1126, INK1117, IPI-145, BKM120, XL147, XL765, Palomid529, GSK1059615, ZSTK474, PWT33597, IC87114, TGI00-115, CAL263, PI-103, GNE-477, CUDC-907, and AEZS-136.

[0263] Examples of AKT inhibitors include Akt-1-1 (inhibits Akt1) (Barnett et al., Biochem.J.2005,385(Pt.2):399-408); Akt-1-1,2 (inhibits Ak1 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); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO05 / 011700); indole-3-carbinol and its derivatives (e.g., U.S. Patent No. 6,656,963; Sarkar and Li J Nutr.2004,134(12)). Examples include, but are not limited to, Suppl):3493S-3498S); Perifosine (e.g., interfering with Akt membrane localization; Dasmahapatra et al. Clin. Cancer Res. 2004, 10(15):5242-52); Phosphatidylinositol ether lipid analogs (e.g., Gills and Dennis Expert. Opin. Investig. Drugs 2004, 13:787-97); and trisirivine (TCN or API-2 or NCI discriminant: NSC154020; Yang et al., Cancer Res. 2004, 64:4394-9).

[0264] mTOR inhibitors include, but are not limited to, ATP-competitive mTORC1 / mTORC2 inhibitors, such as PI-103, PP242, PP30; Torin1; FKBP12 enhancers; 4H-1-benzopyran-4-one derivatives; and rapamycin (also known as sirolimus) and its derivatives, including: temsirolimus (Torisel®); everolimus (Afinitor®). ), WO94 / 09010); sapanasertib, lidafololimus (also known as deforolimus or AP23573); rapalogs, e.g., those disclosed in WO98 / 02441 and WO01 / 14387, e.g., AP23464 and AP23841; 40-(2-hydroxyethyl)rapamycin; 40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin (Also known as CC1779); 40-epi-(tetrazolite)-rapamycin (also known as ABT578); 32-deoxorapamycin; 16-pentinyloxy-32(S)-dihydrolapanisin; derivatives disclosed in WO05 / 005434; U.S. Patents 5,258,389, 5,118,677, 5,118,678, 5,100,883, 5,151,413, and 5,120,842 , and derivatives disclosed in WO94 / 090101, WO92 / 05179, WO93 / 111130, WO94 / 02136, WO94 / 02485, WO95 / 14023, WO94 / 02136, WO95 / 16691, WO96 / 41807, WO96 / 41807, and WO2018204416; as well as phosphorus-containing rapamycin derivatives (e.g., WO05 / 016252). In some embodiments, the mTOR inhibitor is a disteric inhibitor (see, for example, WO2018204416, WO2019212990, and WO2019212991), e.g., RMC-5552.

[0265] Examples of BRAF inhibitors that may be used in combination with the compounds of this disclosure include vemurafenib, dabrafenib, and encorafenib. BRAF may include class 3 BRAF mutations. In some embodiments, class 3 BRAF mutations are 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.

[0266] Examples of MCL-1 inhibitors include, but are not limited to, AMG-176, MIK665, and S63845. The myeloid cell leukemia-1 (MCL-1) protein is one of the major anti-apoptotic members of the B-cell lymphoma-2 (BCL-2) protein family. Overexpression of MCL-1 is closely associated with tumor progression and resistance not only to conventional chemotherapy but also to targeted therapies, including BCL-2 inhibitors such as ABT-263.

[0267] In some embodiments, additional therapeutic agents are selected from the group consisting of MEK inhibitors, HER2 inhibitors, SHP2 inhibitors, CDK4 / 6 inhibitors, mTOR inhibitors, SOS1 inhibitors, and PD-L1 inhibitors. See, for example, Hallin et al., Cancer Discovery, DOI:10.1158 / 2159-8290 (October 28, 2019) and Canon et al., Nature, 575:217 (2019). In some embodiments, the Ras inhibitors of this disclosure are used in combination with MEK inhibitors and SOS1 inhibitors. In some embodiments, the Ras inhibitors of this disclosure are used in combination with PD-L1 inhibitors and SOS1 inhibitors. In some embodiments, the Ras inhibitors of this disclosure are used in combination with PD-L1 inhibitors and SHP2 inhibitors. In some embodiments, the Ras inhibitors of this disclosure are used in combination with MEK inhibitors and SHP2 inhibitors. In some embodiments, the cancer is colorectal cancer, and the treatment comprises administering the Ras inhibitor of this disclosure in combination with a second or third therapeutic agent.

[0268] Examples of proteasome inhibitors include, but are not limited to, carfilzomib (Kyprolis®), bortezomib (Velcade®), and oprozomib.

[0269] Immunotherapy includes, but is not limited to, monoclonal antibodies, immunomodulatory imides (IMiDs), GITR agonists, genetically modified T cells (e.g., CAR-T cells), bispecific antibodies (e.g., BiTE), and anti-PD-1, anti-PD-L1, anti-CTLA4, anti-LAGl, and anti-OX40 agents.

[0270] Immunomodulators (IMiDs) are a class of immunomodulatory drugs (drugs that modulate the immune response) that contain an imide group. Examples of IMiDs include thalidomide and its analogues (lenalidomide, pomalidomide, and apremilast).

[0271] Exemplary anti-PD-1 antibodies and their uses are described in Goldberg et al., Blood 2007, 110(1):186-192; Thompson et al., Clin. Cancer Res. 2007, 13(6):1757-1761; and WO06 / 121168 A1, and further described elsewhere in this Specification.

[0272] Examples of GITR agonists include GITR fusion proteins and anti-GITR antibodies (e.g., bivalent anti-GITR antibodies), e.g., GITR fusion proteins described in U.S. Patent No. 6,111,090, No. 8,586,023, WO2010 / 003118, and WO2011 / 090754; or e.g., U.S. Patent No. 7,025,962, EP1947183, U.S. Patent No. 7,812,135, No. 8,388,967, and No. 8,591 Examples of anti-GITR antibodies include, but are not limited to, those described in No. 886, No. 7,618,632, EP1866339, and WO2011 / 028683, WO2013 / 039954, WO05 / 007190, WO07 / 133822, WO05 / 055808, WO99 / 40196, WO01 / 03720, WO99 / 20758, WO06 / 083289, WO05 / 115451, and WO2011 / 051726.

[0273] Other examples of therapeutic agents that may be used in combination with the compounds of this disclosure are anti-angiogenic agents. Anti-angiogenic agents include, but are not limited to, chemical compositions, antibodies, antigen-binding domains, radionuclides, and combinations and conjugates thereof, which are synthetically prepared in vitro. Anti-angiogenic agents may be agonists, antagonists, allosteric modulators, toxins, or, more generally, may act to inhibit or stimulate their targets (e.g., by activating or inhibiting receptors or enzymes), thereby promoting cell death or halting cell proliferation. In some embodiments, one or more additional therapies include anti-angiogenic agents.

[0274] Anti-angiogenic agents may include MMP-2 (matrix-metalloproteinase 2) inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, and COX-II (cyclooxygenase 11) inhibitors. Non-exclusive examples of anti-angiogenic agents include rapamycin, temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib, and bevacizumab. Examples of useful COX-II inhibitors include arecoxib, valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinase inhibitors include WO96 / 33172, WO96 / 27583, WO98 / 07697, WO98 / 03516, WO98 / 34918, WO98 / 34915, WO98 / 33768, WO98 / 30566, WO90 / 05719, WO99 / 52910, and WO99 / 52 As described in 889, WO99 / 29667, WO99007675, EP0606046, EP0780386, EP1786785, EP1181017, EP0818442, EP1004578, and US20090012085, and U.S. Patents 5,863,949 and 5,861,510. Preferred MMP-2 and MMP-9 inhibitors have little to no activity in inhibiting MMP-1. More preferred are those that selectively inhibit MMP-2 or AMP-9 compared to 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, RO32-3555, and RS13-0830.

[0275] Further exemplary anti-angiogenic agents include KDR (kinase domain receptor) inhibitors (e.g., antibodies and antigen-binding regions that specifically bind to kinase domain receptors), anti-VEGF agents (e.g., VEGF (e.g., bevacizumab), or antibodies or antigen-binding regions that specifically bind to their soluble VEGF receptors or ligand-binding regions), e.g., VEGF-TRAP®, and anti-VEGF receptor agents (e.g., antibodies or antigen-binding regions that specifically bind to them), EGFR inhibitors (e.g., antibodies or antigen-binding regions that specifically bind to them), e.g., Vectibix® (panitumumab), erlotinib (Tarceva®), anti-Ang1 and anti-Ang2 agents (e.g., antibodies or antigen-binding regions that specifically bind to them or their receptors, e.g., Tie2 / Tek), and anti-Tie2 kinase inhibitors (e.g., antibodies or antigen-binding regions that specifically bind to them). Other anti-angiogenic agents include Canas, IL-8, B-FGF, Tek antagonists (US2003 / 0162712, US6,413,932), anti-Tweak agents (e.g., antibodies or antigen-binding domains that specifically bind, or soluble Tweak receptor antagonists; see US6,727,225), ADAM distointegrin domains that antagonize the binding of integrins to their ligands (US2002 / 0042368), and anti-eph receptors or anti-ephrin antibodies or antigen-binding domains that specifically bind. Examples include combined areas (U.S. Patents No. 5,981,245, No. 5,728,813, No. 5,969,110, No. 6,596,852, No. 6,232,447, No. 6,057,124, and their patent family members), anti-PDGF-BB antagonists (e.g., antibody or antigen-binding domains that specifically bind), antibody or antigen-binding domains that specifically bind to PDGF-BB ligands, and PDGFR kinase inhibitors (e.g., antibody or antigen-binding domains that specifically bind to them). Additional anti-angiogenic agents include SD-7784 (Pfizer, USA); sirengitide (Merck KGaA, Germany, EPO0770622); pegaptanib octasodium (Gilead Sciences, USA);Alpha-statin (BioActa, UK); M-PGA (Celgene, USA, US5712291); Ilostat (Arriva, USA, US5892112); Emaxanib (Pfizer, USA, US5792783); Batalanib (Novartis, Switzerland); 2-Methoxyestradiol (EntreMed, USA); TLC ELL-12 (Elan, Ireland); Anecoltab acetate (Alcon, USA); Alpha-D148Mab (Amgen, USA); CEP-7055 (Cephalon, USA); Anti-Vn Mab (Crucell, Netherlands), DAC anti-angioplastic agent (ConjuChem, Canada); Angiocidin (InKine Pharmaceutical, USA); KM-2550 (Kyowa Hakko, Japan); SU-0879 (Pfizer, USA); CGP-79787 (Novartis, Switzerland, EP0970070); ARGENT technology (Ariad, USA); YIGSR-Stealth (Johnson & Johnson, USA); Fibrinogen-E fragment (BioActa, UK); Angiogenesis 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, JP02233610); Platelet factor 4 (RepliGen, USA, EP407122); Vascular endothelial growth factor antagonist (Borean, Denmark); Bevacizumab (pINN) (Genentech, USA); Angiogenesis inhibitor (SUGEN, USA); XL784 (Exelixis, USA); XL647 (Exelixis, USA);MAb, alpha-5 beta-3 integrin, second generation (Applied Molecular Evolution, USA and Medlmmune, USA); Enzastaurin hydrochloride (Lilly, USA); CEP7055 (Cephalon, USA and Sanofi-Synthelabo, France); BC1 (Genoa Institute of Cancer Research, Italy); rBPI21 and BPI-derived anti-angiogenic agents (XOMA, USA); PI88 (Progen, Australia); Silengitide (Merck KGaA, Germany; Munich Technical University, Germany, Scripps Clinic and Research Foundation, USA); AVE8062 (Ajinomoto, Japan); AS1404 (Cancer Research Laboratory, New Zealand); SG292 (Telios, USA); Endostatin (Boston Childrens Hospital, USA); ATN161 (Attenuon, USA); 2-Methoxyestradiol (Boston Childrens Hospital) Hospital, USA); ZD6474 (AstraZeneca, UK); ZD6126 (Angiogene Pharmaceuticals, UK); PPI2458 (Praecis, USA); AZD9935 (AstraZeneca, UK); AZD2171 (AstraZeneca, UK); Batalanib (pINN) (Novartis, Switzerland and Schering AG, Germany); Tissue factor pathway inhibitor (EntreMed, USA); Pegaptanib (Pinn) (Gilead Sciences, USA); Xantolulizole (Yonsei University, South Korea); Vaccine, gene-based, VEGF-2 (Scripps Clinic and Research Foundation, USA); SPV5.2 (Supratek, Canada); SDX103 (University of California at San Diego, USA); PX478 (ProlX, USA);Metastatin (EntreMed, USA); Troponin I (Harvard University, USA); SU6668 (SUGEN, USA); OXI4503 (OXiGENE, USA); o-Guanidine (Dimensional Pharmaceuticals, USA); Motupolamine C (British Columbia University, Canada); CDP791 (Celltech Group, UK); Atiprimod (pINN) (GlaxoSmithKline, UK); E7820 (Eisai, Japan); CYC381 (Harvard University, USA); AE941 (Aeterna, Canada); Vaccine, Angiogenesis (EntreMed, USA); Urokinase Plasminogen Activator Inhibitor (Dendreon, USA); Ogluphanide (pINN) (Melmotte, USA); HIF-Ralfa Inhibitor (Xenova, UK); CEP5214 (Cephalon, USA); BAY RES2622 (Bayer, Germany); Angiocidin (InKine, USA); A6 (Angstrom, USA); KR31372 (Korea Research Institute of Chemical Technology, South Korea); GW2286 (GlaxoSmithKline, UK); EHT0101 (ExonHit, France); CP868596 (Pfizer, USA); CP564959 (OSI, USA); CP547632 (Pfizer, USA); 786034 (GlaxoSmithKline, UK); KRN633 (Kirin Brewery, Japan); Drug delivery systems, intraocular, 2-methoxyestradiol; Anguinex (Maastricht University, Netherlands, and Minnesota) University, USA); ABT510 (Abbott, USA); AAL993 (Novartis, Switzerland); VEGI (ProteomTech, USA); Tumor necrosis factor-alpha inhibitor; SU11248 (Pfizer, USA and SUGEN USA); ABT518 (Abbott, USA); YH16 (Yantai Rongchang, China);S-3APG (Boston Children's Hospital, USA and EntreMed, USA); MAb, KDR (ImClone Systems, USA); MAb, alpha-5 beta (Protein Design, USA); KDR kinase inhibitor (Celltech Group, UK and Johnson & Johnson, USA); GFB116 (South Florida University, USA and Yale University, USA); CS706 (Sankyo, Japan); Comblestatin A4 prodrug (Arizona State University, USA); Chondroitinase AC (IBEX, Canada); BAY RES2690 (Bayer, Germany); AGM1470 (Harvard University, USA, Takeda, Japan and TAP, USA); AG13925 (Agouron, USA); Tetrathiomolybdate (University of Michigan, USA); GCS100 (Wayne State University, USA); CV247 (Ivy Medical, UK); CKD732 (Chong Kun Dang, South Korea; Ilsogladine (Nippon Shinyaku, Japan); RG13577 (Aventis, France); WX360 (Wilex, Germany); Squalamine (Genaera, USA); RPI4610 (Sirna, USA); Heparanase inhibitor (InSight, Israel); KL3106 (Kolon, South Korea); Honokiol (Emory University, USA); ZK CDK (Schering AG, Germany); ZK Angio (Schering AG, Germany); ZK229561 (Novartis, Switzerland, and Schering AG, Germany); XMP300 (XOMA, USA); VGA1102 (Taisho, Japan); VE-Cadherin-2 antagonist (ImClone Systems, USA); Vasostatin (National Institutes of Health, USA);Flk-1 (ImClone Systems, USA); TZ93 (Tsumura, Japan); TumStatin (Beth Israel Hospital, USA); Sclerated soluble FLT1 (vascular endothelial growth factor receptor 1) (Merck & Co, USA); Tie-2 ligand (Regeneron, USA); and thrombospondin 1 inhibitor (Allegheny Health, Educ; The National Institute and Research Foundation (USA) is one example.

[0276] Further examples of therapeutic agents that may be used in combination with the compounds of this disclosure include agents that specifically bind to and inhibit the activity of growth factors, such as antagonists of hepatocyte growth factor (HGF, also known as scatter factor) (e.g., antibodies, antigen-binding domains, or soluble receptors), as well as antibodies or antigen-binding domains that specifically bind to the receptor c-Met.

[0277] Other examples of therapeutic agents that may be used in combination with the compounds of this disclosure are autophagy inhibitors. Examples of autophagy inhibitors include, but are not limited to, chloroquine, 3-methyladenine, hydroxychloroquine (Plaquenil®), bafilomycin A1, 5-amino-4-imidazole carboxamidriboside (AICAR), okadaic acid, autophagy-suppressing algal toxins that inhibit type 2A or type 1 protein phosphatases, cAMP analogs, and drugs that increase cAMP levels, such as adenosine, LY204002, N6-mercaptopurine riboside, and vinblastine. Furthermore, antisense or siRNAs that inhibit the expression of proteins including (but not limited to) ATG5 (which is involved in autophagy) may also be used. In some embodiments, one or more additional therapies include autophagy inhibitors.

[0278] Other examples of therapeutic agents that may be used in combination with the compounds of this disclosure are anti-cancer agents. In some embodiments, one or more additional therapies include anti-cancer agents. Non-limiting examples of anti-cancer agents include acemannan, acralubicin, aldesleukin, alemtuzumab, alitretinoin, altretamine, amiphostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ancer, ancestim, algravin, arsenic trioxide, BAM-002 (Novelos), bexarotene, bicalutamide, bromodeoxyuridine, capecitabine, cermoloukin, cetrorelix, cladribine, clotrimazole, and sita. Rabinocophage, DA3030 (Dong-A), daclizumab, deniroukin difutitox, deslorerin, dexrazoxane, dilazep, docetaxel, docosanol, doxelcalciferol, doxifluridine, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT diclofenac, interferon alpha, daunorubicin, doxorubicin, tretinoin, ederfosine, edrecolomab, eflornithine, emiteflu, epirubicin, epoeth Interferon alpha, etoposide phosphate, exemestane, exislind, fadrozol, filgrastim, finasteride, fludarabine phosphate, formestan, hotemustine, gallium nitrate, gemcitabine, gemtuzumabuzogamicin, gimeracil / oteracil / tegafur combination, glycopine, goserelin, heptaplatin, human chorionic gonadotropin, human fetal alpha-fetoprotein, ibandronate, idarubicin (imiquimod, interferon alpha, interferon alpha, natural type) Interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon alpha-NI, interferon alpha-n3, interferon alphacon-1, interferon alpha, natural type, interferon beta, interferon beta-la, interferon beta-lb, interferon gamma, natural type interferon gamma-la, interferon gamma-lb, interleukin-1 beta, iobenguan, irinotecan,Ilsogladine, Lanreotide, LC9018 (Yakult), Leflunomide, Lenograstim, Lentinan sulfate, Letrozole, Leukocyte alpha interferon, Leuprorelin, Levamisole + Fluorouracil, Rialozol, Lovaplatin, Ronidamin, Lovastatin, Masopropyl, Melalsoprole, Metoclopramide, Mifepristone, Miltefosine, Millimostim, Mispaired double-stranded RNA, Mitoguazone, Mitractol, Mitoxantrone, Morglamostim, Nafarelin, Naloxone + Pentazocine, Naltogra Stim, Nedaplatin, Niltamide, Noscapine, Novel Erythropoiesis-Promoting Protein, NSC631570 Octreotide, Oprelbequin, Osateron, Oxaliplatin, Paclitaxel, Pamidronic Acid, Pegaspargase, Peginterferon Alpha-2b, Pentosan Polysulfate Sodium, Pentostatin, Picibanil, Pirarubicin, Rabbit Antithymocyte Polyclonal Antibody, Polyethylene Glycol Interferon Alpha-2a, Porfimer Sodium, Raloxifene, Larcitrexed, Rasbriem Bodyment (ras) buriembodiment), etidronate rhenium Re186, RII retinamide, rituximab, romultide, samarium (153Sm) lexidonam, salglamostim, schizophyllan, sobuzoxane, sonelmin, strontium-89 chloride, suramin, tasonelmin, tazarotene, tegafur, temoporfin, temozolomide, teniposide, tetrachlorodecaoxide, thalidomide, thymalfacin, thyroid-stimulating hormone alpha, topotecan, toremifene, tositumomab-iodine-131, trastuzumab, treosulfan, Tretinoin, trilostane, trimethrexate, triptorelin, tumor necrosis factor alpha, natural type, ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma solubilizing solution vaccine, barrubicin, verteporfin, vinorelbine, bilirulysine, dinostatin stimulamer, or zoledronic acid; Abarelix; AE941 (Aeterna), ambamustin, antisense oligonucleotide, bcl-2 (Genta), APC8015 (Dendreon), decitabine, dexaaminoglutethimide, diazicon, EL532 (Elan),EM800 (Endorecherche), eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen), fulvestrant, gallocitabine, gastrin-17 immunogene, HLA-B7 gene therapy (Vical), granulocyte-macrophage colony-stimulating factor, histamine dihydrochloride, ibritumomab tiuxetan, ilomastat, IM862 (Cytran), interleukin-2, iproxyfen, LDI200 (Milkhaus), religistim, lintuzumab, CA125 MAb (Biomira), cancer MAb (Japan Pharmaceutical Development), HER-2 and Fc MAb (Medarex), idiotype 105AD7 MAb (CRC Technology), idiotype CEA MAb (Trilex), LYM-1-iodine-131 MAb (Techniclone), polymorphoemic mucin-yttrium-90 MAb (Antisoma), marimast, menogalil, mitsumomab, motexafingadolinium, MX6 (Galderma), nelarabine, noratexed, P30 protein, pegvisomant, pemetrexed, porphyromycin, prinomast, RL0903 (Shire), rubitecan, satraplatin, sodium phenylacetate, sparphosic acid, SRL172 (SR Pharma), SU5416 (SUGEN), TA077 (Tanabe), tetrathiomolybdate, saliblastin, thrombopoietin, tin ethylethiopurine, tirapazamine, cancer vaccine (Biomira), melanoma vaccine (New York University), melanoma vaccine (Sloan Kettering Examples include the melanoma tumor lysis product vaccine (New York Medical College), the viral melanoma cell solubilization vaccine (Royal Newcastle Hospital), or Valspodar.

[0279] Further examples of therapeutic agents that may be used in combination with the compounds of this 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; Atasicept; CP-870893; Lucatumumab; Dasetuzumab; Muromonab-CD3; Ipilumumab; MEDI4736 (Imfinzi(registered trademark)); MSB0010718C; AMP224; Adalimumab (Humira(registered trademark)); ado-trastuzumab emtansine (Kadcyla(registered trademark)); Aflibercept (Eylea(registered trademark)); Alemtuzumab (Campath(registered trademark)); Basiliximab (Simulect(registered trademark)); Belimumab (Benlysta(registered trademark)); Basiliximab (Simulect(registered trademark)); Belimumab (Benlysta(registered trademark)); Brentuximab vedotin (Adcetris(registered trademark)); Canakinumab (Ilaris(registered trademark)); Certolizumab Gol (Cimzia®); Daclizumab (Zenapax®); Daratumumab (Darzalex®); Denosumab (Prolia®); Eculizumab (Soliris®); Ephalizumab (Raptiva®); Gemtuzumab Ozogamicin (Mylotarg®); Golimumab (Simponi®); Ibritumomab Chiuxetan (Zevalin®); Infliximab (Remicade®); Motabizumab (Numax®); Natalizumab (Tysabri®); Obinutuzumab (Gazyva®); Ofatumumab (Arzerra®); Omalizumab (Xolair®); Palivizumab (Synagis®);Examples include pertuzumab (Perjeta®), pertuzumab (Perjeta®), ranibizumab (Lucentis®), laxibakumab (Abthrax®), tocilizumab (Actemra®), tositumomab, tositumomab-i-131, tositumomab and tositumomab-i-131 (Bexxar®), ustekinumab (Stelara®), AMG102, AMG386, AMG479, AMG655, AMG706, AMG745, and AMG951.

[0280] The compounds described herein can be used in combination with other agents disclosed herein or other suitable agents, depending on the condition being treated. Therefore, in some embodiments, one or more compounds of this disclosure are co-administered with other therapies as described herein. When used in combination therapy, the compounds described herein may be administered simultaneously with or separately from a second agent. This combined administration may include simultaneous administration of the two agents in the same dosage form, simultaneous administration in separate dosage forms, and separate administration. That is, the compounds described herein and any agent described herein may be formulated together in the same dosage form and administered simultaneously. Alternatively, the compounds of this disclosure and any therapy described herein may be administered simultaneously, with both agents existing in separate formulations. In other alternative methods, the compounds of this disclosure may be administered, followed by any therapy described herein, or vice versa. In some embodiments of separate administration protocols, the compounds of this disclosure and any therapy described herein are administered at intervals of several minutes, several hours, or several days.

[0281] In some embodiments of any of the methods described herein, the first therapy (e.g., the compounds of this disclosure) and one or more additional therapies are administered simultaneously or sequentially in any order. The first therapeutic agent may be administered immediately before or after one or more additional therapies, or up to 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 24 hours before or after, or up to 1-7, 1-14, 1-21, or 1-30 days before or after.

[0282] The Disclosure also features a kit comprising (a) a pharmaceutical composition comprising an agent described herein (e.g., a compound of the Disclosure), and (b) a package insert containing instructions for performing any of the methods described herein. In some embodiments, the kit comprises (a) a pharmaceutical composition comprising an agent described herein (e.g., a compound of the Invention), (b) one or more additional therapies (e.g., non-pharmacological treatments or therapeutic agents), and (c) a package insert containing instructions for performing any of the methods described herein.

[0283] One aspect of the present disclosure relates to combining separate pharmaceutical compositions in the form of a kit, for the purpose of treating a disease or a related condition using a combination of pharmaceutically active compounds that can be administered separately. The kit may comprise two separate pharmaceutical compositions, i.e., the compounds of the present disclosure, and one or more additional therapies. The kit may comprise a container for housing 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 instructions for using 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), administered at different dosing intervals, or when titration of the individual components of the combination is desired by the prescribing healthcare professional. Embodiment

[0284] Embodiment 1. A method for treating cancer in a subject requiring cancer treatment, comprising administering a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound to the subject, wherein the cancer does not contain a RAS mutation at position 61.

[0285] Embodiment 2. A method for treating cancer in a subject requiring cancer treatment, comprising administering a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound and a RAS(OFF) inhibitor to the subject, wherein the cancer does not contain a RAS mutation at position 61.

[0286] Embodiment 3. A method for treating cancer in a subject requiring cancer treatment, comprising administering a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound and an RTK inhibitor to the subject, wherein the cancer does not contain a RAS mutation at position 61.

[0287] Embodiment 4. A method for treating cancer in a subject requiring cancer treatment, comprising administering a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound and an SHP inhibitor (e.g., an SHP2 inhibitor) to the subject, wherein the cancer does not contain a RAS mutation at position 61.

[0288] Embodiment 5. A method for treating cancer in a subject requiring cancer treatment, comprising administering a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound and an SOS1 inhibitor to the subject, wherein the cancer does not contain a RAS mutation at position 61.

[0289] Embodiment 6. The method according to any one of Embodiments 1 to 5, wherein the cancer includes a RAS mutation.

[0290] Embodiment 7: The method according to Embodiment 6, wherein the RAS mutation is an RAS amplification, or the RAS mutation is located at position 12 or 13.

[0291] Embodiment 8: The method according to any one of Embodiments 1 to 7, wherein the cancer is pancreatic cancer, colorectal cancer, non-small cell lung cancer, gastric cancer, esophageal cancer, ovarian cancer, or uterine cancer.

[0292] Embodiment 9: The method according to any one of Embodiments 1 to 8, wherein the binding of the RAS(ON)GTP hydrolysis-promoting compound to RAS(ON) changes the position of glutamine 61 of RAS(ON) toward the gamma phosphate of GTP bound to the RAS(ON) protein, compared to its position in the absence of the RAS(ON)GTP hydrolysis-promoting compound, thereby increasing the GTP hydrolysis rate compared to the hydrolysis rate of RAS(ON) in the absence of the RAS(ON)GTP hydrolysis-promoting compound.

[0293] Embodiment 10. The method according to any one of Embodiments 1 to 9, wherein the RAS(ON)GTP hydrolysis promoting compound is one of the compounds in Table 1 or a pharmaceutically acceptable salt thereof.

[0294] Embodiment 11. The method according to any one of Embodiments 2 or 6 to 10, wherein the RAS(OFF) inhibitor is a RAS(OFF) inhibitor.

[0295] Embodiment 12. The KRAS(OFF) inhibitor is KRAS G12C The method according to Embodiment 11, wherein the (OFF) inhibitor is used.

[0296] Embodiment 13. The KRAS G12C The method according to Embodiment 12, wherein the (OFF) inhibitor is selected from the group consisting of AMG510 (sotrasib), MRTX849 (adaglasib), MRTX1257, GDC-6036 (divalasib), JDQ443 (opnurasib), ERAS-3490, LY3537982 (olomorasib), BI1823911, BPI-421286, JAB-3312, JAB-21000, JAB-21822 (glesilasib), D-1553, D3S-001, HBI-2438, HS-10370, MK-1084, YL-15293, BBO-8520, FMC-376, GEC255, and GFH925 (IBI351).

[0297] Embodiment 14. The KRAS(OFF) inhibitor is KRAS G12D The method according to Embodiment 11, wherein the (OFF) inhibitor is used.

[0298] Embodiment 15. The KRAS G12D The method according to Embodiment 14, wherein the (OFF) inhibitor is selected from the group consisting of MRTX1133, MRTX282, JAB-22000, ERAS-4, ERAS-5024, HRS-4642, BI-2852, ASP3082, TH-Z827, TH-7835, QTX-3046, GFH375 (VS-7375), INCB161734, and KD-8.

[0299] Embodiment 16. The KRAS(OFF) inhibitor is KRAS G12V The method according to Embodiment 11, wherein the (OFF) inhibitor is used.

[0300] Embodiment 17. The KRAS G12V The method according to Embodiment 16, wherein the (OFF) inhibitor is JAB-23000.

[0301] Embodiment 18. The method according to Embodiment 11, wherein the KRAS(OFF) inhibitor is a pan-RAS(OFF) inhibitor.

[0302] Embodiment 19. The method according to Embodiment 18, wherein the pan-RAS(OFF) inhibitor is JAB-23400, JAB-23425, BI-2493, BI-2865, QTX-3034, QTX3544, ZG2001, BBO-a, BBO-B, or pan KRas-IN-1.

[0303] Embodiment 20. The method according to any one of Embodiments 4 or 6-10, wherein the SHP2 inhibitor is selected from SHP099, TNO155, RMC-4550, RMC-4630, JAB-3068, JAB-3312, RLY-1971, ERAS-601, SH3809, PF-07284892, BBP-398, and any combination thereof.

[0304] Embodiment 21. The method according to any one of Embodiments 5 to 10, wherein the SOS1 inhibitor is selected from RMC-5845, RMC-4948, RMC-0331, BI-1701963, BI-3406, SDR5, MRTX0902, BAY-293, and any combination thereof.

[0305] Embodiment 22: The method according to any one of Embodiments 1 to 21, wherein the RAS(ON)GTP hydrolysis promoting compound and the RAS(OFF) inhibitor, SHP2 inhibitor, RTK inhibitor, or SOS1 inhibitor are administered on the same day.

[0306] Embodiment 23: The method according to any one of Embodiments 1 to 21, wherein the RAS(ON)GTP hydrolysis promoting compound and the RAS(OFF) inhibitor, SHP2 inhibitor, RTK inhibitor, or SOS1 inhibitor are administered simultaneously or sequentially.

[0307] Embodiment 24: The method according to any one of Embodiments 1 to 21, wherein the RAS(ON)GTP hydrolysis-promoting compound and the RAS(OFF) inhibitor, SHP2 inhibitor, RTK inhibitor, or SOS1 inhibitor are administered on different days.

[0308] Embodiment 25: The method according to any one of Embodiments 1 to 22, further comprising administering an additional anticancer therapy.

[0309] Embodiment 26: The method according to Embodiment 25, wherein the additional anticancer therapy is an EGFR inhibitor, SHP2 inhibitor, SOS1 inhibitor, Raf inhibitor, MEK inhibitor, ERK inhibitor, PI3K inhibitor, PTEN inhibitor, AKT inhibitor, mTORC1 inhibitor, BRAF inhibitor, immune checkpoint inhibitor, CDK4 / 6 inhibitor, HER2 inhibitor, RTK inhibitor, or a combination thereof.

[0310] Embodiment 27: The method according to Embodiment 26, wherein the immune checkpoint inhibitor is a PD-L1 inhibitor or a PD-1 inhibitor.

[0311] Embodiment 28: A method for treating RAS protein-related disorders in a subject requiring treatment of RAS protein-related disorders, comprising administering a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound to the subject.

[0312] Embodiment 29. A method for treating RASopathy in a subject requiring treatment of RASopathy, comprising administering a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound to the subject.

[0313] Embodiment 30. The method according to Embodiment 29, wherein the RASopathy is cardiac-facial-cutaneous syndrome, Costello syndrome, Regius syndrome, neurofibromatosis type 1, Noonan syndrome, or capillary malformation-arteriovenous malformation syndrome.

[0314] Embodiment 31. The method according to any one of Embodiments 28 to 30, further comprising administering additional RASopathy therapy.

[0315] Embodiment 32. The method according to Embodiment 31, wherein the additional RASopathy therapy is an EGFR inhibitor, SHP2 inhibitor, SOS1 inhibitor, Raf inhibitor, MEK inhibitor, ERK inhibitor, PI3K inhibitor, PTEN inhibitor, AKT inhibitor, mTORC1 inhibitor, BRAF inhibitor, CDK4 / 6 inhibitor, HER2 inhibitor, RTK inhibitor, or a combination thereof.

[0316] Embodiment 33: A method for inhibiting RAS activity in cells, comprising contacting cells with an effective amount of a RAS(ON)GTP hydrolysis-promoting compound.

[0317] Embodiment 34: A method for increasing the sensitivity of cells to a RAS(OFF) inhibitor, comprising contacting cells with an effective amount of a RAS(ON)GTP hydrolysis-promoting compound, wherein the RAS(ON)GTP hydrolysis-promoting compound synergistically increases the sensitivity of the cells to a RAS(OFF) inhibitor.

[0318] Embodiment 35: Intracellular KRAS, comprising contacting cells with an effective amount of a RAS(ON)GTP hydrolysis-promoting compound. G12C KRAS at the cysteine ​​residue at position 12 G12C A method for increasing the crosslinking rate of (OFF) inhibitors.

[0319] Embodiment 36: A pharmaceutical composition comprising a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound and a RAS(OFF) inhibitor.

[0320] Embodiment 37: A kit comprising a) a RAS(ON)GTP hydrolysis promoting compound and b) a RAS(OFF) inhibitor.

[0321] Embodiment 38: A kit comprising a) a RAS(ON)GTP hydrolysis promoting compound and b) an RTK inhibitor.

[0322] Embodiment 39: A kit comprising a) a RAS(ON)GTP hydrolysis promoting compound and b) an SHP2 inhibitor.

[0323] Embodiment 40: A kit comprising a) a RAS(ON)GTP hydrolysis promoting compound and b) an SOS1 inhibitor.

[0324] Embodiment 41: A kit according to any one of Embodiments 37 to 40, further comprising a package insert containing instructions for the administration of a pharmaceutical composition(s). Other Embodiments

[0325] While this disclosure is described in relation to its specific embodiments, further modifications are possible, and this application is intended to cover any modifications, uses, or adaptations of this disclosure that generally follow the principles of this disclosure, including any deviations from this disclosure that fall within the scope of known or customary practices to which this disclosure is applicable and may apply to the essential features described herein and are subject to the claims. Other embodiments are within the scope of the claims. [Examples]

[0326] This disclosure is further illustrated by the following examples and synthesis examples, which should not be considered to limit the scope or spirit of this disclosure to the specific procedures described herein. It should be understood that the examples are provided to illustrate specific embodiments and are not intended to imply any limitation on the scope of this disclosure. It should also be understood that various other embodiments, modifications, and means to which equivalents thereof may be utilized, without departing from the spirit of this disclosure or the appended claims, which themselves may be suggested to those skilled in the art.

[0327] Example 1. Characterization of RAS(ON)GTP hydrolysis-promoting compounds The rate of RAS GTP hydrolysis was assayed in the presence of various KRAS mutations. To evaluate GTP hydrolysis activity, recombinant KRAS protein (residues 1-169 of KRAS4B) was expressed in E. coli, and a TEV protease-cleaving His6 tag and Ni were used. 2+ Purification was performed using affinity chromatography. The His6 tag was removed by treatment with TEV protease, and the KRAS protein was separated into two Ni 2+ The KRAS protein was isolated by passing it through a column and then performing size exclusion chromatography. The purified KRAS protein was loaded with GTP by incubation on ice for 2 hours in the presence of 2 mM GTP and 10 mM ethylenediaminetetraacetic acid, followed by the addition of 10 mM MgCl2 and incubation on ice for another 1 hour. Excess GTP was removed by dialyzing overnight at 4°C in buffer (12.5 mM HEPES, 75 mM NaCl, pH 7.5). The GTP-loaded KRAS protein was rapidly frozen in liquid nitrogen and then stored at -80°C until use.

[0328] GTP-loaded KRAS protein (1 μM) was mixed with 25 μM recombinant human cyclophylline A and 10 μM of the compound in a reaction buffer (12.5 mM HEPES, 75 mM NaCl, 1 mM MgCl2, 1 mM DTT, 1% DMSO, pH 7.5) preheated to 37°C. Aliquots were taken at a certain time point, quenched by heating to 80°C to denature the protein, and then pelletized by centrifugation to obtain the protein precipitate. The GTP levels of the supernatant were evaluated using Promega GTPase-Glo® according to the manufacturer's instructions. The rate constant of hydrolytic activity was determined by fitting the GTP level as a function of incubation time to a single-step exponential decay.

[0329] Except for those with mutations in the Q61 residue necessary for catalytic hydrolysis activity, all RAS mutants showed increased hydrolysis in the presence of a RAS(ON)GTP hydrolysis-promoting compound (compound E) (Figure 1). Next, RAS GTP hydrolysis activation was characterized using various compounds. Compounds A, B, and C represent potent hydrolysants, respectively. Compound D is a moderate hydrolysant. Compound F represents a class that does not activate GTP hydrolysis by RAS, while the others show varying degrees of RAS GTP hydrolysis activation (Figure 2). The structure of compound F is as follows: [ka] That is correct.

[0330] Example 2. Synergistic effect of combining a RAS(ON)GTP hydrolysis promoting compound with a RAS nucleotide exchange inhibitor. Cell lines were seeded in complete growth medium (RPMI-1640 or DMEM containing 10% fetal bovine serum and 1% penicillin / streptomycin) and incubated overnight in a 37°C, 5% CO2 humidified incubator. In the phospho(Thr202 / Tyr204, Thr185 / Tyr187)-ERK1 / 2 experiment, the compound was added at the indicated concentration the following day and incubated for 4 hours. Cells were lysed, and the amount of phospho(Thr202 / Tyr204, Thr185 / Tyr187)-ERK1 / 2 relative to the total ERK1 / 2 level was assessed using the Meso Scale Diagnostics Kit K15107D according to the manufacturer's instructions. To measure cell viability, the compound was added at the indicated concentration the following day and incubated for a further 5 days. The number of viable cells in each well was assessed using the Promega CellTiter-Glo® reagent according to the manufacturer's instructions. Cell viability was normalized to the DMSO control. The levels or cell viability of phospho(Thr202 / Tyr204, Thr185 / Tyr187)-ERK1 / 2 were plotted as a function of compound concentration using GraphPad Prism. Using the EC50 values ​​obtained from the four-parameter S-shaped concentration-response values, the potency increase factor was calculated as the ratio of the EC50 of the RAS(ON) compound in the absence of RMC-4550 to the EC50 of the RAS(ON) compound in the presence of RMC-4550.

[0331] KRASG12D mutant cell line AsPC-1 was cultured in RPMI-1640 supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. 20,000 cells were seeded in 0.10 mL wells of a tissue culture-treated 96-well plate. After incubation overnight, the compounds were added at the indicated concentrations and incubated for a further 4 hours. The amount of phospho(Thr202 / Tyr204, Thr185 / Tyr187)-ERK1 / 2 relative to total ERK1 / 2 levels was assessed using the Meso Scale Diagnostics Kit K15107D according to the manufacturer's instructions. Relative phosphoERK levels were plotted as a function of the concentration of potent hydrolyzable compound C (Figure 3A) or non-hydrolyzable compound F (Figure 3B) combined with DMSO or 1 μM RMC-4550. KRASG12D mutant cell line AsPC-1 was cultured in RPMI-1640 supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. 4,000 cells were seeded in 0.10 mL wells of tissue-culture-treated 96-well plates. Cell viability was evaluated using Promega CellTiter Glo reagent, and relative cell viability was plotted as a function of the concentration of potent hydrolyzing compound C (Figure 3C) or non-hydrolyzing compound F (Figure 3D) combined with DMSO or 1 μM RMC-4550.

[0332] Example 3. Synergistic effect of combining a RAS(ON)GTP hydrolysis promoting compound with a RAS(OFF) inhibitor. HSA synergistic effect model: Cell lines were seeded in complete growth medium (RPMI-1640 or DMEM containing 10% fetal bovine serum and 1% penicillin / streptomycin) and incubated overnight at 37°C in a 5% CO2 humidified incubator. The compound was added at the indicated concentration the following day, and incubation was continued for a further 5 days. The number of viable cells in each well was assessed using Promega CellTiter-Glo® reagent according to the manufacturer's instructions. Cell viability was normalized to the DMSO control. Cell viability as a function of compound concentration was plotted using GraphPad Prism, which was analyzed using the Combenet software package (Di Veroli GY, Forari C, Wang D, Mollard S, Bramhall JL, Richards FM, Jodrell DI. CombenFit: an interactive platform for the analysis and visualization of drug combinations. Bioinformatics. 2016 Sep 15;32(18):2866-8. doi:10.1093 / bioinformatics / btw230). The best monotherapy (HSA) synergy model (Borisy AA, Elliott PJ, Hurst NW, Lee MS, Lehar J, Price ER, Serbedzija G, Zimmermann GR, Foley MA, Stockwell BR, Keith CT. Systematic discovery of multicomponent therapeutics. Proc Natl Acad Sci USA. 2003 Jun The synergistic effects were evaluated according to (24;100(13):7977-82.doi:10.1073 / pnas.1337088100).

[0333] KRASG12D mutant cell line AsPC-1 was grown in RPMI-1640 supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. 2500 cells were seeded in 0.15 mL wells of a tissue culture-treated 96-well plate. After incubation overnight, the compounds were added at the indicated concentrations and incubated for a further 120 hours. Cell viability was assessed using Promega CellTiter Glo reagent, and relative cell viability was plotted as a function of the concentration of MRTX1133 (Figures 4A and 4G) or MRTX-282 (Figure 4D) in combination with the indicated concentrations of compound A (Figures 4A and 4D) or compound D (Figure 4G), both of which are potent hydrolysants. The synergistic effects between MRTX1133 (Figures 4B and 4H) or MRTX-282 (Figure 4E) and compound A (Figures 4B and 4E) or compound D (Figure 4H) were evaluated using the HAS synergistic effect model. Representative points of synergistic drug combinations between MRTX1133 (Figures 4C and 4I) or MRTX-282 (Figure 4F) and compound A (Figures 4C and 4F) or compound D (Figure 4I) are highlighted.

[0334] KRASG12D mutant cell line HPAC was cultured in RPMI-16 40 supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. 2500 cells were seeded in 0.15 mL wells of a tissue culture-treated 96-well plate. After incubation overnight, the compound was added at the indicated concentration and incubated for a further 120 hours. Cell viability was evaluated using Promega CellTiter Glo reagent, and relative cell viability was plotted as a function of concentration for combinations of MRTX1133 (Figure 5A) or MRTX-282 (Figure 5D) and the indicated concentration for compound A. The synergistic effect between MRTX1133 (Figure 5B) or MRTX-282 (Figure 5E) and compound A was evaluated using the HAS synergy model. The representative aspects of synergistic drug combinations between MRTX1133 (Figure 5C) or MRTX-282 (Figure 5F) and compound A are highlighted.

[0335] KRASG12D mutant cell line Gp2D was cultured in RPMI-16 40 supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. 2500 cells were seeded in 0.15 mL wells of a tissue culture-treated 96-well plate. After incubation overnight, the compound was added at the indicated concentration and incubated for a further 120 hours. Cell viability was evaluated using Promega CellTiter Glo reagent, and relative cell viability was plotted as a function of concentration for combinations of MRTX1133 (Figure 6A) or MRTX-282 (Figure 6D) and the indicated concentration for compound A. The synergistic effect between MRTX1133 (Figure 6B) or MRTX-282 (Figure 6E) and compound A was evaluated using the HAS synergy model. Representative aspects of the synergistic drug combinations between MRTX1133 (Figure 6C) or MRTX-282 (Figure 6F) and compound A are highlighted. For control, compounds D, MRTX1133, and MRTX-282 were used to study HaCat cells (RAS). WT The same experiment was conducted in ). As expected, no synergistic effect was observed (Figures 7A-F). In HaCaT cell lines, a model of normal human dermal keratinocytes, combining RAS(ON) hydrolytic compounds with RAS(OFF) inhibitors did not increase the antiproliferative effect. Conversely, a synergistic antiproliferative effect of the combination of RAS(ON) hydrolytic compounds and RAS(OFF) inhibitors was observed in cancer cell lines, as described above. The increased antiproliferative effect of combination therapy in cancer-derived cells was not seen in similar normal cells, suggesting an increase in the non-toxic therapeutic effect that would be experienced in normal tissues.

[0336] The KRASG12C mutant cell line MiaPaCa2 was cultured in DMEM supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. 2500 cells were seeded in 0.15 mL wells of a tissue culture-treated 96-well plate. After incubation overnight, the compound was added at the indicated concentration and incubated for a further 120 hours. Cell viability was evaluated using Promega CellTiter Glo reagent, and relative cell viability was plotted as a function of concentration for combinations of AMG510 (Figure 8A) or MRTX849 (Figure 8D) and the indicated concentrations of compound A. The synergistic effect between AMG510 (Figure 8B) or MRTX849 (Figure 8E) and compound A was evaluated using the HAS synergy model. Representative points of synergistic drug combinations between AMG510 (Figure 8C) or MRTX849 (Figure 8F) and compound A are highlighted. Furthermore, the crosslinking efficiency was investigated for 1 mM sotrasib alone, in combination with 1 mM compound A, and in combination with 10 mM compound A (Figures 9A-B). As can be seen from the figures, the crosslinking efficiency of sotrasib increases in a concentration-dependent manner in the presence of compound A. KRAS G12C The inhibitor sotracib covalently targets the GDP-bound off state, resulting in a protein-drug adduct observed by reduced mobility via SDS-modified polyacrylamide gel electrophoresis. Treatment of MiaPaca-2 cells with sotracib yields covalently modified KRAS G12C (KRAS G12C The amount of KRAS (-Soto.) increased with increasing incubation time. When the same cells were treated with Solacib combined with 1 or 10 μM of compound A, the amount of KRAS (-Soto.) increased at each time point. G12C and KRAS G12C -Quantify the density of Soto's gel bands, plot them as a function of time, and fit them to a monophase exponential model for KRAS G12C The rate constant for target binding can be obtained. The combination of sotrasib and compound A, compared to sotrasib alone, shows a KRAS G12C This results in faster covalent bonding.

[0337] The complex between the KRAS protein and the RAS-binding domain (RBD) of the RAF1 kinase was measured using a cell assay by nanoluciferase bioluminescence energy transfer (nanoBRET®). U2OS cells were seeded confluence in 3 mL of DMEM supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin (approximately 1 million cells per well). The following day, each well was transfected with 50 ng of plasmid encoding N-terminal nanoluciferase-tagged KRAS4B and 2950 ng of plasmid encoding C-terminal HaloTag® RAF1 RBD (residues 51-149) using FuGENE HD reagent according to the manufacturer's instructions. The following day, each transfected cell pool was transferred to 36 wells of a white 96-well plate in OptiMem phenol-red-free medium supplemented with 4% fetal bovine serum, 1% penicillin / streptomycin, and 100 nM HaloTag® NanoBRET® 618 Ligand. The next day, Vivazine® nanoluciferase substrate and compounds were added at the indicated concentrations. After 4 hours of incubation, synchrotron radiation at 460 nm and 618 nm was measured using a PerkinElmer Envision plate reader. The emission intensity at 618 nm relative to 460 nm is proportional to the number of complexes between the KRAS protein and the RAF1 RBD, which is a surrogate measure of the level of active KRAS.

[0338] Compound E exhibits nearly identical efficacy in inhibiting the RAS-RAF complex across multiple KRAS variants, and KRAS WT It showed somewhat lower efficacy against (Figure 10A). In contrast, pan KRAS-IN-1 (CAS number: 2791263-84-6), an exemplary pan-KRAS(OFF) inhibitor: [ka] RAS WTIt showed maximum efficacy against (Figure 10B). The efficacy against various KRAS mutants generally correlated with the expected hydrolysis rate, and KRAS G12V and KRAS G12R It was the lowest. Co-incubation with RVMD superhydrolyzed agent was KRAS G12V The efficacy of pan-KRAS(OFF) inhibitors used to inhibit this was increased (Figure 10C).

[0339] The KRASG12D mutant cell line AsPC-1 was cultured in RPMI-1640 supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. 2000 cells were seeded in 0.15 mL wells of a tissue culture-treated 96-well plate. After incubation overnight, the compounds were added at the indicated concentrations and incubated for a further 120 hours. Cell viability was evaluated using Promega CellTiter Glo reagent, and relative cell viability was plotted as a function of pan KRAS-IN-1 concentration in combination with the indicated concentrations of compound D (Figure 11A) or compound C (Figure 11D), both of which are potent hydrolysants. The synergistic effect between pan KRAS-IN-1 and compound D (Figure 11B) or compound C (Figure 11E) was evaluated using the HAS synergy model. Representative points of synergistic drug combinations between pan KRAS-IN-1 and compound D (Figure 11C) or compound C (Figure 11F).

[0340] The KRASG12V mutant cell line Capan-2 was cultured in RPMI-1640 supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. 2000 cells were seeded in 0.15 mL wells of a tissue culture-treated 96-well plate. After incubation overnight, the compounds were added at the indicated concentrations and incubated for a further 120 hours. Cell viability was evaluated using Promega CellTiter Glo reagent, and relative cell viability was plotted as a function of pan KRAS-IN-1 concentration against the indicated concentrations of compound D (Figure 12A) or compound C (Figure 12D), both of which are potent hydrolysants. The synergistic effect between pan KRAS-IN-1 and compound D (Figure 12B) or compound C (Figure 12E) was evaluated using the HAS synergy model. Representative points of synergistic drug combinations between pan KRAS-IN-1 and compound D (Figure 12C) or compound C (Figure 12F).

[0341] KRASG12C mutant cell line H358 was cultured in RPMI-1640 supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. 2000 cells were seeded in 0.15 mL wells of a tissue culture-treated 96-well plate. After incubation overnight, the compounds were added at the indicated concentrations and incubated for a further 120 hours. Cell viability was evaluated using Promega CellTiter Glo reagent, and relative cell viability was plotted as a function of pan KRAS-IN-1 concentration in combination with the indicated concentrations of compound D (Figure 13A) or compound C (Figure 13D), both of which are potent hydrolysants. The synergistic effect between pan KRAS-IN-1 and compound D (Figure 13B) or compound C (Figure 13E) was evaluated using the HAS synergy model. Representative points of synergistic drug combinations between pan KRAS-IN-1 and compound D (Figure 13C) or compound C (Figure 13F).

[0342] KRASG12R mutant cell line PSN1 was grown in RPMI-1640 supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin. 2000 cells were seeded in 0.15 mL wells of a tissue culture-treated 96-well plate. After incubation overnight, the compounds were added at the indicated concentrations and incubated for a further 120 hours. Cell viability was evaluated using Promega CellTiter Glo reagent, and relative cell viability was plotted as a function of pan KRAS-IN-1 concentration in combination with the indicated concentrations of compound D (Figure 14A) or compound C (Figure 14D), both of which are potent hydrolysants. The synergistic effect between pan KRAS-IN-1 and compound D (Figure 14B) or compound C (Figure 14E) was evaluated using the HAS synergy model. Representative points of synergistic drug combinations between pan KRAS-IN-1 and compound D (Figure 14C) or compound C (Figure 14F).

Claims

1. A method for treating cancer in a subject requiring cancer treatment, the method comprising administering to the subject a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound and a RAS(OFF) inhibitor, wherein the cancer does not contain an RAS mutation at position 61.

2. A method for treating cancer in a subject requiring cancer treatment, wherein the method comprises administering a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound to the subject.

3. The method according to claim 2, further comprising administering an SHP2 inhibitor or an SOS1 inhibitor.

4. The method according to claim 2, further comprising administering an SHP2 inhibitor, an SOS1 inhibitor, or an RTK inhibitor.

5. The method according to claim 3 or 4, wherein the RAS(ON)GTP hydrolysis promoting compound and the SHP2 inhibitor or SOS1 inhibitor are administered simultaneously or sequentially.

6. The method according to claim 5, further comprising administering a RAS(OFF) inhibitor.

7. The method according to claim 6, wherein the RAS(OFF) inhibitor is administered simultaneously with or consecutively with the RAS(ON)GTP hydrolysis-promoting compound and / or the SHP2 inhibitor, SOS1 inhibitor, or RTK inhibitor.

8. The method according to any one of claims 1 to 7, wherein the cancer includes an RAS mutation.

9. The method according to claim 8, wherein the RAS mutation is located at position 12 or 13.

10. The method according to any one of claims 1 to 9, wherein the cancer is pancreatic cancer, colorectal cancer, non-small cell lung cancer, gastric cancer, esophageal cancer, ovarian cancer, or uterine cancer.

11. The method according to any one of claims 1 to 10, further comprising administering additional anticancer therapy.

12. The method according to claim 11, wherein the additional anticancer therapy is an EGFR inhibitor, a SHP2 inhibitor, a SOS1 inhibitor, a Raf inhibitor, a MEK inhibitor, an ERK inhibitor, a PI3K inhibitor, a PTEN inhibitor, an AKT inhibitor, an mTORC1 inhibitor, a BRAF inhibitor, an immune checkpoint inhibitor, a CDK4 / 6 inhibitor, a HER2 inhibitor, an RTK inhibitor, or a combination thereof.

13. A method for treating RAS protein-related disorders in subjects requiring treatment for RAS protein-related disorders, the method comprising administering to the subject a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound and a RAS(OFF) inhibitor, wherein the RAS does not contain an RAS mutation at position 61.

14. A method for inhibiting RAS activity in cells, the method comprising contacting the cells with an effective amount of a RAS(ON)GTP hydrolysis-promoting compound and a RAS(OFF) inhibitor, wherein the RAS(ON)GTP hydrolysis-promoting compound synergistically increases the sensitivity of the cells to the RAS(OFF) inhibitor.

15. A method for increasing the sensitivity of cells to a RAS(OFF) inhibitor, the method comprising contacting the cells with an effective amount of a RAS(ON)GTP hydrolysis-promoting compound, wherein the RAS(ON)GTP hydrolysis-promoting compound synergistically increases the sensitivity of the cells to the RAS(OFF) inhibitor.

16. The method according to any one of claims 1 and 6 to 18, wherein the RAS(OFF) inhibitor is a KRAS(OFF) inhibitor.

17. The aforementioned KRAS (OFF) inhibitor is KRAS Ｇ１２Ｃ The method according to claim 19, wherein the (OFF) inhibitor is used.

18. The aforementioned KRAS (OFF) inhibitor is KRAS Ｇ１２Ｄ The method according to claim 19, wherein the (OFF) inhibitor is used.

19. The aforementioned KRAS (OFF) inhibitor is KRAS Ｇ１２Ｖ The method according to claim 19, wherein the (OFF) inhibitor is used.

20. The method according to claim 19, wherein the KRAS(OFF) inhibitor is a pan-RAF(OFF) inhibitor.

21. A method for treating RASopathy in a subject requiring treatment for RASopathy, the method comprising administering a therapeutically effective amount of a RAS(ON)GTP hydrolysis-promoting compound to the subject.

22. A method for increasing the crosslinking rate of a KRASG12C(OFF) inhibitor to the cysteine ​​residue at position 12 of KRASG12C in a cell, comprising contacting the cell with an effective amount of a RAS(ON)GTP hydrolysis-promoting compound.

23. The method according to any one of claims 1 to 25, wherein the binding of the RAS(ON)GTP hydrolysis-promoting compound to RAS(ON) changes the position of glutamine 61 of RAS(ON) toward the gamma phosphate of GTP bound to the RAS(ON) protein, compared to the position in the absence of the RAS(ON)GTP hydrolysis-promoting compound, thereby increasing the GTP hydrolysis rate compared to the hydrolysis rate of RAS(ON) in the absence of the RAS(ON)GTP hydrolysis-promoting compound.

24. The method according to any one of claims 1 to 26, wherein the RAS(ON)GTP hydrolysis promoting compound is one of the compounds in Table 1 or a pharmaceutically acceptable salt thereof.

25. A pharmaceutical composition comprising therapeutically effective amounts of a RAS(ON)GTP hydrolysis-promoting compound and a RAS(OFF) inhibitor.

26. The kit includes the following: a) RAS(ON)GTP hydrolysis promoting compounds; and b) RAS (OFF) inhibitors.

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