HDAC inhibitor OKI-179 for the treatment of cancers caused by MAPK pathway mutations

Abstract

OKI-179 alone or in combination with a MAPK pathway inhibitor is indicated for the treatment of cancers caused by one or more mutations in the MAPK pathway.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 408,323, filed September 20, 2022, the disclosure of which is incorporated herein in its entirety for all purposes. [Background technology]

[0002] Background of the Invention It is well known that the RAS pathway is frequently activated in human cancers, potentially offering numerous opportunities for therapeutic intervention. Activated growth factor receptors signal through RAS and other pathways, and RAS mutations occur in over 50% of colorectal and lung cancers, as well as a significant proportion of other cancers (Sanchez-Vega et al., Cell. 173(2): 321-337.e10. doi: 10.1016 / j.cell.2018.03.035. PMID: 29625050; PMCID: PMC6070353 (2018)). Activating RAS pathway mutations are found in over 30% of human tumors and often correlate with poor prognosis. While RAS pathway-targeting drugs have been approved, their activity as single agents is poor, highlighting the need for rational target combinations to improve patient outcomes.

[0003] Several studies have proposed chemo-lethality or synergistic interactions between class I histone deacetylase inhibitors (HDACi) and RAS pathway inhibitors in RAS pathway mutation models (Maertens et al., Cancer Discovery 9, 526-545 (2019); Yamada et al., Mol. Cancer Ther. 17, 17-25 (2018); Faiao-Flores et al., Melanoma Manag. 6(4): MMT29. doi: 10.2217 / mmt-2019-0017 (2019); Ischenko et al., Oncotarget. 6(18), 15814-27 (2015); Wang et al., Cell 173, 1413-25 (2018); Chao et al., Clin Epigenetics 11(1):85. doi: 10.1186 / s13148-019-0681-6 (2019); Bahr et al., Oncotarget 7(43), 69804-69815 (2016)). This chemosynthetic lethality is proposed to result from a drug combination effect, resulting in inhibition of double-stranded (ds) DNA repair and other survival pathways, leading to apoptosis and tumor regression (Maertens et al., Cancer Discovery 9, 526-545 (2019)). The use of HDACi in solid tumors and combination therapy is hampered by low efficacy, lack of selectivity, safety issues, and inconvenient dosing, highlighting the need for better HDACi in the clinic.

[0004] OKI-179 is a novel largazole derivative and a potent, selective, oral Class I HDAC inhibitor that has completed Phase 1 clinical trials as a single agent in patients with solid tumors. The clinical profile of OKI-179 demonstrates the potential to achieve exposure consistent with preclinical activity and robust pharmacodynamic activity at tolerated doses, supporting its development for use in solid tumor combinations. Notably, OKI-179 addresses historical limitations associated with previous HDAC inhibitors by demonstrating improved selectivity, tolerability, and bioavailability, as well as a clear development pathway in molecularly targeted populations. Summary of the Invention

[0005] Summary of the Invention An embodiment of the present invention is a pharmaceutical composition for treating cancer, comprising OKI-179 and a pharmaceutically acceptable carrier, wherein the cancer harbors one or more mutations that lead to activation of the mitogen-activated protein kinase (MAPK) pathway.

[0006] Another aspect of the invention is a pharmaceutical combination for the treatment of cancer, comprising: OKI-179; and One or more mitogen-activated protein kinase (MAPK) pathway inhibitors Including, In combination, the cancer harbors one or more mutations that lead to activation of the MAPK pathway.

[0007] Another aspect of the invention is a pharmaceutical combination comprising: OKI-179; and One or more mitogen-activated protein kinase (MAPK) pathway inhibitors It is a combination including.

[0008] Another aspect of the present invention is a method of treating cancer in a subject, comprising administering a therapeutically effective amount of OKI-179 to a subject in need thereof, wherein the cancer harbors one or more mutations that lead to activation of the MAPK pathway.

[0009] Another aspect of the present invention is a method of treating cancer in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of OKI-179; and One or more mitogen-activated protein kinase (MAPK) pathway inhibitors wherein the cancer harbors one or more mutations that lead to activation of the MAPK pathway.

[0010] In exemplary embodiments, the one or more MAPK pathway inhibitors are selected from an EGFR inhibitor, a RAS inhibitor, a BRAF inhibitor, a pan-RAF inhibitor, a MEK inhibitor, and an ERK inhibitor.

[0011] In an exemplary embodiment, administration of a combination of OKI-179 and one or more MAPK pathway inhibitors to a subject in need thereof results in synergistic inhibition of cancer growth.

[0012] In exemplary embodiments, the mutation occurs in one or more of EGFR, NF1, RAS, BRAF, MAP2K1, GNAQ, and GNA11.

[0013] In an exemplary embodiment, the one or more MAPK pathway inhibitors comprise or consist of an EGFR inhibitor.

[0014] In an exemplary embodiment, the one or more MAPK pathway inhibitors comprise or consist of a RAS inhibitor.

[0015] In an exemplary embodiment, the one or more MAPK pathway inhibitors comprise or consist of a BRAF inhibitor.

[0016] In an exemplary embodiment, the one or more MAPK pathway inhibitors comprise or consist of a pan-RAF inhibitor.

[0017] In an exemplary embodiment, the compound comprises or consists of one or more MAPK pathway inhibitors or MEK inhibitors.

[0018] In an exemplary embodiment, the one or more MAPK pathway inhibitors comprise or consist of an ERK inhibitor.

[0019] In exemplary embodiments, the mutation occurs in two or more of EGFR, NF1, RAS, BRAF, MAP2K1, GNAQ, and GNA11.

[0020] In exemplary embodiments, the mutations occur in three or more of EGFR, NF1, RAS, BRAF, MAP2K1, GNAQ, and GNA11.

[0021] In exemplary embodiments, the mutation occurs in RAS (eg, KRAS, HRAS, or NRAS) alone or in combination with other mutations, including one or more of EGFR, NF1, BRAF, MAP2K1, GNAQ, and GNA11.

[0022] In exemplary embodiments, the mutation occurs in EGFR alone or in combination with other mutations, including one or more of NF1, RAS, BRAF, MAP2K1, GNAQ, and GNA11.

[0023] In exemplary embodiments, the mutation occurs in NF1 alone or in combination with other mutations, including one or more of EGFR, RAS, BRAF, MAP2K1, GNAQ, and GNA11.

[0024] In exemplary embodiments, the mutation occurs in BRAF alone or in combination with other mutations, including one or more of EGFR, NF1, RAS, MAP2K1, GNAQ, and GNA11.

[0025] In exemplary embodiments, the mutation occurs in MAP2K1 alone or in combination with other mutations, including one or more of EGFR, NF1, RAS, BRAF, GNAQ, and GNA11.

[0026] In exemplary embodiments, the mutation occurs in GNAQ alone or in combination with other mutations, including one or more of EGFR, NF1, RAS, BRAF, MAP2K1, and GNA11.

[0027] In exemplary embodiments, the mutation occurs in GNA11 alone or in combination with other mutations, including one or more of EGFR, NF1, RAS, BRAF, MAP2K1, and GNAQ.

[0028] In an exemplary embodiment, the combination is not OKI-179 and binimetinib.

[0029] In an exemplary embodiment, the combination is not OKI-179, binimetinib and encorafenib.

[0030] In an exemplary embodiment, the cancer is colorectal cancer.

[0031] In an exemplary embodiment, the cancer is breast cancer.

[0032] In an exemplary embodiment, the cancer is non-small cell lung cancer (NSCLC).

[0033] In an exemplary embodiment, the cancer is melanoma.

[0034] In an exemplary embodiment, the at least one MAPK pathway inhibitor of the composition and OKI-179 are administered simultaneously.

[0035] In an exemplary embodiment, the at least one MAPK pathway inhibitor and OKI-179 of the composition are administered sequentially. [Brief explanation of the drawings]

[0036] The following diagram is an illustration of a specific embodiment of the present invention and is not intended to otherwise limit the scope of the invention described herein. OKI-005 is an in vitro optimization tool compound. Both OKI-005 and OKI-179 are metabolized to the same active compound, OKI-006. In the diagram, "bini" is synonymous with binimetinib (a MEK1 / 2 inhibitor) and "enco" is synonymous with encorafenib (a BRAF inhibitor).

[0037] [Figure 1] Figures 1A and 1B show how targeting RAS pathway mutations (e.g., BRAF, KRASG12C, and NF1) (Figure 1A) is transforming treatment approaches for cancers such as melanoma, colon, lung, neurofibromatosis, thyroid, cross-organ, and pediatric cancers, along with overall response rate (ORR) and progression-free survival (PFS) values ​​(shown as median overall survival (mOS) vs. control) for the identified drugs used to treat these cancers (Figure 1B).

[0038] [Figure 2] Figure 2 illustrates that, despite the success shown in Figure 1B, RAS pathway targeting has not been particularly effective in certain indications, and even in "sensitive" indications, a significant proportion of patients (represented by the shaded areas) have not received clinical benefit from single-agent RAS pathway inhibition. Illustrative examples include the relative ineffectiveness of the BRAFV600E kinase inhibitor vemurafenib in BRAFV600E melanoma and BRAFV600E colorectal cancer (CRC); and the relative ineffectiveness of the KRASG12C kinase inhibitor sotorasib in KRASG12C pancreatic cancer and KRASG12C colorectal cancer (CRC).

[0039] [Figure 3]Figure 3 illustrates chemosynthetic lethality as an approach to improve RAS pathway targeting by inducing rapid cell death by focusing on pathways required for cell survival after inhibition of mutant RAS pathway signaling. This approach is based on a biological dependency on RAS pathway mutations and rapid initiation of cell death, which circumvents compensatory RAS pathway reactivation and resistance. Example chemical agents include histone deacetylase (HDAC) inhibitors.

[0040] [Figure 4] Figure 4 illustrates the potential synthetic lethal interaction between RAS / mitogen-activated protein kinase (MAPK) inhibition and targeting of nonhomologous end joining (NHEJ) and homologous recombination repair (HRR) in RAS-mutant tumors, as described in Kalimutho et al., Molec. Oncol. 5, 470-90 (2017). The RAS pathway can upregulate HRR as a mechanism to compensate for increased ds-DNA damage. Inhibition of the RAS / MAPK pathway in RAS-mutant cells can downregulate HRR, resulting in a dependency on NHEJ. Thus, there is a potential synthetic lethal interaction between RAS / MAPK inhibition and targeting of NHEJ / HRR in RAS-mutant tumors. In this diagram, DDR is DNA damage response, PARP is poly(ADP-ribose) polymerase, and MYC is myelocytomatosis oncogene.

[0041] [Figure 5] Figure 5 shows that the tolerable AUC (area under the curve) levels of OKI-179 provide effective exposure in animal (mouse) xenograft models, demonstrating the synergistic effects of OKI-179 and binimetinib at physiologically relevant doses. The MTD is the maximum tolerated dose. Nautilus Ph2 doses were clinically determined to reflect the safe and effective range of OKI-179 in combination with binimetinib in patients with NRAS-mutated melanoma, where OKI-179 is administered on a 4-day on / 3-day off schedule.

[0042] [Figure 6] Figures 6A, 6B, and 6C illustrate that OKI-005 in combination with binimetinib exhibits synergistic antiproliferative activity in cell lines harboring RAS mutations. Synergistic effects were observed in models harboring NRAS (Figure 6A - SKMEL2 melanoma cell line) and KRAS (Figure 6B - AGS human gastric adenocarcinoma hyperdiploid cell line and Figure 6C - NCI-H358 non-small cell lung cancer cell line) mutations. Specifically, cells were grown in multiwell plates and incubated with OKI-005 and binimetinib for 72 hours. Viable cell numbers were determined by luminescence readout after addition of CellTiter-Glo (CTG) dye.

[0043] [Figure 7] Figures 7A and 7B illustrate that OKI-005 in combination with binimetinib exhibits synergistic antiproliferative activity in human uveal melanoma cell lines (92-1 and MP46) harboring the GNAQ / 11 Q209L mutation. Specifically, cells are grown in multiwell plates and incubated with OKI-005 and binimetinib for 72 hours. Viable cell numbers are determined by luminescence readout after addition of CellTiter-Glo (CTG) dye.

[0044] [Figure 8] Figure 8 illustrates that in a 72-hour proliferation assay with physiologically relevant concentrations of OKI-005 and binimetinib (where a value of <1 indicates net cell loss), a synergistic effect (i.e., a greater effect than either agent alone) associated with this combination was observed in the RAS mutant pathway compared to the RAS wild-type (WT) pathway from day 3 onwards compared to day 0, as determined by luminescence readout after addition of CellTiter-Glo (CTG) dye. The RAS mutant and RAS WT cell lines involved in this study represent conventional and well-known cell lines for this purpose. See, e.g., Figure 16.

[0045] [Figure 9]Figures 9A, 9B, and 9C illustrate the synergistic effect of combining OKI-005 and binimetinib (at the indicated amounts) in the NRAS-mutated SKMEL2 cell line, which is associated with significant cell growth inhibition and cell death, as measured by PARP cleavage. In particular, in Figures 9A and 9B, cells are grown in multiwell plates and incubated with OKI-005 and binimetinib for 72 hours. Viable cell numbers are determined by luminescence readout after addition of CellTiter-Glo (CTG) dye. SEM is standard error.

[0046] [Figure 10] Figure 10 illustrates the synergistic effect in O6-methylguanine-DNA methyltransferase (MGMT)-positive human tumor cell lines versus MGMT-negative human tumor cell lines ("MGMT Null") when administering a combination of OKI-005 and binimetinib compared to OKI-005 or binimetinib alone.

[0047] [Figure 11] Figures 11A, 11B, and 11C illustrate the administration of OKI-179 and binimetinib at clinically approved or tolerated doses and schedules in NRAS mutant cell line xenografts SKMEL2. As used herein, "5 on / 2 off" refers to a dosing schedule of 5 days on and 2 days off per week. Changes in SKMEL2 tumor pERK (MEKi PD) and Ac-H3K9 (HDACi PD) were observed 2 hours after administration. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a control. Both binimetinib and OKI-179 demonstrated strong on-target PD activity (i.e., increased AC-H3K9 or decreased pERK) at the doses shown. While OKI-179 or binimetinib as single agents delayed tumor growth, regression was modest. OKI-179 in combination with binimetinib showed a significant increase in regression compared with either agent alone after two weeks of dosing.

[0048] [Figure 12]Figures 12A, 12B, and 12C show the results of a combination study of OKI-005 with binimetinib, GDC-0994, and sotorasib in the NCI-H358 (NSCLC KRASG12C) cell line. Figures 12D, 12E, and 12F show the results of a combination study of OKI-005 with binimetinib, GDC-0994, and MRTX1133 (a KRASG12D inhibitor) in the AGS (gastric KRASG12D) cell line. Combination synergy was observed in all cases by Loewe synergy analysis. Specifically, cells were grown in multiwell plates and incubated with OKI-005 and binimetinib for 72 hours. Viable cell numbers were determined by luminescence readout after addition of CellTiter-Glo (CTG) dye.

[0049] [Figure 13] Figure 13A illustrates that OKI-179 combined with binimetinib and encorafenib using the indicated amounts of each agent and dosing regimens (mpk = mg / kg) in HT29 BRAFV600E colorectal (CRC) xenografts showed significantly increased regression after 2 weeks of dosing compared to OKI-179 alone or the combination of binimetinib and encorafenib. Figure 13B illustrates comparative mouse weight changes for OKI-179 alone, the combination of binimetinib and encorafenib, and OKI-179 combined with binimetinib and encorafenib at the indicated doses and up to 15 days after randomization. Greater weight loss was observed with OKI-179 combined with binimetinib and encorafenib than with OKI-179 alone or the combination of binimetinib and encorafenib.

[0050] [Figure 14]Figure 14A shows that OKI-179 in combination with sotorasib in NCI-H358 KRASG12C non-small cell lung cancer xenografts showed a significant increase in regression over a 50-day period compared to OKI-179 or sotorasib alone after dosing at the levels and rates shown. As used herein, "4 on / 3 off" refers to a dosing schedule of 4 days on and 3 days off each week. Figure 14B illustrates the change in mouse weight with OKI-179 and sotorasib alone compared to the combination of OKI-179 and sotorasib.

[0051] [Figure 15] FIG. 15 illustrates the efficacy of the combination of OKI-005 and binimetinib compared to OKI-005 and binimetinib alone at the dose levels shown, as measured by the fold change in CTG signal 3 days after dosing, according to the listed conventional cell line tumor types.

[0052] [Figure 16] FIG. 16 illustrates the efficacy of the combination of OKI-005 and binimetinib compared to OKI-005 and binimetinib alone at the indicated dose levels, as measured by the fold change in CTG signal 3 days after dosing, by MAPK mutations in the listed conventional tumor cell lines.

[0053] [Figure 17] FIG. 17 illustrates the efficacy of the combination of OKI-005 and binimetinib compared to the combination of OKI-005 and the ERK1 / 2 inhibitor GDC-0994 against the listed conventional tumor cell lines sorted by binimetinib response, as measured by fold change in CTG signal 3 days after dosing. DETAILED DESCRIPTION OF THE INVENTION

[0054] Detailed Description of the Invention definition As defined herein, "OKI-179" refers to the compound of the following structure that exists as a benzenesulfonate salt, and is a prodrug of OKI-006. [ka]

[0055] As defined herein, "OKI-005" refers to the compound having the following structure, and is a prodrug of OKI-006. [ka]

[0056] As defined herein, "OKI-006" refers to a compound having the following structure: [ka]

[0057] As used herein, various specific inhibitors have their conventional and well-known meanings, including, but not limited to, MAPK pathway inhibitors, EGFR inhibitors, RAS inhibitors, pan-RAS inhibitors, RAF inhibitors, pan-RAF inhibitors, BRAF inhibitors, MEK inhibitors, and ERK inhibitors.

[0058] As used herein, various specific mutations have their conventional and well-known meanings, such as, but not limited to, a mutation in RAS (e.g., KRAS, HRAS, or NRAS), a mutation in EGFR, a mutation in NF1, a mutation in BRAF, a mutation in MAP2K1, a mutation in GNAQ, and a mutation in GNA11.

[0059] As used herein, various specific cell lines, such as, but not limited to, SKMEL2, AGS, MGMT, NCI-H358, uveal melanoma cell line 92-1 and MP46, have their conventional and well-known meanings.

[0060] As defined herein, "TGI" refers to tumor growth inhibition.

[0061] As defined herein, "R" refers to regression and reflects an observed decrease in tumor size.

[0062] As defined herein, "LOF" refers to loss of function.

[0063] As defined herein, "BID" refers to twice-daily dosing.

[0064] As defined herein, "QD" refers to once daily administration.

[0065] As defined here, "SEM" refers to the standard error of the mean, which is a measure of how far the sample mean (average) of data may be from the true population mean.

[0066] As defined herein, the term "subject" includes, but is not limited to, humans (e.g., men and women of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or elderly adults)) and / or other primates (e.g., monkeys); non-human mammals such as cows, pigs, horses, sheep, mice, goats, cats, and / or dogs; and / or birds such as chickens, ducks, and / or geese.

[0067] As defined herein, "cancer" refers to the physiological condition in mammals that is typically characterized by unregulated cell growth. Examples of cancer include, but are not limited to, carcinoma, lymphoma, leukemia, blastoma, and sarcoma. More specific examples of such cancers include squamous cell carcinoma, small cell lung cancer, non-small cell lung cancer (NSCLC) (including, but not limited to, metastatic non-small cell lung cancer, BRAF mutant NSCLC (e.g., BRAF V600E mutant NSCLC)), glioma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, acute myeloid leukemia (AML), multiple myeloma, gastrointestinal cancer, renal cell carcinoma, kidney cancer (e.g., advanced renal cell carcinoma), ovarian cancer, liver cancer, lymphoblastic leukemia, lymphocytic leukemia, colorectal cancer (including, but not limited to, metastatic colorectal cancer (e.g., microsatellite-stable metastatic colorectal cancer), BRAF V600 mutant colorectal cancer (e.g., BRAF V600E or BRAF V600K mutant colorectal cancer)), endometrial cancer, kidney cancer, prostate cancer, thyroid cancer, melanoma (unresectable or metastatic melanoma, uveal melanoma, BRAF The cancers include, but are not limited to, BRAF V600 mutant melanoma, such as V600E mutant melanoma), chondrosarcoma, neuroblastoma, pancreatic cancer, glioblastoma multiforme, cervical cancer, brain cancer, gastric cancer, urothelial carcinoma (including locally advanced or metastatic urothelial carcinoma), bladder cancer, hepatocellular carcinoma, breast cancer, and head and neck cancer. In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is metastatic colorectal cancer. In some embodiments, the cancer is melanoma. In other embodiments, the cancer is pancreatic cancer. In other embodiments, the cancer is NSCLC. In other embodiments, the cancer is ovarian cancer. In some embodiments, the cancer is a BRAF-mutated melanoma. In some embodiments, the cancer is a BRAF-associated cancer. As used herein, the term "BRAF-associated cancer" refers to a cancer associated with or having a class I, class II, or class III mutation in BRAF (BRAF class I mutations are mutations at the V600 locus). Class II mutations are non-V600 mutations that activate BRAF by signaling through RAS-independent dimers. Class III mutations are "kinase-dead" with reduced kinase activity compared to wild-type BRAF.Non-limiting examples of BRAF-associated cancers are described herein. In some embodiments, the cancer is a KRAS-associated cancer. As used herein, the term "KRAS-associated cancer" refers to cancers associated with or having activating mutations in KRAS, particularly mutations at residues G12, G13, or Q61. Non-limiting examples of KRAS-associated cancers are described herein.

[0068] As defined herein, the term "combination therapy" refers to a dosing regimen of two or more different therapeutically active agents over a period of time, where the therapeutically active agents are administered together or separately. In certain embodiments, the combination therapy is a loose combination.

[0069] The term "non-fixed combination" means that two or more different therapeutic agents are formulated as separate compositions or dosages so that they can be administered separately to a subject in need thereof, either simultaneously or sequentially with varying time limits between each other.

[0070] As defined herein, the term "effective dose" or "effective amount" or "therapeutically effective amount" refers to a specific amount of a pharmaceutical composition containing a therapeutic agent that achieves a clinically beneficial result (i.e., reduction of symptoms, for example). The toxicity and therapeutic efficacy of such a composition can be determined, for example, by the LD 50 (a dose lethal to 50% of the population) and ED 50 The toxic to therapeutic dose ratio is the therapeutic index, which can be determined by standard pharmaceutical procedures in cell cultures or experimental animals to determine the dose therapeutically effective in 50% of the population. The LD 50 / ED 50 Compounds with large therapeutic indices are preferred. The data obtained from these cell culture assays and additional animal studies can be used to formulate a range of dosages for human use. The dosage of such compounds is preferably within the ED range with little or no toxicity. 50The circulating concentration range is within a range including the range of 100-450 mg / day. The dosage varies within this range depending on the dosage form used, the patient's sensitivity, and the route of administration. In certain embodiments, OKI-179 is administered at 100-450 mg / day, with a 4-day on-dose, 3-day off-dose regimen, and the specific dosage depends on several factors associated with the subject. In other specific embodiments, OKI-179 is administered at 100-200 mg / day, with the specific dosage depending on several factors associated with the subject. In most situations, the recommended clinical dose of the RAS pathway inhibitor should be used for administration of the combination.

[0071] As used herein, the term "disease" refers to any disturbance in the normal state of a living animal or part thereof that interferes with or modifies the performance of a vital function. Typically manifested by characteristic signs and symptoms, diseases are usually a response to: i) environmental factors (e.g., nutritional deficiencies, industrial hazards, or climate); ii) specific infectious agents (e.g., parasites, bacteria, or viruses); iii) an inherent defect of the organism (e.g., genetic abnormalities); and / or iv) a combination of these factors.

[0072] The terms "reduce," "inhibit," "reduce," "suppress," "reduce," "prevent," and grammatical equivalents (such as "low," "small"), when used in reference to the manifestation of any symptom in an untreated subject compared to a treated subject, indicate that the amount and / or extent of the symptom in the treated subject is lower than in the untreated subject by any amount recognized as clinically significant by a skilled medical practitioner. In certain embodiments, the amount and / or extent of the symptom in the treated subject is at least 10% lower, at least 25% lower, at least 50% lower, at least 75% lower, and / or at least 90% lower than the amount and / or extent of the symptom in the untreated subject.

[0073] As used herein, the term "inhibitory compound" refers to any compound that can interact with (i.e., attach, bind, etc.) a binding partner under conditions such that the binding partner becomes unresponsive to its natural ligand. Inhibitory compounds can include, but are not limited to, small organic molecules, antibodies, and proteins / peptides.

[0074] As used herein, the term "drug" or "compound" or "pharmaceutical agent" refers to any pharmacologically active substance that can be administered to achieve a desired effect. A drug or compound can be synthetic or naturally occurring, a non-peptide, a protein or peptide, an oligonucleotide or nucleotide, a polysaccharide or sugar.

[0075] As used herein, the term "administration" or "administering" refers to any method of providing a composition to a patient so that the composition has its intended effect in the patient. Examples of methods of administration include local tissue administration (i.e., extravascular administration, e.g., subcutaneous, intramuscular, or intraperitoneal), intravenous, oral ingestion, transdermal patch, topical, inhalation, suppository, and other direct mechanisms.

[0076] As used herein, the term "patient" refers to a human or animal and includes both hospitalized and non-hospitalized subjects. For example, outpatients and nursing home residents are "patients." Patients can be human or non-human animals of any age, and thus include adults and juveniles (i.e., children). The term "patient" is not intended to imply the need for medical treatment. Thus, patients can be willing subjects for experimentation, whether clinical or in support of basic science testing.

[0077] As used herein, the term "synergistic effect" or "synergistic" refers to the phenomenon where the combination of two therapeutic agents in a combination therapy, in terms of measured outcomes, is greater than the sum of the effects of each agent when administered alone.

[0078] As used herein, the term "protein" refers to any of a number of naturally occurring, highly complex substances (e.g., enzymes or antibodies) that contain amino acid residues joined by peptide bonds and contain carbon, hydrogen, nitrogen, oxygen, and typically sulfur. Generally, proteins contain on the order of several hundred amino acids.

[0079] As used herein, the term "peptide" refers to any of a variety of amides derived from two or more amino acids by combining the amino group of one acid with the carboxyl group of another, usually obtained by partial hydrolysis of proteins. Generally, peptides contain several dozen amino acids.

[0080] As used herein, the terms "pharmaceutically acceptable" or "pharmacologically acceptable" refer to molecular entities and compositions that do not produce adverse, allergic or other untoward reactions when administered to animals or humans.

[0081] As used herein, the term "pharmaceutically acceptable carrier" includes any and all solvents or dispersion media including, but not limited to, water, ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, vegetable oils, coatings, isotonic and absorption delaying agents, liposomes, commercially available cleansers, etc. Supplementary biologically active ingredients can also be incorporated into such carriers.

[0082] As used herein, the term "pharmaceutically acceptable salt" refers to a salt that does not adversely affect the biological activity and properties of the compound and is suitable for use in contact with the tissues of a subject without undue toxicity, irritation, and / or allergic response, etc. Pharmaceutically acceptable salts include those derived from suitable inorganic acids and bases, including hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, nitric acid, acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, malonic acid, ascorbic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, benzoic acid, naphthalenesulfonic acid, lactic acid, succinic acid, oxalic acid, stearic acid, and the like. In some cases, pharmaceutically acceptable salts are obtained by reacting a compound having an acidic group, as described herein, with a base to form salts such as ammonium salts, alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., calcium or magnesium salts), salts formed from organic bases, and amino acid salts. Pharmaceutically acceptable salts derived from appropriate bases include alkali metal, alkaline earth metal, and ammonium and quaternary ammonium compounds. Specific metals include, but are not limited to, sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc. Organic bases from which salts can be prepared include, for example, primary, secondary, and tertiary amines.

[0083] As used herein, the term "prodrug" refers to a compound that is converted in vivo to produce a disclosed compound or a pharmaceutically acceptable form of the compound. A prodrug may be inactive when administered to a subject, but is converted to an active compound in vivo. In various cases, a prodrug has improved physicochemical properties (e.g., bioavailability) and / or delivery properties over the parent compound. Prodrugs are typically designed to enhance the pharmaceutical and / or pharmacokinetic properties associated with the parent compound. Prodrug compounds often offer advantages of solubility, tissue compatibility, or delayed release in a subject. Prodrugs include compounds in which a hydroxy, amino, or mercapto group is bonded to any group that cleaves to form a free hydroxy, amino, or mercapto group, respectively, when the prodrug is administered to a subject. It is well known that prodrugs can be prepared, for example, from carboxylic acids in the form of a carboxylic acid ester or thioester.

[0084] As used herein, the term "biologically active" refers to any molecule that has a structural, regulatory, or biochemical function. For example, biological activity can be determined by, for example, restoring wild-type growth in cells lacking protein activity. Cells lacking protein activity can be produced by many methods (i.e., point mutations and frameshift mutations, for example). Complementation can be achieved by transfecting cells lacking protein activity with an expression vector that expresses the protein, its derivatives, or portions thereof.

[0085] As used herein, the term "label" or "detectable label" refers to any composition detectable by spectral, photochemical, biochemical, immunochemical, electrical, optical, or chemical means. Such labels include biotin for staining with labeled streptavidin conjugates, magnetic beads (e.g., Dynabeads (登録商標) ), fluorescent dyes (e.g., fluorescein, Texas Red (登録商標) , rhodamine, green fluorescent protein, etc.), radioactive labels (e.g. 3 H, 125 I, 35 S,14 C or 32 P), enzymes (e.g., horseradish peroxidase, alkaline phosphatase, and others commonly used in ELISA), and calorimetric labels, such as colloidal gold or colored glass or plastic (e.g., polystyrene, polypropylene, latex, etc.) beads. Patents teaching the use of such labels include, but are not limited to, U.S. Patents 3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149; and 4,366,241 (all of which are incorporated herein by reference in their entirety). Labels contemplated by the present invention can be detected by conventional methods. For example, radioactive labels can be detected using photographic film or scintillation counters, and fluorescent markers can be detected with a photodetector that detects emitted light. Enzymatic labels are typically detected by providing the enzyme and a substrate and detecting the reaction product produced by the action of the enzyme on the substrate, whereas calorimetric labels are detected simply by visualization of the colored label.

[0086] The pharmaceutical compositions of the present invention may take any suitable form for the desired route of administration. If the composition is administered orally, any suitable orally deliverable dosage form can be used, including, but not limited to, water, glycols, oils, alcohols, and the like for oral liquid preparations such as suspensions, syrups, elixirs, emulsions, and solutions; or solid carriers such as starches, sugars, kaolin, diluents, lubricants, binders, disintegrants, and the like for powders, pills, capsules, and tablets. Due to ease of administration, tablets and capsules represent the most advantageous oral dosage unit form. Injectable compositions in the form of solutions, suspensions, and emulsions or intravenous infusions are also provided. For parenteral compositions, the carrier usually comprises sterile water, and possibly other ingredients to aid dissolution. Injectable solutions can be used in which the carrier comprises saline solution, glucose solution, or a mixture of saline and glucose solution. Suitable oils include, for example, peanut oil, sesame oil, cottonseed oil, corn oil, soybean oil, synthetic glycerol esters of long-chain fatty acids, and mixtures of these with other oils. In compositions suitable for transdermal administration, the carrier may optionally contain a penetration enhancer and / or a suitable wetting agent, and optionally may contain suitable additives, where the additives may facilitate application of the composition to the skin and / or facilitate the preparation of the composition for delivery. These compositions may be administered in various ways, for example, as a transdermal patch or ointment. Acid or base addition salts of the compounds of the present invention are typically more suitable for the preparation of aqueous compositions due to their increased water solubility over the corresponding neutral forms of the compounds.

[0087] The pharmaceutical compositions of the present invention may contain one or more fillers, diluents, adjuvants, vehicles or other additives to facilitate the preservation and / or administration of the active ingredients contained therein.

[0088] In exemplary embodiments, the pharmaceutical compositions of the invention may contain one or more additional therapeutic agents, e.g., to increase efficacy or reduce unwanted side effects. Examples of such agents include, but are not limited to, agents for treating or preventing cancer, Huntington's disease, cystic fibrosis, liver fibrosis, kidney fibrosis, pulmonary fibrosis, skin fibrosis, rheumatoid arthritis, diabetes, or heart failure.

[0089] In certain embodiments, the additional therapeutic agent that is included is anticancer drug.The example of anticancer drug includes but is not limited to: MAPK pathway inhibitor, alkylating agent such as cyclophosphamide, dacarbazine and cisplatin; antimetabolite such as methotrexate, mercaptopurine, thioguanine, fluorouracil and cytarabine; plant alkaloid such as vinblastine and paclitaxel; antitumor antibiotic such as doxorubicin, bleomycin and mitomycin; hormone / antihormonal agent such as prednisone, tamoxifen and flutamide; other types of anticancer drug such as asparaginase, rituximab, trastuzumab, imatinib, retinoic acid and derivative, colony-stimulating factor, amifostine, camptothecin, topotecan, thalidomide analogue such as lenalidomide, CDK inhibitor and proteasome inhibitor such as velcade.

[0090] In another embodiment, the present invention provides a method for preventing or treating a disease caused by abnormal cell proliferation and / or differentiation in a subject in need of treatment, comprising administering to the subject a therapeutically effective amount of one or more compounds of the present invention. In some embodiments, the method for preventing or treating a disease comprises administering to a subject in need thereof a composition comprising an effective amount of one or more compounds of the present invention and a pharmaceutically acceptable carrier. The administered composition may further comprise a therapeutic agent, such as an anti-cancer agent.

[0091] The present invention includes compounds labeled with various radioactive or non-radioactive isotopes. Examples of atomic isotopes include deuterium ( 2 H), tritium ( 3 H), iodine-125( 125 I), carbon-14( 14 C), nitrogen-15( 15 N), sulfur-35( 35 S) and chlorine-36( 36In exemplary embodiments, one or more hydrogen atoms of the compounds of the invention may be replaced with deuterium. In various embodiments, the compounds of the invention contain at least one deuterium atom, or two or more deuterium atoms, or three or more deuterium atoms, etc. The compounds of the invention described herein may contain tritium ( 3 H), iodine-125( 125 I) and carbon-14( 14 Radiolabeled compounds may also be radiolabeled with radioactive isotopes such as C. Radiolabeled compounds are useful as therapeutic or prophylactic agents, provide research reagents such as in assays, and / or provide diagnostic agents for techniques such as in vivo imaging. Synthetic methods for incorporating isotopes into organic compounds are well known in the art.

[0092] As defined herein, the term "in vivo" refers to an event that takes place within a subject.

[0093] As defined herein, the term "in vitro" refers to an event that takes place outside a subject's body.

[0094] As defined herein, "pharmaceutically acceptable forms" of a compound include, but are not limited to, pharmaceutically acceptable salts, hydrates, solvates, isomers, prodrugs, and isotopically labeled derivatives of the compound.

[0095] As defined herein, the terms "treating" or "treating" cancer refer to administering a compound of the present invention to a subject having or diagnosed with cancer to achieve at least one positive therapeutic effect, such as, for example, a reduction in the number of cancer cells, a reduction in tumor size, a reduction in the rate of cancer cell invasion into peripheral organs, or a reduction in the rate of tumor metastasis or tumor growth, or ameliorating, alleviating, or preventing the progression of the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition. As used herein, the term "treatment," unless otherwise specified, refers to the act of treating, as "treating" is defined immediately above. The term "treating" also includes adjuvant and neoadjuvant treatment of a subject.

[0096] For purposes of this invention, beneficial or desired clinical results include, but are not limited to, one or more of the following: a reduction in the proliferation (or destruction) of neoplastic or cancerous cells; prevention of metastasis or neoplastic cells; a reduction in or decrease in tumor size; an increase in the duration of a subject's response, progression-free survival, or overall survival (e.g., compared to one or more metrics in subjects with a similar cancer who are not receiving treatment or who are receiving a different treatment, or compared to one or more metrics in the same subject prior to treatment); a reduction in symptoms caused by the cancer; an increase in the quality of life of those with cancer; a reduction in the dosage of other medications required to treat the cancer; a delay in the progression of the cancer; a cure for the cancer; overcoming one or more resistance mechanisms of the cancer; and / or an increase in the survival of a cancer patient. Positive therapeutic effects in cancer can be measured in several ways (see, e.g., W.A. Weber, J. Nucl. Med. 50 Suppl. 1:1S-10S (2009)).

[0097] The mitogen-activated protein kinase (MAPK) pathway (also known as the Ras-Raf-MEK-ERK pathway) is a series of proteins in cells that transmit signals from receptors on the cell surface to DNA in the cell's nucleus. The MAPK pathway controls many fundamental cellular processes involved in cell proliferation, differentiation, and survival. In normal cells, RAS activates RAF kinase, which subsequently phosphorylates and activates MEK1 and MEK2, which, upon activation, phosphorylate ERK1 and ERK2. ERK controls the activity and expression of multiple nuclear transcription factors and cytoplasmic proteins required for cell proliferation, differentiation, and survival. In some human cancers, there are several mutations in MAPK pathway proteins that effectively deregulate the pathway and promote cell survival and proliferation of cancer or tumor cells.

[0098] Examples of MAPK pathway inhibitors include, but are not limited to, EGFR inhibitors, MEK inhibitors, ERK inhibitors, BRAF inhibitors, pan-RAF inhibitors, KRAS inhibitors, pan-RAS inhibitors and RAF inhibitors.

[0099] Examples of RAF inhibitors include, but are not limited to, RAF265, sorafenib, dabrafenib (GSK2118436), 5B590885, PLX4720, PLX4032, GDC-0879, and ZM336372.

[0100] Examples of MEK inhibitors include, but are not limited to, trametinib (GSK1120212), selumetinib, binimetinib, cobimetinib, PD-325901, CI-1040 / PD184352, TAK-733, AZD6244, PD318088, PD98059, PD334581, and RDEA119.

[0101] Examples of KRAS inhibitors include, but are not limited to, sotorasib, adagrasib, RMC-6236, RMC-6291, RMC-9805, and RMC-8839.

[0102] Examples of ERK inhibitors include, but are not limited to, ulixertinib, VTX11e, AEZS-131, PD98059, FR180204, and FR148083.

[0103] Mammalian HDACs are divided into four groups depending on sequence homology, subcellular distribution, and catalytic activity. Class I, II, and IV HDACs contain Zn in the catalytic site. 2+ Class III HDACs contain sirtuins 1-7 and NAD +Class I HDACs (HDACs 1, 2, 3, and 8) share homology with Rpd3 and are primarily distributed in the nucleus of normal cells. Class II HDACs can be subdivided into class IIa HDACs and class IIb HDACs. Class IIa enzymes (HDACs 4, 5, 7, and 9) shuttle between the nucleus and cytoplasm and are located in specific tissues such as the brain, heart, and muscle. Class IIb HDACs (HDACs 6 and 10) are primarily located in the cytoplasm. Class IV contains only HDAC11 and is located in the brain, heart, kidney, skeletal muscle, and testis. Below is a table showing the enzyme inhibition of HDACs 1 to 11 by the known HDAC inhibitors vorinostat and romidepsin (FK228) compared to OKI-006, where OKI-006 exhibits high potency as a class I HDAC inhibitor. [Table 1] *OKI-006 is the active form of OKI-179 **Red-FK228 is the active (reduced) form of romidepsin

[0104] Appropriate dosages of OKI-179 alone and in combination with other inhibitors can be determined by those skilled in the art. It is recognized that appropriate dosages may vary from patient to patient. Determining the optimal dosage generally involves balancing the level of therapeutic benefit of the treatments described herein against any risks or adverse side effects. The selected dosage level will depend on various factors, including, but not limited to, the activity of the particular compound, the route of administration, the time of administration, the rate of compound excretion, the duration of treatment, other drugs, compounds, or substances used in combination, and the patient's age, sex, weight, condition, general health, and medical history. The amount of compound administered and the route of administration are ultimately at the discretion of the physician. OKI-179 alone or in combination with other inhibitors may be administered once, continuously, or intermittently (e.g., in divided doses at appropriate intervals) throughout the course of treatment. Below are three examples of OKI-179 administration schedules that achieve the maximum tolerated dose (MTD) in patients with advanced cancer when administered orally. The MTD of OKI-179 in Schedule 1 was 450 mg daily with 4 days on and 3 days off each week, and the recommended phase 2 dose (RP2D) was 300 mg. In Schedule 2, the MTD was 200 mg daily with a continuous dosing regimen, and in Schedule 3, the MTD was 300 mg with 5 days on and 2 days off each week. [Table 2]

[0105] As an example of the mutation frequency associated with various cancers, mutations in NRAS account for 20% of all melanomas, affecting approximately 65,000 patients worldwide. Such melanomas tend to be more aggressive than their wild-type counterparts and often have a poor prognosis. Current treatments for NRAS mutant melanoma remain limited and only modestly effective. No targeted therapies are available beyond PD1i. Preclinical activity of OKI-179 in combination with binimetinib demonstrated regression in multiple RAS pathway mutant models. The following data illustrate the efficacy of the combination of OKI-179 and binimetinib against specific mutant tumor types compared to their wild-type counterparts, where "N" is the conventional model number used for the indicated tumor type and "GI" stands for gastrointestinal. [Table 3]

[0106] The method of the present invention can be carried out to treat patients with cancers that carry mutations that activate the RAS pathway.For example, physicians use available mutation testing methods to test for mutations in genes in the RAS pathway (e.g., EGFR RAS, RAF, MAP2K1, NF1, or GNAQ / 11) as a means of evaluating the suitability of combining OKI-179 with a RAS pathway inhibitor.In an exemplary embodiment, OKI-179 and a RAS pathway inhibitor are co-administered to patients with tumors that have activating mutations in EGFR, RAS, RAF, MAP2K1, NF1, or GNAQ / 11, but this combination is not used for patients that do not have RAS pathway activating mutations.The dose and schedule of OKI-179 and the RAS pathway inhibitor are recognized by physicians as tolerable and effective.The therapeutic effect of each combination can be determined using established methods and approaches to show the impact on tumor shrinkage and / or survival outcomes in patients.

Claims

1. A method of treating cancer in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of OKI-179; and one or more mitogen-activated protein kinase (MAPK) pathway inhibitors wherein the cancer harbors one or more mutations that lead to activation of the MAPK pathway.

2. 10. The method of claim 1, wherein the administration of OKI-179 and one or more MAPK pathway inhibitors in combination results in synergistic inhibition of cancer growth.

3. 2. The method of claim 1, wherein the one or more MAPK pathway inhibitors are selected from an EGFR inhibitor, a RAS inhibitor, a BRAF inhibitor, a pan-RAF inhibitor, a MEK inhibitor, and an ERK inhibitor.

4. 3. The method of claim 1 or claim 2, wherein the mutation occurs in one or more of EGFR, NF1, RAS, BRAF, MAP2K1, GNAQ and GNA11.

5. 3. The method of claim 1 or claim 2, wherein the one or more MAPK pathway inhibitors comprise an EGFR inhibitor.

6. 3. The method of claim 1 or claim 2, wherein the one or more MAPK pathway inhibitors comprise a RAS inhibitor.

7. 3. The method of claim 1 or claim 2, wherein the one or more MAPK pathway inhibitors comprise a BRAF inhibitor.

8. 3. The method of claim 1 or claim 2, wherein the one or more MAPK pathway inhibitors comprise a pan-RAF inhibitor.

9. 3. The method of claim 1 or claim 2, wherein the one or more MAPK pathway inhibitors comprise a MEK inhibitor.

10. 3. The method of claim 1 or claim 2, wherein the one or more MAPK pathway inhibitors comprise an ERK inhibitor.

11. 3. The method of claim 1 or claim 2, wherein the mutation occurs in two or more of EGFR, NF1, RAS, BRAF, MAP2K1, GNAQ and GNA11.

12. 3. The method of claim 1 or claim 2, wherein the mutation occurs in three or more of EGFR, NF1, RAS, BRAF, MAP2K1, GNAQ, and GNA11.

13. 3. The method of claim 1 or claim 2, wherein the at least one MAPK pathway inhibitor and OKI-179 are administered simultaneously.

14. 3. The method of claim 1 or claim 2, wherein the at least one MAPK pathway inhibitor and OKI-179 are administered sequentially.

15. 3. The method of claim 1 or claim 2, wherein the cancer is colorectal cancer.

16. 3. The method of claim 1 or claim 2, wherein the cancer is breast cancer.

17. 3. The method of claim 1 or claim 2, wherein the cancer is non-small cell lung cancer.

18. 3. The method of claim 1 or claim 2, wherein the cancer is melanoma.

19. 1. A pharmaceutical combination for the treatment of cancer, comprising: OKI-179; and one or more mitogen-activated protein kinase (MAPK) pathway inhibitors wherein the cancer harbors one or more mutations that lead to activation of the MAPK pathway.

20. 20. The combination of claim 19, wherein the one or more MAPK pathway inhibitors are selected from an EGFR inhibitor, a RAS inhibitor, a BRAF inhibitor, a pan-RAF inhibitor, a MEK inhibitor, and an ERK inhibitor.

21. 20. The combination of claim 19, wherein administration of a combination of OKI-179 and one or more MAPK pathway inhibitors results in synergistic inhibition of cancer growth.

22. 22. The combination of any of claims 19 to 21, wherein the mutation occurs in one or more of EGFR, NF1, RAS and BRAF, MAP2K1, GNAQ or GNA11.

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