Methods of treating acute myeloid leukemia using combinations of gcn2 modulators, venetoclax, and azacitidine

Abstract

Provided herein are methods of treating acute myeloid leukemia (AML) in a subject in need thereof, comprising administering to the subject combinations of Compound 1, or a pharmaceutically acceptable salt thereof, venetoclax, and 5-azacitidine. Also provided herein are methods of inhibiting / overcoming resistance of AML to venetoclax in a subject in need thereof, and / or improving the efficacy of venetoclax in the treatment of AML in a subject in need thereof, comprising administering to the subject combinations of Compound 1, or a pharmaceutically acceptable salt thereof, venetoclax, and 5-azacitidine.

Description

METHODS OF TREATING ACUTE MYELOID LEUKEMIA USING COMBINATIONS OF GCN2 MODULATORS, VENETOCLAX, AND AZACITIDINECROSS REFERENCE TO RELATED APPLICATION

[0001] This application claims the benefit of, and priority to, U.S. Provisional Application No. 63 / 608,048 filed on December 8, 2023, the contents of which are incorporated herein by referenced in their entirety.BACKGROUND

[0002] Cancer is a leading cause of death worldwide, accounting for nearly 10 million deaths in 2020 (World Health Organization). Targeted therapy and immunotherapy have expanded the horizons for treatment of solid tumors by improving prognosis drastically. However, tumor recurrence, drug resistance, and drug intolerance continue to be major challenges in the management of advanced cancer (Wang et al., “Drug resistance and combating drug resistance in cancer,” Cancer Drug Resistance, 2019, 2(2): 141-160; Chakraborty et al., “The difficulties in cancer treatment” Ecancermedicalscience, 2012, 6:edl6). Cancer cells often experience a variety of stressors in their microenvironment such as hypoxia, low pH, and deficiencies in nutrients. In order to survive harsh tumor microenvironments, cancer cells actively utilize adaptive stress pathways such as the integrated stress response (ISR) (Ye et al., “The GCN2-ATF4 pathway is critical for tumor cell survival and proliferation in response to nutrient deprivation,” EMBO J., 2010, 29(12): 2082-2096; Pakos-Zebrucka et al., “The integrated stress response,” EMBO Rep., 2016, 17(10): 1374-1395). The ISR consists of 4 kinases: protein kinase ribonucleic acid [RNA]-like endoplasmic reticulum kinase, protein kinase double-stranded RNA-dependent, general control nondepressible 2 (GCN2), and heme-regulated inhibitor (Donnelly et al., “The eIF2a kinases: their structures and functions,” Cell Mol Life Sci., 2013, 70(19): 3493-3511). These four kinases sense unique stressors and phosphorylate a-subunit of eukaryotic initiation factor 2 (elF2a) (Albert et al., “Adaptive Protein Translation by the Integrated Stress Response Maintains the Proliferative and Migratory Capacity of Lung Adenocarcinoma Cells,” Mol Cancer Res. , 2019, 17(12): 2343- 2355). The high molecular weight kinase GCN2 senses amino acid deficiency as part of the ISR. Under amino acid starvation, uncharged transfer RNA accumulates and activates GCN2 (Anda et al., “Activation of Gcn2 in response to different stresses,” PLOS ONE, 2017, 12(8): E0182143). Phosphorylation of eIF2a by ISR kinases, such as GCN2, inhibits general protein synthesis during cellular stress but also promotes the translation of select mRNAsincluding activating transcription factor 4 (ATF4) which is a key effector of the ISR (Pakos- Zebrucka et al.). Once translated, ATF4 translocates to the nucleus and drives the expression of genes involved in adaptation to stress such as autophagy, antioxidant response, amino acid biosynthesis, and metabolism (Pakos-Zebrucka et al.; Harding et al., “An integrated stress response regulates amino acid metabolism and resistance to oxidative stress,” Mol Cell, 2003, 11(3): 619-633). Other factors which activate GCN2 include ultraviolet light, viral infection, and oxidative stress (Costa-Mattioli et al., “The integrated stress response: From mechanism to disease,” Science, 2020, 368(6489): eaat5314). ATF4 is important for tumor cells to maintain homeostasis of amino acid metabolism. Activation of the ISR pathway promotes tumor cell survival under nutrient deprivation (Ye et al.). GCN2 / ATF4 expression is elevated in primary human liver, breast, lung, and head and neck tumors and GCN2 activation compared to normal tissue has been observed in colon, breast, and lung tumor samples.

[0003] ISR activation plays a dual role in cell fate decisions. During acute stress conditions, ISR can promote adaptation and during chronic stress conditions this pathway can turn apoptotic which results in increased phosphorylation of eIF2a for an extended time (Wortel et al., “Surviving Stress: Modulation of ATF4-Mediated Stress Responses in Normal and Malignant Cells,” Trends Endocrinol Metabol., 2017, 28(11): 794-806). By reducing protein synthesis or activating apoptotic pathways, prolonged activation of ISR can be harmful to cell growth (Wortel et al.; Harding et al., “Ppplrl4 gene knockout reveals an essential role for translation initiation factor 2 alpha (eIF2alpha) dephosphorylation in mammalian development,” Proc Natl Acad Sci USA, 2009, 106(6); 1832-1837; Miinch, “The different axes of the mammalian mitochondrial unfolded protein response,” BMC Biology, 2018;16(1): 81). Persistent ISR activation as a consequence of mutation of eIF2a phosphatases has been shown to have a deleterious effect on embryogenesis due to inhibition of protein synthesis (Harding et al., “Ppplrl4 gene knockout reveals an essential role for translation initiation factor 2 alpha (eIF2alpha) dephosphorylation in mammalian development,” Proc Natl Acad Sci USA, 2009, 106(6); 1832-1837). GCN2 activation also can have antiproliferative effects through suppression of general protein synthesis and induction of cell cycle arrest preventing cells from growing during times of nutrient scarcity (Lehman et al., “Translation Upregulation of an Individual p21Cipl Transcript Variant by GCN2 Regulates Cell Proliferation and Survival under Nutrient Stress,” PLOS Genetics, 2015, 11(6): el005212). Therefore, continuous activation of the GCN2 pathway could suppress protein synthesis and cell growth, thereby inhibiting tumor proliferation.

[0004] Thus, there remains an unmet need to develop new therapeutic strategies that utilize modulation, either activation or inhibition, of the GCN2 pathway for the treatment of a variety of cancers.SUMMARY

[0005] Provided herein are methods of treating acute myeloid leukemia (AML) in a subject in need thereof; methods of inhibiting / overcoming the resistance of AML to venetoclax therapy in a subject in need thereof; and improving the efficacy of venetoclax therapy in the treatment of AML in a subject in need thereof. The methods described herein generally comprise administering to the subject combinations of Compound 1 , or a pharmaceutically acceptable thereof, venetoclax, and 5-azacitidine.

[0006] In one aspect, provided herein is a method of treating acute myeloid leukemia (AML) in a subject in need thereof, the method comprising administering to the subject an effective amount of Compound 1 , or a pharmaceutically acceptable salt thereof, an effective amount of venetoclax, and an effective amount of 5-azacitidine, wherein Compound 1 is represented by formula (I)

[0007] In some embodiments, administering the effective amounts of Compound 1, or a pharmaceutically acceptable salt thereof, venetoclax, and 5-azacitidine increases activation of the integrated stress response (ISR) in the subject as compared to a subject with AML receiving (a) the effective amount of Compound 1 , or a pharmaceutically acceptable salt thereof, or (b) the effective amounts of Compound 1, or a pharmaceutically acceptable salt thereof, and venetoclax.

[0008] In some embodiments, administering the effective amount of the venetoclax comprises administering about 400 mg venetoclax to the subject daily.

[0009] In another aspect, provided herein is a method of treating acute myeloid leukemia (AML) in a subject receiving venetoclax therapy and 5-azacitidine therapy, the method comprising administering to the subject an effective amount of Compound 1 , or a pharmaceutically acceptable salt thereof, wherein Compound 1 is represented by formula (I)

[0010] In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 75 mg / m25-azacitidine daily. In some embodiments, the 5-azacitidine therapy is administered to the subject parenterally.

[0011] In another aspect, provided herein is a method of inhibiting or overcoming resistance of acute myeloid leukemia (AML) to venetoclax therapy in a subject in need thereof, the method comprising administering to the subject an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, and an effective amount of 5-azacitidine, wherein Compound 1 is represented by formula (I)

[0012] In another aspect, provided herein is a method of improving the efficacy of venetoclax therapy in the treatment of acute myeloid leukemia (AML) in a subject in need thereof, the method comprising administering to the subject an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, and an effective amount of 5-azacitidine, wherein Compound 1 is represented by formula (I)

[0013] In some embodiments, the subject receiving the venetoclax therapy is receiving about 400 mg venetoclax daily. In some embodiments, the venetoclax therapy is administered orally.

[0014] In some embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering about 40 mg to about 75 mg Compound 1 , or an equivalent amount of a pharmaceutically acceptable salt thereof on a free acid equivalent weight basis, to the subject. In some embodiments, administering the effective amount of Compound 1 , or a pharmaceutically acceptable salt thereof, comprises administering about 40 mg Compound 1 , or an equivalent amount of a pharmaceutically acceptable salt thereof on a free acid equivalent weight basis, to the subject. In some embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering about 75 mg Compound 1, or an equivalent amount of a pharmaceutically acceptable salt thereof on a free acid equivalent weight basis, to the subject.

[0015] In some embodiments, administering the effective amount of 5-azacitidine comprises administering about 75 mg / m25-azacitidine to the subject daily.

[0016] In some embodiments, the effective amount of Compound 1 , or a pharmaceutically acceptable salt thereof, is administered orally. In some embodiments, the effective amounts of Compound 1, or a pharmaceutically acceptable salt thereof, and venetoclax are administered orally. In some embodiments, the effective amount of 5-azacitidine is administered to the subject parenterally.

[0017] In some embodiments, the method comprises administering an effective amount of a pharmaceutically acceptable salt of Compound 1 to the subject. In certain embodiments, the pharmaceutically acceptable salt of Compound 1 is a potassium salt of Compound 1.

[0018] In some embodiments, the AML is resistant to venetoclax. In some embodiments, the AML comprises a venetoclax resistance mechanism selected from the group consisting of upregulation of anti-apoptotic proteins, p53 inactivation, activating kinase mutations, and upregulation of oxidative phosphorylation. In some embodiments, the AML is characterized by having a fms like tyrosine kinase 3 (FLT3) internal tandem duplication (ITD) mutation. In some embodiments, the AML is characterized by having a tumor protein p53 (TP53) mutation. In some embodiments, the AML is characterized by upregulation of induced myeloid leukemia cell differentiation protein McLl (MCL-1).BRIEF DESCRIPTION OF THE FIGURES

[0019] FIG. 1A is a graph showing the change in tumor volume over time in a MV4-1 1 mouse xenograft tumor model treated with vehicle control, 0.5 mg / kg HC-7366 twice daily, 2 mg / kg HC-7366 twice daily, 50 mg / kg venetoclax once daily, 50 mg / kg venetoclax once daily + 0.5 mg / kg HC-7366 twice daily, or 50 mg / kg venetoclax once daily + 2 mg / kg HC- 7366 twice daily. PO = by mouth, BID = twice a day, Qd = once a day.

[0020] FIG. IB is a graph showing the percent body weight change over time in a MV4-11 mouse xenograft tumor model treated with vehicle control, 0.5 mg / kg HC-7366 twice daily, 2 mg / kg HC-7366 twice daily, 50 mg / kg venetoclax once daily, 50 mg / kg venetoclax once daily + 0.5 mg / kg HC-7366 twice daily, or 50 mg / kg venetoclax once daily + 2 mg / kg HC- 7366 twice daily. PO = by mouth, BID = twice a day, QD = once a day.

[0021] FIG. 2A is a graph showing the change in tumor volume over time in a MV4-11 mouse xenograft tumor model treated with vehicle control, 0.5 mg / kg HC-7366 twice daily, 1 mg / kg HC-7366 twice daily, 50 mg / kg venetoclax once daily, 50 mg / kg venetoclax once daily + 0.5 mg / kg HC-7366 twice daily, 50 mg / kg venetoclax once daily + 1 mg / kg HC-7366 twice daily, 5 mg / kg 5-azacitidine twice weekly, 50 mg / kg venetoclax once daily + 5 mg / kg 5-azacitidine twice weekly, 50 mg / kg venetoclax once daily + 5 mg / kg 5-azacitidine twice weekly + 0.5 mg / kg HC-7366 twice daily, or 50 mg / kg venetoclax once daily + 5 mg / kg 5- azacitidine twice weekly + 1 mg / kg HC-7366 twice daily. PO = by mouth, IP = intraperitoneal, BID = twice a day, QD = once a day, BIW = twice weekly, 5-Aza = 5- azacitidine, Ven = venetoclax.

[0022] FIG. 2B is a graph showing the change in tumor volume over time in a MV4-11 mouse xenograft tumor model treated with vehicle control, 0.5 mg / kg HC-7366 twice daily, 1mg / kg HC-7366 twice daily, 50 mg / kg venetoclax once daily, 5 mg / kg 5-azacitidine twice weekly, 5 mg / kg 5-azacitidine twice weekly + 0.5 mg / kg HC-7366 twice daily, 5 mg / kg 5- azacitidine twice weekly + 1 mg / kg HC-7366 twice daily, 50 mg / kg venetoclax once daily + 5 mg / kg 5-azacitidine twice weekly, 50 mg / kg venetoclax once daily + 5 mg / kg 5-azacitidine twice weekly + 0.5 mg / kg HC-7366 twice daily, or 50 mg / kg venetoclax once daily + 5 mg / kg 5-azacitidine twice weekly + 1 mg / kg HC-7366 twice daily. PO = by mouth, IP = intraperitoneal, BID = twice a day, QD = once a day, BIW = twice weekly, 5-Aza = 5- azacitidine, Ven = venetoclax.

[0023] FIG 3A is a graph showing the MCL1 expression levels in tumors on day 4 from selected treatment groups described in FIG. 2A, as determined by Simple Western. (Groups treated with vehicle 1 mg / kg HC-7366, 50 mg / kg venetoclax, 5 mg / kg 5-azacitidine, 50 mg / kg venetoclax + 5 mg / kg 5-azacitidine, 50 mg / kg venetoclax + 1 mg / kg HC-7366, and 50 mg / kg venetoclax + 5 mg / kg 5-azacitidine + 1 mg / kg HC-7366.) *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, one-way ANOVA. 5-AZA = 5-azacitidine.

[0024] FIG. 3B is a graph showing the SI 00 calcium-binding protein A8 (S100A8) expression levels in tumors on day 4 from selected treatment groups described in FIG. 2A, as determined by Simple Western. (Groups treated with vehicle, 1 mg / kg HC-7366, 50 mg / kg venetoclax, 5 mg / kg 5-azacitidine, 50 mg / kg venetoclax + 5 mg / kg 5-azacitidine, 50 mg / kg venetoclax + 1 mg / kg HC-7366, and 50 mg / kg venetoclax + 5 mg / kg 5-azacitidine + 1 mg / kg HC-7366.) *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, one-way ANOVA. 5-AZA = 5- azacitidine.

[0025] FIG. 3C is a graph showing the translocase of inner mitochondrial membrane 17 A (TIMM17A) expression levels in tumors on day 4 from selected treatment groups described in FIG. 2A, as determined by Simple Western. (Groups treated with vehicle, 1 mg / kg HC- 7366, 50 mg / kg venetoclax, 5 mg / kg 5-azacitidine, 50 mg / kg venetoclax + 5 mg / kg 5- azacitidine, 50 mg / kg venetoclax + 1 mg / kg HC-7366, and 50 mg / kg venetoclax + 5 mg / kg 5- azacitidine + 1 mg / kg HC-7366.) *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, one-way ANOVA. 5-AZA = 5-azacitidine.

[0026] FIG. 4A is a graph showing the percent viability of KG-1 TP53 mutant cells treated with DMSO, staurosporine, 1 pM 5-azacitidine, 0.04 pM HC-7366, 0.1 pM HC-7366, 0.04 pM HC-7366 + 1 pM 5-azacitidine, 0.1 pM HC-7366 + 1 pM 5-azacitidine, 1 pM 5- azacitidine, 0.04 pM HC-7366, 0.1 pM HC-7366, 0.04 pM HC-7366 + 1 pM 5-azacitidine,or 0.1 pM HC-7366 + 1 pM 5 -azacitidine, at varying concentrations of venetoclax (10-fold dilutions from 0.00003-3 pM). AZA = 5-azacitidine.

[0027] FIG. 4B is a graph showing the percent viability of KG-1 TP53 mutant cells treated with DMSO, staurosporine, 1 pM 5-azacitidine, 0.1 pM HC-7366, 0.1 pM HC-7366 + 1 pM 5-azacitidine, 1 pM 5-azacitidine, 0.1 pM HC-7366, or 0.1 pM HC-7366 + 1 pM 5- azacitidine, at varying concentrations of venetoclax (10-fold dilutions from 0.00003-3 pM). AZA = 5-azacitidine.

[0028] FIG. 4C is a graph showing the percent viability of KG-1 TP53 mutant cells treated with DMSO, staurosporine, 1 pM 5-azacitidine, 0.04 pM HC-7366, 0.04 pM HC-7366 + 1 pM 5-azacitidine, 1 pM 5-azacitidine, 0.04 pM HC-7366, or 0.04 pM HC-7366 + 1 pM 5- azacitidine, at varying concentrations of venetoclax (10-fold dilutions from 0.00003-3 pM). AZA = 5-azacitidine.

[0029] FIG. 4D is a graph showing the percent viability of KASUMI-1 TP53 mutant cells treated with DMSO, staurosporine, 1 pM 5-azacitidine, 0.04 pM HC-7366, 0.1 pM HC-7366, 0.04 pM HC-7366 + 1 pM 5-azacitidine, 0.1 pM HC-7366 + 1 pM 5-azacitidine, 1 pM 5- azacitidine, 0.04 pM HC-7366, 0.1 pM HC-7366, 0.04 pM HC-7366 + 1 pM 5-azacitidine, or 0.1 pM HC-7366 + 1 pM 5-azacitidine, at varying concentrations of venetoclax (10-fold dilutions from 0.00003-3 pM). AZA = 5-azacitidine.

[0030] FIG. 4E is a graph showing the percent viability of KASUMI-1 TP53 mutant cells treated with DMSO, staurosporine, 1 pM 5-azacitidine, 0.1 pM HC-7366, 0.1 pM HC-7366 + 1 pM 5-azacitidine, 1 pM 5-azacitidine, 0.1 pM HC-7366, or 0.1 pM HC-7366 + 1 pM 5- azacitidine, at varying concentrations of venetoclax (10-fold dilutions from 0.00003-3 pM). AZA - 5-azacitidine.

[0031] FIG. 4F is a graph showing the percent viability of KASUMI-1 TP53 mutant cells treated with DMSO, staurosporine, 1 pM 5-azacitidine, 0.04 pM HC-7366, 0.04 pM HC- 7366 + 1 pM 5-azacitidine, 1 pM 5-azacitidine, 0.04 pM HC-7366, or 0.04 pM HC-7366 + 1 pM 5-azacitidine, at varying concentrations of venetoclax (10-fold dilutions from 0.00003-3 pM). AZA = 5-azacitidine.

[0032] FIG. 5A is a graph showing the change in tumor volume over time in a KG- 1 human AML xenograft tumor model treated with vehicle (HC-7366) + vehicle (venetoclax) + vehicle (5-AZA), 0.5 mg / kg HC-7366 BID, 1 mg / kg HC-7366 BID, 50 mg / kg venetoclax Qd, 5-AZA 2 mg / kg BIW, 50 mg / kg venetoclax Qd + 2 mg / kg 5-AZA BIW, 50 mg / kg venetoclax Qd + 0.5 mg / kg HC-7366 BID, 50 mg / kg venetoclax Qd + 1 mg / kg HC-7366 BID, 2mg / kg 5-AZA BIW + 0.5 mg / kg HC-7366 BID, 2mg / kg 5-AZA BIW + 1 mg / kg HC-7366 BID, 50 mg / kg venetoclax Qd + 2 mg / kg 5-AZA BIW + 0.5 mg / kg HC-7366 BID, or 50 mg / kg venetoclax Qd + 2 mg / kg 5-AZA BIW + 0.5 mg / kg HC-7366 BID. PO = by mouth, BID = twice a day, Qd = once a day, BIW = twice weekly.

[0033] FIG. SB is a graph showing the percent body weight change over time in a KG-1 human AML xenograft tumor model treated with vehicle (HC-7366) + vehicle (venetoclax) + vehicle (5-AZA), 0.5 mg / kg HC-7366 BID, 1 mg / kg HC-7366 BID, 50 mg / kg venetoclax Qd, 5-AZA 2 mg / kg BIW, 50 mg / kg venetoclax Qd + 2 mg / kg 5-AZA BIW, 50 mg / kg venetoclax Qd + 0.5 mg / kg HC-7366 BID, 50 mg / kg venetoclax Qd + 1 mg / kg HC-7366 BID, 2mg / kg 5- AZA BIW + 0.5 mg / kg HC-7366 BID, 2mg / kg 5-AZA BIW + 1 mg / kg HC-7366 BID, 50 mg / kg venetoclax Qd + 2 mg / kg 5-AZA BIW + 0.5 mg / kg HC-7366 BID, or 50 mg / kg venetoclax Qd + 2 mg / kg 5-AZA BIW + 0.5 mg / kg HC-7366 BID. PO = by mouth, BID = twice a day, Qd = once a day, BIW = twice weekly.

[0034] FIG. 6A illustrates a graph of tumor volumes in the PK / PD groups prior to tumor collection on Day 8 in KG-1 xenograft tumors treated with HC-7366 administered at 0.5 mg / kg, venetoclax at 50 mg / kg and 5-azacitidine at 2 mg / kg, as single agents or in combination. Tumor volumes were measured after 4 and 7 days of treatment, prior to tumor collection on day 8.

[0035] FIG. 6B illustrates a graph of tumor volumes in the PK / PD groups prior to tumor collection on Day 8 in KG-1 xenograft tumors treated with HC-7366 administered at 1 mg / kg, venetoclax at 50 mg / kg and 5-azacitidine at 2 mg / kg, as single agents or in combination. Tumor volumes were measured after 4 and 7 days of treatment, prior to tumor collection on day 8.

[0036] FIGS. 7A-7L are graphs illustrating SimpleWestem analysis of pharmacodynamic markers and markers of Venetoclax resistance in KG-1 tumors. KG-1 xenograft tumors were treated with HC-7366 administered at 0.5 mg / kg or 1 mg / kg, Venetoclax at 50 mg / kg and 5- azacitidine at 2 mg / kg, as single agents or in combination. Tumors from each group (n=8 per group) were collected on Day 8, lysed, and analyzed for: PSAT1, an ATF4 target gene and pathway engagement marker (FIG. 7A); PUMA, a proapoptotic ATF4 target gene (FIG. 7B); MCL1, an antiapoptotic protein (FIG. 7C); TIMM17A (FIG. 7D); TIMM23 (FIG. 7E), subunits of a mitochondrial import complex, S100A8, a calcium binding protein (FIG. 7F); phospho RB (pS807 / pS811) (FIG. 7G); total RB (FIG. 7H); CDK1 (FIG. 71); cell cycle regulators, cMYC, a transcription factor and proto-oncogene (FIG. 7J); HIFla, a hypoxia-inducible transcription factor (FIG. 7K); and P-actin, a loading control (FIG. 7L). *p<0.05, **p<0.01, ***p<0.001, one-way ANOVA.DETAILED DESCRIPTION

[0037] As generally described herein, the present disclosure provides methods of treating acute myeloid leukemia (AML) in a subject in need thereof. The present disclosure also provides methods of inhibiting / overcoming the resistance of AML to venetoclax therapy in a subject in need thereof and / or improving the efficacy of venetoclax therapy in the treatment of AML in a subject in need thereof. The methods described herein generally comprise administering to the subject combinations of Compound 1, or a pharmaceutically acceptable thereof, venetoclax, and 5 -azacitidine.Definitions

[0038] To facilitate an understanding of the present invention, a number of terms and phrases are defined below.

[0039] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts.

[0040] Throughout the description, where compositions and kits are described as having, including, or comprising specific components, or where processes and methods are described as having, including, or comprising specific steps, it is contemplated that, additionally, there are compositions and kits of the present invention that consist essentially of, or consist of, the recited components, and that there are processes and methods according to the present invention that consist essentially of, or consist of, the recited processing steps.

[0041] In the application, where an element or component is said to be included in and / or selected from a list of recited elements or components, it should be understood that the element or component can be any one of the recited elements or components, or the element or component can be selected from a group consisting of two or more of the recited elements or components.

[0042] Further, it should be understood that elements and / or features of a composition or a method described herein can be combined in a variety of ways without departing from the spirit and scope of the present invention, whether explicit or implicit herein. For example,where reference is made to a particular compound, that compound can be used in various embodiments of compositions of the present invention and / or in methods of the present invention, unless otherwise understood from the context. In other words, within this application, embodiments have been described and depicted in a way that enables a clear and concise application to be written and drawn, but it is intended and will be appreciated that embodiments may be variously combined or separated without parting from the present teachings and invention(s). For example, it will be appreciated that all features described and depicted herein can be applicable to all aspects of the invention(s) described and depicted herein.

[0043] The articles “a” and “an” are used in this disclosure to refer to one or more than one (i.e., to at least one) of the grammatical object of the article, unless the context is inappropriate. By way of example, “an element” means one element or more than one element.

[0044] The term “and / or” is used in this disclosure to mean either “and” or “or” unless indicated otherwise.

[0045] It should be understood that the expression “at least one of’ includes individually each of the recited objects after the expression and the various combinations of two or more of the recited objects unless otherwise understood from the context and use. The expression “and / or” in connection with three or more recited objects should be understood to have the same meaning unless otherwise understood from the context.

[0046] The use of the term “include,” “includes,” “including,” “have,” “has,” “having,” “contain,” “contains,” or “containing,” including grammatical equivalents thereof, should be understood generally as open-ended and non-limiting, for example, not excluding additional unrecited elements or steps, unless otherwise specifically stated or understood from the context.

[0047] Where the use of the term “about” is before a quantitative value, the present invention also includes the specific quantitative value itself, unless specifically stated otherwise. As used herein, the term “about” refers to a ±10%, ±5%, ±3%, ±2%, or ±1% variation from the nominal value unless otherwise indicated or inferred from the context.

[0048] At various places in the present specification, variable or parameters are disclosed in groups or in ranges. It is specifically intended that the description include each and every individual subcombination of the members of such groups and ranges. For example, an integer in the range of 0 to 40 is specifically intended to individually disclose 0, 1, 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, and 40, and an integer in the range of 1 to 20 is specifically intended to individually disclose 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20.

[0049] The use of any and all examples, or exemplary language herein, for example, “such as” or “including,” is intended merely to illustrate better the present invention and does not pose a limitation on the scope of the invention unless claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the present invention.

[0050] As a general matter, compositions specifying a percentage are by weight unless otherwise specified. Further, if a variable is not accompanied by a definition, then the previous definition of the variable controls.

[0051] As used herein, “pharmaceutical composition” or “pharmaceutical formulation” refers to the combination of an active agent with a carrier, inert or active, making the composition especially suitable for diagnostic or therapeutic use in vivo or ex vivo.

[0052] “Pharmaceutically acceptable” means approved or approvable by a regulatory agency of the federal or a state government or the corresponding agency in countries other than the United States, or that is listed in the U.S. Pharmacopoeia or other generally recognized pharmacopoeia for use in animals, and more particularly, in humans.

[0053] As used herein, “pharmaceutically acceptable salt” refers to any salt of an acidic or a basic group that may be present in a compound of the present invention (e.g., Compound 1), which salt is compatible with pharmaceutical administration.

[0054] As is known to those of skill in the art, “salts” of compounds may be derived from inorganic or organic acids and bases. Examples of acids include, but are not limited to, hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, ethanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic and benzenesulfonic acid. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds described herein and their pharmaceutically acceptable acid addition salts.

[0055] Examples of bases include, but are not limited to, alkali metal (e.g., sodium and potassium) hydroxides, alkaline earth metal (e.g., magnesium and calcium) hydroxides, ammonia, and compounds of formula NW4+, wherein W is CM alkyl, and the like.

[0056] Examples of salts include, but are not limited to, acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, flucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, undecanoate, and the like. Other examples of salts include anions of the compounds of the present invention compounded with a suitable cation such as Na+, K+, Ca2+, NH4+, and NW4+(where W can be a Ci -4 alkyl group), and the like.

[0057] For therapeutic use, salts of the compounds of the present invention (e.g., Compound 1) are contemplated as being pharmaceutically acceptable. However, salts of acids and bases that are non-pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound.

[0058] As used herein, “pharmaceutically acceptable excipient” refers to a substance that aids the administration of an active agent to and / or absorption by a subject and can be included in the compositions of the present invention without causing a significant adverse toxicological effect on the patient. Non- limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, such as a phosphate buffered saline solution, emulsions (e.g., such as an oil / water or water / oil emulsions), lactated Ringer’s, normal sucrose, normal glucose, binders, fillers, disin tegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer’s solution), alcohols, oils, gelatins, carbohydrates, fatty acid esters, and colors, and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and / or aromatic substances and the like that do not deleteriously react with the compounds of the invention. For examples of excipients, see Martin, Remington’s Pharmaceutical Sciences, 15th Ed., Mack Publ. Co., Easton, PA (1975).

[0059] A “subject” to which administration is contemplated includes, but is not limited to, humans (i.e., a male or female of any age group, e.g., a pediatric subject (e.g., infant, child, adolescent) or adult subject (e.g., young adult, middle-aged adult or senior adult)) and / or a non-human animal, e.g., a mammal such as primates (e.g., cynomolgus monkeys, rhesus monkeys), cattle, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In certainembodiments, the subject is a human. In certain embodiments, the subject is an adult human. In certain embodiments, the subject is a non-human animal.

[0060] As used herein, “solid dosage form” means a pharmaceutical dose(s) in solid form, e.g., tablets, capsules, granules, powders, sachets, reconstitutable powders, dry powder inhalers and chewables.

[0061] As used herein, “administering” means oral administration, administration as a suppository, topical contact, intravenous administration, parenteral administration, intraperitoneal administration, intramuscular administration, intralesional administration, intrathecal administration, intracranial administration, intranasal administration or subcutaneous administration, or the implantation of a slow-release device, e.g., a mini- osmotic pump, to a subject. Administration is by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, e.g., intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, etc. By “co-administer” it is meant that a composition described herein is administered at the same time, just prior to, or just after the administration of one or more additional therapies (e.g., anti-cancer agent, chemotherapeutic, or immunotherapy). Compound 1, or a pharmaceutically acceptable salt thereof, can be administered alone or can be co-administered to the patient. Co-administration is meant to include simultaneous or sequential administration of the compound individually or in combination (more than one compound or agent). Thus, the preparations can also be combined, when desired, with other active substances (e.g., to reduce metabolic degradation).

[0062] As used herein, “fasting state” means at least 1 hour before food or at least 2 hours after food is consumed by a subject.

[0063] The terms “disease,” “disorder,” and “condition” are used interchangeably herein.

[0064] As used herein, and unless otherwise specified, the terms “treat,” “treating” and “treatment” contemplate an action that occurs while a subject is suffering from the specified disease, disorder or condition, which reduces the severity of the disease, disorder or condition, or retards or slows the progression of the disease, disorder or condition (e.g., “therapeutic treatment”).

[0065] In general, an “effective amount” of a compound (e.g., Compound 1, or a pharmaceutically acceptable salt thereof) refers to an amount sufficient to elicit the desiredbiological response, e.g., to treat AML. As will be appreciated by those of ordinary skill in this art, the effective amount of a compound of the disclosure may vary depending on such factors as the desired biological endpoint, the pharmacokinetics of the compound, the disease being treated, the mode of administration, and the age, weight, health, and condition of the subject.CompoundsCompound 1

[0066] Compound 1 , as represented by formula (I) shown below, is a selective modulator of general control nonderepressible 2 (GCN2), e.g., activation or inhibition of GCN2, and is also known as 6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6-difluorophenyl)-N- methylimidazo[l ,5-a]pyrazine- 1 -carboxamide:

[0067] Compound 1 may also be referred to as HC-7366 throughout the present disclosure.A method of chemically synthesizing Compound 1 is described in Example 1.

[0068] PCT International Application No. PCT / US2022 / 013383, published as WO 2022 / 159746, which is incorporated herein by reference, describes the synthesis of Compound 1 and also discloses the GCN2 modulatory activity of Compound 1.

[0069] In one aspect, provided herein is a method of administering effective amounts of Compound 1, or a pharmaceutically acceptable salt thereof, venetoclax, and 5 -azacitidine for the treatment of AML in a subject in need thereof.

[0070] In various embodiments, provided herein is a method of administering effective amounts of a pharmaceutically acceptable salt of Compound 1, venetoclax, and 5-azacitidine, for the treatment AML in a subject in need thereof.

[0071] In certain embodiments, the pharmaceutically acceptable salt of Compound 1 is a potassium salt. In certain embodiments, the potassium salt of Compound 1 is a hydrate. In certain embodiments, the potassium salt of Compound 1 is a monohydrate. A method of preparing a potassium salt of Compound 1 is described in Example 2.Venetoclax

[0072] Venetoclax is a BH3-mimetic which blocks the anti- apop to tic B-cell lymphoma-2 protein and is represented by the following structural formula:

[0073] Venetoclax is currently approved for the treatment of chronic lymphocytic leukemia (CLL) or small lymphocytic lymphoma (SLL) in adult patients. Venetoclax is also indicated as part of a combination therapy with azacitidine, decitabine, or low-dose cytarabine for the treatment of AML in newly diagnosed adults aged 75 years or older or with comorbidities that preclude use of intensive induction chemotherapy. Venetoclax and its methods of use and administration are disclosed in, for example, U.S. Patent Nos. 8,546,399; 8,722,657; 9,174,982; 9,539,251 ; 10,730,873; 10,993,942; 11,110,087; 11,369,599; 11,413,282; and 11 ,590, 128, which are incorporated herein by reference. Venetoclax may also be referred to as GDC-0199, ABT-199, RG-7601, or under the trade names Venclexta® or Venclyxto®.

[0074] In certain embodiments, provided herein is a method of administering effective amounts of venetoclax, Compound 1, or a pharmaceutically acceptable salt thereof, and 5- azacitidine for the treatment of AML in subject in need thereof 5 -Azacitidine

[0075] 5 -Azacitidine is a chemical analogue of the nucleoside cytidine which has antineoplastic activity and is represented by the following structural formula:

[0076] 5-Azacitidine is approved for the treatment of myelodysplastic syndrome and myeloid leukemia in adult patients, and juvenile myelomonocytic leukemia in pediatric patients. 5- Azacitidine and its methods of use and administration are disclosed in, for example, “Biological Effects of 5-azacytidine in Eukaryotes: A Review” (Oncology (1974) 30 (5): 405- 422), and U.S. Patent Nos. 8,846,628 and 11,571,436, which are incorporated herein by reference. 5-Azacitidine may also be referred to as 5-azacytidine, azacytidine, ladakamycin, 4-amino-l-P-D-ribofuranosyl-S-triazin-2(177)-one, U-18496, CC-486, or under the trade names Vidaza®, Azadine, or Onureg®.

[0077] In certain embodiments, provided herein is a method of administering effective amounts of 5 -azacitidine, Compound 1, or a pharmaceutically acceptable salt thereof, and venetoclax for the treatment of AML in a subject in need thereofPharmaceutical Compositions

[0078] Provided herein are pharmaceutical compositions generally comprising Compound 1 , or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

[0079] In one aspect, provided herein is a method of administering a pharmaceutical composition comprising Compound 1, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients in combination with venetoclax and 5- azacitidine for the treatment of AML.

[0080] In another aspect, provided herein is a method of administering a pharmaceutical composition comprising a pharmaceutically acceptable salt of Compound 1 and one or more pharmaceutically acceptable excipients in combination with venetoclax and 5 -azacitidine, for the treatment of AML.

[0081] In various embodiments, provided herein is a pharmaceutical composition comprising an effective amount of Compound 1 , or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

[0082] In various embodiments, provided herein is a pharmaceutical composition comprising an effective amount of a pharmaceutically acceptable salt of Compound 1 and one or more pharmaceutically acceptable excipients.

[0083] In certain embodiments, the amount of Compound 1, or a pharmaceutically acceptable salt thereof, in a pharmaceutical composition described herein is about 10 mg to about 150 mg, about 20 mg to about 150 mg, about 40 mg to about 150 mg, about 75 mg to about 150 mg, about 125 mg to about 150 mg, about 10 mg to about 125 mg, about 10 mg to about 75 mg, about 10 mg to about 40 mg, about 10 mg to about 20 mg, about 20 mg to about 125 mg, about 20 mg to about 75 mg, about 20 mg to about 40 mg, about 40 mg to about 125 mg, about 40 mg to about 75 mg, or about 75 mg to about 125 mg, on a free acid equivalent weight basis. In certain embodiments, the amount of Compound 1, or a pharmaceutically acceptable salt thereof, in a pharmaceutical composition described herein is about 10 mg to about 150 mg on a free acid equivalent weight basis.

[0084] In certain embodiments, the amount of Compound 1, or a pharmaceutically acceptable salt thereof, in a pharmaceutical composition described herein is about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, or about 150 mg, on a free acid equivalent weight basis. In certain embodiments, the amount of Compound 1 , or a pharmaceutically acceptable salt thereof, in a pharmaceutical composition described herein is about 10 mg on a free acid equivalent weight basis. In certain embodiments, the amount of Compound 1, or a pharmaceutically acceptable salt thereof, in a pharmaceutical composition described herein is about 20 mg on a free acid equivalent weight basis. In certain embodiments, the amount of Compound 1, or a pharmaceutically acceptable salt thereof, in a pharmaceutical composition described herein is about 40 mg on a free acid equivalent weight basis. In certain embodiments, the amount of Compound 1, or a pharmaceutically acceptable salt thereof, in a pharmaceutical composition described herein is about 75 mg on a free acid equivalent weight basis. In certain embodiments, the amount of Compound 1 , or a pharmaceutically acceptable salt thereof, in a pharmaceutical composition described herein is about 125 mg on a free acid equivalent weight basis. In certain embodiments, the amount of Compound 1 , or a pharmaceutically acceptable salt thereof, in a pharmaceutical composition described herein is about 150 mg on a free acid equivalent weight basis.

[0085] In certain embodiments, the amount of the pharmaceutically acceptable salt of Compound 1 in a pharmaceutical composition described herein is about 10 mg to about 150 mg, about 20 mg to about 150 mg, about 40 mg to about 150 mg, about 75 mg to about 150 mg, about 125 mg to about 150 mg, about 10 mg to about 125 mg, about 10 mg to about 75 mg, about 10 mg to about 40 mg, about 10 mg to about 20 mg, about 20 mg to about 125 mg, about 20 mg to about 75 mg, about 20 mg to about 40 mg, about 40 mg to about 125 mg, about 40 mg to about 75 mg, or about 75 mg to about 125 mg, on a free acid equivalent weight basis. In certain embodiments, the amount of the pharmaceutically acceptable salt of Compound 1 in a pharmaceutical composition described herein is about 10 mg to about 150 mg on a free acid equivalent weight basis.

[0086] In certain embodiments, the amount of the pharmaceutically acceptable salt of Compound 1 in a pharmaceutical composition described herein is about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, or about 150 mg, on a free acid equivalent weight basis. In certain embodiments, the amount of the pharmaceutically acceptable salt of Compound 1 in a pharmaceutical composition described herein is about 10 mg on a free acid equivalent weight basis. In certain embodiments, the amount of the pharmaceutically acceptable salt of Compound 1 in a pharmaceutical composition described herein is about 20 mg on a free acid equivalent weight basis. In certain embodiments, the amount of the pharmaceutically acceptable salt of Compound 1 in a pharmaceutical composition described herein is about 40 mg on a free acid equivalent weight basis. In certain embodiments, the amount of the pharmaceutically acceptable salt of Compound 1 in a pharmaceutical composition described herein is about 75 mg on a free acid equivalent weight basis. In certain embodiments, the amount of the pharmaceutically acceptable salt of Compound 1 in a pharmaceutical composition described herein is about 125 mg on a free acid equivalent weight basis. In certain embodiments, the amount of the pharmaceutically acceptable salt of Compound 1 in a pharmaceutical composition described herein is about 150 mg on a free acid equivalent weight basis.

[0087] In various embodiments, provided herein are pharmaceutical compositions comprising:(i) about 10 mg to about 150 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis; and(ii) one or more pharmaceutically acceptable excipients.

[0088] In various embodiments, provided herein are pharmaceutical compositions comprising:(i) about 10 mg to about 150 mg of a pharmaceutically acceptable salt of Compound 1, on a free acid equivalent weight basis; and(ii) one or more pharmaceutically acceptable excipients.

[0089] In another aspect, provided herein are pharmaceutical compositions comprising about 10 mg to about 150 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis, and one or more pharmaceutically acceptable excipients, for the treatment of AML in a subject in need thereof.

[0090] In another aspect, provided herein are pharmaceutical compositions comprising about 10 mg to about 150 mg of a pharmaceutically acceptable salt of Compound 1, on a free acid equivalent weight basis, and one or more pharmaceutically acceptable excipients, for the treatment of AML in a subject in need thereof.

[0091] In another aspect, provided herein are pharmaceutical compositions comprising about 40 mg to about 75 mg Compound 1, or an equivalent amount of a pharmaceutically acceptable salt thereof on a free acid equivalent weight basis, and one or more pharmaceutically acceptable excipients, for the treatment of AML in a subject in need thereof.

[0092] In another aspect, provided herein are pharmaceutical compositions comprising about 40 mg Compound 1 , or an equivalent amount of a pharmaceutically acceptable salt thereof on a free acid equivalent weight basis, and one or more pharmaceutically acceptable excipients, for the treatment of AML in a subject in need thereof.

[0093] In another aspect, provided herein are pharmaceutical compositions comprising about 75 mg Compound 1, or an equivalent amount of a pharmaceutically acceptable salt thereof on a free acid equivalent weight basis, and one or more pharmaceutically acceptable excipients, for the treatment of AML in a subject in need thereof.

[0094] In certain embodiments, the pharmaceutically acceptable salt of Compound 1 is a potassium salt. In certain embodiments, the potassium salt of Compound 1 is a hydrate. In certain embodiments, the potassium salt of Compound 1 is a monohydrate.

[0095] The pharmaceutical compositions described herein can be administered by a variety of routes including, but not limited to, oral (enteral) administration, parenteral (by injection) administration, rectal administration, transdermal administration, intradermal administration, intrathecal administration, subcutaneous (SC) administration, intravenous (IV)administration, intramuscular (IM) administration, and intranasal administration. In certain embodiments, the pharmaceutical compositions described herein are administered orally.

[0096] The pharmaceutical compositions described herein may also be administered chronically (“chronic administration”). Chronic administration refers to administration of a compound or pharmaceutical composition thereof over an extended period of time, e.g., for example, over 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc., or may be continued indefinitely, for example, for the rest of the subject’s life. In certain embodiments, the chronic administration is intended to provide a constant level of the compound in the blood, e.g., within the therapeutic window over the extended period of time.

[0097] The pharmaceutical compositions described herein may be presented in unit dosage forms to facilitate accurate dosing. The term “unit dosage forms” refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, premeasured ampules or syringes of the liquid compositions or pills, tablets, capsules or the like in the case of solid compositions.

[0098] In certain embodiments, the pharmaceutical compositions provided herein are administered to the patient as a solid dosage form. In certain embodiments, the solid dosage form is a capsule.

[0099] In certain embodiments, the pharmaceutical composition is an immediate release capsule formulation of HC-7366 potassium salt monohydrate. In some embodiments, the capsule is a hard gelatin capsule. In some embodiments, the capsule comprises one or more of lactose monohydrate, microcrystalline cellulose, croscarmellose sodium, colloidal silicon dioxide, and magnesium stearate. In some embodiments, the capsule comprises each of lactose monohydrate, microcrystalline cellulose, croscarmellose sodium, colloidal silicon dioxide, and magnesium stearate. In some embodiments, the capsule comprises 10 mg, 25 mg, or 100 mg of HC-7366 potassium salt monohydrate on a free acid equivalent weight basis.

[0100] Although the descriptions of pharmaceutical compositions provided herein are principally directed to pharmaceutical compositions which are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts. Modification of pharmaceutical compositions suitable for administration to humans in order to render the compositionssuitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and / or perform such modification with ordinary experimentation. General considerations in the formulation and / or manufacture of pharmaceutical compositions can be found, for example, in Remington: The Science and Practice of Pharmacy 21sted., Lippincott Williams & Wilkins, 2005.Methods of Use and Treatment

[0101] Provided herein are methods of treating acute myeloid leukemia (AML) in a subject in need thereof. Also provided herein are methods of inhibiting / overcoming resistance of AML to venetoclax therapy, and / or improving the efficacy of venetoclax therapy in the treatment of AML. The methods generally comprise administering to the subject an effective amount of Compound 1 , or a pharmaceutically acceptable salt thereof, an effective amount of venetoclax, and an effective amount of 5 -azacitidine.

[0102] In one aspect, provided herein is a method of treating acute myeloid leukemia (AML) in a subject in need thereof, the method comprising administering to the subject an effective amount of Compound 1 , or a pharmaceutically acceptable salt thereof, an effective amount of venetoclax, and an effective amount of 5 -azacitidine, wherein Compound 1 is represented by formula (I)

[0103] In some embodiments, administering the effective amounts of Compound 1, or a pharmaceutically acceptable salt thereof, venetoclax, and 5-azacitidine increases activation of the integrated stress response (ISR) in the subject as compared to a subject with AML receiving (a) the effective amount of Compound 1 , or a pharmaceutically acceptable salt thereof, or (b) the effective amounts of Compound 1, or a pharmaceutically acceptable salt thereof, and venetoclax.

[0104] In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 100 mg to about 600 mg, about 150 mg to about 600 mg,about 200 mg to about 600 mg, about 250 mg to about 600 mg, about 300 mg to about 600 mg, about 350 mg to about 600 mg, about 400 mg to about 600 mg, about 100 mg to about 550 mg, about 100 mg to about 500 mg, about 100 mg to about 450 mg, about 100 mg to about 400 mg, about 100 mg to about 350 mg, about 100 mg to about 300 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 150 mg to about 550 mg, about 200 mg to about 500 mg, about 250 mg to about 450 mg, or about 300 mg to about 400 mg venetoclax to the subject.

[0105] In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 100 mg to about 600 mg, about 150 mg to about 600 mg, about 200 mg to about 600 mg, about 250 mg to about 600 mg, about 300 mg to about 600 mg, about 350 mg to about 600 mg, about 400 mg to about 600 mg, about 100 mg to about 550 mg, about 100 mg to about 500 mg, about 100 mg to about 450 mg, about 100 mg to about 400 mg, about 100 mg to about 350 mg, about 100 mg to about 300 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 150 mg to about 550 mg, about 200 mg to about 500 mg, about 250 mg to about 450 mg, or about 300 mg to about 400 mg venetoclax to the subject daily.

[0106] In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 20 mg, about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, or about 600 mg venetoclax to the subject. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 20 mg venetoclax to the subject. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 50 mg venetoclax to the subject. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 100 mg venetoclax to the subject. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 200 mg venetoclax to the subject. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 300 mg venetoclax to the subject. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 400 mg venetoclax to the subject. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 500 mg venetoclax to the subject. In some embodiments,administering the effective amount of venetoclax to the subject comprises administering about 600 mg venetoclax to the subject.

[0107] In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 20 mg, about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, or about 600 mg venetoclax to the subject daily. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 20 mg venetoclax to the subject daily. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 50 mg venetoclax to the subject daily. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 100 mg venetoclax to the subject daily. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 200 mg venetoclax to the subject daily. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 300 mg venetoclax to the subject daily. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 400 mg venetoclax to the subject daily. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 500 mg venetoclax to the subject daily. In some embodiments, administering the effective amount of venetoclax to the subject comprises administering about 600 mg venetoclax to the subject daily.

[0108] In some embodiments, the effective amounts of Compound 1, or a pharmaceutically acceptable salt thereof, and venetoclax are administered orally.

[0109] In another aspect, provided herein is a method of treating acute myeloid leukemia (AML) in a subject receiving venetoclax therapy and 5-azacitidine therapy, the method comprising administering to the subject an effective amount of Compound 1 , or a pharmaceutically acceptable salt thereof, wherein Compound 1 is represented by formula (I)

[0110] In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 50 mg / m2to about 100 mg / m2, about 60 mg / m2to about 100 mg / m2, about 70 mg / m2to about 100 mg / m2, about 75 mg / m2to about 100 mg / m2, about 50 mg / m2to about 90 mg / m2, about 50 mg / m2to about 80 mg / m2, about 50 mg / m2to about 75 mg / m2, about 60 mg / m2to about 90 mg / m2, or about 70 mg / m2to about 80 mg / m25-azacitidine.

[0111] In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 50 mg / m2to about 100 mg / m2, about 60 mg / m2to about 100 mg / m2, about 70 mg / m2to about 100 mg / m2, about 75 mg / m2to about 100 mg / m2, about 50 mg / m2to about 90 mg / m2, about 50 mg / m2to about 80 mg / m2, about 50 mg / m2to about 75 mg / m2, about 60 mg / m2to about 90 mg / m2, or about 70 mg / m2to about 80 mg / m25-azacitidine daily.

[0112] In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 50 mg / m2, about 55 mg / m2, about 60 mg / m2, about 65 mg / m2, about 70 mg / m2, about 75 mg / m2, about 80 mg / m2, about 85 mg / m2, about 90 mg / m2, about 95 mg / m2, or about 100 mg / m25-azacitidine. In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 50 mg / m25-azacitidine. In some embodiments, the subject receiving the 5- azacitidine therapy is receiving about 60 mg / m25-azacitidine. In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 70 mg / m25-azacitidine. In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 75 mg / m25-azacitidine. In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 80 mg / m25-azacitidine. In some embodiments, the subject receiving the 5- azacitidine therapy is receiving about 90 mg / m25-azacitidine. In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 100 mg / m25-azacitidine.

[0113] In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 50 mg / m2, about 55 mg / m2, about 60 mg / m2, about 65 mg / m2, about 70 mg / m2, about 75 mg / m2, about 80 mg / m2, about 85 mg / m2, about 90 mg / m2, about 95 mg / m2, or about 100mg / m25-azacitidine daily. In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 50 mg / m25-azacitidine daily. In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 60 mg / m25-azacitidine daily. In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 70 mg / m25- azacitidine daily. In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 75 mg / m25-azacitidine daily. In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 80 mg / m25-azacitidine daily. In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 90 mg / m25- azacitidine daily. In some embodiments, the subject receiving the 5-azacitidine therapy is receiving about 100 mg / m25-azacitidine daily.

[0114] In some embodiments, the 5-azacitidine therapy is administered to the subject parenterally.

[0115] In another aspect, provided herein is a method of inhibiting or overcoming resistance of acute myeloid leukemia (AML) to venetoclax therapy in a subject in need thereof, the method comprising administering to the subject an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, and an effective amount of 5-azacitidine, wherein Compound 1 is represented by formula (I)

[0116] In another aspect, provided herein is a method of improving the efficacy of venetoclax therapy in the treatment of acute myeloid leukemia (AML) in a subject in need thereof, the method comprising administering to the subject an effective amount of Compound 1 , or a pharmaceutically acceptable salt thereof, and an effective amount of 5-azacitidine, wherein Compound 1 is represented by formula (I)

[0117] In some embodiments, the subject receiving the venetoclax therapy is receiving about 100 mg to about 600 mg, about 150 mg to about 600 mg, about 200 mg to about 600 mg, about 250 mg to about 600 mg, about 300 mg to about 600 mg, about 350 mg to about 600 mg, about 400 mg to about 600 mg, about 100 mg to about 550 mg, about 100 mg to about 500 mg, about 100 mg to about 450 mg, about 100 mg to about 400 mg, about 100 mg to about 350 mg, about 100 mg to about 300 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 150 mg to about 550 mg, about 200 mg to about 500 mg, about 250 mg to about 450 mg, or about 300 mg to about 400 mg venetoclax.

[0118] In some embodiments, the subject receiving the venetoclax therapy is receiving about 100 mg to about 600 mg, about 150 mg to about 600 mg, about 200 mg to about 600 mg, about 250 mg to about 600 mg, about 300 mg to about 600 mg, about 350 mg to about 600 mg, about 400 mg to about 600 mg, about 100 mg to about 550 mg, about 100 mg to about 500 mg, about 100 mg to about 450 mg, about 100 mg to about 400 mg, about 100 mg to about 350 mg, about 100 mg to about 300 mg, about 100 mg to about 250 mg, about 100 mg to about 200 mg, about 150 mg to about 550 mg, about 200 mg to about 500 mg, about 250 mg to about 450 mg, or about 300 mg to about 400 mg venetoclax daily.

[0119] In some embodiments, the subject receiving the venetoclax therapy is receiving about 20 mg, about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, or about 600 mg venetoclax. In some embodiments, the subject receiving the venetoclax therapy is receiving about 20 mg venetoclax. In some embodiments, the subject receiving the venetoclax therapy is receiving about 50 mg venetoclax. In some embodiments, the subject receiving the venetoclax therapy is receiving about 100 mg venetoclax. In some embodiments, the subject receiving the venetoclax therapy is receiving about 200 mg venetoclax. In some embodiments, the subject receiving the venetoclax therapy is receivingabout 300 mg venetoclax. In some embodiments, the subject receiving the venetoclax therapy is receiving about 400 mg venetoclax. In some embodiments, the subject receiving the venetoclax therapy is receiving about 500 mg venetoclax. In some embodiments, the subject receiving the venetoclax therapy is receiving about 600 mg venetoclax.

[0120] In some embodiments, the subject receiving the venetoclax therapy is receiving about 20 mg, about 50 mg, about 100 mg, about 150 mg, about 200 mg, about 250 mg, about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, or about 600 mg venetoclax daily. In some embodiments, the subject receiving the venetoclax therapy is receiving about 20 mg venetoclax daily. In some embodiments, the subject receiving the venetoclax therapy is receiving about 50 mg venetoclax daily. In some embodiments, the subject receiving the venetoclax therapy is receiving about 100 mg venetoclax daily. In some embodiments, the subject receiving the venetoclax therapy is receiving about 200 mg venetoclax daily. In some embodiments, the subject receiving the venetoclax therapy is receiving about 300 mg venetoclax daily. In some embodiments, the subject receiving the venetoclax therapy is receiving about 400 mg venetoclax daily. In some embodiments, the subject receiving the venetoclax therapy is receiving about 500 mg venetoclax daily. In some embodiments, the subject receiving the venetoclax therapy is receiving about 600 mg venetoclax daily.

[0121] In some embodiments, the venetoclax therapy is administered orally.

[0122] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 10 mg to about 150 mg, about 15 mg to about 150 mg, about 20 mg to about 150 mg, about 25 mg to about 150 mg, about 30 mg to about 150 mg, about 35 mg to about 150 mg, about 40 mg to about 150 mg, about 45 mg to about 150 mg, about 50 mg to about 150 mg, about 55 mg to about 150 mg, about 60 mg to about 150 mg, about 65 mg to about 150 mg, about 70 mg to about 150 mg, about 75 mg to about 150 mg, about 80 mg to about 150 mg, about 85 mg to about 150 mg, about 90 mg to about 150 mg, about 95 mg to about 150 mg, about 100 mg to about 150 mg, about 105 mg to about 150 mg, about 110 mg to about 150 mg, about 115 mg to about 150 mg, about 120 mg to about 150 mg, about 125 mg to about 150 mg, about 130 mg to about 150 mg, about 135 mg to about 150 mg, about 140 mg to about 150 mg, or about 145 mg to about 150 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis.

[0123] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 10 mg to about 150 mg, about 15 mg to about 145 mg, about 20 mg to about 140 mg, about 25 mg to about 135 mg, about 30 mg to about 135 mg, about 35 mg to about 130 mg, about 40 mg to about 125 mg, about 45 mg to about 120 mg, about 50 mg to about 115 mg, about 55 mg to about 110 mg, about 60 mg to about 105 mg, about 65 mg to about 100 mg, about 70 mg to about 95 mg, about 75 mg to about 90 mg, or about 80 to about 85 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis.

[0124] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 10 mg to about 75 mg, about 15 mg to about 75 mg, about 20 mg to about 75 mg, about 25 mg to about 75 mg, about 30 mg to about 75 mg, about 35 mg to about 75 mg, about 40 mg to about 75 mg, about 45 mg to about 75 mg, about 50 mg to about 75, about 55 mg to about 75 mg, about 60 mg to about 75 mg, about 65 mg to about 75 mg, or about 70 mg to about 75 mg of Compound 1 , or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis.

[0125] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg, about 125 mg, about 130 mg, about 135 mg, about 140 mg, about 145 mg, or about 150 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis.

[0126] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 10 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis. In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 20 mg of Compound 1 , or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis. In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 40 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalentweight basis. In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 75 mg of Compound 1 , or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis. In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 125 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis. In certain embodiments, administering the effective amount of Compound I, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 150 mg of Compound 1 , or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis.

[0127] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject at least about 10 mg, at least about 15 mg, at least about 20 mg, at least about 25 mg, at least about 30 mg, at least about 35 mg, at least about 40 mg, at least about 45 mg, at least about 50 mg, at least about 55 mg, at least about 60 mg, at least about 65 mg, at least about 70 mg, at least about 75 mg, at least about 80 mg, at least about 85 mg, at least about 90 mg, at least about 95 mg, at least about 100 mg, at least about 105 mg, at least about 110 mg, at least about 115 mg, at least about 120 mg, at least about 125 mg, at least about 130 mg, at least about 135 mg, at least about 140 mg, or at least about 145 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis.

[0128] In some embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, to the subject comprises administering about 40 mg to about 75 mg Compound 1, or an equivalent amount of a pharmaceutically acceptable salt thereof on a free acid equivalent weight basis, to the subject.

[0129] In some embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, to the subject comprises administering about 40 mg Compound 1, or an equivalent amount of a pharmaceutically acceptable salt thereof on a free acid equivalent weight basis, to the subject.

[0130] In some embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, to the subject comprises administering about 75 mg Compound 1, or an equivalent amount of a pharmaceutically acceptable salt thereof on a free acid equivalent weight basis, to the subject.

[0131] In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 50 mg / m2to about 100 mg / m2, about 60 mg / m2to about 100 mg / m2, about 70 mg / m2to about 100 mg / m2, about 75 mg / m2to about 100 mg / m2, about 50 mg / m2to about 90 mg / m2, about 50 mg / m2to about 80 mg / m2, about 50 mg / m2to about 75 mg / m2, about 60 mg / m2to about 90 mg / m2, or about 70 mg / m2to about 80 mg / m25-azacitidine to the subject.

[0132] In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 50 mg / m2to about 100 mg / m2, about 60 mg / m2to about 100 mg / m2, about 70 mg / m2to about 100 mg / m2, about 75 mg / m2to about 100 mg / m2, about 50 mg / m2to about 90 mg / m2, about 50 mg / m2to about 80 mg / m2, about 50 mg / m2to about 75 mg / m2, about 60 mg / m2to about 90 mg / m2, or about 70 mg / m2to about 80 mg / m25-azacitidine to the subject daily.

[0133] In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 50 mg / m2, about 55 mg / m2, about 60 mg / m2, about 65 mg / m2, about 70 mg / m2, about 75 mg / m2, about 80 mg / m2, about 85 mg / m2, about 90 mg / m2, about 95 mg / m2, or about 100 mg / m25-azacitidine to the subject. In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 50 mg / m25-azacitidine to the subject. In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 60 mg / m25- azacitidine to the subject. In some embodiments, administering the effective amount of 5- azacitidine to the subject comprises administering about 70 mg / m25-azacitidine to the subject. In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 75 mg / m25-azacitidine to the subject. In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 80 mg / m25-azacitidine to the subject. In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 90 mg / m25-azacitidine to the subject. In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 100 mg / m25- azacitidine to the subject.

[0134] In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 50 mg / m2, about 55 mg / m2, about 60 mg / m2, about 65 mg / m2, about 70 mg / m2, about 75 mg / m2, about 80 mg / m2, about 85 mg / m2, about 90 mg / m2, about 95 mg / m2, or about 100 mg / m25-azacitidine to the subject daily. In someembodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 50 mg / m25-azacitidine to the subject daily. In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 60 mg / m25-azacitidine to the subject daily. In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 70 mg / m25- azacitidine to the subject daily. In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 75 mg / m25-azacitidine to the subject daily. In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 80 mg / m25-azacitidine to the subject daily. In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 90 mg / m25-azacitidine to the subject daily. In some embodiments, administering the effective amount of 5-azacitidine to the subject comprises administering about 100 mg / m25-azacitidine to the subject daily.

[0135] In some embodiments, the effective amount of 5-azacitidine is administered to the subject parenterally. In certain embodiments, the effective amount of 5-azacitidine is administered to the subject intravenously. In certain embodiments, the effective amount of 5- azacitidine is administered to the subject subcutaneously.

[0136] In some embodiments, the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, is administered orally.

[0137] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered orally to the subject once daily for 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 consecutive days. In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered orally to the subject once daily for 21 consecutive days.

[0138] In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered orally to the subject once daily for at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, or 36 consecutive days. In certain embodiments, Compound 1, or a pharmaceutically acceptable salt thereof, is administered orally to the subject once daily for at least 21 consecutive days.

[0139] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 10 mg to about 150 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis.

[0140] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering orally to the subject about 10 mg to about 150 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis.

[0141] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering orally to the subject about 10 mg to about 150 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis, daily.

[0142] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering orally to the subject about 10 mg to about 150 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis, once daily.

[0143] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering orally to the subject about 10 mg to about 150 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis, once daily for 21 consecutive days.

[0144] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, to the subject comprises administering about 40 mg to about 75 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis, to the subject.

[0145] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, to the subject comprises administering orally about 40 mg to about 75 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis, to the subject.

[0146] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, to the subject comprises administering orally about 40 mg to about 75 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis, to the subject, daily.

[0147] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, to the subject comprises administering orally about 40 mg to about 75 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis, to the subject, once daily.

[0148] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, to the subject comprises administering orally about 40 mg to about 75 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis, to the subject, once daily for 21 consecutive days.

[0149] In certain embodiments, the subject is in a fasting state. In certain embodiments, the subject is not in a fasting state. In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject the effective amount about 1 hour before a meal or about 2 hours after a meal.

[0150] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 10 mg to about 150 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis, about 1 hour before a meal or about 2 hours after a meal.

[0151] In certain embodiments, administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, comprises administering to the subject about 40 mg to about 75 mg of Compound 1, or a pharmaceutically acceptable salt thereof, on a free acid equivalent weight basis, about 1 hour before a meal or about 2 hours after a meal.

[0152] In certain embodiments, the methods comprise administering an effective amount of a pharmaceutically acceptable salt of Compound 1 to the subject. In certain embodiments, the pharmaceutically acceptable salt of Compound 1 is a potassium salt of Compound 1. In certain embodiments, the potassium salt is a hydrate. In certain embodiments, the potassium salt is a monohydrate.

[0153] In some embodiments, the AML is resistant to B-cell lymphoma inhibitors. In some embodiments, the AML is resistant to venetoclax. In some embodiments, the AML comprises a venetoclax resistance mechanism selected from the group consisting of upregulation of anti-apoptotic proteins, p53 inactivation, activating kinase mutations, and upregulation of oxidative phosphorylation.

[0154] In some embodiments, the AML is characterized by having a FLT3 ITD mutation. In some embodiments, the AML is characterized by having a TP53 mutation. In some embodiments, the AML is characterized by upregulation of MCL-1.

[0155] In certain embodiments, the subject has previously been administered at least one prior line of therapy. In certain embodiments, the subject has previously been administered fewer than five prior lines of therapy. In certain embodiments, the subject has previouslybeen administered one, two, three, or 4 prior lines of therapy. In certain embodiments, the subject has not been administered a prior line of therapy.

[0156] In certain embodiments, the subject has previously been administered at least one and no more than 5 prior lines of therapy.

[0157] Prior lines of therapy include, but are not limited to, surgery, radiation therapy (e.g., external beam radiation therapy or internal radiation therapy), chemotherapy (e.g., alkylating agents, nitrosoureas, anti-metabolites, plant alkaloids and natural products, anti-tumor antibiotics, hormonal agents, and biological response modifiers), gene therapy, DNA therapy, viral therapy (e.g., oncolytic virus therapy), RNA therapy, adjuvant therapy, and immunotherapy (e.g., immune checkpoint inhibition, adoptive cell therapies, (e.g., tumorinfiltrating lymphocyte therapy, engineered T-cell receptor therapy, CAR T-cell therapy, natural killer cell therapy), or monoclonal antibodies).

[0158] In another aspect, provided herein are methods of treating AML in a subject in need thereof, comprising administering to the subject any one of the pharmaceutical compositions described herein to the subject.

[0159] In certain embodiments, the subject is a human. In certain embodiments, the subject is an adult human.EXAMPLES

[0160] In order that the disclosure described herein may be more fully understood, the following examples are set forth. The synthetic and biological examples described in this application are offered to illustrate the compounds, pharmaceutical compositions, and methods provided herein and are not to be construed in any way as limiting their scope.Example 1: Synthesis of 6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6- difluorophenyI)-N-methyIimidazo[l,5-a]pyrazine-l-carboxamide (Compound 1)Synthesis of 1-a: ethyl 2-(5-bromopyrazin-2-yl)-2-[(diphenylmethylidene) aminol acetate

[0161] 2,5 -Dibromopyrazine (10 g, 42 mmol, 1 equiv.), ethyl 2- [(diphenylmethylidene)amino]acetate (11.8 g, 44 mmol, 1.05 equiv.), tetrabutylammonium bromide (TBAB) (13.6 g, 42 mmol, 1 equiv.) and K2CO3 (17.4 g, 126 mmol, 3 equiv.) in (N- methyl-2-pyrrolidone) NMP (200 mL) were stirred overnight at 100 °C in an oil bath. The reaction mixture was cooled and filtered. The filtrate was diluted with 200 mL of water. The resulting solution was extracted with 2 x 200 mL of ethyl acetate and the organic layerscombined. The resulting mixture was washed with 2 x 200 mL of water. The mixture was dried over anhydrous sodium sulfate and concentrated. The residue was applied to a silica gel column, eluting with ethyl acetate / petroleum ether (PE) (1 / 10). The collected fractions were combined and concentrated to give ethyl 2-(5-bromopyrazin-2-yl) -2- [(diphenylmethylidene) amino] acetate (8 g, 45% yield) as a yellow solid.LCMS (ES, m / z): [M+H]+: 424Synthesis of 1-b: ethyl 2-amino-2-(5-bromopyrazin-2-yl) acetate

[0162] Into a 250 mL round-bottom flask, was placed ethyl 2-(5-bromopyrazin-2-yl)-2- [(diphenylmethylidene)amino] acetate (8 g, 18.8 mmol, 1 equiv.), tetrahydrofuran (THF) (10 mL) and HC1 (aqueous, 1 M) (20 mL). The resulting solution was stirred for 30 min at 25 °C. The solution formed was diluted with 50 mL of water and extracted with 2 x 50 mL of dichloromethane. The aqueous layers were adjusted to pH 8 with NH3.H2O and further extracted with 3 x 50 mL of dichloromethane. The combined organic layers were dried over anhydrous sodium sulfate and concentrated. Ethyl 2-amino-2-(5-bromopyrazin-2-yl)acetate (4.7 g, 96% yield) was isolated as a yellow solid which was used in next step directly without further purification.LCMS (ES, m / z): [M+H]+: 260Synthesis of 1-c: ethyl 6-bromoimidazo IT, 5-a]pyrazine-l -carboxylate

[0163] Into a 50 mL round-bottom flask, was placed ethyl 2-amino-2-(5-bromopyrazin-2-yl) acetate (4.2 g, 0.02 mol, 1 equiv.) and triethyl orthoformate (20 mL). The resulting solution was stirred for 2 h at 80 °C in an oil bath. The reaction mixture was cooled, and the solids collected by filtration. Air drying gave ethyl 6-bromoimidazo [l,5-a]pyrazine-l -carboxylate (2.2 g, 50% yield) as a brown solid.LCMS (ES, m / z): [M+H]+: 270Synthesis of 1-d: 2,4-difluoro-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)aniline

[0164] 3 -Bromo-2,4-difluoroaniline (10 g, 48 mmol, 1 equiv.), [1,1 '- Bis(diphenylphosphino)ferrocene]dichloropalladium(II) (Pd(dppf)Ch) (3.5 g, 4.8 mmol, 0.1 equiv.), bis(pinacolato)diboron (18.3 g, 72 mmol, 1.5 equiv.) and potassium acetate (KOAc) (14.2 g, 144.2 mmol, 3 equiv.) were dissolved in dioxane (240 mL). The resulting solutionwas stirred overnight at 100 °C in an oil bath. The reaction mixture was cooled, and the solids removed by filtration. The filtrate was concentrated and diluted with dichloromethane (DCM) (100 mL), then washed with 2 x 100 mL of water and 100 mL of brine. The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was applied to a silica gel column, eluting with ethyl acetate / petroleum ether (1 / 10). 2,4-Dilluoro-3-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)aniline (8 g, 65% yield) was isolated as a yellow solid.LCMS (ES, m / z): [M+H]+: 256Synthesis of 1-e: ethyl 6-(3-amino-2,6-difluorophenyl)imidazo|T,5-a]pyrazine-l-carboxylate

[0165] Ethyl 6-bromoimidazo[l,5-a]pyrazine-l-carboxylate (500 mg, 1.9 mmol, 1 equiv.), 2,4-difluoro-3- (4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2-yl)aniline (708 mg, 2.8 mmol, 1.5 equiv.), Pd(dppf)CL (135 mg, 0.2 mmol, 0.1 equiv.), K2CO3 (767 mg, 5.6 mmol, 3 equiv.) in dioxane (10 mL) and H2O (2 mL) were stirred for 1 h at 60 °C in an oil bath. The reaction mixture was cooled, diluted with water (20 mL) and extracted with 3 x 20 mL of dichloromethane. The organic layers were dried over anhydrous sodium sulfate and concentrated. The residue was applied to a silica gel column and eluted with ethyl acetate / PE (1 / 2). Ethyl 6-(3-amino-2,6-difluorophenyl)imidazo[l,5-a]pyrazine-l-carboxylate (200 mg 34% yield) was isolated as a brown solid.LCMS (ES, m / z): [M+H]+: 319Synthesis of 1-f: ethyl 6-[3-(5-chloro-2-methoxypyridine-3-sulfonamido)-2,6- difluorophenyl]imidazo[l,5-a]pyrazine-l-carboxylate

[0166] Ethyl 6-(3-amino-2, 6-difluorophenyl)imidazo[l,5-a]pyrazine-l- carboxylate (150 mg, 0.5 mmol, 1 equiv.) in DCM (5 mL) was treated with pyridine (186 mg, 2.3 mmol, 5 equiv.), then 5 -chloro-2-methoxypyridine-3 -sulfonyl chloride (137 mg, 0.6 mmol, 1.2 equiv.). The resulting solution was stirred overnight. The resulting mixture was concentrated and purified by Flash- Prep- HPLC with the following conditions: Column, WelFlashTM Cl 8- I, Spherical Cl 8 20-40 pm; mobile phase: 0.1% Formic Acid / 5-70% MeCN over 15 min; Detector, 254 & 220 nm. Ethyl 6-[3-(5-chloro-2- methoxypyridine-3-sulfonamido)-2,6- difluorophenyl]imidazo[l,5-a]pyrazine-l-carboxylate (320 mg 97% yield) was isolated as a yellow solid.LCMS (ES, m / z): [M+H]+: 524Synthesis of 1-g: 6-[3-(5-chloro-2-methoxypyridine-3-sulfonamido)-2,6- difluorophenyl]imidazo[l,5-a] pyrazine- 1 -carboxylic acid

[0167] Ethyl 6-[3-(5-chloro-2-methoxypyridine-3-sulfonamido)-2,6- difluorophenyl]imidazo[l,5-a]pyrazine-l -carboxylate (200 mg, 0.4 mmol, 1 equiv.), methanol (MeOH) (2 mL), THF (2 mL) , H2O (2 mL) and LiOH (27 mg, 1.1 mmol, 3 equiv.) were stirred for 1 h at 60 °C in an oil bath. After concentration, the crude product was purified by Flash-Prep-high performance liquid chromatography (HPLC) with the following conditions: Column, WelFlashTM C 18-1, Spherical C18 20-40 pm; mobile phase: 5-60% acetonitrile (MeCN) / 0.1% ammonia over 15 min; Detector, 254 nm. 6-[3-(5-Chloro-2- methoxypyridine-3-sulfonamido)-2,6- difluorophenyl] imidazo [ 1 ,5-a] pyrazine- 1 -carboxylic acid (170 mg, 90% yield) was isolated as a yellow solid.LCMS (ES, m / z): [M+H]+: 496Synthesis of 6-[3-(5-chloro-2-methoxypyridine- 3-sulfonamido)-2,6-difluorophenyl]-N- methyli midazol 1 ,5-a]pyrazine- 1 -carboxamide

[0168] 6-[3-(5-Chloro-2-methoxypyridine-3-sulfonamido)-2,6- difluorophenyl]imidazo[l,5- a]pyrazine-l -carboxylic acid (170 mg, 0.3 mmol, 1 equiv.) in N,N-dimethylformamide (DMF) (4 mL) was treated with diisopropylethylamine (DIEA) (133 mg, 1 mmol, 3 equiv), methylamine hydrochloride (16 mg, 0.5 mmol, 1.5 equiv.) and 1- [Bis(dimethylamino)methylene]- 1H- 1 ,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU) (195 mg, 0.5 mmol, 1.5 equiv.). The resulting solution was stirred for 1 hr, then concentrated. The crude product was purified by Flash-Prep-HPLC with the following conditions: Column, WelFlashTM Cl 8-1, Spherical Cl 8 20-40 pm, 120 g; mobile phase: 5-60% MeCN / 0.1% formic acid over 20 min. 6-[3-(5-Chloro-2- methoxypyridine-3-sulfonamido)-2,6-difluorophenyl]-N-methylimidazo[l,5-a]pyrazine-l- carboxamide (43 mg, 25% yield) was isolated as an off-white solid.Liquid chromatography / mass spectrometry (LCMS) (ES, m / z): |M+H|+: 509 'H nuclear magnetic resonance spectroscopy (NMR) (300 MHz, DMSO-tfc) 3 10.46 (s, 1H), 9.51 (d, J= 1.6 Hz, 1H), 8.66 (d, 7= 5.0 Hz, 2H), 8.51 (d, 7 = 2.6 Hz, 1H), 8.43 (d, 7 = 4.9 Hz, 1H), 8.09 (d, 7 = 2.6 Hz, 1H), 7.42 (td, 7 = 8.8, 5.8 Hz, 1H), 7.25 (td, 7 = 9.3, 1.4 Hz, 1H), 3.92 (s, 3H), 2.84 (d, 7= 4.7 Hz, 3H).Example 2: Preparation of Compound 1 Potassium Salt

[0169] To a stirred mixture of 6-(3-((5-chloro-2-methoxypyridine)-3-sulfonamido)-2,6- difluorophenyl)-N-methylimidazo[l,5-a]pyrazine-l-carboxamide (as prepared in Example 1) in aqueous isopropyl alcohol (IPA) was slowly added 1.1 equivalents of an aqueous KOH solution and the solution heated. The resulting mixture was cooled, the solid collected, washed with IPA / water, and then dried under heat and vacuum to afford potassium ((5- chloro-2-methoxypyridin-3-yl)sulfonyl)(2,4-difluoro-3-(l-(methylcarbamoyl)imidazo[l,5- a]pyrazin-6-yl)phenyl)amide.19F NMR (400 MHz, d6-DMSO): -129.62 and -127.72 ppm.Example 3: In Vivo Tumor Growth Inhibition Study - HC-7366 and Venetoclax Combination Therapy

[0170] The effect of combination treatment with HC-7366 and venetoclax on tumor growth inhibition was investigated in a MV4-11 mouse xenograft tumor model.Study DesignAnimalsSpecies & Strain: BALB / c nude miceSupplier: Beijing AniKeeper Biotech Co., Ltd.Age: 6 to 8 weeksTotal Number: 120 mice including spare, study used 60 miceSex: FemaleBody Weight: 18 to 24 gAnimal MaintenanceQuarantine: Animals were quarantined for 7 days before the study. The general health of the animals was evaluated by a veterinarian, and complete health checks were performed. Animals with abnormalities were excluded prior to the study.Housing: General procedures for animal care and housing were in accordance with the standard, Commission on Life Sciences, National Research Council, Standard operating procedures (SOPs) of Pharmaron, Inc. The mice were kept in laminar flow rooms at constant temperature and humidity with 3-5 mice in each cage. Animals were housed in polycarbonate cage which is in the size of 300 x 180 x 150 mm3and in an environmentally monitored, well- ventilated room maintained at a temperature of (22 ± 3 °C) and a relative humidity of 40%-80%. Fluorescent lighting provided illumination approximately 12 hours per day. The bedding material is soft wood, which is changed once per week.Diet: Animals had free access to irradiation sterilized dry granule food during the entire study period except for time periods specified by the protocol.Water: Sterile drinking water in a bottle was available to all animals ad libitum during the quarantine and study periods. The bottle and the stopper with attached sipper tube were autoclaved prior to use. Samples of water from the animal facility were analyzed and results of water analysis were retained in the facility records and were reviewed by the veterinarian, or designee, to assure that no known contaminants were present that could interfere with or affect the outcome of studies.Groups and Treatments

[0171] Groups and treatments were started when the mean tumor volume reached about 154 mm3on Day 13 after cell inoculation. Based on the tumor volume and body weight, mice were assigned to respective groups such that the average starting tumor size is the same for each treatment group. The study groups and number of animals per group are shown in Table 1.Table 1: Groups and TreatmentsPO - by mouth, BID - twice a day, QD - once a dayVehicle ControlVehicle for HC-7366: 5% (v / v) DMSO and 95% (v / v) Captisol (20% w / v) in PBS (final pH7.4)Vehicle for Venetoclax: 0.5% carboxymethylcellulose (CMC)FormulationThe formulations were prepared according to Table 2.Table 2: FormulationExperimental Method and Measurement ParametersMethod for Tumor InoculationEach mouse was inoculated subcutaneously on the right flank with MV4-11 tumor cells (1 x 107) in IMEM and Matrigel mixture (1 :1) without serum for tumor development. The treatments were started when the mean tumor size reached approximately 154 mm3. Mice then were assigned to groups such that the mean tumor volume was the same for each treatment group. The treatments were administered to the tumor-bearing mice according to the study design shown in Table 1.Measurement ParametersFor routine monitoring, all study animals were monitored not only for tumor growth but also behavior such as mobility, food, and water consumption (by cage side checking only), body weight (BW), eye / hair matting and any other abnormal effect. Any mortality and / or abnormal clinical signs were recorded. Sponsors were notified immediately if abnormal clinical signs, or if tolerability issues were observed.Body WeightBody weights of all animals were measured and recorded three times weekly throughout the study. Body weight change, expressed in %, was calculated using the following formula:BW change (%) = (BWDayx / BWDayo) x 100Tumor MeasurementsThe measurements of tumor size were conducted three times weekly with a caliper and recorded. The tumor volume (mm3) was estimated using the formula: TV = a x b2 / 2, where “a” and “b” were long and short diameters of a tumor, respectively. The TVs were used for calculation of the tumor growth inhibition (TGI, an indicator of antitumor effectiveness) value using the formula: TGI = (1-T / C) x 100%, where “T” and “C” was the mean relative volumes (% tumor growth) of the tumors in the treated and the control groups, respectively, on a given day after tumor inoculation.Samples CollectionThe detailed sample collection and processing are shown in Table 3.Table 3: Samples collectionData Acquisition and Statistical AnalysisStatistical Analysis: All statistical tests were conducted, and the level of significance was set at 5% or P < 0.05. The group means and standard deviation were calculated for allmeasurement parameters as study designed. One-way analysis of variance (ANOVA) followed by Dunnett T3 test were applied among the groups.The antitumor efficacy of HC-7366 alone or in combination with Venetoclax in MV4-11 human AML xenograft model is summarized in Table 4 and displayed graphically in FIG. 1AHC-7366 alone at 0.5 mg / kg or 2 mg / kg BID did not significantly inhibit tumor growth with TGI values of 0.9% or 0.2% (P values > 0.05 vs. vehicle control group), respectively. Venetoclax alone at 50 mg / kg QD, and in combination with HC-7366 at 0.5 mg / kg or 2 mg / kg BID showed significant antitumor efficacy with TGI values of 44.9%, 33.6%, and 55.2% ( values < 0.05 vs. vehicle control group), respectively. There was a transient combination benefit observed with venetoclax + HC-7366 at 2 mg / kg, but no combination benefit was observed when 0.5 mg / kg HC-7366 was combined with venetoclax.Table 4. Antitumor activity of treatment groupsNote: a. Mean ± SEM; c. vs. Vehicle control, via Dunnett T3 ; b. TGI = (1-T / C) x 100%, T / C = 100*(Vt relative volumes (% tumor growth) / VC relative volumes)Treatments were well-tolerated during the treatment period, and no obvious BW loss or clinical abnormalities were observed in any of the groups tested (FIG. IB).Example 4: In Vivo Tumor Growth Inhibition Study - HC-7366 with Venetoclax and / or 5-Azacitidine (AZA)

[0173] The effect of combination treatment of HC-7366 with venetoclax or venetoclax and 5- azacitidine on tumor growth was investigated in a MV4-11 mouse xenograft tumor model.Study DesignAnimalsSpecies & Strain: BALB / c nude miceSupplier: Beijing AniKeeper Biotech Co., Ltd.Age: 6 to 8 weeksTotal Number: 384 mice including spare, study used 192 miceSex: FemaleBody Weight: 17 to 22 gAnimal MaintenanceQuarantine: Animals were quarantined for 7 days before the study. The general health of the animals was evaluated by a veterinarian, and complete health checks were performed.Animals with abnormalities were excluded prior to the study.Housing: General procedures for animal care and housing were in accordance with the standard, Commission on Life Sciences, National Research Council, Standard operating procedures (SOPs) of Pharmaron, Inc. The mice were kept in laminar flow rooms at constant temperature and humidity with 3-5 mice in each cage. Animals were housed in polycarbonate cage which is in the size of 300 x 180 x 150 mm3and in an environmentally monitored, well- ventilated room maintained at a temperature of (22 ± 3 °C) and a relative humidity of 40%-80%. Fluorescent lighting provided illumination approximately 12 hours per day. The bedding material is soft wood, which is changed once per week.Diet: Animals had free access to irradiation sterilized dry granule food during the entire study period except for time periods specified by the protocol.Water: Sterile drinking water in a bottle was available to all animals ad libitum during the quarantine and study periods. The bottle and the stopper with attached sipper tube were autoclaved prior to use. Samples of water from the animal facility were analyzed and results of water analysis were retained in the facility records and were reviewed by the veterinarian, or designee, to assure that no known contaminants were present that could interfere with or affect the outcome of studies.Groups and Treatments

[0174] Groups and treatments were started when the mean tumor volume reached about 158 mm3on Day 12 after cell inoculation. Based on the tumor volume and body weight, mice were assigned to respective groups such that the average starting tumor size is the same for each treatment group. The study groups and number of animals per group are shown in Table 5 and Table 6.Table 5: Groups and Treatments for Efficacy StudyPO - by mouth, IP - intraperitoneal , BID - twice a day, QD - once a day, BIW - twice weeklyTable 6: Groups and Treatments for PK / PD StudyPO - by mouth, IP - intraperitoneal, BID - twice a day, QD - once a day, BIW - twice weekly VehiclesVehicle for HC-7366: 5% (v / v) DMSO and 95% (v / v) Captisol (20% w / v) in PBS (pH 7.4)Vehicle for Venetoclax: 0.5% CMCVehicle for 5-AZA: 20% NMP + 40% PEG400 + 40% salineFormulationThe formulations were prepared according to Table 7 and Table 8.Table 7: Formulation for Efficacy StudyTable 8: Formulation for PK / PD StudyExperimental Method and Measurement ParametersMethod for Tumor InoculationEach mouse was inoculated subcutaneously on the right flank with MV4-11 tumor cells (1 x 107) in IMDM and Matrigel mixture (1 :1) without serum for tumor development. The treatments were started when the mean tumor size reached approximately 158 mm3. Mice then were assigned to groups such that the mean tumor volume was the same for each treatment group. The treatments were administered to the tumor-bearing mice according to the study design shown in Table 5 or Table 6.Measurement ParametersFor routine monitoring, all study animals were monitored not only tumor growth but also behavior such as mobility, food, and water consumption (by cage side checking only), body weight (BW), eye / hair matting and any other abnormal effect. Any mortality and / or abnormal clinical signs were recorded. Sponsor were notified immediately if abnormal clinical signs, or if tolerability issues were observed.Body WeightBody weights of all animals were measured and recorded three times weekly throughout the study. Body weight change, expressed in %, was calculated using the following formula:BW change (%) = (BWDayx / BWDayo) x 100Tumor MeasurementsThe measurements of tumor size were conducted three times weekly with a caliper and recorded. The tumor volume (mm3) was estimated using the formula: TV = a x b2 / 2, where “a” and “b” were long and short diameters of a tumor, respectively. The TVs were used for calculation of the tumor growth inhibition (TGI, an indicator of antitumor effectiveness) value using the formula: TGI = (1-T / C) x 100%, where “T” and “C” was the mean relative volumes (% tumor growth) of the tumors in the treated and the control groups, respectively, on a given day after tumor inoculation.Samples CollectionThe detailed sample collection and processing are shown in Table 9.Table 9: Samples collection for PK / PD studyData Acquisition and Statistical AnalysisStatistical Analysis: All statistical tests were conducted, and the level of significance was set at 5% or P < 0.05. The group means and standard deviation were calculated for all measurement parameters as study designed. One-way analysis of variance (ANOVA) followed by Dunnett T3 test or T-test were applied among the groups.The antitumor efficacy of HC-7366 alone or in combination with Venetoclax and / or 5-AZA in MV4-11 human AML xenograft model is summarized in Table 10 and displayed graphically in FIG. 2 A and FIG. 2B.HC-7366 alone at 0.5 mg / kg or 1 mg / kg BID was not efficacious in this model with TGI values of -1.0% or -1.7% (P values > 0.05 vs. vehicle control group), respectively.Venetoclax alone at 50 mg / kg QD and in combination with HC-7366 at 0.5 mg / kg or 1 mg / kg BID showed similar significant antitumor effects with TGI values of 61.8%, 61.4%, and 61.6% (P values < 0.001 vs. vehicle control group), respectively. Combination of Venetoclax and HC-7366 did not improve the effects of venetoclax monotherapy.5-AZA alone at 5 mg / kg BIW and in combination with HC-7366 at 0.5 mg / kg or 1 mg / kg BID showed significant antitumor effects with TGI values of 62.6%, 57.4%, and 45.2% (P values < 0.001 vs. vehicle control group), respectively. The combination of 5-AZA and HC- 7366 did not improve the effect of 5-AZA monotherapy.Venetoclax at 50 mg / kg in combination with 5-AZA at 5 mg / kg showed significant antitumor effect with TGI value of 89.9% (P value < 0.001 vs. vehicle control group) and after adding HC-7366 at 0.5 mg / kg or 1 mg / kg BID produced significant anti-tumor effects with TGI values of 92.9% and 95.6% (P values < 0.001 vs. vehicle control group), respectively. In addition, at the end of study (Day 40 post tumor inoculation), the triple combination of HC-7366 at 1 mg / kg significantly improved the effect of Venetoclax + 5-AZA.Table 10. Antitumor activity of treatment groupsNote: a. Mean ± SEM; b. TGI = (1-T / C) x 100%, T / C = 100*(Vt relative volumes (% tumor growth) / VC relative volumes); c. vs. Vehicle control, via Dunnett T3.Some animals in Venetoclax + 5-AZA, Venetoclax + 5-AZA + HC-7366 combination groups showed body weight loss. One animal in Venetoclax + 5-AZA group and two animals in Venetoclax 50 mg / kg + 5-AZA 5 mg / kg + HC-7366 0.5 mg / kg group were found dead during the late stage of treatment. No other clinical abnormalities were observed in any of the groups tested.Example 5: Venetoclax Resistance- Associated Markers Protein Analysis

[0175] The combination benefit of HC-7366 with venetoclax and / or 5-AZA was associated with reduction in venetoclax resistance-associated markers as evaluated using the study protocol described below.Simple Western Protein Analysis

[0176] Methods describing protein extraction and analysis have been described elsewhere (Stokes et al., Pharmaceutics, 2022, 14(10), 2233; Stokes et al., Clin Cancer Res, 2023, 29(23), 4870-4882). Total protein was extracted from tumor tissues and homogenized by polytron in lysis buffer consisting of 2x Laemmli SDS sample buffer (Novex), supplemented with 10% BME (Gibco), IX benzonase (EMD Millipore Sigma), phosphatase inhibitors (Roche) and Mini protease inhibitor tablet (Roche). The homogenate was incubated at room temperature for 10 min, then boiled for 10 min, followed by centrifugation for 10 min at max speed on a benchtop centrifuge. Protein detection was performed on the Jess SimpleWestern high-throughput protein analysis platform (ProteinS imple) according to manufacturer’s protocol using either a 12-230 kDa Separation Module (ProteinS imple, SM-W004) or a 2-40 kDa Separation Module (ProteinSimple, SM-W009) and Total Protein Detection Module (ProteinSimple, DM-TP01 ). Antibodies: MCL1 (ProteinTech, 1 :50, Cat. #16225-1 -AP), S100A8 (ProteinTech, 1 :50, Cat. # 15792-1 -AP). S100A9 (ProteinTech, 1 :50, Cat. # 26992-1-AP), TIMM17A (Cell Signaling Technologies, 1:50, Cat. # 45948S).

[0177] Tumors from the groups defined in the PK / PD component of Example 4 were collected on Day 4, lysed, and analyzed for the presence of venetoclax resistance-associated markers: MCL1, an anti-apoptotic protein (FIG. 3A), S100A8 (FIG. 3B), and TIMM17A, a translocase of inner mitochondrial membrane (FIG. 3C) using Simple Western. Changes in S100A9 expression were nearly identical to S100A8 (not shown). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, one-way ANOVA. A significant reduction in all tested venetoclax resistance-associated markers was observed in the triple combination treatment (HC-7366 at 1 mg / kg, venetoclax, and 5-AZA).Example 6: In Vitro Studies - Combination Benefit of HC-7366 and Venetoclax and / or 5-AZA in TP53 Mutant AML Cell Lines

[0178] KG-1 (p4) cells were cultured in Iscove’s Modified Dulbecco’s Medium (ATCC, 30- 2005) supplemented with 20% fetal bovine serum, and Kasumi-1 (p8) cells were cultured in RPMI-1640 (ATCC, 30-2001) supplemented with 20 % fetal bovine serum. For viability experiments, cells were seeded at 5xl06cells per well in 12-well plates in single, double or triple combinations of: HC-7366 (0.04 and 0.1 pM), Azacitidine (1 |iM, Sigma, A2385), and Venetoclax (10-fold dilutions from 0.00003-3 pM, ABT-199, GDC-0199). After 72 hours of treatment, viability was assessed using Annexin V (Biolegend, 640918 at 1:200 dilution) and 7AAD (Biolegend, 420404 at 1:25 dilution) staining which was analyzed with a Cytek Aurora flow cytometer.

[0179] Cell viability curves for treated KG-1 cells are provided in FIGs. 4A-4C. Cell viability curves for treated Kasumi-1 cells are provided in FIGs. 4D-4F. Venetoclax + HC- 7366 + 5-azacitidine showed triplet combination benefit in both KG-1 and Kasumi-1 TP53 mutant cell lines in vitro.Example 7: In Vivo Tumor Growth Inhibition and PKPD Study - HC-7366 alone or combination with Venetoclax and / or 5-AZA in KG-1 human AML xenograft model Study DesignAnimalsSpecies & Strain: NOD SCID miceSupplier: Beijing AniKeeper Biotech Co., Ltd.Age: 6 to 8 weeksTotal Number: 288 mice including spare, study used 192 miceSex: FemaleBody Weight: 18 to 22 gAnimal MaintenanceQuarantine: Animals were quarantined for 7 days before the study. The general health of the animals was evaluated by a veterinarian, and complete health checks were performed.Animals with abnormalities were excluded prior to the study.Housing: General procedures for animal care and housing were in accordance with the standard, Commission on Life Sciences, National Research Council, Standard operating procedures (SOPs) of Pharmaron, Inc. The mice were kept in laminar flow rooms at constant temperature and humidity with 3-5 mice in each cage. Animals were housed in polycarbonate cage which is in the size of 300 x 180 x 150 mm3and in an environmentally monitored, well- ventilated room maintained at a temperature of (22 ± 3 °C) and a relative humidity of 40%-80%. Fluorescent lighting provided illumination approximately 12 hours per day. The bedding material is soft wood, which is changed once per week.Diet: Animals had free access to irradiation sterilized dry granule food during the entire study period except for time periods specified by the protocol.Water: Sterile drinking water in a bottle was available to all animals ad libitum during the quarantine and study periods. The bottle and the stopper with attached sipper tube were autoclaved prior to use. Samples of water from the animal facility were analyzed and results of water analysis were retained in the facility records and were reviewed by the veterinarian, or designee, to assure that no known contaminants were present that could interfere with or affect the outcome of studies.Groups and Treatments

[0180] Groups and treatments were started when the mean tumor volume reached about 201 mm3on Day 21 after cell inoculation for Efficacy Study and PKPD study. Based on the tumor volume and body weight, mice were assigned to respective groups such that the average starting tumor size is the same for each treatment group. The study groups and number of animals per group are shown in Table 11.Table 11: Groups and Treatments for Efficacy StudyPO - by mouth, BID - twice a day, QD - once a dayTable 12: Groups and Treatments for PKPD StudyPO - by mouth, BID - twice a day, QD - once a dayVehicle ControlVehicle for HC-7366: 5% (v / v) DMSO and 95% (v / v) Captisol (20% w / v) in PBS (final pH7.4)Vehicle for Venetoclax: 0.5% carboxymethylcellulose (CMC)Vehicle for 5-AZA: 20% NMP + 40% PEG400 + 40% salineFormulationThe formulations were prepared according to Table 13.Table 13: Formulation for Efficacy StudyTable 14: Formulation for PKPD StudyExperimental Method and Measurement ParametersMethod for Tumor InoculationEach mouse was inoculated subcutaneously on the right flank with KG-1 tumor cells (5 x 106) in 100 pl RPMI 1640 for tumor development. The treatments were started when the mean tumor size reached approximately 201 mm3. Mice then were assigned to groups such that the mean tumor volume was the same for each treatment group. The treatments were administered to the tumor-bearing mice according to the study design shown in Table 11. Measurement ParametersFor routine monitoring, all study animals were monitored not only for tumor growth but also behavior such as mobility, food, and water consumption (by cage side checking only), body weight (BW), eye / hair matting and any other abnormal effect. Any mortality and / or abnormal clinical signs were recorded. Sponsors were notified immediately if abnormal clinical signs, or if tolerability issues were observed.Body WeightBody weights of all animals were measured and recorded three times weekly throughout the study. Body weight change, expressed in %, was calculated using the following formula:BW change (%) = (BWDayx / BWDayo) x 100Tumor MeasurementsThe measurements of tumor size were conducted three times weekly with a caliper and recorded. The tumor volume (mm3) was estimated using the formula: TV = a x b2 / 2, where “a” and “b” were long and short diameters of a tumor, respectively. The TVs were used for calculation of the tumor growth inhibition (TGI, an indicator of antitumor effectiveness) value using the formula: TGI = (1-T / C) x 100%, where “T” and “C” was the mean relative volumes (% tumor growth) of the tumors in the treated and the control groups, respectively, on a given day after tumor inoculation.Sample CollectionThe detailed sample collection and processing data are shown in Table 15 and Table 16.Table 15: Sample Collection for Efficacy StudyTable 16: Sample Collection for PKPD StudyData Acquisition and Statistical AnalysisStatistical Analysis: All statistical tests were conducted, and the level of significance was set at 5% or P < 0.05. The group means and standard deviation were calculated for all measurement parameters as study designed. One-way analysis of variance (ANOVA) followed by Dunnett T3 test or T-test were applied among the groups.The antitumor efficacy of HC-7366 alone or in combination with Venetoclax and / or 5- AZA in KG-1 human AML xenograft model is summarized in Table 17 and displayed graphically in FIG. 5A.HC-7366 alone at 0.5 mg / kg or 1 mg / kg BID showed significant antitumor effects with TGI value of 69.5% and TR value of 27.9% (P values < 0.05 vs. vehicle control group), respectively.Venetoclax alone at 50 mg / kg QD showed minor antitumor trend with TGI value of 30.2% (P value > 0.05 vs. vehicle control group). Venetoclax at 50 mg / kg QD in combination with HC- 7366 at 0.5 mg / kg or 1 mg / kg BID produced significant antitumor effects with TR values of 96.0% and 100.0% (P values < 0.001 vs. vehicle control group), respectively.5- AZA alone at 2 mg / kg BIW showed significant antitumor effect with TGI value of 45.4% (P value < 0.05 vs. vehicle control group), when in combination with HC-7366 at 0.5 mg / kgor 1 mg / kg BID, more significant antitumor effects were observed with TR values of 51.5% and 62.5% (P values < 0.001 vs. vehicle control group), respectively.Venetoclax at 50 mg / kg in combination with 5-AZA at 2 mg / kg showed significant antitumor activity with TGI value of 66.4% (P value < 0.05 vs. vehicle control group) and after adding HC-7366 at 0.5 mg / kg or 1 mg / kg BID produced greater significant antitumor effects with TR values of 95.5% and 100.0% (P values < 0.001 vs. vehicle control group), respectively.Table 17. Antitumor activity of treatment groups in KG-1 tumor modelNote: a. Mean ± SEM; b. TGI = (1-T / C) x 100%, T / C = 100*(Vt relative volumes (% tumorr growth) / Vc relative volumes); c. vs. Vehicle control, via Dunnett T3; d. Tumor Regression= (l-(Td / T0)) x 100%.Treatments were well-tolerated during the treatment period and no clinical abnormalities were observed in any of the groups tested. The antitumor effects of compound HC-7366 alone or in combination with Venetoclax and / or 5-AZA in KG-1 human AML xenograft model were evaluated. A combination of Venetoclax and HC-7366 demonstrated improved effects over treatment with Venetoclax or HC-7366 alone. A combination of 5-AZA and HC-7366 demonstrated improved effects over treatment with 5-AZA or HC-7366 alone. Finally, the triple combination of Venetoclax + 5-AZA + HC-7366 demonstrated significantly improved effects over treatment with Venetoclax + 5-AZA. Regarding the safety profile, animals treated with HC-7366 + 5-AZA, or Venetoclax + 5-AZA + HC-7366 showed nominal 10%-l 5% body weight loss (FIG. 5B).PKPD Study Methods and ResultsAn analysis of tumor lysates was performed by SimpleWestem to evaluate the effect of HC- 7366 combination with 5-AZA and venetoclax on protein expression of pharmacodynamic markers in tumors collected on Day 8 of treatment. Markers that correlate with efficacy in this model include reductions in the antiapoptotic protein MCL-1, inhibition of TIMM17A and TIMM23, subunits of a mitochondrial protein import complex, repression of cell cycle regulators phosphorylated (pS807 / pS811) retinoblastoma protein (RB), total RB, and cyclin dependent kinase 1 (CDK1), as well as reductions in the proto-oncogenes cMYC and HIFla. Methods for SimpleWestem Protein AnalysisTotal protein was extracted from tumor tissues and homogenized by polytron in lysis buffer consisting of 2x Laemmli SDS sample buffer (Novex), supplemented with 10% BME (Gibco), IX benzonase (HMD Millipore Sigma), phosphatase inhibitors (Roche) and Mini protease inhibitor tablet (Roche). The homogenate was incubated at room temperature for 10 min, then boiled for 10 min, followed by centrifugation for 10 min at max speed on a benchtop centrifuge. Protein detection was performed on the Jess SimpleWestem high- throughput protein analysis platform (ProteinSimple) according to manufacturer’s protocol using either a 12-230 kDa Separation Module (ProteinSimple, SM-W004) or a 2-40 kDa Separation Module (ProteinSimple, SM-W009) and Total Protein Detection Module (ProteinSimple, DM-TP01). Antibodies: PSAT1 (ProteinTech, 1 :500, Cat. # 10501-1-AP), PUMA (Cell Signaling Technologies, 1 :50, Cat. #98672), MCL-1 (ProteinTech, 1:50, Cat. #16225-1-AP), S100A8 (ProteinTech, 1 :50, Cat. #15792-1-AP), S100A9 (ProteinTech, 1:50, Cat. # 26992-1-AP), TIMM17A (Cell Signaling Technologies, 1:50, Cat. # 45948), TIMM23 (Cell Signaling Technologies, 1 :2000, Cat. # 34822), total RB (Cell Signaling Technologies, 1: 100, Cat. # 9309), phospho RB S807 / S811 (Cell Signaling Technologies, 1:50, Cat. # 8516), CDK1 (Abclonal, 1:5000, Cat. # A22347), cMYC (Cell Signaling Technologies, 1 :50, Cat. # 18583), HIFla (Cell Signaling Technologies, 1:500, Cat. # 36169), |3-actin (R&D Systems, 1 :50, Cat. # MAB8929).PKPD Study Results and DiscussionProtein expression of PSAT1, phosphoserine aminotransferase 1 and PUMA, p53 upregulated modulator of apoptosis, was measured by SimpleWestem high-throughput protein analysis. PS ATI is induced by the transcription factor ATF4 during ISR. PUMA is a proapoptotic protein and is also induced by ATF4. Induction of both PSAT1 and PUMA was observed in tumors treated with HC-7366 as a monotherapy, and in HC-7366+5-AZA treated tumors (FIGS. 7A and 7B). In HC-7366+Venetoclax and triplet groups, PSAT1 and PUMA were induced when HC-7366 was used at 0.5 mg / kg, but not in the HC-7366 1 mg / kg doublet and triplet groups. At the time of tumor sampling on Day 8, the tumors in the HC- 7366+Venetoclax and triplet groups have already begun to regress (FIGS. 6A and 6B). The decrease in PSAT1 and PUMA detection in these efficacious groups may be due to induction of cell death pathways in cells highly expressing these markers, leading to their elimination from the tumor.Upregulation of the antiapoptotic protein MCL- 1 is a common mechanism by which AML tumor cells become resistant to Venetoclax. Single agent HC-7366, 5-AZA, or Venetoclax each had little effect on MCL-1 levels relative to the vehicle control (FIG. 7C). However, significant decreases in MCL- 1 expression were observed in the groups with greatest efficacy, the HC-7366 doublet with Venetoclax, and the triplet tumor group, with HC-7366 at 1 mg / kg. These data indicate that HC-7366 combined with 5-AZA and Venetoclax may overcome Venetoclax resistance that is mediated by MCL-1 upregulation.TIMM17A and TIMM23 are subunits that make up a mitochondrial protein import complex, functioning to maintain mitochondrial proteostasis. Decreases in TIMM17A protein levels have been shown in response to stress signaling through eIF2a. While no significant changes in TIMM17A expression were observed in the tumors treated with HC-7366 monotherapy, the combination of HC-7366 with Venetoclax or HC-7366 with Venetoclax and 5-AZA lead to modest decreases in TIMM17A and TIMM23 at 0.5 mg / kg and more significant decreases in the doublet and triplet groups when HC-7366 was used at 1 mg / kg (FIGS. 7D and 7E). Inhibition of TIMM17A and TIMM23 by the combination of HC-7366 and Venetoclax suggests mitochondrial disruption that could also oppose Venetoclax resistance.S 100 proteins are calcium binding proteins that function in a number of cellular pathways. Single agent HC-7366, or HC-7366 in combination with Venetoclax or 5-AZA shows a trend toward reduction of S100A8 expression, although the reduction does not reach statistical significance (FIG. 7F). Analysis of S100A9, which heterodimerizes with S100A8, produced similar results. Elevated expression of S100A8 / A9 correlates with reduced sensitivity andresistance to Venetoclax, so the ability of HC-7366 to inhibit expression of these proteins represents another mechanism by which HC-7366 opposes Venetoclax resistance.Decreases in cell cycle regulators correlate with efficacy in the KG-1 model. Expression of phospho RB (pS807 / pS811), total RB, and CDK1 each show modest decreases in the groups with HC -7366+5 -AZA combination (FIGS. 7G, 7H and 71) However, more significant, dosedependent decreases were observed when HC-7366 was combined with Venetoclax as a doublet or triplet with Venetoclax and 5-AZA.5- AZA as a single agent or in combination, modestly reduced protein levels of the protooncogene c-MYC, while the HC-7366 combination with Venetoclax as a doublet or triplet produced more significant reductions in c-MYC (FIG. 7J). Similarly, the proto-oncogene HfFla was reduced by the combination of HC-7366 with 5-AZA and more significantly by HC-7366 and Venetoclax in the doublet and triplet groups (FIG. 7K).Analysis of a control protein, fj-actin, did not reveal differences in detection levels for any of the groups (FIG. 7L). These data confirm that the changes in the levels of pharmacodynamic markers and markers of Venetoclax resistance can be attributed to biological effects of the treatments rather than sample quality or integrity.INCORPORATION BY REFERENCE

[0181] This application refers to various issued patents, published patent applications, journal articles, and / or other publications, all of which are incorporated herein by reference. If there is a conflict between any of the incorporated references and the instant specification, the specification shall control. In addition, any particular embodiment of the present disclosure that falls within the prior art may be explicitly excluded from any one or more of the claims. Because such embodiments are deemed to be known to one of ordinary skill in the art, they may be excluded even if the exclusion is not set forth explicitly herein. Any particular embodiment of the disclosure can be excluded from any claim, for any reason, whether or not related to the existence of prior art.EQUIVALENTS

[0182] The invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The foregoing embodiments are therefore to be considered in all respects illustrative rather than limiting the invention described herein. Scope of the invention is thus indicated by the appended claims rather than by the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein.

Claims

CLAIMSWHAT IS CLAIMED:

1. A method of treating acute myeloid leukemia (AML) in a subject in need thereof, the method comprising administering to the subject an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, an effective amount of venetoclax, and an effective amount of 5-azacitidine, wherein Compound 1 is represented by formula (I)2. The method of claim 1 , wherein administering the effective amounts of Compound 1 , or a pharmaceutically acceptable salt thereof, venetoclax, and 5-azacitidine increases activation of the integrated stress response (ISR) in the subject as compared to a subject with AML receiving (a) the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, or (b) the effective amounts of Compound 1 , or a pharmaceutically acceptable salt thereof, and venetoclax.

3. The method of claim 1 or 2, wherein administering the effective amount of venetoclax to the subject comprises administering about 400 mg venetoclax to the subject daily.

4. A method of treating acute myeloid leukemia (AML) in a subject receiving venetoclax therapy and 5-azacitidine therapy, the method comprising administering to the subject an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, wherein Compound 1 is represented by formula (I)5. The method of claim 4, wherein the subject receiving the 5-azacitidine therapy is receiving about 75 mg / m25-azacitidine daily.

6. The method of claim 4 or 5, wherein the 5-azacitidine therapy is administered to the subject parenterally.

7. A method of inhibiting or overcoming resistance of acute myeloid leukemia (AML) to venetoclax therapy in a subject in need thereof, the method comprising administering to the subject an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, and an effective amount of 5-azacitidine, wherein Compound 1 is represented by formula (I)8. A method of improving the efficacy of venetoclax therapy in the treatment of acute myeloid leukemia (AML) in a subject in need thereof, the method comprising administering to the subject an effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, and an effective amount of 5-azacitidine, wherein Compound 1 is represented by formula (I)9. The method of any one of claims 4-8, wherein the subject receiving the venetoclax therapy is receiving about 400 mg venetoclax daily.

10. The method of any one of claims 4-9, wherein the venetoclax therapy is administered orally.

11. The method of any one of claims 1-10, wherein administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, to the subject comprises administering about 40 mg to about 75 mg Compound 1, or an equivalent amount of a pharmaceutically acceptable salt thereof on a free acid equivalent weight basis, to the subject.

12. The method of any one of claims 1-11, wherein administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, to the subject comprises administering about 40 mg Compound 1 , or an equivalent amount of a pharmaceutically acceptable salt thereof on a free acid equivalent weight basis, to the subject.

13. The method of any one of claims 1-11 , wherein administering the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, to the subject comprises administering about 75 mg Compound 1, or an equivalent amount of a pharmaceutically acceptable salt thereof on a free acid equivalent weight basis, to the subject.

14. The method of any one of claims 1-3 and 7-13, wherein administering the effective amount of 5-azacitidine to the subject comprises administering about 75 mg / m25-azacitidine to the subject daily.

15. The method of any one of claims 1-14, wherein the effective amount of Compound 1, or a pharmaceutically acceptable salt thereof, is administered orally.

16. The method of any one of claims 1-3, wherein the effective amounts of Compound 1, or a pharmaceutically acceptable salt thereof, and venetoclax are administered orally.

17. The method of any one of claims 1-3 and 7-16, wherein the effective amount of 5- azacitidine is administered to the subject parenterally.

18. The method of any one of claims 1-17, wherein the method comprises administering an effective amount of a pharmaceutically acceptable salt of Compound 1 to the subject.

19. The method of claim 18, wherein the pharmaceutically acceptable salt of Compound 1 is a potassium salt of Compound 1.

20. The method of any one of claims 1-19, wherein the AML is resistant to venetoclax.

21. The method of any one of claims 1-20, wherein the AML comprises a venetoclax resistance mechanism selected from the group consisting of up-regulation of anti-apoptotic proteins, p53 inactivation, activating kinase mutations, and upregulation of oxidative phosphorylation.

22. The method of any one of claims 1-21 , wherein the AML is characterized by having a FLT3 ITD mutation.

23. The method of any one of claims 1-21 , wherein the AML is characterized by having a TP53 mutation.

24. The method of any one of claims 1-21 , wherein the AML is characterized by upregulation of MCL-1.