Crystalline solid forms of bet inhibitor

Abstract

To provide crystalline solid forms of an inhibitor of BET proteins, such as BRD2, BRD3, BRD4 and BRD-t, useful in the treatment of diseases such as cancer.SOLUTION: The invention provides crystalline solid forms of the compound having the formula in the figure.SELECTED DRAWING: None

Description

[Technical Field]

[0001] This application relates to a crystalline solid form of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one, an inhibitor of BET proteins such as BRD2, BRD3, BRD4, and BRD-t, which is useful in the treatment of diseases such as cancer. This application includes methods for its preparation and intermediates in its preparation. [Background technology]

[0002] The eukaryotic genome is highly organized within the cell nucleus. DNA is packaged in chromatin by wrapping around a core of histone proteins to form nucleosomes. These nucleosomes are further compacted by aggregation and folding to form a highly condensed chromatin structure. While various distinct condensation states are possible, the cohesiveness of this structure changes during the cell cycle, reaching its most compact state during cell division. Chromatin structure plays an important role in regulating gene transcription by regulating protein access to DNA. Chromatin structure is controlled by a series of post-translational modifications to histone proteins, primarily within the tails of histones H3 and H4, which extend beyond the core nucleosome structure. These reversible modifications include acetylation, methylation, phosphorylation, ubiquitination, and sumoylation. Specific enzymes modifying specific residues within the histone tails write and erase these epigenetic marks, forming the epigenetic code. Other nuclear proteins bind to these marks and exert their specified output through the regulation of chromatin structure and gene transcription. Increasing evidence links genetic alterations to genes encoding epigenetic modifiers and regulators that lead to abnormalities in histone marks in diseases such as neurodegenerative disorders, metabolic diseases, inflammation, and cancer.

[0003] Histone acetylation is commonly associated with the activation of gene transcription because the modification weakens the interaction between DNA and histone proteins, increasing access to DNA by the transcriptional machinery. Specific proteins "read" the epigenetic code by binding to acetylated lysine residues in histones. Highly conserved protein modules called bromodomains bind to acetylated lysine residues on histones and other proteins. There are more than 60 bromodomain-containing proteins in the human genome.

[0004] The BET (Bromodomain and Extra-Terminal) family of bromodomain-containing proteins includes four proteins (BRD2, BRD3, BRD4, and BRD-t) that share a conserved structural organization, containing tandem N-terminal bromodomains that can bind to acetylated lysine residues on histones and other proteins. While BRD2, BRD3, and BRD4 are ubiquitously expressed, BRD-t is restricted to germ cells. BRD proteins play essential, but non-redundant, roles in regulating gene transcription and controlling cell proliferation. BET proteins associate with large protein complexes, including Mediator, PAFc, and the super-elongation complex, which control many aspects of gene transcription. BRD2 and BRD4 proteins have been shown to remain complexed with chromosomes during mitosis and are required to promote the transcription of key genes, including cyclin D and c-Myc, which initiate the cell cycle (Mochizuki J Biol. Chem. 2008 283:9040-9048). BRD4 is essential for recruiting the protein translation elongation factor B complex to the promoters of inducible genes, resulting in the phosphorylation of RNA polymerase II and stimulating the transcription and elongation of proliferative genes (Jang et al. Mol. Cell 2005 19:523-534). In some cases, the kinase activity of BRD4 can directly phosphorylate and activate RNA polymerase II (Devaiah et al. PNAS 2012 109:6927-6932). BRD4-deficient cells exhibit impaired cell cycle progression. BRD2 and BRD3 have been reported to bind to histones around transcriptionally active genes, potentially contributing to the promotion of transcription elongation (Leroy et al. Mol. Cell 2008 30:51-60).In addition to acetylated histones, BET proteins have been shown to selectively bind to acetylated transcription factors, including NF-kB and the RelA subunit of GATA1, thereby directly modulating the transcriptional activity of these proteins and controlling the expression of genes involved in inflammation and hematopoietic differentiation (Huang et al., Mol. Cell. Biol. 2009 29:1375-1387; Lamonica Proc. Nat. Acad. Sci. 2011 108:E159-168).

[0005] Recurrent translocations involving NUT (nuclear protein of the testis) and BRD3 or BRD4 to form a novel fusion oncogene, BRD-NUT, are found in highly malignant forms of epithelial neoplasia (French et al., Cancer Research 2003 63:304-307; French et al., Journal of Clinical Oncology 2004 22:4135-4139). Selective ablation of this oncogene restores normal cell differentiation and reverses the tumorigenic phenotype (Filippakopoulos et al., Nature 2010 468:1068-1073). Genetic knockdown of BRD2, BRD3, and BRD4 has been shown to reduce proliferation and survival of a wide range of hematologic and solid tumor cells (Zuber et al., Nature 2011 478:524-528; Delmore et al., Cell 2011 146:904-917). Apart from their role in cancer, BET proteins regulate the inflammatory response to bacterial challenge, exhibiting significant reductions in inflammatory cytokine levels and protection from obesity-induced diabetes in a BRD2 hypomorphic mouse model (Wang et al., Biochem J. 2009 425:71-83; Belkina et al., J. Immunol 2013). Furthermore, some viruses utilize these BET proteins to bind their genomes to host cell chromatin as part of the viral replication process, or to promote viral gene transcription and repression (You et al., Cell 2004 117:349-60; Zhu et al., Cell Reports 2012 2:807-816).

[0006] Inhibitors of BET proteins are currently under development. Exemplary BET protein inhibitors are described, for example, in U.S. Patent Application Publication Nos. 2014 / 0275030; 2015 / 0011540; 2015 / 0148375; 2015 / 0148342; 2015 / 0148372; 2015 / 0175604; and 2016 / 007572. Specifically, the BET inhibitor compound 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one is described in U.S. Patent Application Publication No. US2015 / 0307493. For drug development, it is usually advantageous to use a form of the drug that has desirable properties with respect to its preparation, purification, reproducibility, stability, bioavailability, and other characteristics. Thus, the solid crystalline forms of the compounds provided herein help meet the ongoing need for the development of BET inhibitors for the treatment of diseases. Summary of the Invention

[0007] In particular, the present application provides a crystalline solid form of a BET protein inhibitor, which is 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one.

[0008] The present application also provides a pharmaceutical composition comprising 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one in crystalline solid form and at least one pharmaceutically acceptable carrier.

[0009] The present application also provides methods of using crystalline solid forms of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one for the treatment of diseases and disorders associated with the activity of BET proteins.

[0010] Additionally, the present application provides methods for preparing 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one and its crystalline solid forms.

[0011] Furthermore, the present application provides intermediate compounds and methods for preparing the same that are useful in the synthesis of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one.

[0012] The details of one or more embodiments are set forth in the description below. Other features, objects, and advantages will become apparent from the specification and claims. [Brief explanation of the drawings]

[0013] [Figure 1] 1 is an X-ray powder diffraction (XRPD) pattern of Form I of Compound 1. [Figure 2] 1 is a differential scanning calorimetry (DSC) thermogram of Form I of Compound 1. [Figure 3] 1 is a thermogravimetric analysis (TGA) thermogram of Form I of Compound 1. [Figure 4] 1 is an XRPD pattern of Form II of Compound 1. [Figure 5] 1 is a DSC thermogram of Compound 1 Form II. [Figure 6] 1 is a TGA thermogram of Compound 1 Form II. [Figure 7] 1 is an XRPD pattern of Form Ia of Compound 1. [Figure 8] 1 is an XRPD pattern of Form III of Compound 1. [Figure 9] 1 is an XRPD pattern of Form IV of Compound 1. [Figure 10] 1 is an XRPD pattern of Form V of Compound 1. [Figure 11] 1 is an XRPD pattern of Form Va of Compound 1. [Figure 12] 1 is an XRPD pattern of Form VI of Compound 1. [Figure 13] 1 is an XRPD pattern of Form VII of Compound 1. [Figure 14] 1 is an XRPD pattern of Form VIII of Compound 1. [Figure 15] 1 is an XRPD pattern of Form IX of Compound 1. [Figure 16] 1 is an XRPD pattern of Form X of Compound 1. [Figure 17] 1 is an XRPD pattern of Form XI of Compound 1. [Figure 18] 1 is an XRPD pattern of Form XII of Compound 1. [Figure 19] 1 is an XRPD pattern of Form XIII of Compound 1. [Figure 20] 1 is an XRPD pattern of Form XIV of Compound 1. [Figure 21] 1 is an XRPD pattern of Form XV of Compound 1. DETAILED DESCRIPTION OF THE INVENTION

[0014] Crystalline forms and their preparation methods In particular, the present application provides a crystalline solid form of a BET protein inhibitor, which is 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (see below), referred to herein as "Compound 1." [ka]

[0015] Typically, different crystalline forms of the same substance have different bulk properties, e.g., with respect to hygroscopicity, solubility, stability, etc. Forms with higher melting points often have superior thermodynamic stability and are advantageous for extending the shelf life of drug formulations containing the solid form. Forms with lower melting points often have inferior thermodynamic stability but are advantageous for increasing water solubility, leading to improved drug bioavailability. For heat and humidity stability, weakly hygroscopic forms are desirable and are less likely to decompose during long-term storage. Anhydrous forms are often desirable because they allow for consistent manufacturing without worrying about weight or composition variations due to changes in solvent or water content. On the other hand, hydrated or solvated forms may be advantageous in that they are less hygroscopic and may exhibit improved humidity stability under storage conditions.

[0016] The crystalline solid forms of the present invention may contain solvents such as water (e.g., hydrated forms) or may be substantially free of water and solvents (e.g., anhydrous forms). In some embodiments, the crystalline solid form is anhydrous. In further embodiments, the crystalline solid form is hydrated.

[0017] Compound 1 can be obtained in a solid crystalline form designated Form I, as described below and in the Examples. Experimental data indicate that Form I is anhydrous. Form I is characterized by its XRPD pattern and other solid-state characteristics. In some embodiments, Form I has a characteristic XRPD peak at about 12.7° in 2-theta. In some embodiments, Form I has one or more characteristic XRPD peaks selected from about 8.7°, about 9.8°, and about 12.7° in 2-theta. In some embodiments, Form I has one or more characteristic XRPD peaks selected from about 8.7°, about 9.8°, about 12.7°, about 21.4°, and about 23.3° in 2-theta.

[0018] In some embodiments, Form I has two or more characteristic XRPD peaks selected from about 8.7°, about 9.8°, about 12.7°, about 21.4°, and about 23.3° two-theta.

[0019] In some embodiments, Form I has two or more characteristic XRPD peaks selected from about 8.7°, about 9.8°, about 11.6°, about 12.7°, about 14.7°, about 15.7°, about 20.0°, about 21.4°, about 23.3°, and about 27.1° two-theta.

[0020] In some embodiments, Form I has three or more characteristic XRPD peaks selected from about 8.7°, about 9.8°, about 11.6°, about 12.7°, about 14.7°, about 15.7°, about 20.0°, about 21.4°, about 23.3°, and about 27.1° two-theta.

[0021] In some embodiments, Form I has four or more characteristic XRPD peaks selected from about 8.7°, about 9.8°, about 11.6°, about 12.7°, about 14.7°, about 15.7°, about 20.0°, about 21.4°, about 23.3°, and about 27.1° two-theta.

[0022] In some embodiments, Form I has an XRPD pattern substantially as shown in FIG.

[0023] In some embodiments, Form I has a DSC thermogram characterized by an endothermic peak at a temperature of about 266° C. In some embodiments, Form I has a DSC thermogram substantially as shown in FIG.

[0024] In some embodiments, Form I has a TGA thermogram substantially as shown in FIG.

[0025] Form I can generally be prepared by precipitating Form I from a solution comprising Compound 1 and a solvent. In some embodiments, the solvent comprises methanol, acetone, n-heptane, or a mixture thereof. For example, Form I can be prepared by precipitating Form I from a solution comprising Compound 1 and acetone. Preparation of Form I can include adding Compound 1 to a saturated acetone solution of Compound 1 and stirring the resulting solution at about 25° C. for about 3 days.

[0026] In some embodiments, precipitation of Form I is achieved by (1) reducing the temperature of a solution of Compound 1 (e.g., a hot solution of Compound 1), (2) concentrating the solution of Compound 1, (3) adding an anti-solvent to a solution of Compound 1, or any combination thereof. In some embodiments, precipitation is achieved by adding an anti-solvent to a solution of Compound 1, wherein the solution of Compound 1 comprises a protic solvent and an aprotic solvent. In some embodiments, the protic solvent is methanol, the aprotic solvent is acetone, and the anti-solvent is n-heptane. In some embodiments, precipitation of Form I is achieved by adding n-heptane to a solution of Compound 1, wherein the solution of Compound 1 comprises methanol and acetone.

[0027] In some embodiments, preparing Form I includes: (ia) heating a solution of Compound 1 to a temperature of about 50°C to about 60°C; (iia) reducing the volume of a solution of Compound 1 at a temperature of about 50°C to about 60°C to produce a low volume solution of Compound 1; (iiia) adding an anti-solvent to a low volume solution of Compound 1 while maintaining the temperature at about 55°C to about 65°C to form a warm solution of Compound 1; and (iva) cooling a hot solution of Compound 1 to a temperature of about 15°C to about 30°C to precipitate Form I.

[0028] In some embodiments, preparing Form I includes: (ib) heating a solution of Compound 1 (the solution containing methanol and acetone as solvents) to a temperature of about 50°C to about 60°C; (iib) reducing the volume of a solution of Compound 1 at a temperature of about 50°C to about 60°C to produce a low volume solution of Compound 1; (iiib) adding n-heptane to a low volume solution of Compound 1 while maintaining the temperature at about 55°C to about 65°C to form a warm solution of Compound 1; and (ivb) cooling the hot solution of Compound 1 to a temperature of about 15° C. to about 30° C. to precipitate Form I.

[0029] Compound 1 can also be obtained as a crystalline form designated Form II, as described below and in the Examples. Experimental data indicate that Form II is anhydrous. Form II is characterized by its XRPD pattern and other solid-state characteristics. In some embodiments, Form II has a characteristic XRPD peak at about 17.0° in 2-theta. In some embodiments, Form II has one or more characteristic XRPD peaks selected from about 17.0° and about 19.3° in 2-theta. In some embodiments, Form II has one or more characteristic XRPD peaks selected from about 16.2°, about 17.0°, and about 19.3° in 2-theta.

[0030] In some embodiments, Form II has two or more characteristic XRPD peaks selected from about 6.7°, about 9.5°, about 10.5°, about 14.8°, about 16.2°, about 17.0°, about 18.8°, and about 19.3° two-theta.

[0031] In some embodiments, Form II has three or more characteristic XRPD peaks selected from about 6.7°, about 9.5°, about 10.5°, about 14.8°, about 16.2°, about 17.0°, about 18.8°, and about 19.3° two-theta.

[0032] In some embodiments, Form II has four or more characteristic XRPD peaks selected from about 6.7°, about 9.5°, about 10.5°, about 14.8°, about 16.2°, about 17.0°, about 18.8°, and about 19.3° two-theta.

[0033] In some embodiments, Form II has an XRPD pattern substantially as shown in FIG.

[0034] In some embodiments, Form II has a DSC thermogram characterized by an endothermic peak at a temperature of about 268° C. In some embodiments, Form II has a DSC thermogram substantially as shown in FIG.

[0035] In some embodiments, Form II has a TGA thermogram substantially as shown in FIG.

[0036] Form II can generally be prepared by precipitating Form II from a solution comprising Compound 1 and a solvent. In some embodiments, the solvent comprises tetrahydrofuran (THF), acetone, n-heptane, or a mixture thereof. In some embodiments, precipitating Form II is accomplished by (1) lowering the temperature of a solution of Compound 1, (2) concentrating a solution of Compound 1, (3) adding an anti-solvent to a solution of Compound 1, or any combination thereof. In some embodiments, precipitating Form II is accomplished by adding an anti-solvent to a solution of Compound 1, wherein the solution comprises an ethereal solvent and an aprotic solvent. In some embodiments, the ethereal solvent is THF, the aprotic solvent is acetone, and the anti-solvent is n-heptane. In some embodiments, precipitating Form II is accomplished by adding n-heptane to a solution of Compound 1, wherein the solution of Compound 1 comprises THF and acetone.

[0037] In some embodiments, preparing Form II includes: (ic) heating the solution of Compound 1 to a temperature of about 50°C to about 60°C; (iic) reducing the volume of a solution of Compound 1 at a temperature of about 50°C to about 60°C to produce a low volume solution of Compound 1; (iiic) adding an anti-solvent to a low volume solution of Compound 1 while maintaining the temperature at about 55°C to about 65°C to form a warm solution of Compound 1; and (ivc) cooling the hot solution of Compound 1 to a temperature of about 15° C. to about 30° C. to precipitate Form II.

[0038] In some embodiments, preparing Form II includes: (id) heating a solution of Compound 1 (the solution containing THF and acetone as solvents) to a temperature of about 50°C to about 60°C; (iid) reducing the volume of a solution of Compound 1 at a temperature of about 50°C to about 60°C to produce a low volume solution of Compound 1; (iiid) adding n-heptane to a low volume solution of Compound 1 while maintaining the temperature at about 55°C to about 65°C to form a warm solution of Compound 1; and (ivd) cooling the warm solution of Compound 1 to a temperature of about 15°C to about 30°C to precipitate Form II.

[0039] Compound 1 can also be obtained in solid crystalline forms designated Forms Ia, III, IV, V, Va, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, and XV, which are described below and in the Examples. Forms Ia, III, IV, V, Va, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, and XV are characterized by their XRPD patterns and other solid-state characteristics.

[0040] In some embodiments, Form Ia has one or more characteristic XRPD peaks selected from about 8.8°, about 10.0°, about 11.7°, about 12.8°, and about 13.5° in two-theta. In some embodiments, Form Ia has one or more characteristic XRPD peaks selected from about 8.8°, about 10.0°, about 11.7°, about 12.8°, about 13.5°, about 20.0°, about 21.5°, about 22.6°, and about 23.3° in two-theta. In some embodiments, Form Ia has an XRPD pattern substantially as shown in Figure 7.

[0041] In some embodiments, Form III has one or more characteristic XRPD peaks selected from about 7.8°, about 12.4°, about 13.1°, about 15.2°, and about 15.5° in 2-theta. In some embodiments, Form III has one or more characteristic XRPD peaks selected from about 7.8°, about 12.4°, about 13.1°, about 15.2°, about 15.5°, about 16.9°, about 17.5°, and about 20.3° in 2-theta. In some embodiments, Form III has an XRPD pattern substantially as shown in Figure 8.

[0042] In some embodiments, Form IV has one or more characteristic XRPD peaks selected from about 11.2°, about 16.3°, about 18.7°, and about 22.1° 2-theta, hi some embodiments, Form IV has an XRPD pattern substantially as shown in FIG.

[0043] In some embodiments, Form V has one or more characteristic XRPD peaks selected from about 8.2°, about 8.5°, about 14.1°, about 16.3°, and about 17.1° in 2-theta. In some embodiments, Form V has one or more characteristic XRPD peaks selected from about 8.2°, about 8.5°, about 14.1°, about 16.3°, about 17.1°, about 18.9°, about 19.8°, about 21.8°, and about 22.7° in 2-theta. In some embodiments, Form V has an XRPD pattern substantially as shown in Figure 10.

[0044] In some embodiments, Form Va has one or more characteristic XRPD peaks selected from about 8.7°, about 16.5°, about 17.3°, about 19.9°, and about 21.6° two-theta. In some embodiments, Form Va has an XRPD pattern substantially as shown in Figure 11. In some embodiments, Form Va has a DSC thermogram characterized by an endothermic peak at a temperature of about 133°C, an endothermic peak at a temperature of about 267°C, or a combination thereof.

[0045] In some embodiments, Form VI has one or more characteristic XRPD peaks selected from about 8.5°, about 9.6°, about 11.4°, and about 12.1° in 2-theta. In some embodiments, Form VI has one or more characteristic XRPD peaks selected from about 8.5°, about 9.6°, about 11.4°, about 12.1°, about 13.5°, about 14.5°, about 15.2°, about 17.1°, about 17.7°, about 18.1°, about 19.2°, and about 20.7° in 2-theta. In some embodiments, Form VI has an XRPD pattern substantially as shown in Figure 12.

[0046] In some embodiments, Form VII has one or more characteristic XRPD peaks selected from about 9.9°, about 12.2°, about 14.8°, and about 15.7° in 2-theta. In some embodiments, Form VII has one or more characteristic XRPD peaks selected from about 9.9°, about 12.2°, about 14.8°, about 15.7°, about 17.0°, about 17.5°, and about 18.8° in 2-theta. In some embodiments, Form VII has an XRPD pattern substantially as shown in Figure 13. In some embodiments, Form VII has a DSC thermogram characterized by an endothermic peak at a temperature of about 126°C, an endothermic peak at a temperature of about 256°C, an exothermic peak at a temperature of about 260°C, an endothermic peak at a temperature of about 267°C, or a combination thereof.

[0047] In some embodiments, Form VIII has one or more characteristic XRPD peaks selected from about 8.1°, about 8.5°, about 16.2°, and about 17.0° in 2-theta. In some embodiments, Form VIII has one or more characteristic XRPD peaks selected from about 8.1°, about 8.5°, about 16.2°, about 16.6°, about 17.0°, about 17.5°, about 18.0°, about 18.9°, about 19.6°, and about 20.1° in 2-theta. In some embodiments, Form VIII has an XRPD pattern substantially as shown in Figure 14. In some embodiments, Form VIII has a DSC thermogram characterized by an endothermic peak at a temperature of about 145°C, an endothermic peak at a temperature of about 265°C, or a combination thereof.

[0048] In some embodiments, Form IX has one or more characteristic XRPD peaks selected from about 8.6°, about 9.1°, about 11.4°, about 13.4°, and about 15.2° in two-theta. In some embodiments, Form IX has one or more characteristic XRPD peaks selected from about 8.6°, about 9.1°, about 11.4°, about 13.4°, about 15.2°, about 18.2°, about 22.1°, about 22.8°, and about 23.9° in two-theta. In some embodiments, Form IX has an XRPD pattern substantially as shown in Figure 15.

[0049] In some embodiments, Form X has one or more characteristic XRPD peaks selected from about 14.9°, about 15.3°, about 15.8°, and about 17.0° in 2-theta. In some embodiments, Form X has one or more characteristic XRPD peaks selected from about 14.9°, about 15.3°, about 15.8°, about 17.0°, about 17.7°, about 18.3°, and about 19.7° in 2-theta. In some embodiments, Form X has an XRPD pattern substantially as shown in Figure 16. In some embodiments, Form X has a DSC thermogram characterized by an endothermic peak at a temperature of about 121°C, an endothermic peak at a temperature of about 267°C, or a combination thereof.

[0050] In some embodiments, Form XI has one or more characteristic XRPD peaks selected from about 8.9°, about 12.8°, about 18.0°, about 21.5°, about 22.6°, and about 23.3° 2-theta, hi some embodiments, Form XI has an XRPD pattern substantially as shown in Figure 17.

[0051] In some embodiments, Form XII has one or more characteristic XRPD peaks selected from about 5.6°, about 11.7°, about 13.8°, and about 14.5° in 2-theta. In some embodiments, Form XII has one or more characteristic XRPD peaks selected from about 5.6°, about 11.7°, about 13.8°, about 14.5°, about 16.9°, about 17.7°, and about 18.7° in 2-theta. In some embodiments, Form XII has one or more characteristic XRPD peaks selected from about 5.6°, about 11.7°, about 13.8°, about 14.5°, about 16.9°, about 17.7°, about 18.7°, about 23.5°, about 24.6°, about 34.3°, about 44.2°, and 44.6° in 2-theta. In some embodiments, Form XII has an XRPD pattern substantially as shown in FIG. 18. In some embodiments, Form XII has a DSC thermogram characterized by an endothermic peak at a temperature of about 264°C.

[0052] In some embodiments, Form XIII has one or more characteristic XRPD peaks selected from about 5.7°, about 8.6°, about 9.8°, and about 11.8° in 2-theta. In some embodiments, Form XIII has one or more characteristic XRPD peaks selected from about 5.7°, about 8.6°, about 9.8°, about 11.8°, about 12.6°, about 13.4°, about 14.1°, about 14.8°, about 16.6°, and about 19.1° in 2-theta. In some embodiments, Form XIII has an XRPD pattern substantially as shown in Figure 19. In some embodiments, Form XIII has a DSC thermogram characterized by an endothermic peak at a temperature of 267°C.

[0053] In some embodiments, Form XIV has one or more characteristic XRPD peaks selected from about 4.0°, about 11.2°, about 11.9°, about 14.1°, about 14.8°, and about 15.9° two-theta. In some embodiments, Form XIV has an XRPD pattern substantially as shown in Figure 20. In some embodiments, Form XIV has a DSC thermogram characterized by an endothermic peak at a temperature of 267°C.

[0054] In some embodiments, Form XV has one or more characteristic XRPD peaks selected from about 7.4°, about 9.6°, about 12.4°, about 13.4°, and about 15.5° in 2-theta. In some embodiments, Form XV has one or more characteristic XRPD peaks selected from about 7.4°, about 9.6°, about 12.4°, about 13.4°, about 15.5°, about 16.9°, about 17.7°, about 19.0°, about 19.5°, about 20.6°, and about 22.5° in 2-theta. In some embodiments, Form XV has an XRPD pattern substantially as shown in Figure 21. In some embodiments, Form XV has a DSC thermogram characterized by an endothermic peak at a temperature of about 85°C, an endothermic peak at a temperature of about 172°C, an exothermic peak at a temperature of about 192°C, an endothermic peak at a temperature of about 268°C, or a combination thereof.

[0055] As used herein, the phrase "solid form" refers to a compound provided herein in either an amorphous state or a crystalline state ("crystalline form" or "crystalline solid" or "crystalline solid form"). Thus, a compound provided herein in a crystalline state may optionally contain a solvent or water within the crystal lattice, forming, for example, a solvated or hydrated crystal form. As used herein, the term "hydrated" is intended to refer to a crystal form that contains water molecules within the crystal lattice. Examples of "hydrated" crystal forms include hemihydrate, monohydrate, dihydrate, etc. Other hydrated forms, such as channel hydrates, are also included within the meaning of this term.

[0056] The various crystalline forms of the compounds provided herein (e.g., Compound 1) are characterized by X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), and / or thermogravimetric analysis (TGA). The X-ray powder diffraction (XRPD) pattern of reflections (peaks) is typically considered a fingerprint of a particular crystalline form. It is well known that the relative intensities of XRPD peaks can vary significantly depending on, among other things, the sample preparation technique, crystal size distribution, various filters used, sample mounting procedures, and the specific instrument used. In some cases, new peaks may be observed or existing peaks may disappear depending on the type or settings of the instrument (e.g., whether or not a Ni filter is used). As used herein, the term "peak" or "characteristic peak" refers to a reflection whose relative height / intensity is at least about 3% of the maximum peak height / intensity. Furthermore, instrumental differences and other factors may affect the 2-theta value. Thus, peak designations as reported herein may vary above or below about 0.2° (2 theta), and the terms "substantially" or "about" as used herein in connection with XRPD are intended to refer to such variations.

[0057] Similarly, temperature measurements associated with DSC, TGA, or other thermal experiments can vary by approximately ±3° C. depending on the instrument, the specific setup, sample preparation, etc. Thus, crystalline forms reported herein having DSC thermograms "substantially" as shown in any of the figures are understood to take into account such variations.

[0058] The term "crystalline form" is intended to refer to a crystalline material with a particular lattice arrangement. Different crystalline forms of the same substance typically have different crystal lattices (e.g., unit cells) and typically have different physical properties and sometimes different water or solvent content due to the differences in the crystal lattices. Differences in the crystal lattice can be distinguished by solid-state characterization methods such as X-ray powder diffraction (XRPD). Additionally, other characterization methods such as differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and dynamic vapor sorption (DVS) are useful in distinguishing between crystalline forms, as well as determining stability and solvent / water content.

[0059] Different crystalline forms of a particular substance, such as Compound 1, can include both anhydrous forms of the substance and solvated / hydrated forms of the substance, with each anhydrous and solvated / hydrated form being distinguished from the other by differences in XRPD patterns or other solid-state characterization methods showing differences in the crystal lattice. In some cases, a single crystalline form (e.g., identified by a unique XRPD pattern) can have varying water or solvent content, where the lattice (as well as the XRPD pattern) remains substantially unchanged as the composition with respect to water and / or solvent changes.

[0060] In some embodiments, the compounds of the present application (or hydrates and solvates thereof) are prepared in batches, which are referred to as batches, samples, or preparations. A batch, sample, or preparation may contain a compound provided herein in any of the crystalline or amorphous forms described herein, including hydrated and non-hydrated forms, and mixtures thereof.

[0061] The compounds disclosed herein may include all isotopes of atoms present therein. Isotopes include atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium.

[0062] In some embodiments, a compound provided herein (e.g., Compound 1), or a salt thereof, or a crystalline form thereof, is substantially isolated. The term "substantially isolated" means that the compound or salt is at least partially or substantially separated from the environment in which it was formed or detected. A partial isolate can include, for example, a composition enriched for a compound, salt, or crystalline form provided herein. A substantial isolate can include a composition containing at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% by weight of a compound, salt, or crystalline form provided herein.

[0063] The phrase "pharmaceutically acceptable," as used herein, means that such compounds, materials, compositions, and / or dosage forms are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, and are commensurate with a reasonable benefit / risk ratio.

[0064] Preparation of Compound 1 The present application further provides a method for preparing compound 1, which is adaptable to scale-up. The method for preparing compound 1 is described in US2015 / 0307493, which is incorporated herein by reference in its entirety. Compared to the method described in US2015 / 0307493, the method provided herein has the distinct advantage of being adaptable to scale-up. For example, the method provided herein can produce a high-quality product with a high yield, even using less hazardous reagents. Furthermore, the method provided herein can produce compound 7 in situ without isolating compound 7 (see below), improving the efficiency of large-scale production.

[0065] In some embodiments, the method for preparing Compound 1 includes the step of preparing Compound 8: [ka] with B1, where B1 is a base.

[0066] In some embodiments, B1 is an alkali metal hydroxide base, such as sodium hydroxide. The reaction of compound 8 with B1 can be carried out in a solvent. In some embodiments, the solvent includes an ether solvent, such as 1,4-dioxane. Ether solvents, such as 1,4-dioxane, can provide compound 1 in high yield and good quality. In some embodiments, the reaction of compound 8 with B1 is carried out at an elevated temperature, for example, at a temperature of about 50°C to about 85°C (e.g., about 60°C to about 80°C or about 65°C to about 75°C). In some embodiments, the temperature is about 70°C. In some embodiments, B1 is provided in molar excess relative to the amount of compound 8. In some embodiments, about 3 to about 4 equivalents or about 3.5 equivalents of B1 are used, based on 1 equivalent of compound 8.

[0067] In some embodiments, the method further comprises compound 7: [ka] Compound 9: [ka] in the presence of P2 and B2 to produce compound 8, where P2 is a transition metal catalyst and B2 is a base.

[0068] In some embodiments, P2 is a transition metal catalyst such as a palladium catalyst. Examples of palladium catalysts include [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (Pd(dppf)Cl2, e.g., Pd(dppf)Cl2-CH2Cl2), dichloro(bis{di-tert-butyl[4-(dimethylamino)phenyl]-phosphoranyl})palladium (Pd-132), Pd(PPh3)4, and tetrakis(tri(o-tolyl)phosphine)palladium(0). In some embodiments, P2 is Pd(dppf)Cl2. In some embodiments, B2 is an alkali metal bicarbonate base such as sodium bicarbonate. In some embodiments, B2 is an alkali metal carbonate base such as K2CO3. The reaction of compound 7 with compound 9 can be carried out in a solvent. In some embodiments, the solvent comprises a protic solvent, an ethereal solvent, or a mixture thereof. In some embodiments, the solvent comprises water, 1,4-dioxane, or a mixture thereof. In some embodiments, the reaction of compound 7 with compound 9 is carried out at an elevated temperature, e.g., a temperature of about 80°C to about 100°C (e.g., about 85°C to about 95°C). In some embodiments, the temperature is about 90°C. In some embodiments, about 1 equivalent of compound 9 is used based on 1 equivalent of compound 7 or compound 6 (having the structure shown below). In some embodiments, a sufficient catalytic amount of P2 is provided. For example, about 0.01 to about 0.05 equivalents or about 0.03 equivalents of P2 are used based on 1 equivalent of compound 7. In some embodiments, a molar excess of B2 is provided relative to the amount of compound 9. In some embodiments, about 2 to about 3 equivalents or about 2.5 equivalents of B2 are used based on 1 equivalent of compound 9.

[0069] In some embodiments, the method further comprises compound 6: [ka] with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) in the presence of P3 and B3 to produce compound 7, where P3 is a transition metal catalyst and B3 is a base.

[0070] In some embodiments, P3 is a transition metal catalyst such as a palladium catalyst. Examples of palladium catalysts include [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (Pd(dppf)Cl2, e.g., Pd(dppf)Cl2-CHCl2), dichloro(bis{di-tert-butyl[4-(dimethylamino)phenyl]-phosphoranyl})palladium (Pd-132), Pd(PPh3)4, and tetrakis(tri(o-tolyl)phosphine)palladium(0). In some embodiments, P3 is Pd(dppf)Cl2. In some embodiments, B3 is an alkali metal acetate base such as potassium acetate. The reaction of compound 6 with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) can be carried out in a solvent. In some embodiments, the solvent comprises an ether solvent such as 1,4-dioxane. In some embodiments, the reaction of compound 6 with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) is carried out at elevated temperatures, e.g., from about 70°C to about 90°C (e.g., 75°C to about 85°C). In some embodiments, the temperature is about 80°C. In some embodiments, a molar excess of the reagent 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) is provided relative to the amount of compound 6. In some embodiments, about 2 to about 2.5 equivalents of 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) are used, based on 1 equivalent of compound 6. In some embodiments, a molar excess of B3 is provided relative to the amount of compound 6. In some embodiments, about 3 to about 3.5 equivalents of B3 are used, based on 1 equivalent of compound 6. In some embodiments, a sufficient catalytic amount of P3 is provided. In some embodiments, about 0.01 to about 0.05 equivalents or about 0.03 equivalents of P3 are used, based on 1 equivalent of compound 6.

[0071] In some embodiments, after compound 7 is formed, the subsequent reaction to form compound 8 is carried out in the same reaction vessel without isolating compound 7. When compound 7 is formed and then the reaction to form compound 8 is carried out in the same reaction vessel (without isolating compound 7), compound 8 can be formed from compounds 7 and 9 without the addition of P2, for example, by forming compound 7 using P3 (a transition metal catalyst) in the same reaction vessel. Alternatively, the coupling reaction to form compound 8 from compound 6 can be carried out in two steps, in which compound 7 is isolated and P2 is used in the reaction to form compound 8 from compound 7.

[0072] Alternatively, compound 8 can be prepared by converting compound 6 to compound 15: [ka] in the presence of P4 and B4, where P4 is a transition metal catalyst and B4 is a base.

[0073] In some embodiments, P4 is a transition metal catalyst such as a palladium catalyst. Examples of palladium catalysts include 4-(di-tert-butylphosphino)-N,N-dimethylaniline-dichloropalladium (2:1), Pd(dppf)Cl (e.g., Pd(dppf)Cl-CHCl), dichloro(bis{di-tert-butyl[4-(dimethylamino)phenyl]-phosphoranyl})palladium (Pd-132), Pd(PPh), and tetrakis(tri(o-tolyl)phosphine)palladium(0). In some embodiments, P4 is 4-(di-tert-butylphosphino)-N,N-dimethylaniline-dichloropalladium (2:1). In some embodiments, P4 is Pd(dppf)Cl (e.g., Pd(dppf)Cl-CHCl). In some embodiments, B4 is a base such as cesium fluoride. In another embodiment, B4 is an alkali metal carbonate, such as K2CO3. The reaction of compound 6 with compound 15 can be carried out in a solvent. In some embodiments, the solvent includes a protic solvent, an ethereal solvent, or a mixture thereof. In some embodiments, the reaction is carried out in a solvent including 1,4-dioxane, water, or a mixture thereof. In some embodiments, the reaction of compound 6 with compound 15 is carried out at an elevated temperature (e.g., above room temperature), such as near reflux. In some embodiments, about 1 equivalent of compound 15 is used based on 1 equivalent of compound 6. In some embodiments, a molar excess of B4 is provided relative to compound 6. In some embodiments, about 3 to about 4 equivalents, or about 3.5 equivalents, of B4 are used based on 1 equivalent of compound 6. P4 is typically provided in a sufficient catalytic amount. In some embodiments, about 0.01 to about 0.1 equivalents, or about 0.05 equivalents, of P4 are used based on 1 equivalent of compound 6.

[0074] In some embodiments, compound 15 can be prepared by a method comprising reacting compound 9 with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) in the presence of P8 and B8, wherein P8 is a transition metal catalyst and B8 is a base.

[0075] In some embodiments, P8 is a transition metal catalyst, such as a palladium catalyst. Examples of palladium catalysts include tris(dibenzylideneacetone)dipalladium(0) (Pd2(dba)3, 4-(di-tert-butylphosphino)-N,N-dimethylaniline-dichloropalladium (2:1), Pd(dppf)Cl2 (e.g., Pd(dppf)Cl2-CH2Cl2), dichloro(bis{di-tert-butyl[4-(dimethylamino)phenyl]-phosphoranyl})palladium (Pd-132), Pd(PPh3)4, and tetrakis(tri(o-tolyl)phosphino). In some embodiments, P8 is tris(dibenzylideneacetone)dipalladium(0) (Pd2(dba)3 (e.g., dicyclohexyl(2',4',6'-triisopropylbiphenyl-2-yl)phosphine (Xphos) can be added as a ligand). In some embodiments, B8 is an alkali metal acetate base, such as potassium acetate. Compound 9 and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi( The reaction of compound 9 with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) can be carried out in a solvent. In some embodiments, the solvent includes an ethereal solvent such as 1,4-dioxane. In some embodiments, the reaction of compound 9 with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) is carried out at a temperature of about 75°C to about 95°C. In some embodiments, the temperature is about 80°C to about 90°C or about 80°C to about 85°C. In some embodiments, 1 equivalent of compound 9 is reacted with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane). Based on 9, about 2 equivalents of 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) are used. In some embodiments, a molar excess of B8 is provided relative to compound 9. In some embodiments, about 2 to about 3 equivalents of B8 are used based on 1 equivalent of compound 9. P8 is typically provided in a sufficient catalytic amount. In some embodiments, about 0.01 to about 0.1 equivalents or about 0.025 equivalents of P8 are used based on 1 equivalent of compound 9.

[0076] In some embodiments, compound 6 can be prepared according to the procedures of US2015 / 0307493, which is incorporated herein by reference in its entirety.

[0077] In some embodiments, compound 6 is compound 5: [ka] with a methylating agent and B5, wherein B5 is a base. In some embodiments, the methylating agent is methyl iodide (MeI), dimethyl sulfate, dimethyl carbonate, or tetramethylammonium chloride. In some embodiments, the methylating agent is methyl iodide. In some embodiments, B5 is an alkali metal carbonate base, such as potassium carbonate (K2CO3). In some embodiments, the reaction of compound 5 with the methylating agent can be carried out in a solvent, including, for example, an aprotic solvent, such as N,N-dimethylformamide (DMF). In some embodiments, the reaction of compound 5 with the methylating agent can be carried out at a temperature of about 10°C to about 20°C or about 15°C to about 20°C.

[0078] In some embodiments, compound 5 is compound 4: [ka] The compound 4 can be prepared by a method comprising reacting 4 with 2-bromo-2-methylpropanoyl bromide and B6, where B6 is a base. In some embodiments, B6 is an alkali metal carbonate, such as potassium carbonate (K2CO3). The reaction of compound 4 with 2-bromo-2-methylpropanoyl bromide can be carried out in the presence of a solvent. For example, the solvent can include acetonitrile, water, or a mixture thereof. The reaction of compound 4 with 2-bromo-2-methylpropanoyl bromide can be carried out at an elevated temperature, for example, a temperature of about 60°C to about 90°C. In some embodiments, the temperature is about 75°C.

[0079] In some embodiments, compound 4 is compound 3: [ka] with a reducing agent. In some embodiments, the reducing agent is sodium hydrosulfite or H2 / Raney Ni. The reaction of compound 3 with the reducing agent can be carried out in the presence of a solvent. In some embodiments, the solvent includes a protic solvent (e.g., water and methanol), an ethereal solvent (tetrahydrofuran), or a mixture thereof. In some embodiments, the reaction of compound 3 with sodium hydrosulfite is carried out in water, tetrahydrofuran, or a mixture thereof. In some embodiments, the reaction of compound 3 with H2 / Raney Ni is carried out in methanol. In some embodiments, the reaction of compound 3 with the reducing agent is carried out at room temperature. In some embodiments, sodium hydrosulfite is used in combination with sodium bicarbonate. The reaction of compound 3 with sodium hydrosulfite and sodium bicarbonate can produce compound 4 under milder process conditions than H2 / Raney Ni, which can be dangerous in large-scale production.

[0080] In some embodiments, compound 3 is compound 2: [ka] with N-bromosuccinimide (NBS). The use of NBS allows for high yields and good quality of product in large scale production, e.g., on the kilogram scale. In some embodiments, the reaction is carried out in a solvent comprising an aprotic solvent such as N,N-dimethylformamide (DMF). In some embodiments, the reaction is carried out at room temperature.

[0081] In some embodiments, compound 2 is compound 1a: [ka] with nitric acid and acetic acid. In some embodiments, the reaction is carried out at a temperature of about 60°C to about 90°C or about 75°C to about 80°C.

[0082] In some embodiments, compound 9 can be prepared according to the procedures of US2015 / 0307493 and WO2013 / 097601, each of which is incorporated by reference herein in its entirety.

[0083] In some embodiments, compound 9 is compound 14: [ka] with methyl iodide and sodium hydride. In some embodiments, the reaction is carried out in a solvent comprising an aprotic solvent such as N,N-dimethylformamide (DMF).

[0084] In some embodiments, compound 14 is compound 13: [ka] with an acid. In some embodiments, the acid is a strong aqueous acid such as HCl. In some embodiments, the reaction is carried out in a solvent comprising an ethereal solvent such as 1,4-dioxane.

[0085] In some embodiments, compound 13 is compound 12: [ka] with p-toluenesulfonyl chloride (p-TsCl) and sodium hydride (NaH). In some embodiments, the reaction is carried out in a solvent comprising an aprotic solvent such as N,N-dimethylformamide (DMF).

[0086] In some embodiments, compound 12 is compound 11: [ka] with iron (Fe) and acetic acid (HOAc). In some embodiments, the reaction is carried out in a solvent, including an ethereal solvent such as tetrahydrofuran (THF). The combination of iron and acetic acid can be used as a reducing agent, and may be a safe alternative to reducing agents such as H2 / Raney Ni, which can be dangerous in large-scale production.

[0087] In some embodiments, compound 11 is selected from the group consisting of compound 10: [ka] with 1,1-diethoxy-N,N-dimethylmethanamine and further with B7, where B7 is a base. In some embodiments, B7 is an alkali metal alkoxide, such as lithium methanolate. In some embodiments, the reaction is carried out in a solvent, including an aprotic solvent, such as N,N-dimethylformamide (DMF).

[0088] In some embodiments, the method for preparing compound 6 includes: (i) reacting compound 1a with nitric acid and acetic acid to produce compound 2; (ii) reacting compound 2 with N-bromosuccinimide (NBS) to form compound 3; (iii) reacting compound 3 with a reducing agent to form compound 4; (iv) reacting compound 4 with 2-bromo-2-methylpropanoyl bromide and B6 to form compound 5; and (v) reacting compound 5 with a methylating agent and B5 to form compound 6.

[0089] In some embodiments, the method for preparing compound 9 includes: (i) reacting compound 10 with 1,1-diethoxy-N,N-dimethylmethanamine and B7 to produce compound 11; (ii) reacting compound 11 with iron (Fe) and acetic acid (HOAc) to produce compound 12; (iii) reacting compound 12 with p-toluenesulfonyl chloride (p-TsCl) and sodium hydride (NaH) to produce compound 13; (iv) reacting compound 13 with an acid to form compound 14; and (v) Reacting compound 14 with methyl iodide and sodium hydride to form compound 9.

[0090] In some embodiments, the method for preparing Compound 1 or a salt thereof includes: (i) reacting compound 6 with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) in the presence of P3 and B3 to produce compound 7; (ii) reacting compound 7 with compound 9 in the presence of P2 and B2 to form compound 8; and (iii) reacting compound 8 with B1 to produce compound 1 or a salt thereof.

[0091] In some embodiments, the method for preparing Compound 1 or a salt thereof includes: (i) reacting compound 6 with compound 15 in the presence of P4 and B4 to form compound 8; and (ii) reacting compound 8 with B1 to produce compound 1 or a salt thereof.

[0092] In some embodiments, the method for preparing Compound 1 or a salt thereof includes: (i) reacting compound 9 with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) in the presence of P8 and B8 to produce compound 15; (ii) reacting compound 6 with compound 15 in the presence of P4 and B4 to form compound 8; and (iii) reacting compound 8 with B1 to produce compound 1 or a salt thereof.

[0093] In some embodiments, the method for preparing Compound 1 includes: (i) reacting compound 1a with nitric acid and acetic acid to produce compound 2; (ii) reacting compound 2 with N-bromosuccinimide (NBS) to form compound 3; (iii) reacting compound 3 with a reducing agent to form compound 4; (iv) reacting compound 4 with 2-bromo-2-methylpropanoyl bromide and B6 to form compound 5; (v) reacting compound 5 with a methylating agent and B5 to produce compound 6; (vi) reacting compound 6 with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) in the presence of P3 and B3 to produce compound 7; (vii) reacting compound 7 with compound 9 in the presence of P2 and B2 to form compound 8; and (viii) reacting compound 8 with B1 to form compound 1.

[0094] In some embodiments, the method for preparing Compound 1 includes: (i) reacting compound 1a with nitric acid and acetic acid to produce compound 2; (ii) reacting compound 2 with N-bromosuccinimide (NBS) to form compound 3; (iii) reacting compound 3 with a reducing agent to form compound 4; (iv) reacting compound 4 with 2-bromo-2-methylpropanoyl bromide and B6 to form compound 5; (v) reacting compound 5 with a methylating agent and B5 to produce compound 6; (vi) reacting compound 6 with compound 15 in the presence of P4 and B4 to form compound 8; and (vii) reacting compound 8 with B1 to form compound 1.

[0095] In some embodiments, the method for preparing Compound 1 includes: (i) reacting compound 10 with 1,1-diethoxy-N,N-dimethylmethanamine and B7 to produce compound 11; (ii) reacting compound 11 with iron (Fe) and acetic acid (HOAc) to produce compound 12; (iii) reacting compound 12 with p-toluenesulfonyl chloride (p-TsCl) and sodium hydride (NaH) to produce compound 13; (iv) reacting compound 13 with an acid to form compound 14; (v) reacting compound 14 with methyl iodide and sodium hydride to produce compound 9; (vi) reacting compound 7 with compound 9 in the presence of P2 and B2 to form compound 8; and (vii) reacting compound 8 with B1 to form compound 1.

[0096] In some embodiments, the method for preparing Compound 1 includes: (i) reacting compound 10 with 1,1-diethoxy-N,N-dimethylmethanamine and B7 to produce compound 11; (ii) reacting compound 11 with iron (Fe) and acetic acid (HOAc) to produce compound 12; (iii) reacting compound 12 with p-toluenesulfonyl chloride (p-TsCl) and sodium hydride (NaH) to produce compound 13; (iv) reacting compound 13 with an acid to form compound 14; (v) reacting compound 14 with methyl iodide and sodium hydride to produce compound 9; (vi) reacting compound 9 with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) in the presence of P8 and B8 to produce compound 15; (vii) reacting compound 6 with compound 15 in the presence of P4 and B4 to form compound 8; and (viii) reacting compound 8 with B1 to form compound 1.

[0097] In some embodiments, the present invention provides [ka] or a salt thereof.

[0098] In some embodiments, provided herein is a method of reacting compound 6 with 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) in the presence of P3 and B3 to produce compound 7.

[0099] It will be understood that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment (provided that the embodiments are intended to be combined as if set forth in multiple dependent form). Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination.

[0100] In some embodiments, the term "high-temperature solution of Compound 1" used herein refers to a solution at a temperature above room temperature. For example, a high-temperature solution of Compound 1 is at a temperature above about room temperature, such as above about 20°C, above about 30°C, above about 40°C, above about 50°C, above about 60°C, above about 70°C, above about 80°C, above about 90°C, or above about 100°C.

[0101] In some embodiments, concentrating a solution as described herein refers to reducing the volume of the solution by evaporating the solvent, heating the solution, applying a vacuum to the solution, or any combination thereof.

[0102] The phrase "alkali metal bicarbonate base," used alone or in combination with other terms, as used herein, refers to a base having the formula M(HCO), where M is an alkali metal (e.g., lithium, sodium, or potassium). Examples of alkali metal bicarbonate bases include, but are not limited to, lithium bicarbonate, sodium bicarbonate, and potassium bicarbonate.

[0103] The phrase "alkali metal carbonate base," used alone or in combination with other terms, as used herein, refers to a base having the formula MCO, where M refers to an alkali metal (e.g., lithium, sodium, or potassium). Examples of alkali metal carbonate bases include, but are not limited to, lithium carbonate, sodium carbonate, and potassium carbonate.

[0104] The phrase "alkali metal hydroxide base," used alone or in combination with other terms, as used herein, refers to a base having the formula MOH, where M refers to an alkali metal (e.g., lithium, sodium, or potassium). Examples of alkali metal hydroxide bases include, but are not limited to, lithium hydroxide, sodium hydroxide, and potassium hydroxide.

[0105] The phrase "alkali metal acetate base," used alone or in combination with other terms, as used herein, refers to a base having the formula M(OC(O)CH), where M is an alkali metal (e.g., lithium, sodium, or potassium). Examples of alkali metal acetate bases include, but are not limited to, lithium acetate, sodium acetate, and potassium acetate.

[0106] As used herein, the phrase "transition metal catalyst" refers to a metal catalyst (e.g., a palladium catalyst or a nickel catalyst) suitable for catalyzing carbon-carbon coupling reactions. Examples of transition metal catalysts include, but are not limited to, PdCl(PPh), Pd(PPh), dichloro(bis{di-tert-butyl[4-(dimethylamino)phenyl]-phosphoranyl})palladium (Pd-132), NiCl(dppf), and NiCl(dppp), where (dppf) refers to 1,1'-bis(diphenylphosphino)ferrocene and (dppp) refers to 1,3-bis(diphenylphosphino)propane.

[0107] Examples of palladium catalysts include, but are not limited to, PdCl(PPh), Pd(PPh), dichloro(bis{di-tert-butyl[4-(dimethylamino)phenyl]-phosphoranyl})palladium (Pd-132), palladium on carbon, PdCl, Pd(OAc), PdCl(MeCN), tris(dibenzylideneacetone)dipalladium(0) (Pd(dba), 4-(di-tert-butylphosphino)-N,N-dimethylaniline-dichloropalladium (2:1), Pd(dppf)Cl (e.g., Pd(dppf)Cl-CHCl), and tetrakis(tri(o-tolyl)phosphine)palladium(0).

[0108] As used herein, the term "reacting" is used as known in the art and generally refers to combining chemical reagents in a manner that allows interaction at a molecular level to occur, to effect a chemical or physical transformation. In some embodiments, the reaction involves two reagents, where one or more equivalents of the second reagent relative to the first reagent are used. The reaction steps of the methods described herein can be carried out for times and under conditions suitable to prepare the specified product.

[0109] In some embodiments, an antisolvent as used herein refers to a solvent in which Compound 1 is less soluble than a solution of another solvent or solvent mixture. For example, antisolvents can include, but are not limited to, benzene, cyclohexane, pentane, hexane, heptane (e.g., n-heptane), toluene, cycloheptane, methylcyclohexane, heptane, ethylbenzene, m-xylene, o-xylene, or p-xylene, octane, indane, nonane, or naphthalene.

[0110] The reactions of the methods described herein can be carried out in a suitable solvent, which can be readily selected by one skilled in the art of organic synthesis. A suitable solvent can be substantially non-reactive with the starting materials (reactants), intermediates, or products at the temperature at which the reaction is carried out, which can range, for example, from the freezing temperature of the solvent to the boiling point of the solvent. A given reaction can be carried out in one solvent or a mixture of multiple solvents. Suitable solvents for each reaction step can be selected depending on the particular reaction step. In some embodiments, the reaction can be carried out in the absence of a solvent, such as when at least one of the reagents is a liquid or gas.

[0111] Suitable solvents may include halogenated solvents such as carbon tetrachloride, bromodichloromethane, dibromochloromethane, bromoform, chloroform, bromochloromethane, dibromomethane, butyl chloride, dichloromethane (methylene chloride), tetrachloroethylene, trichloroethylene, 1,1,1-trichloroethane, 1,1,2-trichloroethane, 1,1-dichloroethane, 2-chloropropane, α,α,α-trifluorotoluene, 1,2-dichloroethane, 1,2-dibromoethane, hexafluorobenzene, 1,2,4-trichlorobenzene, 1,2-dichlorobenzene, chlorobenzene, fluorobenzene, mixtures thereof, and the like.

[0112] Suitable ether solvents include dimethoxymethane, tetrahydrofuran, 1,3-dioxane, 1,4-dioxane, furan, tetrahydrofuran (THF), diethyl ether, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, diethylene glycol dimethyl ether (diglyme), diethylene glycol diethyl ether, triethylene glycol dimethyl ether, anisole, tert-butyl methyl ether, mixtures thereof, and the like.

[0113] By way of example and not limitation, suitable protic solvents may include water, methanol, ethanol, 2-nitroethanol, 2-fluoroethanol, 2,2,2-trifluoroethanol, ethylene glycol, 1-propanol, 2-propanol, 2-methoxyethanol, 1-butanol, 2-butanol, isobutyl alcohol, tert-butyl alcohol, 2-ethoxyethanol, diethylene glycol, 1-pentanol, 2-pentanol, or 3-pentanol, neopentyl alcohol, tert-pentyl alcohol, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, cyclohexanol, benzyl alcohol, phenol, or glycerol.

[0114] By way of example and not limitation, suitable aprotic solvents may include N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA), 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU), 1,3-dimethyl-2-imidazolidinone (DMI), N-methylpyrrolidinone (NMP), formamide, N-methylacetamide, N-methylformamide, acetonitrile, dimethyl sulfoxide, propionitrile, ethyl formate, methyl acetate, hexachloroacetone, acetone, ethyl methyl ketone, ethyl acetate, sulfolane, N,N-dimethylpropionamide, tetramethylurea, nitromethane, nitrobenzene, or hexamethylphosphoramide.

[0115] Suitable hydrocarbon solvents include benzene, cyclohexane, pentane, hexane, toluene, cycloheptane, methylcyclohexane, heptane, ethylbenzene, m-xylene, o-xylene, or p-xylene, octane, indane, nonane, or naphthalene.

[0116] The reactions of the processes described herein can be carried out in air or under an inert atmosphere. Reactions involving reagents or products that are substantially air-reactive can generally be carried out using techniques for the synthesis of air-labile materials well known to those skilled in the art.

[0117] As used herein, the expressions "ambient temperature" and "room temperature" are understood in the art and generally refer to a temperature near, for example, the reaction temperature, i.e., the temperature of the room in which the reaction is carried out, e.g., from about 20°C to about 30°C.

[0118] How to use Compound 1 or its salts is a BET protein inhibitor and is therefore useful in the treatment of diseases and disorders associated with the activity of BET proteins. Any form of Compound 1, including any of the embodiments described herein, can be used for the uses described herein.

[0119] Compound 1 can inhibit one or more of the BET proteins BRD2, BRD3, BRD4, and BRD-t. In some embodiments, compound 1 selectively inhibits one or more BET proteins over another protein. By "selective," it is meant that the compound binds to the BET protein with higher affinity or inhibits the BET protein with higher potency compared to a reference substance, such as another BET protein. For example, the compound may be selective for BRD2 over BRD3, BRD4, and BRD-t; for BRD3 over BRD2, BRD4, and BRD-t; for BRD4 over BRD2, BRD3, and BRD-t; or for BRD-t over BRD2, BRD3, and BRD-t. In some embodiments, the compound inhibits more than one BET protein or all BET proteins. In general, selectivity can be at least about 5-fold, at least about 10-fold, at least about 20-fold, at least about 50-fold, at least about 100-fold, at least about 200-fold, at least about 500-fold, or at least about 1000-fold.

[0120] Compound 1 is therefore useful for treating BET protein-mediated disorders. The terms "BET protein-mediated disorder" or "BET-mediated disorder" refer to any disorder, disease, or condition in which one or more BET proteins, such as BRD2, BRD3, BRD4, and / or BRD-t, or variants thereof, are implicated, or where the disease or condition is associated with the expression or activity of one or more BET proteins. Thus, compound 1, as an inhibitor of BET proteins, can be used to treat or lessen the severity of diseases and conditions in which BET proteins, such as BRD2, BRD3, BRD4, and / or BRD-t, or variants thereof, are known to be involved.

[0121] Diseases and conditions treatable with Compound 1 include, but are not limited to, cancer and other proliferative disorders, autoimmune diseases, chronic inflammatory diseases, acute inflammatory diseases, sepsis, and viral infections. Diseases can be treated by administering a therapeutically effective amount or dosage of Compound 1 or any of its embodiments, or a pharmaceutical composition thereof to an individual (e.g., a patient) in need of treatment. The present disclosure also provides a solid form of Compound 1 or any of its embodiments, or a pharmaceutical composition comprising the solid form, for use in treating a BET-mediated disease or disorder. Also provided is the use of a solid form of Compound 1 or any of its embodiments, or a pharmaceutical composition comprising the solid form, in the manufacture of a medicament for treating a BET-mediated disease or disorder.

[0122] Diseases treatable with Compound 1 include cancers, including adrenal gland carcinoma, acinic cell carcinoma, acoustic neuroma, acral lentiginous melanoma, acral hidradenoma, acute eosinophilic leukemia, acute erythroleukemia, acute lymphocytic leukemia, acute megakaryoblastic leukemia, acute monocytic leukemia, acute promyelocytic leukemia, adenocarcinoma, adenoid cystic carcinoma, adenoma, adenoid odontogenic tumor, adenosquamous carcinoma, adipose tissue tumors, adrenocortical carcinoma, adult T-cell leukemia / lymphoma, aggressive NK-cell leukemia, AIDS-related lymphoma, alveolar rhabdomyosarcoma, alveolar soft part sarcoma, ameloblastic fibroma, anaplastic large cell lymphoma, anaplastic thyroid carcinoma, angioimmunoblastic T-cell lymphoma, angiomyolipoma, angiosarcoma, astrocytic leukemia, and steroid hormone receptor agonist (SHR) receptor agonist (SHR). Cell tumor, atypical teratoid rhabdoid tumor, B-cell chronic lymphocytic leukemia, B-cell prolymphocytic leukemia, B-cell lymphoma, basal cell carcinoma, biliary tract cancer, bladder cancer, blastoma, bone cancer, Brenner tumor, pheochromocytoma, Burkitt lymphoma, breast cancer, brain tumor, carcinoma, carcinoma in situ, carcinosarcoma, cartilage tumor, cementoma, myeloid sarcoma, chondroma, chordoma, choriocarcinoma, choroid plexus papilloma, clear cell sarcoma of the kidney, craniopharyngioma, cutaneous T-cell lymphoma, cervical cancer, colorectal cancer, Degos disease, desmoplastic small round cell tumor, diffuse large B-cell lymphoma, dysembryoplastic neuroepithelial tumor, dysgerminoma, embryonal carcinoma, Endocrine tumors, endodermal sinus tumors, enteropathy-type T-cell lymphoma, esophageal cancer, fetal in utero, fibroma, fibrosarcoma, follicular lymphoma, follicular thyroid carcinoma, ganglioneuroma, gastrointestinal cancer, germ cell tumors, gestational choriocarcinoma, giant cell fibroblastoma, giant cell tumor of bone, glioblastoma multiforme, glioma, gliomatosis cerebri, glucagonoma, gonadoblastoma, granulosa cell tumor, ginandroblastoma, gallbladder cancer, gastric cancer, hairy cell leukemia, hemangioblastoma, head and neck cancer, hemangiopericytoma, hematologic malignancies, hepatoblastoma, hepatosplenic T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, invasive lobular carcinoma, intestinal cancer, kidney Pancreatic cancer, laryngeal cancer, lentigo maligna, lethal midline carcinoma, leukemia, Leydig cell tumor, liposarcoma, lung cancer, lymphangioma, lymphangiosarcoma, lymphoepithelioma, lymphoma, acute lymphocytic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, liver cancer, small cell lung cancer, non-small cell lung cancer, MALT lymphoma, malignant fibrous histiocytoma, malignant peripheral nerve sheath tumor, malignant Triton tumor, mantle cell lymphoma, marginal zone B-cell lymphoma, mast cell leukemia, mediastinal germ cell tumor, medullary breast carcinoma, medullary thyroid carcinoma, medulloblastoma, melanoma, meningioma, Merkel cell carcinoma, mesothelioma, metastatic urothelial carcinoma, mixed Mullerian tumor,Mucinous tumor, multiple myeloma, muscle tissue tumor, mycosis fungoides, myxoid liposarcoma, myxoma, myxosarcoma, nasopharyngeal carcinoma, schwannoma, neuroblastoma, neurofibroma, neuroma, nodular melanoma, ocular carcinoma, oligoastrocytoma, oligodendroglioma, oncocytoma, optic nerve sheath meningioma, optic nerve tumor, oral cancer, osteosarcoma, ovarian cancer, Pancoast tumor, papillary thyroid carcinoma, paraganglioma, pineoblastoma, pineocytoma, pituitary cell tumor, pituitary adenoma, pituitary tumor, plasmacytoma, polyembryomatous tumor, precursor T-lymphoblastic lymphoma, primary central nervous system lymphoma, primary effusion lymphoma, primary peritoneal carcinoma, prostate cancer, pancreatic cancer, pharyngeal cancer, pseudomyxoma peritonei, renal cell carcinoma, renal medullary carcinoma, retinoblastoma, rhabdomyoma, striated muscle The tumor may include, but is not limited to, sarcoma, Richter's transformation, rectal cancer, sarcoma, schwannomatosis, seminoma, Sertoli cell tumor, sex cord-stromal tumor, signet ring cell carcinoma, skin cancer, small round blue cell tumor, small cell carcinoma, soft tissue sarcoma, somatostatinoma, sooty warts, spinal tumor, splenic marginal zone lymphoma, squamous cell carcinoma, synovial sarcoma, Sezary's disease, small intestine cancer, squamous cell carcinoma, gastric cancer, T-cell lymphoma, testicular cancer, thecoma, thyroid cancer, transitional cell carcinoma, throat cancer, urachal carcinoma, genitourinary cancer, urothelial carcinoma, uveal melanoma, uterine cancer, verrucous carcinoma, optic pathway glioma, vulvar cancer, vaginal cancer, Waldenstrom's macroglobulinemia, Warthin's tumor, and Wilms' tumor. In some embodiments, the cancer is selected from the group consisting of adenocarcinoma, adult T-cell leukemia / lymphoma, bladder cancer, blastoma, bone cancer, breast cancer, brain tumor, carcinoma, myeloid sarcoma, cervical cancer, colon cancer, esophageal cancer, gastrointestinal cancer, glioblastoma multiforme, glioma, gallbladder cancer, stomach cancer, head and neck cancer, Hodgkin's lymphoma, non-Hodgkin's lymphoma, intestinal cancer, kidney cancer, laryngeal cancer, leukemia, lung cancer, lymphoma, liver cancer, small cell lung cancer, non-small cell lung cancer, mesothelioma, multiple myeloma, The cancer may be acute myeloid leukemia (AML), diffuse large B-cell lymphoma (DLBCL), eye cancer, optic nerve tumor, oral cancer, ovarian cancer, pituitary tumor, primary central nervous system lymphoma, prostate cancer, pancreatic cancer, pharyngeal cancer, renal cell carcinoma, rectal cancer, sarcoma, skin cancer, spinal tumor, small intestine cancer, gastric cancer, T-cell lymphoma, testicular cancer, thyroid cancer, throat cancer, genitourinary cancer, urothelial carcinoma, uterine cancer, vaginal cancer, or Wilms' tumor.

[0123] In some embodiments, the cancer is a hematological cancer.

[0124] In some embodiments, the cancer is multiple myeloma, acute myeloid leukemia (AML), or diffuse large B-cell lymphoma (DLBCL).

[0125] Diseases treatable with Compound 1 also include MYC-dependent cancers, which are cancers associated with at least one of myc RNA expression or MYC protein expression. Patients eligible for such treatment can be identified by measuring myc RNA expression or MYC protein expression in cancer tissues or cells.

[0126] Diseases treatable with Compound 1 also include non-cancerous proliferative disorders. Examples of treatable proliferative diseases include, but are not limited to, benign soft tissue tumors, bone tumors, brain and spinal cord tumors, eyelid and orbital tumors, granulomas, lipomas, meningiomas, multiple endocrine neoplasia, nasal polyps, pituitary tumors, prolactinomas, pseudotumor cerebri, seborrheic keratosis, gastric polyps, thyroid nodules, cystic pancreatic tumors, hemangiomas, vocal cord nodules, polyps, and cysts, Castleman's disease, chronic pilonidal disease, dermatofibromas, pilonidal cysts, pyogenic granulomas, and juvenile polyposis syndrome.

[0127] Diseases and conditions treatable with Compound 1 also include chronic autoimmune and inflammatory conditions. Examples of treatable autoimmune and inflammatory conditions include acute, hyperacute, or chronic organ transplant rejection, acute gout, acute inflammatory reactions (such as acute respiratory distress syndrome and ischemia / reperfusion injury), Addison's disease, agammaglobulinemia, allergic rhinitis, allergies, alopecia, Alzheimer's disease, appendicitis, atherosclerosis, asthma, osteoarthritis, juvenile arthritis, psoriatic arthritis, rheumatoid arthritis, and atopic dermatitis. inflammation, autoimmune alopecia, autoimmune hemolytic and thrombocytopenic states, autoimmune hypopituitarism, autoimmune polyendocrine disorders, Behçet's disease, bullous skin diseases, cholecystitis, chronic idiopathic thrombocytopenic purpura, chronic obstructive pulmonary disease (COPD), cirrhosis, osteoarthritis, depression, dermatitis, dermatomyositis, eczema, enteritis, encephalitis, gastritis, glomerulonephritis, giant cell arteritis, Goodpasture's syndrome, Guillain-Barré syndrome, gingivitis Inflammation, Graves' disease, Hashimoto's thyroiditis, hepatitis, hypophysitis, inflammatory bowel disease (Crohn's disease and ulcerative colitis), inflammatory pelvic disease, irritable bowel syndrome, Kawasaki disease, LPS-induced endotoxin shock, meningitis, multiple sclerosis, myocarditis, myasthenia gravis, mycosis fungoides, myositis, nephritis, osteomyelitis, pancreatitis, Parkinson's disease, pericarditis, pernicious anemia, pneumonia, primary biliary sclerosing cholangitis, polyarteritis nodosa, psoriasis, retinitis, scleritis, sclera These include scleracierma, scleroderma, sinusitis, Sjogren's disease, sepsis, septic shock, sunburn, systemic lupus erythematosus, tissue transplant rejection, thyroiditis, type 1 diabetes, Takayasu's arteritis, urethritis, uveitis, vasculitis, vasculitis including giant cell arteritis, vasculitis with organ involvement such as glomerulonephritis, vitiligo, Waldenstrom's macroglobulinemia, and Wegener's granulomatosis.

[0128] Diseases and conditions treatable with Compound 1 also include diseases and conditions involving an inflammatory response to infection with bacteria, viruses, fungi, parasites, or their toxins, such as sepsis, sepsis syndrome, septic shock, endotoxemia, systemic inflammatory response syndrome (SIRS), multiple organ dysfunction syndrome, toxic shock syndrome, acute lung injury, ARDS (adult respiratory distress syndrome), acute renal failure, fulminant hepatitis, burns, acute pancreatitis, postoperative syndrome, sarcoidosis, Herxheimer reaction, encephalitis, myelitis, meningitis, malaria, and SIRS associated with viral infections such as influenza, herpes zoster, herpes simplex, and coronavirus.

[0129] Other diseases treatable with Compound 1 include viral infections. Examples of treatable viral infections include Epstein-Barr virus, hepatitis B virus, hepatitis C virus, herpes virus, human immunodeficiency virus, human papillomavirus, adenovirus, poxvirus, and other episomal DNA viruses. Thus, Compound 1 can be used to treat diseases and conditions such as herpes simplex infection and reactivation, herpes simplex, shingles infection and reactivation, chickenpox, shingles, human papillomavirus, cervical tumors, adenovirus infections associated with acute respiratory disease, and poxvirus infections such as cowpox and smallpox, as well as African swine fever. In some embodiments, Compound 1 can be used to treat human papillomavirus infections of the skin or cervical epithelium.

[0130] Diseases and conditions treatable with Compound 1 also include conditions associated with ischemia-reperfusion injury, including, but not limited to, myocardial infarction, cerebrovascular ischemia (stroke), acute coronary syndrome, renal reperfusion injury, organ transplantation, coronary artery bypass grafting, cardiopulmonary bypass surgery, and pulmonary, renal, hepatic, gastrointestinal, or peripheral limb embolism.

[0131] Compound 1 is also useful for the treatment of lipid metabolism disorders through modulation of APO-A1, such as hypercholesterolemia, atherosclerosis, and Alzheimer's disease.

[0132] Compound 1 is also useful in the treatment of fibrotic conditions such as idiopathic pulmonary fibrosis, renal fibrosis, post-operative stenosis, keloid formation, scleroderma, and myocardial fibrosis.

[0133] Compound 1 can also be used to treat ophthalmic indications such as dry eye.

[0134] Compound 1 can also be used to treat cardiac diseases such as heart failure.

[0135] As used herein, the term "contacting" refers to bringing together the specified components in an in vitro or in vivo system. For example, "contacting" a BET protein with Compound 1 (e.g., Compound 1 in a solid form, such as a crystalline solid form) encompasses administering Compound 1 to an individual or patient, e.g., a human, having a BET protein, as well as introducing, e.g., a solid form of a compound provided herein, into a sample containing a cell preparation or purified preparation containing a BET protein.

[0136] As used herein, the terms "individual" or "patient" are used interchangeably and refer to any animal, including a mammal, preferably a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse, or primate, and most preferably a human.

[0137] As used herein, the phrase "therapeutically effective amount" refers to that amount of an active compound or pharmaceutical agent that elicits the biological or medicinal response desired by a researcher, veterinarian, physician, or other clinician in a tissue, system, animal, individual, or human.

[0138] As used herein, the term "treating" or "treatment" refers to inhibiting a disease, e.g., inhibiting a disease, condition, or disorder in an individual experiencing or developing a lesion or symptom of the disease, condition, or disorder (i.e., preventing further development of the lesion and / or symptom), or ameliorating a disease, e.g., ameliorating a disease, condition, or disorder in an individual experiencing or developing a lesion or symptom of the disease, condition, or disorder (i.e., reversing the lesion and / or symptom), e.g., reducing the severity of the disease.

[0139] As used herein, the term "prevent" or "prevention" refers to preventing a disease, e.g., preventing a disease, condition, or disorder in an individual who may be susceptible to the disease, condition, or disorder but who has not yet experienced or developed the lesions or symptoms of the disease.

[0140] Combination therapy Compound 1 can be used in combination therapy, in which Compound 1 is administered in conjunction with other therapies, such as the administration of one or more additional therapeutic agents. The additional therapeutic agents are typically those commonly used to treat the particular condition being treated. Additional therapeutic agents for the treatment of BET protein-associated diseases, disorders, or conditions can include, for example, chemotherapeutic agents, anti-inflammatory agents, steroids, immunosuppressants, and Bcr-Abl, Flt-3, RAF, FAK, and JAK kinase inhibitors. The one or more additional pharmaceutical agents can be administered to the patient simultaneously or sequentially.

[0141] In some embodiments, Compound 1 can be used in combination with a therapeutic agent that targets epigenetic regulators. Examples of epigenetic regulators include histone lysine methyltransferase, histone arginine methyltransferase, histone demethylase, histone deacetylase, histone acetylase, and DNA methyltransferase. Examples of histone deacetylase inhibitors include vorinostat.

[0142] To treat cancer and other proliferative diseases, Compound 1 can be used in combination with chemotherapeutic agents or other antiproliferative agents. Compound 1 can also be used in combination with medical therapies such as surgery or radiation therapy, e.g., gamma irradiation, neutron radiation therapy, electron beam radiation therapy, proton therapy, brachytherapy, and unsealed brachytherapy. Examples of suitable chemotherapeutic agents include abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, bleomycin, bortezomib, intravenous busulfan, oral busulfan, calsterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, and clofala. cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, denileukin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolone propionate, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib mesylate, interferon alfa-2a, irinotecan, lapatinib nitrile, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, mechlorethamine, megestrol acetate, melphalan, mercaptopril methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone phenpropionate, nelarabine, nofetumomab, oxaliplatin, paclitaxel, pamidronate, panitumumab, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobroman, plicamycin, procarbazine, quinacrine, rasburicase, rituximab, ruxolitinib, sorafenib, streptozocin,sunitinib, sunitinib maleate, tamoxifen, temozolomide, teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, and zoledronic acid.

[0143] Compound 1 can be used in combination with ruxolitinib to treat cancer and other proliferative diseases.

[0144] Compound 1 can be used in combination with one or more immune checkpoint inhibitors. Exemplary immune checkpoint inhibitors include inhibitors of immune checkpoint molecules such as CD27, CD28, CD40, CD122, CD96, CD73, CD47, OX40, GITR, CSF1R, JAK, PI3K delta, PI3K gamma, TAM, arginase, CD137 (also known as 4-1BB), ICOS, A2AR, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, VISTA, PD-1, PD-L1, and PD-L2. In some embodiments, the immune checkpoint molecule is a stimulatory checkpoint molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, and CD137. In some embodiments, the immune checkpoint molecule is an inhibitory checkpoint molecule selected from A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3, and VISTA. In some embodiments, the compounds provided herein can be used in combination with one or more agents selected from a KIR inhibitor, a TIGIT inhibitor, a LAIR1 inhibitor, a CD160 inhibitor, a 2B4 inhibitor, and a TGFRbeta inhibitor.

[0145] In some embodiments, the inhibitor of an immune checkpoint molecule is an anti-PD1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

[0146] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of PD-1, such as an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab, pembrolizumab (also known as MK-3475), pidilizumab, SHR-1210, PDR001, or AMP-224. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab or pembrolizumab. In some embodiments, the anti-PD1 antibody is pembrolizumab.

[0147] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of PD-L1, such as an anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-L1 monoclonal antibody is BMS-935559, MEDI4736, MPDL3280A (also known as RG7446), or MSB0010718C. In some embodiments, the anti-PD-L1 monoclonal antibody is MPDL3280A or MEDI4736.

[0148] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CTLA-4, e.g., an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab.

[0149] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of LAG3, e.g., an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016 or LAG525.

[0150] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of GITR, e.g., an anti-GITR antibody. In some embodiments, the anti-GITR antibody is TRX518 or MK-4166.

[0151] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of OX40, such as an anti-OX40 antibody or an OX40L fusion protein. In some embodiments, the anti-OX40 antibody is MEDI0562. In some embodiments, the OX40L fusion protein is MEDI6383.

[0152] Compound 1 can be used in combination with one or more drugs for the treatment of diseases such as cancer. In some embodiments, the drug is an alkylating agent, a proteasome inhibitor, a corticosteroid, or an immunomodulatory agent. Examples of alkylating agents include cyclophosphamide (CY), melphalan (MEL), and bendamustine. In some embodiments, the proteasome inhibitor is carfilzomib. In some embodiments, the corticosteroid is dexamethasone (DEX). In some embodiments, the immunomodulatory agent is lenalidomide (LEN) or pomalidomide (POM).

[0153] To treat autoimmune or inflammatory conditions, Compound 1 can be administered in conjunction with a corticosteroid, such as triamcinolone, dexamethasone, fluocinolone, cortisone, prednisolone, or flumetholone.

[0154] To treat autoimmune or inflammatory conditions, Compound 1 can be administered in conjunction with immunosuppressants such as fluocinolone acetonide (Retisert®), rimexolone (AL-2178, Vexol, Alcon), or cyclosporine (Restasis®).

[0155] Compound 1 may be administered in combination with other steroids, including Dehydrex™ (Holles Labs), Civamide (Opko), sodium hyaluronate (Vismed, Lantibio / TRB Chemedia), cyclosporine (ST-603, Sirion Therapeutics), ARG101(T) (testosterone, Argentis), AGR1012(P) (Argentis), ecabet sodium (Senju-Ista), gefarnate (Santen), 15-(s)-hydroxyeicosatetraenoic acid (15(S)-HETE), cevilemine, doxycycline (ALTY-0501, Alacrity), minocycline, iDestrin™ (NP50301, Nascent), steroids, and steroids for the treatment of autoimmune or inflammatory conditions. Pharmaceuticals), cyclosporine A (Nova22007, Novagali), oxytetracycline (Duramycin, MOLI1901, Lantibio), CF101(2S,3S,4R,5R)-3,4-dihydroxy-5-[6-[(3-iodophenyl)methylamino]purin-9-yl]-N-methyl-oxolane-2-carbamyl, Can-Fite Biopharma), voclosporin (LX212 or LX214, Lux Biosciences), ARG103 (Agentis), RX-10045 (synthetic resolvin analog, Resolvyx), DYN15 (Dyanmis Therapeutics), rivoglitazone (DE011, Daiichi Sanko), TB4 (RegeneRx), OPH-01 (Ophtalmis Monaco), PCS101 (Pericor Science), REV1-31 (Evolutec), lacritin (Senju), rebamipide (Otsuka-Novartis), OT-551 (Othera), PAI-2 (University of Pennsylvania and Temple University) It may be administered in conjunction with one or more additional agents selected from: pilocarpine, tacrolimus, pimecrolimus (AMS981, Novartis), loteprednol etabonate, rituximab, diquafosol tetrasodium (INS365, Inspire), KLS-0611 (Kissei Pharmaceuticals), dehydroepiandrosterone, anakinra, efalizumab, mycophenolate sodium, etanercept (Embrel®), hydroxychloroquine, NGX267 (TorreyPines Therapeutics), or thalidomide.

[0156] In some embodiments, Compound 1 can be administered in combination with one or more drugs selected from antibiotics, antivirals, antifungals, anesthetics, anti-inflammatory drugs, including steroidal and non-steroidal anti-inflammatory drugs, and anti-allergy drugs. Suitable drugs include aminoglycosides such as amikacin, gentamicin, tobramycin, streptomycin, netilmicin, and kanamycin; fluoroquinolones such as ciprofloxacin, norfloxacin, ofloxacin, trovafloxacin, lomefloxacin, levofloxacin, and enoxacin; naphthyridine; sulfonamides; polymyxins; chloramphenicol; neomycin; paramomycin; colistimethate; bacitracin; vancomycin; and the like. comycin; tetracyclines; rifampin and its derivatives ("rifampins"); cycloserine; beta-lactams; cephalosporins; amphotericins; fluconazole; flucytosine; natamycin; miconazole; ketoconazole; corticosteroids; diclofenac; flurbiprofen; ketorolac; suprofen; cromolyn; lodoxamide; levocabastine; naphazoline; antazoline; pheniramine; or azalide antibiotics.

[0157] Other examples of one or more agents that may be used in combination with the provided compounds include drugs for treating Alzheimer's disease, such as donepezil and rivastigmine; drugs for treating Parkinson's disease, such as L-DOPA / carbidopa, entacapone, ropinirole, pramipexole, bromocriptine, pergolide, trihexyphenidyl, and amantadine; drugs for treating multiple sclerosis (MS), such as beta interferons (e.g., Avonex® and Rebif®), glatiramer acetate, and mitoxantrone; drugs for treating asthma, such as albuterol and montelukast; drugs for treating schizophrenia, such as Zyprexa, Risperdal, Seroquel, and haloperidol; corticosteroids, such as dexamethasone or prednisone, TNF blockers, IL-1 anti-inflammatory agents such as RA, azathioprine, cyclophosphamide, and sulfasalazine; immunomodulators including immunosuppressants such as cyclosporine, tacrolimus, rapamycin, mycophenolate mofetil, interferon, corticosteroids, cyclophosphamide, azathioprine, and sulfasalazine; neurotrophic factors such as acetylcholinesterase inhibitors, MAO inhibitors, interferons, anticonvulsants, ion channel blockers, riluzole, or antiparkinsonian drugs; drugs to treat cardiovascular disease such as beta-blockers, ACE inhibitors, diuretics, nitrates, calcium channel blockers, or statins; drugs to treat liver disease such as corticosteroids, cholestyramine, interferons, and antivirals; drugs to treat blood disorders such as corticosteroids, anti-leukemia drugs, or growth factors; or drugs to treat immune deficiency disorders such as gamma globulin.

[0158] In some embodiments, compound 1 is administered in combination with a JAK kinase inhibitor (e.g., ruxolitinib, tofacitinib, baricitinib, CYT387, GLPG0634, lestaurtinib, pacritinib, TG101348, or a JAK1-selective inhibitor), a Pim kinase inhibitor (including one or more inhibitors of PIM1, PIM2, and PIM3), a PI3 kinase inhibitor including a PI3K delta-selective inhibitor and a broad-spectrum PI3K inhibitor, a MEK inhibitor, a cyclin-dependent kinase inhibitor, a b-RAF inhibitor, an mTOR inhibitor, a proteasome inhibitor (e.g., bortezomib, carfilzomib), an HDAC inhibitor (e.g., panobinostat, vorinostat), a DNA methyltransferase inhibitor, dexamethasone, melphalan, or an immunomodulatory agent (e.g., lenolidomide, pomalidomide).

[0159] Formulation, Dosage Form, and Administration When used as a pharmaceutical, Compound 1 (e.g., Compound 1 in a solid form, such as a crystalline solid form) can be administered in the form of a pharmaceutical composition. These compositions can be prepared by methods well known in the pharmaceutical art and can be administered by a variety of routes, depending on whether local or systemic treatment is desired and the area to be treated.

[0160] Administration can be topical (transdermal, epithelial, intraocular, and transmucosal, including intranasal, vaginal, and rectal delivery), pulmonary (e.g., by inhalation or inhalation of powders or aerosols, including with a nebulizer; intratracheal or intranasal), oral, or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, or intramuscular injection or infusion; or intracranial, e.g., intrathecal or intraventricular, administration. Parenteral administration can be in the form of a single bolus dose or can be, for example, via a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration can include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Conventional pharmaceutical carriers, aqueous, powder, or oily bases, thickeners, and the like may be necessary or desirable.

[0161] The present application also includes pharmaceutical compositions containing Compound 1 or a pharmaceutically acceptable salt thereof as an active ingredient in combination with one or more pharmaceutically acceptable carriers (excipients). In some embodiments, the compositions are suitable for topical administration. In preparing the compositions described herein, the active ingredient is typically mixed with an excipient, diluted with an excipient, or enclosed within such a carrier, for example, in the form of a capsule, sachet, paper, or other container. When the excipient serves as a diluent, it can be a solid, semi-solid, or liquid substance that acts as a vehicle, carrier, or medium for the active ingredient. Thus, the compositions can be in the form of tablets, pills, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solids or in liquid vehicles), ointments containing, for example, up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

[0162] When preparing a formulation, Compound 1 can be milled to an appropriate particle size before being combined with other ingredients. If Compound 1 is substantially insoluble, it can be milled to a particle size of less than 200 mesh. If Compound 1 is substantially water-soluble, it can be milled to a particle size of, for example, about 40 mesh, allowing it to be dispersed substantially uniformly in the formulation.

[0163] Compound 1 may be milled using known milling procedures, such as wet milling, to produce a particle size suitable for tablet formulations and other formulation types. Micronized (nanoparticulate) preparations of Compound 1 can be prepared by methods known in the art (see, e.g., International Application No. WO 2002 / 000196).

[0164] Some examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup and methylcellulose.Preparation can also include lubricants such as talc, magnesium stearate and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methylbenzoate and propylhydroxybenzoate; sweeteners; and flavoring agents.Compositions provided herein can be formulated by using procedures known in the art to obtain rapid release, sustained release or delayed release of active ingredient after administration to patients.

[0165] The composition can be formulated in a unit dosage form containing a desired amount of the active ingredient. The term "unit dosage form" refers to a physically discrete unit suitable as a unitary dosage for human subjects and other mammals, each unit containing a predetermined amount of active material calculated to obtain a desired therapeutic effect in the presence of a suitable pharmaceutical excipient.

[0166] The active compound may be effective over a wide dosage range, but is generally administered in a pharmaceutically effective amount, although it will be understood that the amount of compound actually administered will usually be determined by the physician depending on the relevant circumstances, including the condition being treated, the selected route of administration, the actual compound being administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, etc.

[0167] When preparing solid compositions such as tablets, the main active ingredient is mixed with pharmaceutical excipients to form a solid pre-formulation composition containing a homogeneous mixture of the compounds provided herein.When such a pre-formulation composition is referred to as homogeneous, the active ingredient is generally evenly dispersed throughout the composition so that the composition can be easily subdivided into unit dosage forms of equal potency, such as tablets, pills, and capsules.This solid pre-formulation is then subdivided into the above-mentioned types of unit dosage forms.

[0168] The tablets or pills described herein can be coated or otherwise compounded to provide a dosage form that offers the advantage of prolonged action. For example, the tablet or pill can be in the form of an outer coating containing an inner and outer dosage component. The two components are separated by an enteric layer that resists disintegration in the stomach, allowing the inner component to pass intact into the duodenum or to be delayed in release. A variety of materials can be used for such enteric layers or coatings, including numerous polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

[0169] Liquid forms into which Compound 1 and compositions described herein may be incorporated when administered orally or by injection include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and flavored emulsions with edible oils (such as cottonseed oil, sesame oil, coconut oil, or peanut oil), as well as elixirs and similar pharmaceutical vehicles.

[0170] Compositions for inhalation or inhalation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents, or mixtures thereof, as well as powders. Liquid or solid compositions can contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, the compositions are administered via the oral or nasal respiratory route for local or systemic effects. The compositions can be nebulized using an inert gas. Nebulized solutions can be inhaled directly from the nebulizing device, or the nebulizing device can be attached to a face mask, tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions can be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0171] Topical formulations may contain one or more conventional carriers. In some embodiments, ointments may contain water and one or more hydrophobic carriers (e.g., selected from liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum, etc.). The carrier composition of creams may be an aqueous component in combination with glycerol and one or more other ingredients, such as glycerol monostearate, PEG-glycerol monostearate, and cetylstearyl alcohol. Gels may be formulated using isopropyl alcohol and water, appropriately combined with other ingredients, such as glycerol, hydroxyethylcellulose, etc. Topical formulations may be suitably packaged, for example, in 100g tubes, optionally accompanied by instructions for the treatment of a selected indication, such as psoriasis or other skin conditions.

[0172] The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of the administration, such as prophylaxis or treatment, the condition of the patient, the method of administration, etc. For therapeutic use, the composition can be administered to a patient already suffering from a disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective amount will be determined by the judgment of the attending clinician depending on factors such as the pathology of the disease being treated and the severity of the disease, the age, weight, and general condition of the patient, etc.

[0173] The compositions administered to patients may be in the form of pharmaceutical compositions described above. These compositions may be sterilized by conventional sterilization techniques or may be sterile filtered. Aqueous solutions may be packaged for ready use or lyophilized, with the lyophilized preparation being combined with a sterile aqueous carrier prior to administration. The pH of the compound preparations is generally 3-11, more preferably 5-9, and most preferably 7-8. It will be appreciated that the use of certain of the above-mentioned excipients, carriers, or stabilizers may result in the formation of pharmaceutical salts.

[0174] The therapeutic dosage of Compound 1 can vary according to, for example, the particular application being treated, the method of administration of the compound, the patient's health and condition, and the judgment of the prescribing physician. The proportion or concentration of a compound provided herein in a pharmaceutical composition can vary depending on several factors, including dosage, chemical characteristics (e.g., hydrophobicity), and route of administration. The dosage can depend on variables such as the type and progression of the disease or disorder, the overall health of the particular patient, the relative biological availability of the selected compound, the excipient formulation, and its route of administration. Effective amounts can be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0175] The compositions provided herein can further comprise one or more additional pharmaceutical agents, such as a chemotherapeutic agent, a steroid, an anti-inflammatory compound, or an immunosuppressant, examples of which are listed above.

[0176] The present invention will be further described in detail by way of specific examples. The following examples are provided for illustrative purposes and are not intended to limit the present invention in any way. Those skilled in the art will readily recognize various non-critical parameters that can be changed or modified to achieve essentially the same results. As described below, the compounds of the examples have been found to be inhibitors of one or more BET proteins. [Example]

[0177] Example 1. Synthesis of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (Compound 1) The synthesis of intermediate compound 5 was carried out according to Scheme 1. [ka]

[0178] Step 1a. 4-(Methylsulfonyl)-2-nitrophenol (Compound 2) To a stirred solution of 4-(methylsulfonyl)-phenol (Compound 1a, 10 g, 58.1 mmol) in acetic acid (HOAc, 91 mL) was added nitric acid (69%, 4.2 mL, 70 mmol, 1.2 equiv.) over 1 min at room temperature. When an exotherm was observed, the reaction was heated to 70 °C. The reaction mixture was stirred at 75–80 °C for 3 h. Nitric acid (69%, 0.3 mL, 5.0 mmol, 0.086 equiv.) was added, and the mixture was stirred for an additional 1 h. The reaction mixture was cooled to 15 °C, and water (230 mL) was added. After stirring for 30 min, the resulting solid was collected by filtration, rinsed with water (2 × 45 mL), and dried under vacuum at 45 °C for 5 h to give the desired crude product, 4-(methylsulfonyl)-2-nitrophenol (Compound 2, 11.0 g). Crude compound 2 was then dissolved in tetrahydrofuran (THF, 110 mL) at 55°C, and warm water (45°C, 275 mL) was slowly added. The solution was gradually cooled to room temperature and stirred overnight at room temperature, then further cooled to 9°C and stirred at 9°C for 1 hour. The solid was collected by filtration and dried under vacuum at 50°C overnight to give 4-(methylsulfonyl)-2-nitrophenol (compound 2, 10.15 g, theoretical 12.6 g, 80.6% yield) as a yellow powder. Compound 2: CHNOS (M+H) + LCMS calculated for: 218.0, found: 218.1; 1 H NMR (300 MHz, DMSO-d6) δ 12.20 (br s, 1H), 8.34 (d, J = 2.4 Hz, 1H), 8.00 (dd, J = 8.8 Hz, J = 2.4 Hz, 1H), 7.30 (d, J = 8.8 Hz, 1H), 3.22 (s, 3H) ppm.

[0179] Step 2a. 2-Bromo-4-(methylsulfonyl)-6-nitrophenol (Compound 3) To a solution of 4-(methylsulfonyl)-2-nitrophenol (compound 2, 825 g, 3.8 mol) in DMF (5.9 L) was added N-bromosuccinimide (NBS, 680 g, 3.82 mol, 1.0 equiv.) at 0 °C. After 10 min, the cold bath was removed and the reaction mixture was stirred at room temperature for 2 h. When the reaction was complete as determined by LCMS, water (5.9 L) was added and the mixture was stirred at room temperature for 1 h. The solid was filtered, washed with water (3 × 2.5 L), and dried under vacuum at 45 °C overnight to give 2-bromo-4-(methylsulfonyl)-6-nitrophenol (compound 3, 1085 g, theoretical 1131.1 g, 95.9% yield) as a yellow powder, which was used in the next reaction without further purification. Compound 3: CHBrNOS(MH) - LCMS calculated for: 293.9, found: 294.0; 1 H NMR (300 MHz, DMSO-d6) δ 8.33 (d, J = 2.0 Hz, 1H), 8.31 (d, J = 2.0 Hz, 1H), 3.27 (s, 3H) ppm.

[0180] Step 3a. 2-Amino-6-bromo-4-(methylsulfonyl)phenol (Compound 4) To a solution of 2-bromo-4-(methylsulfonyl)-6-nitrophenol (compound 3, 1037 g, 3.5 mol) and sodium hydrosulfite (NaSO, technical grade 85%, 3.15 kg, 15.4 mol, 4.4 equiv.) in a 1:1 mixture of tetrahydrofuran (THF, 10 L) and water (10 L), sodium bicarbonate (NaHCO, 2.6 kg, 30.95 mol, 8.8 equiv.) was added portionwise over 1 h. The resulting reaction mixture was stirred at room temperature for 2 h. Upon completion of the reaction as determined by LCMS, the reaction mixture was extracted with ethyl acetate (EtOAc, 2 × 10 L). The combined organic layers were concentrated under reduced pressure. The residue was dissolved in ethyl acetate (EtOAc, 13 L), and the insoluble material was removed by filtration. The filtrate was evaporated under reduced pressure to give crude 2-amino-6-bromo-4-(methylsulfonyl)phenol (compound 4, 736.5 g, theoretical 931.4 g, 79% yield) as a beige powder, which was used in the next reaction without further purification. Compound 4: C7H8BrNO3S (M+H) + LCMS calculated for: 265.9, found: 266.1; 1 H NMR (300 MHz, DMSO-d6) δ 7.15 (d, J = 2.4 Hz, 1H), 7.10 (d, J = 2.4 Hz, 1H), 6.8 (br s, 2H), 3.4 (br s, 1H), 3.09 (s, 3H) ppm.

[0181] Step 4a. 8-Bromo-2,2-dimethyl-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (Compound 5) To a solution of 2-amino-6-bromo-4-(methylsulfonyl)phenol (compound 4, 391 g, 1.47 mol) in acetonitrile (8 L) was added a solution of potassium carbonate (KCO, 842 g, 6.1 mol, 4.15 equiv.) in water (2.8 L) at room temperature. 2-Bromo-2-methylpropanoyl bromide (466 mL, 864 g, 3.76 mol, 2.56 equiv.) was then added to the reaction mixture over 20 minutes at room temperature, and the resulting reaction mixture was stirred overnight at room temperature. After LCMS confirmed the formation of the corresponding ring-opened intermediate, the reaction mixture was heated to 75 °C for 6 hours. The reaction mixture was concentrated to half its volume under reduced pressure. Water (4 L) and 1N aqueous hydrochloric acid (HCl, 2.24 L) were added, and the mixture was stirred for 15 minutes. The solid was collected by filtration, washed with water (1.2 L), and dried overnight under vacuum at 50 °C to give the desired crude product (compound 5, 404 g). The crude product was then triturated with a 5:1 mixture of heptane and MTBE (1.2 L) at room temperature for 3 hours. The solid was collected by filtration, washed with heptane (1 L), and dried under vacuum to give 8-bromo-2,2-dimethyl-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (compound 5, 401 g, theoretical 491.3 g, yield 81.6%, purity 98%) as a yellow to brown powder. Compound 5:C 11 H 12 BrNO4S(M+H) + LCMS calculated for: 334.0, found: 333.9; 1 H NMR (300 MHz, DMSO-d6) δ 11.10 (s, 1H), 7.74 (d, J = 2.0 Hz, 1H), 7.38 (d, J = 2.0 Hz, 1H), 3.22 (s, 3H), 1.46 (s, 6H) ppm.

[0182] Step 5a. 8-Bromo-2,2,4-trimethyl-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (Compound 6) A 200 L glass reactor was equipped with an overhead stirrer, thermocouple, addition funnel, and nitrogen inlet, and the apparatus was purged with nitrogen. DMF (30.0 L) and 8-bromo-2,2-dimethyl-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (compound 5, 3000 g, 8.98 mol) were added to the reactor, and the mixture was stirred at ambient temperature until a solution was formed. Potassium carbonate (K2CO3, 1371 g, 9.92 mol, 1.11 equiv.) and methyl iodide (MeI, 1536 g, 0.67 L, 10.83 mol, 1.21 equiv.) were then added to the reactor, maintaining the internal temperature at approximately 17 °C. The resulting reaction mixture was stirred for approximately 4 h until the methylation reaction was complete as determined by HPLC. While maintaining an internal temperature of about 19° C., potable water (60.0 L) was charged to the reactor, and the mixture was stirred at ambient temperature for about 2.5 hours. The solid was collected by filtration, and the wet cake was washed with potable water (30.0 L), air-dried for about 15.5 hours, and then vacuum-dried at about 45° C. to give crude 8-bromo-2,2,4-trimethyl-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (Compound 6, 2834 g, theoretical 3127 g, 90.6% yield) as an off-white to yellow powder, which was used in the next reaction without further purification. Compound 6: 1 H NMR (400 MHz, DMSO-d6) δ 7.83 (d, J = 1.9 Hz, 1H), 7.59 (d, J = 1.9 Hz, 1H), 3.37 (s, 3H), 3.31 (d, J = 3.4 Hz, 3H), 1.49 (s, 6H) ppm; 13 C NMR (101 MHz, DMSO-d6) δ 167.47 (s), 144.14 (s), 136.03 (s), 131.46 (s), 126.07 (s), 113.71 (s), 111.25 (s), 79.80 (s), 43.98 (s), 29.42 (s), 24.28 (s) ppm.

[0183] The synthesis of intermediate compound 9 was carried out according to Scheme 2. [ka]

[0184] Step 1b. (E)-2-(5-Bromo-2-methoxy-3-nitropyridin-4-yl)-N,N-dimethylethenamine (Compound 11) A solution of lithium methanolate (11.5 g, 0.303 mol, 0.147 equiv.) in methanol (300 mL) was added to a solution of 5-bromo-2-methoxy-4-methyl-3-nitropyridine (compound 10, 508 g, 2.057 mol) in DMF (5.0 L). The reaction mixture was heated to 90 °C, and 1,1-dimethoxy-N,N-dimethylmethanamine (2180 mL, 8.0 equiv.) was added over 10 min. The reaction mixture was stirred at 90–95 °C overnight. Upon completion of the reaction as determined by LCMS, the reaction mixture was cooled to 5 °C, and ice-cold water (12.2 L) was added via the addition funnel. The mixture was stirred in the cooling bath for 1 h, and the precipitated solid was collected by filtration. The solid was washed with ice-cold water (2 L), sucked dry for 2 hours, and then dried under vacuum at 40° C. overnight to give crude (E)-2-(5-bromo-2-methoxy-3-nitropyridin-4-yl)-N,N-dimethylethenamine (compound 11, 506 g, theoretical 619.2 g, yield 81.7%) as a red solid, which was used in the next reaction without further purification. 1 H NMR (300 MHz, DMSO-d6) δ 8.22 (s, 1H), 7.03 (d, J = 3.5 Hz, 1H), 4.79 (d, J = 3.5 Hz, 1H), 3.86 (s, 3H), 2.89 (s, 6H) ppm.

[0185] Step 2b. 4-Bromo-7-methoxy-1H-pyrrolo[2,3-c]pyridine (Compound 12) Iron powder (Fe, 1085 g, 19.5 mol, 10 equiv.) and acetic acid (HOAc, 4380 mL, 4595 g, 76.5 mol, 39.3 equiv.) were added sequentially to a solution of (E)-2-(5-bromo-2-methoxy-3-nitropyridin-4-yl)-N,N-dimethyletheamine (Compound 11, 587 g, 1.95 mol) in tetrahydrofuran (THF, 5.25 L). The reaction mixture was heated to 40 °C, causing a gentle, steady exotherm that reached 77 °C over 1 h. After stirring at 75 °C for an additional 2 h, LCMS confirmed the reaction was complete. The reaction mixture was cooled to 50 °C, diluted with ethyl acetate (EtOAc, 4 L), and stirred at room temperature overnight. The solids were removed by filtration through Celite, and the Celite was rinsed with ethyl acetate (EtOAc, 6 L). The combined filtrates were concentrated under reduced pressure. The residue was dissolved in ethyl acetate (16 L of EtOAc), and the solution was washed with a solution of sodium carbonate (900 g) in water (12 L) and saturated brine (2 L). The combined aqueous layers were extracted with ethyl acetate (4 L of EtOAc). The combined organic layers were evaporated under reduced pressure. Heptane (4 L) was added, and the solvent was removed under reduced pressure to quantitatively obtain crude 4-bromo-7-methoxy-1H-pyrrolo[2,3-c]pyridine (compound 12, 450 g) as a dark solid, which was used in the next reaction without further purification. Compound 12: CHBrNO (M+H) + LCMS calculated value for: 227.0, found value: 227.1; 1 H NMR (300 MHz, DMSO-d6) δ 7.73 (s, 1H), 7.53 (d, J = 3.0 Hz, 1H), 6.40 (d, J = 3.0 Hz, 1H), 3.99 (s, 3H) ppm.

[0186] Step 3b. 4-Bromo-7-methoxy-1-tosyl-1H-pyrrolo[2,3-c]pyridine (Compound 13) A 60% dispersion of sodium hydride in mineral oil (NaH, 120 g, 3 mol, 1.5 equiv.) was added in portions over 15 min to a solution of crude 4-bromo-7-methoxy-1H-pyrrolo[2,3-c]pyridine (compound 12, 450 g, 1.95 mol) in DMF (4.5 L). The reaction mixture reached a temperature of 38 °C. The reaction mixture was stirred for 10 min and then cooled to 20 °C. p-Toluenesulfonyl chloride (p-TsCl, 562 g, 2.95 mol, 1.5 equiv.) was added all at once, and the mixture was stirred at room temperature for 2 h. When the reaction was complete as determined by LCMS, water (9 L) was added. The solid was collected by filtration, rinsed with water (2.5 L), and then dissolved in ethyl acetate (EtOAc, 5 L). The solution was washed with water (3 L). The aqueous layer was back-extracted with ethyl acetate (EtOAc, 3 L). The combined organic layers were concentrated under reduced pressure to give crude 4-bromo-7-methoxy-1-tosyl-1H-pyrrolo[2,3-c]pyridine (compound 13, 801 g) quantitatively as a dark solid, which was used in the next reaction without further purification. 15 H 13 BrN2O3S(M+H) + LCMS calculated for: 381.0; Found: 381.0; 1 H NMR (300 MHz, DMSO-d6) δ 8.15 (d, J = 3.8 Hz, 1H), 7.97 (s, 1 H), 7.83 (d, J = 8.5 Hz, 2H), 7.43 (d, J = 8.5 Hz, 2H), 6.78 (d, J = 3.8 Hz, 1H), 3.80 (s, 3H), 2.36 (s, 3H) ppm.

[0187] Step 4b. 4-Bromo-1-tosyl-1H-pyrrolo[2,3-c]pyridin-7-ol (Compound 14) Crude 4-bromo-7-methoxy-1-tosyl-1H-pyrrolo[2,3-c]pyridine (compound 13, 801 g, 1.95 mol) was dissolved in 4 M HCl in 1,4-dioxane (5.6 L, 22.4 mol, 11.5 equiv.) and stirred at 40–45°C for 12 h. The reaction mixture was concentrated under reduced pressure, and the residue was suspended in ethyl ether (EtO, 1.5 L). The solid was filtered, washed sequentially with ethyl ether (EtO, 0.5 L) and heptane (1 L), and then dried overnight in vacuo at 40°C to give crude 4-bromo-1-tosyl-1H-pyrrolo[2,3-c]pyridin-7-ol (compound 14, 648 g, theoretical 716 g, 90.5% yield over three steps) as a yellow powder, which was used in the next reaction without further purification. Compound 14:C 14 H 11 BrN2O3S(M+H) + LCMS calculated value for: 367.0, found value: 366.9; 1 H NMR (300 MHz, DMSO-d6) δ 11.46 (s, 1H), 8.01 (d, J = 3.5 Hz, 1H), 7.92 (d, J = 8.2 Hz, 2H), 7.38 (d, J = 8.2 Hz, 2H), 7.33 (s, 1 H), 6.57 (d, J = 3.5 Hz, 1H), 2.36 (s, 3H) ppm.

[0188] Step 5b. 4-Bromo-6-methyl-1-tosyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one (Compound 9) A 60% dispersion of sodium hydride in mineral oil (NaH, 132 g, 3.3 mol, 1.2 equiv.) was added in portions over 15 min to a solution of 4-bromo-1-tosyl-1H-pyrrolo[2,3-c]pyridin-7-ol (compound 14, 1000 g, 2.72 mol) in DMF (5 L). The reaction mixture reached a temperature of 39 °C. After stirring for 30 min, the reaction mixture was cooled to 20 °C. Iodomethane (MeI, 205 mL, 467 g, 3.3 mol, 1.2 equiv.) was added, and the reaction mixture was stirred at room temperature for 2.5 h. After LCMS showed the reaction was complete, water (13 L) was added, and the reaction mixture was stirred for 30 min. The solid was filtered and washed sequentially with water (2.5 L) and heptane (4 L). The solid was then dissolved in dichloromethane (DCM, 9 L), and the solution was transferred to a separatory funnel. Residual water (approximately 200 mL) was removed. The dichloromethane solution was treated with a mixture of sodium sulfate (NaSO, 200 g), silica gel (SiO, 170 g), and activated carbon (20 g) for 1 hour. The solids were removed by filtration through a pad of Celite (750 g), and the Celite pad was washed with dichloromethane (DCM, 3 L). Toluene (1.2 L) was added to the combined filtrate. The dichloromethane was removed under reduced pressure. The resulting solid in toluene was collected by filtration, washed successively with toluene (1.2 L) and heptane (1.2 L), and dried under vacuum at 40° C. for 2 hours to give crude 4-bromo-6-methyl-1-tosyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one (compound 9, 728 g, theoretical 1036.9 g, yield 70.2%, purity 99.3%), which was used in the next reaction without further purification. 15 H 13 BrN2O3S(M+H) + LCMS calculated value for: 381.0, found value: 381.0; 1 H NMR (300 MHz, DMSO-d6) δ 8.03 (m, 1H), 7.93 (m, 2H), 7.78 (s, 1H), 7.41 (m, 2H), 6.58 (m, 1H), 3.37 (s, 3H), 2.36 (s, 3H) ppm.

[0189] The synthesis of compound 1 was carried out according to Scheme 3. [ka]

[0190] Steps 1 and 2. 2,2,4-trimethyl-8-(6-methyl-7-oxo-1-tosyl-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (Compound 8) A 100 L glass reactor equipped with an overhead stirrer, thermocouple, addition funnel, and nitrogen inlet was placed in a 22 L glass reactor equipped with an overhead stirrer, condenser, thermocouple, addition funnel, and nitrogen inlet, and each equipment was purged with nitrogen. 1,4-Dioxane (15.8 L), 8-bromo-2,2,4-trimethyl-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (compound 6, 1008 g, 2.90 mol, 1.05 equiv.), bis(pinacolato)diboron (1472 g, 5.80 mol, 2.11 equiv.), and potassium acetate (KOAc, 854 g, 8.70 mol, 3.16 equiv.) were added to the 100 L reactor. Nitrogen was bubbled through the reaction mixture for 22 minutes, and Pd(dppf)Cl-CHCl (60.08 g, 0.07 mol, 0.03 eq) was rinsed into a 100 L reactor with 1,4-dioxane (0.5 L). Nitrogen was bubbled through the reaction mixture again for 22 minutes. The resulting reaction mixture was heated to a gentle reflux (approximately 81 °C) and stirred at reflux for approximately 19 hours until the first coupling reaction was complete as determined by HPLC. The reaction mixture was then cooled to approximately 28 °C. Separately, a degassed aqueous sodium bicarbonate solution was prepared by thoroughly mixing sodium bicarbonate (NaHCO, 578 g, 6.89 mol, 2.50 eq) and drinking water (8.3 L) until a solution was formed, and then bubbled nitrogen through the solution for approximately 34 minutes. Degassed aqueous sodium bicarbonate and 4-bromo-6-methyl-1-tosyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one (compound 9, 1050 g, 2.75 mol) were sequentially charged to a 100 L reactor at ambient temperature. The resulting reaction mixture in the 100 L reactor was heated to a gentle reflux (approximately 89 °C) and stirred at reflux for approximately 2.5 hours until the second coupling reaction was confirmed to be complete by HPLC. The reaction mixture was cooled to approximately 29 °C and then charged with drinking water (26.3 L) and ethyl acetate (EtOAc, 39.4 L). The mixture was stirred at ambient temperature for approximately 19 minutes and then filtered through a bed of Celite (1050 g). The filter cake was washed with ethyl acetate (EtOAc, 4.2 L). The filtrate and washings were returned to the 100 L reactor, the phases were separated, and the organic phase was retained in the reactor.Separately, an aqueous sodium bisulfite solution was prepared by thoroughly mixing sodium bisulfite (17,052 g) with drinking water (41.0 L). Approximately one-third of the aqueous sodium bisulfite solution (15.6 L) was added to the organic solution in the 100 L reactor, and the resulting mixture was heated to approximately 50°C and stirred at approximately 54°C for approximately 1 hour. The mixture was cooled to approximately 39°C and filtered through the same Celite pad as before, and the filter cake was washed with ethyl acetate (4.2 L). The combined filtrate and washes were returned to the 100 L reactor, the phases were separated, and the organic phase was retained in the reactor. Approximately one-third of the aqueous sodium bisulfite solution (15.6 L) was added to the organic solution in the 100 L reactor, and the resulting mixture was heated to approximately 50°C and stirred at approximately 52°C for approximately 1 hour. The reaction mixture was cooled to approximately 40°C, the phases were separated, and the organic phase was retained in the reactor. The remainder of the aqueous sodium bisulfite solution (15.6 L) was added to the organic solution in the 100 L reactor, and the resulting mixture was heated to approximately 50 °C and stirred at approximately 50 °C for approximately 1 hour. The mixture was cooled to approximately 40 °C, the phases were separated, and the organic phase was left in the reactor. The organic phase was washed sequentially with potable water (10.5 L) and an aqueous sodium chloride solution prepared separately from 2100 g of sodium chloride and 10.5 L of potable water. The organic phase was concentrated under reduced pressure at approximately 42 °C to a target volume of 11 L (10–12 L per kg of compound 9 charged). The residue was transferred to a 22 L reactor. The organic phase was further concentrated under reduced pressure at approximately 52 °C to a target volume of 5 L (5–6 L per kg of compound 9 charged). The residue was cooled to approximately 24 °C and stirred at approximately 19 °C for approximately 11.5 hours. The solid was collected by filtration, and the filter cake was washed with n-heptane (4.2 L) and air-dried for about 4 hours, followed by further drying under vacuum at about 15-17°C to give crude 2,2,4-trimethyl-8-(6-methyl-7-oxo-1-tosyl-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (compound 8, 1232 g, theoretical 1566.5 g, 78.6% yield) as a yellow to brown powder, which was combined with another batch of crude compound 8 produced by the same procedure and further purified as described below.

[0191] A 100 L glass reactor was equipped with an overhead stirrer, condenser, thermocouple, addition funnel, and nitrogen inlet, and the apparatus was purged with nitrogen. Methylene chloride (34 L) and crude 2,2,4-trimethyl-8-(6-methyl-7-oxo-1-tosyl-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (Compound 8, 3400 g) were added to the reactor, and the mixture was stirred at about 17°C until a solution was formed. Si-thiol (850 g) was added to the resulting solution, and the mixture was heated to about 31°C and stirred at about 31°C for about 2.5 hours. The mixture was then cooled to about 20°C and filtered. The filter cake was washed with methylene chloride (14 L), and the combined filtrate and washings were concentrated under vacuum at about 32° C. to give purified 2,2,4-trimethyl-8-(6-methyl-7-oxo-1-tosyl-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (Compound 8, 3728 g) as a yellow to brown powder, which was used directly in the next reaction with the organic solvents without further drying. 1 H NMR (400 MHz, DMSO-d6) δ 7.99 (dd, J = 5.9, 2.3 Hz, 3H), 7.65 (d, J = 2.0 Hz, 1H), 7.59 (d, J = 2.0 Hz, 1H), 7.56 (s, 1H), 7.44 (d, J = 8.2 Hz, 2H), 6.46 (d, J = 3.5 Hz, 1H), 3.48 (s, 3H), 3.42 (s, 3H), 3.30 (s, 3H), 2.39 (s, 3H), 1.38 (s, 6H) ppm; 13C NMR (101 MHz, DMSO-d6) δ 167.50 (s), 152.60 (s), 145.55 (s), 144.64 (s), 136.22 (s), 135.96 (s), 134.83 (s), 131.27 (s), 130.86 (s), 130.07 (s), 128.88 (s), 125.37 (s), 124.56 (s), 121.93 (s), 113.72 (s), 108.32 (s), 106.83 (s), 79.01 (s), 60.21 (s), 44.17 (s), 36.95 (s), 29.46 (s), 24.28 (s), 21.59 (s), 21.22 (s), 14.55 (s) ppm.

[0192] Step 3. 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (Compound 1) A 50 L glass reactor was equipped with an overhead stirrer, distillation apparatus, thermocouple, addition funnel, and nitrogen inlet, and the reactor was purged with nitrogen. 1,4-Dioxane (10.2 L) and 2,2,4-trimethyl-8-(6-methyl-7-oxo-1-tosyl-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (compound 8, 3724 g of solvent from the previous step, 3400 g dry equivalent, 5.97 mol) were added to the reactor with stirring, and the reaction mixture was heated to approximately 62 °C. Separately, an aqueous sodium hydroxide solution was prepared by thoroughly mixing sodium hydroxide (NaOH, 860 g, 21.49 mol, 3.60 equivalents) with drinking water (21.5 L). Aqueous sodium hydroxide was charged to the reactor over approximately 26 minutes, maintaining the internal temperature below 70°C. The reaction mixture was heated to approximately 84°C and stirred at approximately 84°C for approximately 2.5 hours until the deprotection reaction was complete as determined by HPLC. The reaction mixture was distilled under vacuum at approximately 70°C to a target volume of 17 L (5 L per kg of compound 8 charged). Potable water (13.6 L) was charged and vacuum distillation continued at approximately 76°C until an additional 7 L (2 L per kg of compound 8 charged) was collected. The residual mixture was cooled to approximately 25°C and stirred at approximately 18°C ​​for approximately 11 hours. The solid was collected by filtration, and the filter cake was washed with water (34 L), dried on the filter for about 1 hour, and then air-dried for about 5 days to give crude 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (compound 1, 1728 g, theoretical 2480 g, yield 69.7%), which was purified according to the procedure described below.

[0193] A 50 L glass reactor was equipped with an overhead stirrer, thermocouple, and nitrogen inlet, and the apparatus was purged with nitrogen. Acetonitrile (17.2 L) and crude 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (crude Compound 1, 1726 g, 4.25 mol) were added to the reactor with stirring. The resulting mixture was heated to approximately 72 °C and stirred at 70–75 °C for approximately 1.5 hours. The mixture was then cooled to approximately 25 °C and stirred at ambient temperature for approximately 1 hour. The solid was collected by filtration, and the filter cake was washed with acetonitrile (9 L) and then returned to the reactor with acetonitrile (17 L). The mixture was heated to about 39°C and stirred at about 39°C for about 1.5 hours. The mixture was cooled to about 17°C and stirred at about 17°C for about 15 hours. The solid was collected by filtration, and the filter cake was washed with methylene chloride (9 L). The product was dried on the filter for about 2 hours and then air-dried for about 1 day to provide purified 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (Compound 1, 1458 g, theoretical 1726 g, 84.5% yield), which was recrystallized to obtain the desired crystalline form according to the procedure described below.

[0194] Step 4. Recrystallization of 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (Compound 1) A 100 L glass reactor was equipped with an overhead stirrer, thermocouple, addition funnel, and nitrogen inlet, and a 50 L glass reactor was equipped with an overhead stirrer, condenser, thermocouple, addition funnel, and nitrogen inlet, and each apparatus was purged with nitrogen. Methanol (18.9 L), compound 1 (1454 g), and acetone (18.9 L) were sequentially charged to the 100 L reactor with stirring. The resulting mixture was heated to approximately 57°C and stirred at approximately 57°C for approximately 1.25 hours until a clear solution was obtained. The mixture was transferred to a clean 50 L reactor through an in-line filter. Methanol (2.9 L) was rinsed through the 100 L reactor and filter and then passed through the filter into the 50 L reactor. The mixture in the 50 L reactor was heated to approximately 52°C and stirred at approximately 56°C for approximately 7 minutes until a clear solution was obtained. The solution in the reactor was then concentrated under reduced pressure at approximately 58°C to a target volume of 38 L. Filtered n-heptane (37.7 L) was charged in portions to the reactor, maintaining the internal temperature below 60 °C. Vacuum distillation continued at approximately 59 °C until a target volume of 22 L remained. The residual mixture was cooled to approximately 24 °C and stirred at approximately 17 °C for approximately 6.75 hours. The solid was collected by filtration, and the filter cake was washed with filtered n-heptane (7.3 L), dried on the filter for approximately 1 hour, and then vacuum dried at 60-65 °C to give 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (Compound 1, 1404 g, theoretical 1454 g, 96.6%) as a white to off-white crystalline (Form I) powder. Compound 1:mp 266.4℃; 1 H NMR (400 MHz, DMSO-d6) δ 12.13 (s, 1H), 7.67 (d, J = 1.9 Hz, 1H), 7.62 (d, J = 1.9 Hz, 1H), 7.33 (s, 2H), 6.19 (s, 1H), 3.59 (s, 3H), 3.43 (s, 3H), 3.31 (s, 3H), 1.41 (s, 6H) ppm; 13C NMR (101 MHz, DMSO-d6) δ 167.66 (s), 154.57 (s), 144.55 (s), 134.74 (s), 130.96 (s), 130.33 (s), 129.68 (s), 127.40 (s), 126.96 ppm.

[0195] Crystalline Compound 1 drug substance Form II was obtained by recrystallization in a mixture of tetrahydrofuran (THF), acetone, and n-heptane using a procedure similar to that described above. Both Forms I and II have very distinct melting endothermic peaks in DSC, with a peak melting temperature difference of approximately 1°C between the two forms: 266.4°C for Form I and 267.5°C for Form II. However, despite their very different XRD patterns, both Forms I and II are stable in aqueous suspension. Studies have shown that Form I is the most stable form in MeOH and acetone, whereas Form II is more stable in IPA. In mixtures of methanol, acetone, and n-heptane, Forms I and II are interconvertible depending on conditions such as solvent ratio, temperature, and time. Forms I and II of crystalline Compound 1 have similar solubility in organic solvents and water.

[0196] Form I can also be obtained by adding about 30 mg of Compound 1 to about 2 mL of a saturated or cloudy solution of Compound 1 in acetone, followed by stirring at 25±1° C. for 3 days.

[0197] An alternative synthesis of compound 8 was carried out according to Scheme 4. [ka]

[0198] Step 1x. 6-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-tosyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one (Compound 15) A 500 mL three-neck round-bottom flask comprising a T-tube assembly connected to a mineral oil bubbler was fitted with a condenser and nitrogen inlet. A flask was charged with 4-bromo-6-methyl-1-[(4-methylphenyl)sulfonyl]-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one (compound 9, 10.0 g, 26.2 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (13 g, 52 mmol, 2.0 equiv.), dicyclohexyl(2',4',6'-triisopropylbiphenyl-2-yl)phosphine (Xphos, 1.2 g, 2.6 mmol, 0.1 equiv.), potassium acetate (5.66 g, 57.7 mmol, 2.2 equiv.), and 1,4-dioxane (110 mL). After degassing the mixture with nitrogen for 5 minutes, tris(dibenzylideneacetone)dipalladium(0) (Pd2(dba)3, 600 mg, 0.656 mmol, 0.025 equiv.) was added to the mixture, and nitrogen degassing was continued for 1-2 minutes. The reaction mixture was then heated to 80°C and stirred at 80-86°C for 19 hours. After HPLC showed the reaction was complete, the reaction mixture was cooled to room temperature. 2-Methoxy-2-methylpropane (MTBE, 50 mL) and silica gel (SiO2, 8 g) were added, and the mixture was stirred at room temperature for 30 minutes. The mixture was filtered through a silica gel pad, and the silica gel pad was washed with MTBE. The combined filtrates were concentrated under reduced pressure, and the residue was purified by flash column (silica gel, gradient 0-80% EtOAc in hexanes) to give 6-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-tosyl-1H-pyrrolo[2,3-c]pyridin-7(6H)-one (compound 15, 9.5 g, theoretical 11.22 g, 84.7%) as a brown to red oil, which solidified upon standing at room temperature under vacuum. Compound 15:C 21 H 25 BN2O5S(M+H) + , (2M+Na) +LCMS calculated for: m / z 429.3, 879.3; found: 429.1, 879.3.

[0199] Step 2x. 2,2,4-trimethyl-8-(6-methyl-7-oxo-1-tosyl-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (Compound 8) 8-Bromo-2,2,4-trimethyl-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one (compound 6, 22.4 g, 64.5 mmol) and 6-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-tosyl-1H-pyrrolo[2,3-c A solution of ]pyridin-7(6H)-one (compound 15, 29.0 g, 67.7 mmol, 1.05 equiv.) was treated with cesium fluoride (CsF, 33.9 g, 223 mmol, 3.46 equiv.) and 4-(di-tert-butylphosphino)-N,N-dimethylaniline-dichloropalladium (2:1) (2.0 g, 2.8 mmol, 0.043 equiv.) at ambient temperature. The resulting reaction mixture was then degassed three times, each time by filling with a steady stream of nitrogen gas. The reaction mixture was then heated to reflux for 2–3 h. After HPLC showed the coupling reaction was complete, the reaction mixture was gradually cooled to 30 °C, followed by the addition of water (300 mL) and 2-methoxy-2-methylpropane (MTBE, 300 mL). The mixture was then stirred at ambient temperature for 15 min, after which the two layers were separated. The aqueous layer was extracted with methoxy-2-methylpropane (MTBE, 100 mL). The combined extracts were treated with a solution of sodium hydrogen bisulfate (40 g) in water (200 mL), and the resulting mixture was stirred at ambient temperature for 2 hours. The solid was collected by filtration, washed with water, and dried overnight in a vacuum oven to give the desired product, 2,2,4-trimethyl-8-(6-methyl-7-oxo-1-tosyl-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (compound 8, 20.0 g, theoretical 36.74 g, yield 54.4%), as an off-white to yellow powder, which was used directly in the next reaction without further purification.

[0200] The two layers of the filtrate were separated, and the organic layer was dried over MgSO and concentrated under reduced pressure. The residue was then purified by column chromatography (SiO, gradient elution of 40–100% EtOAc in hexanes) to afford a second crop of the desired compound, 2,2,4-trimethyl-8-(6-methyl-7-oxo-1-tosyl-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-benzo[b][1,4]oxazin-3(4H)-one (compound 8, 13.8 g, theoretical 36.74 g, 37.5% yield; total 33.8 g, 91.9% yield), as a pink oil, which solidified under vacuum at room temperature and was used directly in the next reaction without further purification.

[0201] The batch of compound 8 produced by this alternative synthetic process was confirmed to be identical to the material produced by the original synthesis described in Scheme 3. This material was then converted to compound 1 following the same procedure as described in Scheme 3.

[0202] Example 2. X-ray powder diffraction (XRPD) study of Form I and Form II Forms I and II of Compound 1 were characterized by XRPD. XRPD was obtained from a Bruker D2 PHASER X-ray Powder Diffractometer instrument. The general experimental procedure for XRPD was as follows: (1) K β X-rays were irradiated from a copper source at 1.054056 Å using a filter and a LYNXEYE™ detector; (2) the X-ray power was 30 kV, 10 mA; and (3) the sample powder was dispersed on a background-free sample holder. The general measurement conditions for XRPD were as follows: start angle 5°; stop angle 30°; sampling = 0.015°; and scan rate = 2° / min.

[0203] The XRPD pattern of Form I is shown in FIG. 1 and the XRPD data is shown in Table 1. [Table 1]

[0204] The XRPD pattern of Form II of Compound 1 is shown in FIG. 4, and the XRPD data is shown in Table 2. [Table 2]

[0205] Example 3. Differential Scanning Calorimetry (DSC) Study of Form I and Form II Forms I and II of Compound 1 were characterized by DSC. DSC was obtained from a TA Instruments differential scanning calorimeter, Model Q2000, equipped with an autosampler. DSC instrument conditions were as follows: 25-300°C at 10°C / min; Tzero aluminum sample pan and lid; and a nitrogen gas flow rate of 50 mL / min.

[0206] The DSC thermogram of Form I is shown in Figure 2. The DSC thermogram of Form I showed a large endothermic event with an onset temperature of 264.7°C and a peak temperature of 266.4°C, which is believed to be the melting / decomposition of the compound.

[0207] The DSC thermogram of Form II is shown in Figure 5. The DSC thermogram of Form II showed a large endothermic event with an onset temperature of 266.7°C and a peak temperature of 267.5°C, which is believed to be the melting / decomposition of the compound.

[0208] Example 4. Thermogravimetric Analysis (TGA) Study of Forms I and II Forms I and II of Compound 1 were characterized by TGA. The TGA was obtained from a PerkinElmer thermogravimetric analyzer, Model Pyris 1. The general experimental conditions for the TGA were as follows: 25°C to 350°C ramp at 10°C / min; nitrogen purge gas flow rate of 60 mL / min; ceramic crucible sample holder.

[0209] The TGA thermogram of Form I is shown in Figure 3. A weight loss of approximately 0.4% was observed up to 150°C, which is believed to be associated with a loss of water or residual solvent. Significant weight loss was observed above 250°C, which is believed to be associated with decomposition of the compound.

[0210] The TGA thermogram of Form II is shown in Figure 6. Significant weight loss is observed above 250°C, which is believed to be associated with decomposition of the compound.

[0211] Example 5. Preparation of Forms Ia, III, IV, V, Va, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, and XV, and Amorphous Compound 1 Forms Ia, III, IV, V, Va, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, and XV of Compound 1, and amorphous Compound 1, were prepared according to the procedures in Table 3 below. These forms were analyzed by XRPD (see Example 6), DSC (see Example 7), and TGA (see Example 8). [Table 3]

[0212] Example 6. XRPD of Forms Ia, III, IV, V, Va, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, and XV, and Amorphous XRPD testing of Example 5 was performed on various forms. X-ray powder diffraction (XRPD) data were obtained from a Rigaku MiniFlex X-ray powder diffractometer (XRPD). The general experimental procedure for XRPD was as follows: (1) K β (1) X-rays were irradiated using a copper source at 1.054056 Å using a filter; (2) the X-ray power was 30 KV, 15 mA; and (3) the sample powder was dispersed on a background-free sample holder. The general measurement conditions for XRPD were as follows: start angle 3°; stop angle 45°; sampling = 0.02°; and scan rate = 2° / min.

[0213] Figures 7-21 are XRPD patterns for Forms Ia, III, IV, V, Va, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, and XV, respectively. Tables 4-18 provide peak listings for Forms Ia, III, IV, V, Va, VI, VII, VIII, IX, X, XI, XII, XIII, XIV, and XV, respectively. The amorphous solid of Example 6 was analyzed using XRPD and determined to be amorphous. [Table 4] [Table 5] [Table 6] [Table 7] [Table 8] [Table 9] [Table 10] [Table 11] [Table 12] [Table 13-1] [Table 13-2] [Table 14] [Table 15] [Table 16] [Table 17] [Table 18]

[0214] Example 7. DSC and TGA studies of polymorphs DSC studies were performed on Forms Va, VII, VIII, X, XII, XIII, XIV, and XV. DSC was obtained from a TA Instruments Differential Scanning Calorimeter, Model Q200, equipped with an autosampler. DSC instrument conditions were as follows: 30-300°C at 10°C / min; Tzero aluminum sample pan and lid; and a nitrogen gas flow rate of 50 mL / min.

[0215] TGA studies were performed on Forms Va, VII, VIII, X, XIII, and XV. The TGA was obtained from a TA Instrument Thermogravimetric Analyzer, Model Q500. The general experimental conditions for the TGA were as follows: 20°C to 600°C at 20°C / min; nitrogen purge, gas flow rate 40 mL / min followed by equilibration of purge flow rate; sample purge flow rate 60 mL / min; platinum sample pan.

[0216] Table 19 below shows the DSC and TGA results. [Table 19]

[0217] From the foregoing description, various modifications of the present invention in addition to those described herein will be apparent to those skilled in the art. Such modifications are intended to fall within the scope of the appended claims. Each reference cited in this application, including all patents, patent applications, and publications, is hereby incorporated by reference in its entirety.

Claims

1. formula: 【Chemistry 1】 and a pharmaceutically acceptable excipient, the pharmaceutical composition is formulated in a unit dosage form of a tablet suitable for oral administration; The solid form is crystalline The pharmaceutical composition.

2. 10. The pharmaceutical composition of claim 1, wherein the solid form is anhydrous.

3. 2. The pharmaceutical composition of claim 1, wherein the solid form is Form I having three or more characteristic XRPD peaks selected from about 8.7°, about 9.8°, about 11.6°, about 12.7°, about 14.7°, about 15.7°, about 20.0°, about 21.4°, about 23.3°, and about 27.1° in 2-theta.

4. 4. The pharmaceutical composition of claim 3, wherein the solid form has an XRPD pattern substantially as shown in FIG.

5. 5. The pharmaceutical composition of claim 3 or 4, wherein the solid form has a DSC thermogram characterized by an endothermic peak at a temperature of about 266°C.

6. 5. The pharmaceutical composition of claim 3 or 4, wherein the solid form has a DSC thermogram substantially as shown in Figure 2.

7. 5. The pharmaceutical composition of claim 3 or 4, wherein the solid form has a TGA thermogram substantially as shown in Figure 3.

8. 2. The pharmaceutical composition of claim 1, wherein the solid form is Form II having three or more characteristic XRPD peaks selected from about 6.7°, about 9.5°, about 10.5°, about 14.8°, about 16.2°, about 17.0°, about 18.8°, and about 19.3° in 2-theta.

9. 9. The pharmaceutical composition of claim 8, wherein the solid form has an XRPD pattern substantially as shown in Figure 4.

10. 9. The pharmaceutical composition of claim 8, wherein the solid form has a DSC thermogram characterized by an endothermic peak at a temperature of about 268°C.

11. 10. The pharmaceutical composition of claim 8 or 9, wherein the solid form has a DSC thermogram substantially as shown in Figure 5.

12. 10. The pharmaceutical composition of claim 8 or 9, wherein the solid form has a TGA thermogram substantially as shown in Figure 6.

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