Pik3a inhibitors and methods of making and using the same

Abstract

This disclosure relates to PI3Kα inhibitors, their crystalline forms, solvates, compositions thereof, and methods of use. For example, this disclosure describes compounds in solid form, including crystalline solid forms of the following compounds: N-((1S,2R,4S)-4-(((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)carbamoyl)-2-hydroxycyclopentyl)pyrimidin-5-carboxamide; and (1S,3S,4R)-3-acetamido-N-((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)-4-hydroxycyclopentane-1-carboxamide.

Description

Cross-reference of related applications

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 599,157, filed November 15, 2023, the entire contents of which are incorporated herein by reference. Background Technology

[0002] Phosphatidylinositol 3-kinase (PI3K) belongs to the lipid kinase family and catalyzes the transfer of phosphate groups to the D-3' position of inositol lipids to produce phosphoinositol-3-phosphate (PIP), phosphoinositol-3,4-bisphosphate (PIP2), and phosphoinositol-3,4,5-triphosphate (PIP3). These products then act as second messengers in the signal cascade, docking proteins containing pleckstrin-homology domains, FYVE, Phox, and other phospholipid-binding domains to various signal transduction complexes typically located at the plasma membrane (Vanhaesebroeck et al., Annu. Rev. Biochem 70:535 (2001); Katso et al., Annu. Rev. Cell Dev. Biol. 17:615 (2001)). In the two class 1 PI3K subclasses, class 1A PI3K is a heterodimer composed of a catalytic p110 subunit (α, β, or δ isoform) constitutively bound to a regulatory subunit, which may be p85α, p55α, p50α, p85β, or p55γ. Class 1B has a family member, namely a heterodimer composed of a catalytic p110γ subunit bound to one of the two regulatory subunits p101 or p84 (Fruman et al., Annual Review of Biochemistry 67:481 (1998); Suire et al., Current Biology 15:566 (2005)). The modular domains of the p85 / 55 / 50 subunit include the Src homology (SH2) domain, which, under specific sequence conditions, binds to phosphotyrosine residues on activating receptors and cytoplasmic tyrosine kinases, leading to the activation and localization of class 1A PI3Ks. Class 1B PI3Ks are directly activated by G protein-coupled receptors that bind to peptide and non-peptide ligands across a variety of lineages (Stephens et al., Cell 89:105 (1997); Kazo et al., Annual Reviews of Cell and Developmental Biology 17:615-675 (2001)).

[0003] Therefore, the resulting class I PI3K phospholipid products link upstream receptors to downstream cellular activities, including proliferation, survival, chemotaxis, cell migration, motility, metabolism, inflammation and allergic responses, transcription and translation (Cantley et al., Cell 64:281 (1991); Escobedo and Williams, Nature 335:85 (1988); Fantl et al., Cell 69:413 (1992)). In many cases, PIP2 and PIP3 recruit Aid (the product of a human homolog of the viral oncogene v-Akt) to the plasma membrane, where Aid acts as a node in many intracellular signaling pathways that are essential for growth and survival (Vanter et al., Cell 69:413-423 (1992); Bader et al., Nature Review Cancer 5:921 (2005); Vivanco and Sawyer, Nature Review Cancer 2:489 (2002)).

[0004] Aberrant regulation of PI3K, which typically enhances survival through Aid activation, is one of the most prevalent phenomena in human cancer and has been shown to occur at multiple levels. The tumor suppressor gene PTEN, which dephosphorylates phosphatidylinositol at the 3' position of the inositol ring and thus antagonizes PI3K activity, is functionally lost in various tumors. In other tumors, the genes for p110α isoforms, PIK3CA, and Akt are amplified, and increased protein expression of their gene products has been observed in several human cancers. Furthermore, mutations and translocations of p85α that regulate the p85-p110 complex have been described in human cancers. Finally, somatic missense mutations in PIK3CA, which has been described with significant frequency in a wide range of human cancers, activating downstream signaling pathways (Kang et al., *Proceedings of the National Academy of Sciences*, USA 102:802 (2005); Samuels et al., *Science* 304:554 (2004); Samuels et al., *Cancer Cell* 7:561-573 (2005)). These observations suggest that dysregulation of phosphoinositol-3 kinase and its upstream and downstream components is one of the most common dysregulations associated with human cancers and proliferative diseases (Parsons et al., *Nature* 436:792 (2005); Hennessey et al., *Nature Rev. Drug Disc.* 4:988-1004 (2005)).

[0005] Given the above, PI3Kα inhibitors will have particular value in the treatment of proliferative diseases and other conditions. While various PI3K inhibitors have been developed (e.g., taselisib, alpelisib, buparlisib, etc.), these molecules inhibit multiple class 1A PI3K isoforms. Inhibitors active against multiple class 1A PI3K isoforms are called "pan-PI3K" inhibitors. A major obstacle to the clinical development of existing PI3K inhibitors is the inability to achieve the desired target inhibitory levels in tumors while avoiding toxicity to cancer patients. Pan-PI3K inhibitors share certain target-related toxicities, including diarrhea, rash, fatigue, and hyperglycemia. The toxicity of PI3K inhibitors depends on their isoform selectivity profile. Inhibition of PI3Kα is associated with hyperglycemia and rash, while inhibition of PI3Kδ or PI3Kγ is associated with diarrhea, bone marrow suppression, and elevated transaminase levels (Hanker et al., Cancer Discovery (2019) PMID:30837161).

[0006] Polymorphism is the ability of a substance to crystallize in more than one lattice arrangement. Crystallinity or polymorphism can affect many aspects of the solid-state properties of a pharmaceutical substance. Crystalline forms can differ significantly from amorphous forms, and different crystalline forms of a substance can differ significantly from each other in many ways, including solubility, dissolution rate, and / or bioavailability. Generally, it is difficult to predict whether a given compound will form various crystalline solid forms. Predicting the physical properties of these crystalline solid forms is even more difficult. Furthermore, for certain formulations, such as those suitable for subcutaneous application, the crystalline form of the therapeutic agent can be advantageous. Summary of the Invention

[0007] This disclosure generally relates to compounds I and II and their solvates as well as their solid forms.

[0008] In some embodiments, this disclosure provides a compound in solid form, wherein the compound is compound I:

[0009]

[0010] Or its solvates.

[0011] In some embodiments, this disclosure provides a compound in solid form, wherein the compound is compound II:

[0012]

[0013] Or its solvates.

[0014] On the other hand, this article provides a pharmaceutical composition comprising a compound described herein or a pharmaceutically acceptable salt thereof, or a solvate thereof or in solid form thereof, and a pharmaceutically acceptable excipient.

[0015] On the other hand, this document provides a method for inhibiting PI3Kα activity and treating the conditions, diseases and / or symptoms described herein using compounds or solvates thereof or solid forms thereof or pharmaceutical compositions described herein. Attached Figure Description

[0016] Figure 1 Plot the XRPD diagram of compound I, form A.

[0017] Figure 2 Plot the XRPD diagram of compound I form B.

[0018] Figure 3 Plot the XRPD diagram of compound I in form C.

[0019] Figure 4 Plot the XRPD diagram of compound II form A.

[0020] Figure 5 Plot the XRPD diagram of compound II form B.

[0021] Figure 6 DSC and TGA thermograms of compound I form A were plotted.

[0022] Figure 7 DSC thermogram of compound I form B.

[0023] Figure 8 DSC and TGA thermograms of compound I form C were plotted.

[0024] Figure 9 DSC and TGA thermograms of compound II form A were plotted.

[0025] Figure 10 DSC and TGA thermograms of compound II form B were plotted. Detailed Implementation

[0026] General description of certain embodiments of this disclosure

[0027] In some embodiments, compounds I and II are PI3Kα inhibitors and are suitable for treating conditions, diseases, and / or symptoms, such as “PI3Kα-mediated” conditions, diseases, and / or symptoms as described herein. In some embodiments, this disclosure provides compounds in solid form (e.g., as a free base or salt or solvate), which imparts, for example, improved water solubility, stability, and ease of formulation.

[0028] solid form of compound I

[0029] In some embodiments, this document provides a compound in solid form, wherein the compound is compound I:

[0030]

[0031] Or its solvates.

[0032] Considering this, compound I (i.e., (N-((1S,2R,4S)-4-(((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)carbamoyl)-2-hydroxycyclopentyl)pyrimidine-5-carboxamide) can exist in a variety of physical forms. For example, compound I can be in solution, suspension or solid form. In some embodiments, compound I is in solid form. When compound I is in solid form, the compound can be amorphous, crystalline or a mixture thereof. Exemplary solid forms are described in more detail below.

[0033] In some embodiments, compound I is an amorphous solid. In some embodiments, compound I is a crystalline solid. In some embodiments, compound I is a mixture of one or more crystalline forms. In some embodiments, compound I is a mixture of an amorphous solid and one or more crystalline forms.

[0034] In some embodiments, compound I is anhydrous. In some embodiments, compound I is in hydrated form. In some embodiments, compound I is in hemihydrate form.

[0035] In some embodiments, this disclosure provides a substantially impurity-free form of Compound I. As used herein, the term "substantially impurity-free" means that the compound is free of significant amounts of foreign matter. Such foreign matter may include different forms of Compound I, residual solvents, or any other impurities that may arise from the preparation and / or isolation of Compound I.

[0036] In some embodiments, the solid form of Compound I or its solvate is present in an amount of at least about 95, 95.5, 96, 96.5, 97, 97.5, 98.0, 98.5, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, or 99.9% by weight, wherein the percentage is based on the total weight of the composition. In some embodiments, the solid form of Compound I or its solvate contains no more than about 0.40, no more than about 0.35, no more than about 0.3, no more than about 0.25, no more than about 0.2, no more than about 0.15, no more than about 0.10, or no more than about 0.05% by weight of any single impurity, wherein the percentage is based on the total weight of the composition. In some embodiments, the impurity is selected from those impurities described in the examples herein.

[0037] In some embodiments, the solid form of compound I or its solvation is present in an amount of at least about 95, 95.5, 96, 96.5, 97, 97.5, 98.0, 98.5, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, or 99.9 HPLC area% relative to the total area of ​​the HPLC chromatogram. In some embodiments, the solid form of compound I or its solvation contains any single impurity not exceeding about 0.4, about 0.35, about 0.3, about 0.25, about 0.2, about 0.15, about 0.10, or about 0.05 HPLC area% relative to the total area of ​​the HPLC chromatogram. In some embodiments, the impurity is selected from those impurities described in the examples herein. In some embodiments, the HPLC method is selected from the HPLC methods described in the examples herein.

[0038] The structures described for compounds I are intended to include all tautomers. Furthermore, the structures described herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, in addition to hydrogen replaced by deuterium or tritium or carbon replaced by... 13 C or 14 Compounds having the structure of this invention, other than those enriched by carbon replacement, are within the scope of this disclosure.

[0039] It has been found that compound I can exist in a variety of solid forms. Such exemplary forms include polymorphs, such as those described herein. In some embodiments, compound I is crystalline in form A, form B, or form C as described herein.

[0040] In some embodiments, compound I is in crystalline form A. In some embodiments, form A of compound I has a generally similar Figure 1 The X-ray powder diffraction pattern depicted in the figure.

[0041] In some embodiments, compound I is in crystalline form B. In some embodiments, form B of compound I has a generally similar Figure 2 The X-ray powder diffraction pattern depicted in the figure.

[0042] In some embodiments, compound I is in crystalline form C. In some embodiments, form C of compound I has a generally similar... Figure 3 The X-ray powder diffraction pattern depicted in the figure.

[0043] Form A of compound I

[0044] In some embodiments, compound I is in solid form as form A. In some embodiments, form A of compound I can be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group consisting of: about 17.4 2θ, about 13.3 2θ, and about 16.5 2θ. In some embodiments, form A of compound I can be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group consisting of: about 17.4 2θ, about 13.3 2θ, about 16.5 2θ, about 24.2 2θ, about 19.8 2θ, about 14.5 2θ, and about 18.2 2θ. In some embodiments, form A of compound I may be characterized by an X-ray powder diffraction pattern having at least three characteristic peaks (in ° 2θ), each selected from the group consisting of: about 17.4 2θ, about 13.3 2θ, about 16.5 2θ, about 24.2 2θ, about 19.8 2θ, about 14.5 2θ, and about 18.2 2θ. In some embodiments, form A of compound I may be characterized by an X-ray powder diffraction pattern having at least four characteristic peaks (in ° 2θ), each selected from the group consisting of: about 17.4 2θ, about 13.3 2θ, about 16.5 2θ, about 24.2 2θ, about 19.8 2θ, about 14.5 2θ, and about 18.2 2θ. In some embodiments, form A of compound I may be characterized by an X-ray powder diffraction pattern having at least five characteristic peaks (in ° 2θ), each selected from the group consisting of: about 17.4 2θ, about 13.3 2θ, about 16.5 2θ, about 24.2 2θ, about 19.8 2θ, about 14.5 2θ, and about 18.2 2θ. In some embodiments, form A of compound I may be characterized by an X-ray powder diffraction pattern having at least six characteristic peaks (in ° 2θ), each selected from the group consisting of: about 17.4 2θ, about 13.3 2θ, about 16.5 2θ, about 24.2 2θ, about 19.8 2θ, about 14.5 2θ, and about 18.2 2θ. In some embodiments, compound I in form A can be characterized by an X-ray powder diffraction pattern containing characteristic peaks at about 17.4 2θ, about 13.3 2θ, about 16.5 2θ, about 24.2 2θ, about 19.8 2θ, about 14.5 2θ and about 18.2 2θ.

[0045] In some embodiments, compound I in form A has a generally similar Figure 1The X-ray powder diffraction pattern depicted herein is an X-ray powder diffraction pattern. In some embodiments, form A of compound I may be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.1. In some embodiments, form A of compound I may be characterized by an X-ray powder diffraction pattern having at least three characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.1. In some embodiments, form A of compound I may be characterized by an X-ray powder diffraction pattern having at least four characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.1. In some embodiments, form A of compound I may be characterized by an X-ray powder diffraction pattern having at least five characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.1. In some embodiments, form A of compound I may be characterized by an X-ray powder diffraction pattern having at least six characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.1. In some embodiments, form A of compound I may be characterized by an X-ray powder diffraction pattern having at least seven characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.1.

[0046] Table 1.1 List of XRPD peaks for compound I form A (each peak is within ±0.2° 2θ).

[0047]

[0048]

[0049] As used herein, in the case of peaks in ° 2θ units, the term “approximately” means that the peak can be a given 2θ value ±0.2, or a given 2θ value ±0.1, or a given value. For example, a peak “approximately 12.0 2θ” means that the peak can be 11.8 2θ, 11.92θ, 12.0 2θ, 12.1 2θ, or 12.2 2θ.

[0050] In some embodiments, compound I in form A has a generally similar Figure 6 The TGA plot depicted in the figure. In some embodiments, the form A of compound I may be characterized by being highly similar to two or more plots simultaneously (e.g., Figure 1 and Figure 6 ).

[0051] Form B of compound I

[0052] In some embodiments, compound I is in solid form form B. In some embodiments, form B of compound I can be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group consisting of: about 18.5 2θ, about 11.4 2θ, and about 17.5 2θ. In some embodiments, form B of compound I can be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group consisting of: about 18.5 2θ, about 11.4 2θ, about 17.5 2θ, about 5.7 2θ, about 13.6 2θ, about 17.3 2θ, and about 10.7 2θ. In some embodiments, form B of compound I may be characterized by an X-ray powder diffraction pattern having at least three characteristic peaks (in ° 2θ), each selected from the group consisting of: about 18.5 2θ, about 11.4 2θ, about 17.5 2θ, about 5.7 2θ, about 13.6 2θ, about 17.3 2θ, and about 10.7 2θ. In some embodiments, form B of compound I may be characterized by an X-ray powder diffraction pattern having at least four characteristic peaks (in ° 2θ), each selected from the group consisting of: about 18.5 2θ, about 11.4 2θ, about 17.5 2θ, about 5.7 2θ, about 13.6 2θ, about 17.3 2θ, and about 10.7 2θ. In some embodiments, form B of compound I may be characterized by an X-ray powder diffraction pattern having at least five characteristic peaks (in ° 2θ), each selected from the group consisting of: about 18.5 2θ, about 11.4 2θ, about 17.5 2θ, about 5.7 2θ, about 13.6 2θ, about 17.3 2θ, and about 10.7 2θ. In some embodiments, form B of compound I may be characterized by an X-ray powder diffraction pattern having at least six characteristic peaks (in ° 2θ), each selected from the group consisting of: about 18.5 2θ, about 11.4 2θ, about 17.5 2θ, about 5.7 2θ, about 13.6 2θ, about 17.3 2θ, and about 10.7 2θ. In some embodiments, form B of compound I can be characterized by an X-ray powder diffraction pattern containing characteristic peaks at about 18.5 2θ, about 11.4 2θ, about 17.5 2θ, about 5.7 2θ, about 13.6 2θ, about 17.3 2θ, and about 10.7 2θ.

[0053] In some embodiments, compound I in form B has a generally similar Figure 2The X-ray powder diffraction pattern depicted herein is an X-ray powder diffraction pattern. In some embodiments, form B of compound I may be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.2. In some embodiments, form B of compound I may be characterized by an X-ray powder diffraction pattern having at least three characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.2. In some embodiments, form B of compound I may be characterized by an X-ray powder diffraction pattern having at least four characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.2. In some embodiments, form B of compound I may be characterized by an X-ray powder diffraction pattern having at least five characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.2. In some embodiments, form B of compound I may be characterized by an X-ray powder diffraction pattern having at least six characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.2. In some embodiments, form B of compound I may be characterized by an X-ray powder diffraction pattern having at least seven characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.2.

[0054] Table 1.2 List of XRPD peaks for compound I form B (each peak is within ±0.2° 2θ).

[0055]

[0056]

[0057]

[0058] In some embodiments, compound I in form B has a generally similar Figure 7 The TGA plot depicted in the figure. In some embodiments, the form B of compound I may be characterized by being highly similar to two or more plots simultaneously (e.g., Figure 2 and Figure 7 ).

[0059] The form of compound I, C

[0060] In some embodiments, compound I is in solid form as form C. In some embodiments, form C of compound I can be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group consisting of: about 19.3 2θ, about 19.9 2θ, and about 20.0 2θ. In some embodiments, form C of compound I can be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group consisting of: about 19.3 2θ, about 19.9 2θ, about 20.0 2θ, about 10.1 2θ, about 19.6 2θ, about 15.2 2θ, and about 22.0 2θ. In some embodiments, compound I in form C may be characterized by an X-ray powder diffraction pattern having at least three characteristic peaks (in ° 2θ), each selected from the group consisting of: about 19.3 2θ, about 19.9 2θ, about 20.0 2θ, about 10.1 2θ, about 19.6 2θ, about 15.2 2θ, and about 22.0 2θ. In some embodiments, compound I in form C may be characterized by an X-ray powder diffraction pattern having at least four characteristic peaks (in ° 2θ), each selected from the group consisting of: about 19.3 2θ, about 19.9 2θ, about 20.0 2θ, about 10.1 2θ, about 19.6 2θ, about 15.2 2θ, and about 22.0 2θ. In some embodiments, compound I in form C may be characterized by an X-ray powder diffraction pattern having at least five characteristic peaks (in ° 2θ), each selected from the group consisting of: about 19.3 2θ, about 19.9 2θ, about 20.0 2θ, about 10.1 2θ, about 19.6 2θ, about 15.2 2θ, and about 22.0 2θ. In some embodiments, compound I in form C may be characterized by an X-ray powder diffraction pattern having at least six characteristic peaks (in ° 2θ), each selected from the group consisting of: about 19.3 2θ, about 19.9 2θ, about 20.0 2θ, about 10.1 2θ, about 19.6 2θ, about 15.2 2θ, and about 22.0 2θ. In some embodiments, form C of compound I can be characterized by an X-ray powder diffraction pattern containing characteristic peaks at about 19.3 2θ, about 19.9 2θ, about 20.0 2θ, about 10.1 2θ, about 19.6 2θ, about 15.2 2θ, and about 22.0 2θ. In some embodiments, form C of compound I has a generally similar Figure 3 The X-ray powder diffraction pattern depicted in Figure A.

[0061] In some embodiments, compound I in form C has a generally similar form to Figure 3The X-ray powder diffraction pattern depicted herein is an X-ray powder diffraction pattern. In some embodiments, form C of compound I may be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.3. In some embodiments, form C of compound I may be characterized by an X-ray powder diffraction pattern having at least three characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.3. In some embodiments, form C of compound I may be characterized by an X-ray powder diffraction pattern having at least four characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.3. In some embodiments, form C of compound I may be characterized by an X-ray powder diffraction pattern having at least five characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.3. In some embodiments, form C of compound I may be characterized by an X-ray powder diffraction pattern having at least six characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.3. In some embodiments, form C of compound I may be characterized by an X-ray powder diffraction pattern having at least seven characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.3.

[0062] Table 1.3 List of XRPD peaks for compound I (each peak is within ±0.2° 2θ).

[0063]

[0064]

[0065] In some embodiments, compound I in form C has a generally similar form to Figure 8 The TGA diagram depicted in the figure. In some embodiments, the form C of compound I may be highly similar to two or more of these diagrams (e.g., Figure 3 and Figure 8 ).

[0066] solid form of compound II

[0067] In some embodiments, this document provides a compound in solid form, wherein the compound is compound II:

[0068] ,

[0069] Or its solvates.

[0070] Consideration has been given that compound II (i.e., (1S,3S,4R)-3-acetamido-N-((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)-4-hydroxycyclopentane-1-carboxamide) can exist in a variety of physical forms. For example, compound II can be in the form of a solution, suspension, or solid. In some embodiments, compound II is in the form of a solid. When compound II is in the form of a solid, the compound can be amorphous, crystalline, or a mixture thereof. Exemplary solid forms are described in more detail below.

[0071] In some embodiments, compound II is an amorphous solid. In some embodiments, compound II is a crystalline solid. In some embodiments, compound II is a mixture of one or more crystalline forms. In some embodiments, compound II is a mixture of an amorphous solid and one or more crystalline forms.

[0072] In some embodiments, compound II is in anhydrous form. In some embodiments, compound II may be in hydrate form. In some embodiments, compound II may be in hemihydrate form.

[0073] In some embodiments, this disclosure provides a solid form of compound II that is substantially free of impurities. As used herein, the term "substantially free of impurities" means that the compound is free of significant amounts of foreign matter. Such foreign matter may include different forms of compound II, residual solvents, or any other impurities that may arise from the preparation and / or separation of compound II.

[0074] In some embodiments, the solid form of compound II or its solvation is present in an amount of at least about 95, 95.5, 96, 96.5, 97, 97.5, 98.0, 98.5, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, or 99.9% by weight, wherein the percentage is based on the total weight of the composition. In some embodiments, the solid form of compound II or its solvation contains no more than about 0.40, no more than about 0.35, no more than about 0.3, no more than about 0.25, no more than about 0.2, no more than about 0.15, no more than about 0.10, or no more than about 0.05% by weight of any single impurity, wherein the percentage is based on the total weight of the composition. In some embodiments, the impurity is selected from those impurities described in the examples herein.

[0075] In some embodiments, the solid form of compound II or its solvation is present in an amount of at least about 95, 95.5, 96, 96.5, 97, 97.5, 98.0, 98.5, 99, 99.1, 99.2, 99.3, 99.4, 99.5, 99.6, 99.7, 99.8, or 99.9 HPLC area% relative to the total area of ​​the HPLC chromatogram. In some embodiments, the solid form of compound II or its solvation contains any single impurity not exceeding about 0.4, about 0.35, about 0.3, about 0.25, about 0.2, about 0.15, about 0.10, or about 0.05 HPLC area% relative to the total area of ​​the HPLC chromatogram. In some embodiments, the impurity is selected from those impurities described in the examples herein. In some embodiments, the HPLC method is selected from the HPLC methods described in the examples herein.

[0076] The structures described for compounds II are intended to include all tautomers. Furthermore, the structures described herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, in addition to hydrogen replaced by deuterium or tritium or carbon replaced by... 13 C or 14 Compounds having the structure of this invention, other than those enriched by carbon replacement, are within the scope of this disclosure.

[0077] It has been found that compound II can exist in a variety of solid forms. Such exemplary forms include crystalline polymorphs, such as those described herein. In some embodiments, compound II is crystalline in form A or form B as described herein.

[0078] In some embodiments, compound II is in crystalline form A. In some embodiments, form A of compound II has a generally similar Figure 4 The X-ray powder diffraction pattern depicted in the figure.

[0079] In some embodiments, compound II is in crystalline form B. In some embodiments, form B of compound II has a generally similar Figure 5 The X-ray powder diffraction pattern depicted in the figure.

[0080] Form A of compound II

[0081] In some embodiments, compound II is in solid form as form A. In some embodiments, form A of compound II can be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group consisting of: about 15.7 2θ, about 14.7 2θ, and about 20.0 2θ. In some embodiments, form A of compound II can be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group consisting of: about 15.7 2θ, about 14.7 2θ, about 20.0 2θ, about 21.0 2θ, about 23.2 2θ, about 16.5 2θ, about 17.5 2θ, and about 13.3 2θ. In some embodiments, form A of compound II may be characterized by an X-ray powder diffraction pattern having at least three characteristic peaks (in ° 2θ), each selected from the group consisting of: about 15.7 2θ, about 14.7 2θ, about 20.0 2θ, about 21.0 2θ, about 23.2 2θ, about 16.5 2θ, about 17.5 2θ, and about 13.3 2θ. In some embodiments, form A of compound II may be characterized by an X-ray powder diffraction pattern having at least four characteristic peaks (in ° 2θ), each selected from the group consisting of: about 15.7 2θ, about 14.7 2θ, about 20.0 2θ, about 21.0 2θ, about 23.2 2θ, about 16.5 2θ, about 17.5 2θ, and about 13.3 2θ. In some embodiments, form A of compound II may be characterized by an X-ray powder diffraction pattern having at least five characteristic peaks (in ° 2θ), each selected from the group consisting of: about 15.7 2θ, about 14.7 2θ, about 20.0 2θ, about 21.0 2θ, about 23.2 2θ, about 16.5 2θ, about 17.5 2θ, and about 13.3 2θ. In some embodiments, form A of compound II may be characterized by an X-ray powder diffraction pattern having at least six characteristic peaks (in ° 2θ), each selected from the group consisting of: about 15.7 2θ, about 14.7 2θ, about 20.0 2θ, about 21.0 2θ, about 23.2 2θ, about 16.5 2θ, about 17.5 2θ, and about 13.3 2θ. In some embodiments, compound II in form A can be characterized by an X-ray powder diffraction pattern containing characteristic peaks at about 15.7 2θ, about 14.7 2θ, about 20.0 2θ, about 21.0 2θ, about 23.2 2θ, about 16.5 2θ, about 17.5 2θ and about 13.3 2θ.

[0082] In some embodiments, compound II in form A has a generally similar Figure 4The X-ray powder diffraction pattern depicted herein is an X-ray powder diffraction pattern. In some embodiments, form A of compound II may be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.4. In some embodiments, form A of compound II may be characterized by an X-ray powder diffraction pattern having at least three characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.4. In some embodiments, form A of compound II may be characterized by an X-ray powder diffraction pattern having at least four characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.4. In some embodiments, form A of compound II may be characterized by an X-ray powder diffraction pattern having at least five characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.4. In some embodiments, form A of compound II may be characterized by an X-ray powder diffraction pattern having at least six characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.4. In some embodiments, form A of compound II may be characterized by an X-ray powder diffraction pattern having at least seven characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.4.

[0083] Table 1.4 List of XRPD peaks for compound II form A (each peak is within ±0.2° 2θ).

[0084]

[0085] In some embodiments, compound II in form A has a generally similar Figure 9 The TGA plot depicted in the figure. In some embodiments, form A of compound II may be highly similar to two or more plots simultaneously (e.g., Figure 4 and Figure 9 ).

[0086] Form B of compound II

[0087] In some embodiments, compound II is in solid form form B. In some embodiments, form B of compound II can be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group consisting of: about 23.6 2θ, about 10.9 2θ, and about 16.9 2θ. In some embodiments, form B of compound II can be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group consisting of: about 23.6 2θ, about 10.9 2θ, about 16.9 2θ, about 15.1 2θ, about 26.1 2θ, about 12.9 2θ, and about 19.4 2θ. In some embodiments, form B of compound II may be characterized by an X-ray powder diffraction pattern having at least three characteristic peaks (in ° 2θ), each selected from the group consisting of: about 23.6 2θ, about 10.9 2θ, about 16.9 2θ, about 15.1 2θ, about 26.1 2θ, about 12.9 2θ, and about 19.4 2θ. In some embodiments, form B of compound II may be characterized by an X-ray powder diffraction pattern having at least four characteristic peaks (in ° 2θ), each selected from the group consisting of: about 23.6 2θ, about 10.9 2θ, about 16.9 2θ, about 15.1 2θ, about 26.1 2θ, about 12.9 2θ, and about 19.4 2θ. In some embodiments, form B of compound II may be characterized by an X-ray powder diffraction pattern having at least five characteristic peaks (in ° 2θ), each selected from the group consisting of: about 23.6 2θ, about 10.9 2θ, about 16.9 2θ, about 15.1 2θ, about 26.1 2θ, about 12.9 2θ, and about 19.4 2θ. In some embodiments, form B of compound II may be characterized by an X-ray powder diffraction pattern having at least six characteristic peaks (in ° 2θ), each selected from the group consisting of: about 23.6 2θ, about 10.9 2θ, about 16.9 2θ, about 15.1 2θ, about 26.1 2θ, about 12.9 2θ, and about 19.4 2θ. In some embodiments, form B of compound II can be characterized by an X-ray powder diffraction pattern containing characteristic peaks at about 23.6 2θ, about 10.9 2θ, about 16.9 2θ, about 15.1 2θ, about 26.1 2θ, about 12.9 2θ, and about 19.4 2θ.

[0088] In some embodiments, compound II in form B has a generally similar Figure 5The X-ray powder diffraction pattern depicted herein is an X-ray powder diffraction pattern. In some embodiments, form B of compound II may be characterized by an X-ray powder diffraction pattern having at least two characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.5. In some embodiments, form B of compound II may be characterized by an X-ray powder diffraction pattern having at least three characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.5. In some embodiments, form B of compound II may be characterized by an X-ray powder diffraction pattern having at least four characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.5. In some embodiments, form B of compound II may be characterized by an X-ray powder diffraction pattern having at least five characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.5. In some embodiments, form B of compound II may be characterized by an X-ray powder diffraction pattern having at least six characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.5. In some embodiments, form B of compound II may be characterized by an X-ray powder diffraction pattern having at least seven characteristic peaks (in ° 2θ), each selected from the group of peaks listed in Table 1.5.

[0089] Table 1.5 List of XRPD peaks for compound II form B (each peak is within ±0.2° 2θ).

[0090]

[0091]

[0092] In some embodiments, compound II in form B has a generally similar Figure 10 The TGA plot depicted in the figure. In some embodiments, form B of compound II may be highly similar to two or more plots simultaneously (e.g., Figure 5 and Figure 10 ).

[0093] Composition

[0094] Another aspect of this disclosure provides pharmaceutical compositions comprising the compounds disclosed herein formulated with a pharmaceutically acceptable carrier. Specifically, this disclosure provides pharmaceutical compositions comprising the compounds disclosed herein formulated with one or more pharmaceutically acceptable carriers. These formulations include those suitable for oral, topical (e.g., percutaneous), buccal, ocular, parenteral (e.g., subcutaneous, intramuscular, intradermal, or intravenous), rectal, vaginal, or aerosol administration, but the most suitable form of administration in any given case will depend on the extent and severity of the condition being treated and the nature of the particular compound used. For example, the disclosed compositions may be formulated in unit dose form and / or may be formulated for oral, subcutaneous, or intravenous administration.

[0095] The exemplary pharmaceutical compositions of this disclosure may be used in pharmaceutical formulations, such as solid, semi-solid, or liquid forms, comprising one or more of the compounds of this disclosure as active ingredients, mixed with organic or inorganic carriers or excipients suitable for external, enteral, or parenteral administration. The active ingredient may be compounded with, for example, commonly used, non-toxic, pharmaceutically acceptable carriers for use in tablets, pills, capsules, suppositories, solutions, emulsions, suspensions, and any other suitable forms. The active target compound is included in the pharmaceutical composition in an amount sufficient to produce the desired effect on the course or symptoms of a disease.

[0096] In some embodiments, the pharmaceutically acceptable composition may contain the disclosed compound and / or its pharmaceutically acceptable salt in concentrations ranging from about 0.01 to about 2.0 wt%, for example, from 0.01 to about 1 wt% or from about 0.05 to about 0.5 wt%. The composition may be formulated as a solution, suspension, ointment, or capsule, etc. The pharmaceutical composition may be prepared as an aqueous solution and may contain additional components, such as preservatives, buffers, tension agents, antioxidants, stabilizers, viscosity modifiers, etc.

[0097] For the preparation of solid compositions, such as tablets, the main active ingredient may be mixed with a pharmaceutical carrier, such as a conventional tablet-forming ingredient (e.g., corn starch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate, or gum), and other pharmaceutical diluents, such as water, to form a solid preformed composition containing a homogeneous mixture of the compounds of this disclosure or their non-toxic, pharmaceutically acceptable salts. When these preformed compositions are referred to as homogeneous compositions, it means that the active ingredient is uniformly dispersed throughout the composition, such that the composition can be readily further divided into equally effective unit dosage forms, such as tablets, pills, and capsules.

[0098] Pharmaceutically acceptable carriers are well known to those skilled in the art and include, for example, adjuvants, diluents, excipients, fillers, lubricants, and mordants. In some embodiments, the carrier is a diluent, adjuvant, excipient, or mordant. In some embodiments, the carrier is a diluent, adjuvant, or excipient. In some embodiments, the carrier is a diluent or adjuvant. In some embodiments, the carrier is an excipient. Generally, pharmaceutically acceptable carriers are chemically inert to the active compound and are non-toxic under the conditions of use. Examples of pharmaceutically acceptable carriers may include, for example, aqueous solutions of water or physiological saline, polymers such as polyethylene glycol, carbohydrates and their derivatives, oils, fatty acids, or alcohols. Non-limiting examples of oils as pharmaceutical carriers include petroleum, animal, plant, or synthetic oils, such as peanut oil, soybean oil, mineral oil, sesame oil, etc. Pharmaceutical carriers may also be physiological saline, gum arabic, gelatin, starch paste, talc, keratin, colloidal silica, urea, etc. Additionally, adjuvants, stabilizers, thickeners, lubricants, and colorants may be used. Other examples of suitable pharmaceutical carriers are described in, for example, the following literature: *Remington's: The Science and Practice of Pharmacy*, 22nd edition (edited by Allen and Loyd V., Jr., Pharmaceutical Press (2012)); *Modern Pharmaceutics*, 5th edition (Alexander T. Florence and Juergen Siepmann, CRC Press (2009)); *Handbook of Pharmaceutical Excipients*, 7th edition (Rowe and Raymond C.; Sheskey and Paul J.; Cook and Walter G.; Fenton, Marian E. (Fenton, Marian E., ed., Medical Publishing House (2012)) (each of which is incorporated herein by reference in its entirety).

[0099] In some embodiments, the compounds of this disclosure are formulated into pharmaceutical compositions for administration to a subject in a biocompatible form suitable for in vivo administration. According to another aspect, this disclosure provides a pharmaceutical composition comprising a mixture of the disclosed compounds with a pharmaceutically acceptable diluent and / or carrier. A pharmaceutically acceptable carrier is "acceptable" in the sense that it is compatible with the other components of the composition and does not harm the recipient. Pharmaceutically acceptable carriers as used herein may be selected from a variety of organic or inorganic materials used as materials in pharmaceutical formulations and incorporated as analgesics, buffers, binders, disintegrants, diluents, emulsifiers, excipients, extenders, lubricants, solubilizers, stabilizers, suspending agents, tension agents, mediators, and viscosity increasers. Pharmaceutical additives, such as antioxidants, flavorings, colorings, flavor enhancers, preservatives, and sweeteners, may also be added. Examples of acceptable pharmaceutical carriers include carboxymethyl cellulose, crystalline cellulose, glycerin, gum arabic, lactose, magnesium stearate, methylcellulose, powders, physiological saline, sodium alginate, sucrose, starch, talc, and water. In some embodiments, the term "pharmaceutically acceptable" means approved by a federal or state regulatory agency, or listed in the United States Pharmacopeia or other recognized pharmacopoeia, for use in animals and, more particularly, in humans.

[0100] Surfactants, such as detergents, are also suitable for use in formulations. Specific examples of surfactants include polyvinylpyrrolidone, polyvinyl alcohol, vinyl acetate and copolymers of vinylpyrrolidone, polyethylene glycol, benzyl alcohol, mannitol, glycerol, sorbitol or dehydrated sorbitol polyoxyethylene esters; lecithin or sodium carboxymethyl cellulose; or acrylic acid derivatives, such as methacrylates; anionic surfactants, such as basic stearates, especially sodium stearate, potassium stearate or ammonium stearate; calcium stearate or triethanolamine stearate; alkyl sulfates, especially sodium lauryl sulfate and sodium cetyl sulfate; sodium dodecylbenzene sulfonate or sodium dioctyl sulfosuccinate; or fatty acids, especially fatty acids derived from coconut oil; cationic surfactants, such as those having the formula N + R'R''R'''R'''Y - A water-soluble quaternary ammonium salt, wherein the R groups are the same or different, optionally hydroxylated hydrocarbon groups, and Y - It is an anion of strong acids, such as halogen, sulfate, and sulfonate anions; cetyltrimethylammonium bromide is one of the usable cationic surfactants, which has the formula N +An amine salt of R'R'R''', wherein the R groups are the same or different, optionally being hydroxylated hydrocarbon groups; octadecylamine hydrochloride is one of the cationic surfactants that can be used; a nonionic surfactant, such as optionally polyoxyethylene esters of sorbitol, especially polysorbate 80 or polyoxyethylene alkyl ethers; polyethylene glycol stearate, polyoxyethylene derivatives of castor oil, polyglycerol esters, polyoxyethylene fatty alcohols, polyoxyethylene fatty acids, or copolymers of ethylene oxide and propylene oxide; an amphoteric surfactant, such as a substituted lauryl compound of betaine.

[0101] When administered to subjects, the disclosed compounds and pharmaceutically acceptable carriers may be sterile. Suitable pharmaceutical carriers may also include excipients such as starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skim milk powder, glycerin, propylene, ethylene glycol, polyethylene glycol 300, water, ethanol, polysorbate 20, etc. Where necessary, the compositions disclosed herein may also contain small amounts of wetting agents or emulsifiers, or pH buffers.

[0102] The pharmaceutical formulations disclosed herein are prepared using methods well known in pharmaceutical technology. Optionally, one or more auxiliary ingredients (e.g., buffers, flavoring agents, surfactants, etc.) are also added. The choice of carrier is determined by the solubility and chemical properties of the compound, the chosen route of administration, and standard pharmaceutical practice.

[0103] Additionally, the compounds and / or compositions of this disclosure are administered to human or animal subjects via known procedures including oral, sublingual, or buccal administration. In some embodiments, the compounds and / or compositions are administered orally.

[0104] In solid dosage forms (capsules, tablets, pills, sugar-coated pills, powders, granules, etc.) intended for oral administration, the subject composition is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and / or gum arabic. (3) Humectants, such as glycerin; (4) Disintegrants, such as agar-agar, calcium carbonate, potato or cassava starch, alginic acid, certain silicates and sodium carbonate; (5) Solution blockers, such as paraffin; (6) Absorption promoters, such as quadratic ammonium compounds; (7) Wetting agents, such as acetyl alcohol and glyceryl monostearate; (8) Adsorbents, such as kaolin and bentonite; (9) Lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate and mixtures thereof; and (10) Colorants. In the case of capsules, tablets and pills, the composition may also contain buffers. Similar types of solid compositions may also be used as fillers in soft-filled and hard-filled gelatin capsules when using excipients such as lactose and high molecular weight polyethylene glycol.

[0105] For oral administration, formulations of the compounds disclosed herein may be provided in dosage forms such as capsules, tablets, powders, granules, or in the form of suspensions or solutions. Capsule formulations may be gelatin, soft capsules, or solids. Tablet and capsule formulations may further contain one or more adjuvants, binders, diluents, disintegrants, excipients, fillers, or lubricants, each known in the art. Examples of such agents include carbohydrates (e.g., lactose or sucrose), anhydrous dicalcium hydrogen phosphate, corn starch, mannitol, xylitol, cellulose or derivatives thereof, microcrystalline cellulose, gelatin, stearates, silica, talc, sodium glycolate starch, gum arabic, flavoring agents, preservatives, buffers, disintegrants, and coloring agents. Orally administered compositions may contain one or more optional agents, such as sweeteners, e.g., fructose, aspartame, or saccharin; flavoring agents, e.g., peppermint, wintergreen oil, or cherry oil; coloring agents; and preservatives, to provide a pharmaceutically palatable formulation.

[0106] Tablets can be manufactured by compression or molding, optionally together with one or more adjunct ingredients. Compressed tablets can be prepared using binders (e.g., gelatin or hydroxypropyl methylcellulose), lubricants, inert diluents, preservatives, disintegrants (e.g., sodium glycolate starch or croscarmellose sodium), surfactants, or dispersants. Molded tablets can be prepared by molding a mixture of the subject composition wetted with an inert liquid diluent in a suitable machine. Tablets and other solid dosage forms, such as sugar-coated pills, capsules, pellets, and granules, can optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in pharmaceutical compounding techniques.

[0107] Compositions for inhalation or inhalation include solutions and suspensions in the form of pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, as well as powders. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the subject composition, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents; solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, methyl benzoate, propylene glycol, 1,3-butanediol, oils (specifically, cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerin, tetrahydrofuranyl ethanol, fatty acid esters of polyethylene glycol and sorbitol, cyclodextrins, and mixtures thereof.

[0108] In addition to the main composition, the suspension may also contain suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitol esters, microcrystalline cellulose, aluminum hydroxide, bentonite, agar-agar and astragalus, and mixtures thereof.

[0109] Formulations for rectal or vaginal administration may be presented in suppository form, which can be prepared by mixing the subject composition with one or more suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol, suppository wax, or salicylates, which are solid at room temperature but liquid at body temperature and thus will melt and release the active agent in the body cavity.

[0110] Dosage forms for transdermal application of the subject composition include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches, and inhalers. The active ingredient may be mixed under aseptic conditions with a pharmaceutically acceptable carrier and with any preservatives, buffers, or propellants that may be required.

[0111] In addition to the main composition, ointments, pastes, creams and gels may also contain excipients such as animal and vegetable fats, oils, waxes, paraffins, starches, astragalus, cellulose derivatives, polyethylene glycol, silicones, bentonite, silicic acid, talc and zinc oxide or mixtures thereof.

[0112] In addition to the main composition, powders and sprays may also contain excipients such as lactose, talc, silica, aluminum hydroxide, calcium silicate, and polyamide powder, or mixtures of these substances. Sprays may additionally contain conventional propellants such as chlorofluorocarbons and unsubstituted volatile hydrocarbons such as butane and propane.

[0113] The compositions and compounds disclosed herein can alternatively be administered via aerosols. This is achieved by preparing aqueous aerosols, liposome formulations, or solid particles containing the compounds. Non-aqueous suspensions (e.g., fluorocarbon propellants) can be used. Sonic sprayers can be used because they minimize the exposure of the agent to shear, which can cause degradation of the compounds contained in the subject composition. Typically, aqueous aerosols are manufactured by formulating an aqueous solution or suspension of the subject composition with conventionally pharmaceutically acceptable carriers and stabilizers. Carriers and stabilizers vary depending on the requirements of the specific subject composition but typically include nonionic surfactants (Tweens, Pluronics, or polyethylene glycol); harmless proteins such as serum albumin; sorbitol esters; oleic acid; lecithin; amino acids such as glycine; buffers; salts; sugars or sugar alcohols. Aerosols are typically prepared from isotonic solutions.

[0114] The pharmaceutical compositions of this disclosure suitable for parenteral administration comprise the subject composition and one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted into sterile injectable solutions or dispersions just before use, and may contain antioxidants, buffers, antibacterial agents, solutes or suspending agents or thickeners that make the formulation isotonic with the blood of the intended recipient.

[0115] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of this disclosure include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils (e.g., olive oil), and injectable organic esters (e.g., ethyl oleate and cyclodextrin). Appropriate flowability can be maintained, for example, by using a coating material such as lecithin, by maintaining the desired particle size in the case of a dispersion, and by using a surfactant. For example, the crystalline forms provided herein can be milled to obtain a specific particle size, and in at least some embodiments, such crystalline forms remain substantially stable after milling.

[0116] For example, this document provides a composition suitable for subcutaneous administration comprising a suspension in the disclosed crystalline form. Subcutaneous administration may be superior to intravenous administration, which typically requires a physician's consultation and can be more painful and invasive. When administered to a patient, a typical dose of the crystalline compound may be from about 1 mg to about 8 mg of the compound. In one embodiment, this document discloses a pharmaceutically acceptable composition formed from the disclosed crystalline form, for example, by mixing the crystalline form with excipients and / or solvents.

[0117] In one embodiment, this document provides a composition comprising the disclosed crystalline form, suitable for subcutaneous administration at doses sufficient to deliver about 0.001 mg / kg to about 100 mg / kg, about 0.01 mg / kg to about 50 mg / kg, about 0.1 mg / kg to about 40 mg / kg, about 0.5 mg / kg to about 30 mg / kg, about 0.001 mg / kg to about 4 mg / kg, about 0.1 mg / kg to about 10 mg / kg, or about 1 mg / kg to about 25 mg / kg of a subject's body weight, daily, once or more daily, every other day, every three or four days, weekly, every two weeks, every three weeks, or every four weeks. In some embodiments, the desired dose may be delivered using multiple administrations (e.g., two, three, four, five, six, seven, eight, nine, or ten administrations). In some embodiments, administration may be performed once, twice, or three times per week.

[0118] Treatment may be continued for as long or as short a period of time as needed. The composition may be administered, for example, once to four or more times daily. Suitable treatment periods may be, for example, at least about one week, at least about two weeks, at least about one month, at least about six months, at least about one year, or indefinite. Treatment may be terminated when the desired outcome (e.g., weight loss target) is achieved. Treatment regimens may include a corrective period during which a dose sufficient to cause weight loss is administered, followed by a maintenance period during which a lower dose sufficient to cause weight gain is administered. Suitable maintenance doses may be found in the lower part of the dose ranges provided herein, but based on the disclosure herein, the corrective and maintenance doses for individual subjects can be readily determined by those skilled in the art without extensive experimentation. Maintenance doses may be used to maintain the weight of subjects whose weight has been previously controlled by other means, including diet and exercise, obesity treatment procedures (e.g., bypass surgery or banding surgery), or treatment with other pharmacological agents.

[0119] In some embodiments, this document provides a pharmaceutical composition comprising a crystalline form of compound I or II described herein, or a solvation thereof. In some embodiments, this document provides a pharmaceutical composition comprising a crystalline form of compound I described herein, including, for example, form A, form B, or form C, or a solvation thereof. In some embodiments, this document provides a pharmaceutical composition comprising a crystalline form of compound II described herein, including, for example, form A or form B, or a solvation thereof. In some embodiments, the pharmaceutical compositions provided herein comprise one or more pharmaceutically acceptable excipients described herein.

[0120] Reagent test kit

[0121] In one embodiment, a kit is provided for treating or alleviating a disease or condition of consideration. For example, the disclosed kit comprises a disclosed crystalline compound, such as a crystalline form of compound I, disposed in a first container. In some embodiments, the kit may further comprise a pharmaceutically acceptable excipient disposed in a second container. Such a kit may include written instructions describing the preparation of a pharmaceutical composition suitable for administration to a patient from the crystalline form. For example, the written instructions may describe the preparation of a pharmaceutically acceptable form for patient administration by mixing an excipient with the crystalline compound disclosed herein. The disclosed kit may further include written instructions describing how to administer the resulting composition to a patient.

[0122] method

[0123] In some embodiments, a method for preparing a crystalline form of the disclosed compound I is described herein, comprising: (a) preparing a solution of compound I; (b) adjusting the temperature such that a solid crystalline form of compound I precipitates from the solution; and (c) separating the solid crystalline form. In some embodiments, the solution of compound (I) comprises a solvent selected from the group consisting of water, methanol, ethanol, acetone, methyl ethyl ketone, ethyl acetate, isopropyl acetate, acetonitrile, tert-butyl methyl ether, dichloromethane, tetrahydrofuran, 1,4-dioxane, benzyl alcohol, 2-MeTHF, IPAc, and MtBE. In some embodiments, the solution of compound I comprises a solvent selected from the solvents described in the examples herein.

[0124] In some embodiments, a method for preparing the crystalline form of the disclosed compound I is described herein, comprising: (a) preparing a solution of compound I in a solvent; (b) adjusting the temperature; (c) adding heptane to the solution; and (d) separating the solid crystalline form. In some embodiments, the solution of compound I comprises a solvent selected from the group consisting of water, methanol, ethanol, acetone, methyl ethyl ketone, ethyl acetate, isopropyl acetate, acetonitrile, tert-butyl methyl ether, dichloromethane, tetrahydrofuran, 1,4-dioxane, benzyl alcohol, 2-MeTHF, IPAc, and MtBE. In some embodiments, the solution of compound I comprises a solvent selected from the solvents described in the examples herein.

[0125] In some embodiments, a method for preparing a crystalline form of the disclosed compound II is contemplated herein, comprising: (a) preparing a solution of compound II; (b) adjusting the temperature such that a solid crystalline form of compound II precipitates from the solution; and (c) separating the solid crystalline form. In some embodiments, the solution of compound II comprises a solvent selected from the group consisting of water, methanol, ethanol, acetone, methyl ethyl ketone, ethyl acetate, isopropyl acetate, acetonitrile, tert-butyl methyl ether, dichloromethane, tetrahydrofuran, 1,4-dioxane, benzyl alcohol, 2-MeTHF, IPAc, and MtBE. In some embodiments, the solution of compound II comprises a solvent selected from the solvents described in the examples herein.

[0126] In some embodiments, a method for preparing the crystalline form of the disclosed compound II is contemplated herein, comprising: (a) preparing a solution of compound II in a solvent; (b) adjusting the temperature; (c) adding heptane to the solution; and (d) separating the solid crystalline form. In some embodiments, the solution of compound II comprises a solvent selected from the group consisting of water, methanol, ethanol, acetone, methyl ethyl ketone, ethyl acetate, isopropyl acetate, acetonitrile, tert-butyl methyl ether, dichloromethane, tetrahydrofuran, 1,4-dioxane, benzyl alcohol, 2-MeTHF, IPAc, and MtBE. In some embodiments, the solution of compound II comprises a solvent selected from the solvents described in the examples herein.

[0127] In some embodiments, the step of adjusting the temperature includes heating the solution. In some embodiments, heating the solution includes heating the solution to about 50°C. In some embodiments, adjusting the temperature includes cooling the solution. In some embodiments, cooling the solution includes cooling the solution to about 0°C, about 5°C, or about 25°C.

[0128] In other embodiments, the disclosed method further includes the step of coupling compound A6 with compound A7 to form compound I:

[0129] .

[0130] In other embodiments, the disclosed method further includes the step of converting compound A5 into compound A6:

[0131] .

[0132] In other embodiments, the disclosed method further includes the step of coupling compound A4 with compound S1 to form compound A5:

[0133] .

[0134] In other embodiments, the disclosed method further includes the step of converting compound SM1 into compound S1:

[0135] .

[0136] In other embodiments, the disclosed method further includes the step of converting compound 4 into compound SM1:

[0137] .

[0138] In some embodiments, the step of converting compound 4 into compound SM1 further includes the step of adding an enzyme catalyst.

[0139] In other embodiments, the disclosed method further includes the step of converting compound S2 into compound II:

[0140] .

[0141] method

[0142] The compounds and compositions described herein can generally be used to inhibit kinases or mutants thereof. In some embodiments, the kinase inhibited by the compounds and compositions described herein is phosphatidylinositol 3-kinase (PI3K). In some embodiments, the kinase inhibited by the compounds and compositions described herein is one or more of PI3Kα, PI3Kδ, and PI3Kγ. In some embodiments, the kinase inhibited by the compounds and compositions described herein is PI3Kα. In some embodiments, the kinase inhibited by the compounds and compositions described herein is PI3Kα containing at least one of the following mutations: E542X, E545X, Q546X, H1047X, and G1049X, where X is any amino acid other than its wild type. In some embodiments, the kinase inhibited by the compounds and compositions described herein is a PI3Kα containing at least one of the following mutations: E542K, E542Q, E545A, E545G, E545K, E545Q, Q546E, Q546K, Q546L, Q546P, Q546R, H1047R, H1047L, H1047Y, G1049R, and G1049S. In some embodiments, the kinase inhibited by the compounds and compositions described herein is a PI3Kα containing at least one of the following mutations: E542K, E545K, and H1047R.

[0143] The compounds or compositions disclosed herein can be used in applications benefiting from the inhibition of PI3K enzymes. For example, the PI3K inhibitors of this disclosure are generally used to treat cell proliferation disorders. The compounds or compositions disclosed herein can also be used in applications benefiting from the inhibition of PI3Kα enzymes. For example, the PI3Kα inhibitors of this disclosure are generally suitable for treating cell proliferation disorders.

[0144] Aberrant regulation of PI3K, which typically enhances survival through Aid activation, is one of the most prevalent phenomena in human cancer and has been shown to occur at multiple levels. The tumor suppressor gene PTEN, which dephosphorylates phosphatidylinositol at the 3' position of the inositol ring and thus antagonizes PI3K activity, is lost in a variety of tumors. In other tumors, the genes for p110α isoforms, PIK3CA, and Akt are amplified, and increased protein expression of their gene products has been demonstrated in several human cancers. Furthermore, mutations and translocations of p85α that upregulate the p85-p110 complex have been described in human cancers. Finally, somatic missense mutations in PIK3CA that activate downstream signaling pathways have been described with significant frequency in numerous human cancers (Kang et al., *Proceedings of the National Academy of Sciences* USA 102:802 (2005); Samuels et al., *Science* 304:554 (2004); Samuels et al., *Cancer Cell* 7:561-573 (2005)). These observations suggest that dysregulation of phosphoinositol-3 kinase and its upstream and downstream components is one of the most common dysregulations associated with human cancer and proliferative diseases (Parsons et al., Nature 436:792 (2005); Hennessy et al., Nature Reviews Drug Discovery 4:988-1004 (2005)).

[0145] Treatment of the disease

[0146] The provided compounds are PI3Kα inhibitors and are therefore suitable for treating one or more conditions associated with the activity of PI3Kα or its mutants. Therefore, in some embodiments, this disclosure provides a method of treating a subject with a PI3Kα-mediated condition comprising administering to a subject in need a therapeutically effective amount of the disclosed compound or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable composition of any of the foregoing. In some embodiments, this disclosure provides a method of treating a subject with a PI3Kα-mediated condition comprising administering to a subject in need a therapeutically effective amount of the disclosed compound or a pharmaceutically acceptable composition thereof. In some embodiments, the subject has a mutated PI3Kα. In some embodiments, the subject has a PI3Kα containing at least one of the following mutations: H1047R, E542K, and E545K. In some embodiments, the subject has PI3Kα containing at least one of the following mutations: E542K, E542Q, E545A, E545G, E545K, E545Q, Q546E, Q546K, Q546L, Q546P, Q546R, H1047R, H1047L, H1047Y, G1049R, and G1049S.

[0147] As used herein, the term "PI3Kα-mediated" condition, disease, and / or symptom refers to any disease or other harmful symptom in which PI3Kα or its mutants are known to play a role. Therefore, another embodiment of this disclosure relates to treating or reducing the severity of one or more diseases in which PI3Kα or its mutants are known to play a role. Such PI3Kα-mediated conditions include, but are not limited to, proliferative conditions (e.g., cancer). In some embodiments, PI3Kα-mediated conditions are conditions mediated by mutated PI3Kα. In some embodiments, PI3Kα-mediated conditions are conditions mediated by PI3Kα containing at least one of the following mutations: H1047R, E542K, and E545K. In some embodiments, the subject has PI3Kα containing at least one of the following mutations: E542K, E542Q, E545A, E545G, E545K, E545Q, Q546E, Q546K, Q546L, Q546P, Q546R, H1047R, H1047L, H1047Y, G1049R, and G1049S. In some embodiments, the subject has PI3Kα containing at least one of the mutations in Table A:

[0148] Table A.

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

[0171] In some embodiments, this disclosure provides a method for treating proliferative diseases, the method comprising administering to a patient in need a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable composition of any of the foregoing. In some embodiments, this disclosure provides a method for treating proliferative diseases, the method comprising administering to a patient in need a therapeutically effective amount of a compound of the present disclosure or a pharmaceutically acceptable composition thereof.

[0172] In some embodiments, the treatment method comprises the steps of: (i) identifying a subject in need of such treatment; (ii) providing the disclosed compound or a pharmaceutically acceptable salt thereof; and (iii) administering the provided compound in a therapeutically effective amount to treat, suppress, and / or prevent a disease state or symptom in the subject in need of such treatment. In some embodiments, the subject has a mutated PI3Kα. In some embodiments, the subject has a PI3Kα containing at least one of the following mutations: H1047R, E542K, and E545K. In some embodiments, the subject has a PI3Kα containing at least one of the following mutations: E542K, E542Q, E545A, E545G, E545K, E545Q, Q546E, Q546K, Q546L, Q546P, Q546R, H1047R, H1047L, H1047Y, G1049R, and G1049S.

[0173] In some embodiments, the treatment method comprises the steps of: (i) identifying a subject in need of such treatment; (ii) providing a composition comprising the disclosed compound or a pharmaceutically acceptable salt thereof; and (iii) administering the composition in a therapeutically effective amount to treat, suppress, and / or prevent a disease state or symptom in the subject in need of such treatment. In some embodiments, the subject has a mutated PI3Kα. In some embodiments, the subject has a PI3Kα containing at least one of the following mutations: H1047R, E542K, and E545K. In some embodiments, the subject has a PI3Kα containing at least one of the following mutations: E542K, E542Q, E545A, E545G, E545K, E545Q, Q546E, Q546K, Q546L, Q546P, Q546R, H1047R, H1047L, H1047Y, G1049R, and G1049S.

[0174] Another aspect of this disclosure provides a compound, or a pharmaceutically acceptable salt thereof, as defined herein, or a pharmaceutical composition thereof, for the treatment of the conditions described herein. Another aspect of this disclosure provides the use of a compound, or a pharmaceutically acceptable salt thereof, as defined herein, or a pharmaceutical composition thereof, for the treatment of the conditions described herein. Similarly, this disclosure provides the use of a compound, or a pharmaceutically acceptable salt thereof, as defined herein, for the preparation of a medicament for the treatment of the conditions described herein.

[0175] Cellular proliferative diseases

[0176] In some embodiments, the condition is a proliferative disorder. In some embodiments, the proliferative disorder is cancer. In some embodiments, cancer is a tumor. In some embodiments, cancer is a solid tumor. In some embodiments, the proliferative disorder is the growth of tumors and / or cancer cells. In some embodiments, the proliferative disorder is a tumor. In some embodiments, the proliferative disorder is a solid tumor. In some embodiments, the proliferative disorder is the growth of cancer cells.

[0177] In some embodiments, the solid tumor has a PI3Kα containing at least one of the following mutations: E542X, E545X, Q546X, H1047X, and G1049X, where X is any amino acid other than its wild type. In some embodiments, the solid tumor has a PI3Kα containing at least one of the following mutations: E542K, E542Q, E545A, E545G, E545K, E545Q, Q546E, Q546K, Q546L, Q546P, Q546R, H1047R, H1047L, H1047Y, G1049R, and G1049S. In some embodiments, the solid tumor has a PI3Kα containing at least one of the following mutations: H1047R, E542K, and E545K.

[0178] In some embodiments, the cancer is selected from sarcoma; lung cancer; bronchial cancer; prostate cancer; breast cancer (including sporadic breast cancer and Cowden disease); pancreatic cancer; gastrointestinal cancer; colon cancer; rectal cancer; carcinoma; colon cancer tumor; adenoma; colorectal adenoma; thyroid cancer; liver cancer; intrahepatic bile duct cancer; hepatocellular carcinoma; adrenal cancer; stomach / gastric cancer; glioma; glioblastoma; endometrial cancer; melanoma; kidney cancer; renal pelvis cancer; bladder cancer; uterine corpus cancer; cervical cancer; vaginal cancer; ovarian cancer (including clear cell ovarian cancer); multiple myeloma; esophageal cancer; leukemia; acute myeloid leukemia; chronic myeloid leukemia; lymphocytic leukemia; myeloid leukemia; brain cancer; brain tumor; oral and pharyngeal cancer; laryngeal cancer; small intestine cancer; non-Hodgkin's lymphoma. Lymphoma; villous colonic adenoma; neoplasm formation; epithelial neoplasm formation; lymphoma; breast cancer; basal cell carcinoma; squamous cell carcinoma; actinic keratosis; neck cancer; head cancer; polycythemia vera; essential thrombocythemia; myelofibrosis with myeloid metaplasia; and Waldenstrom macroglobulinemia.

[0179] In some embodiments, the cancer is selected from lung cancer; bronchial cancer; prostate cancer; breast cancer (including sporadic breast cancer and Cowden's disease); pancreatic cancer; gastrointestinal cancer; colon cancer; rectal cancer; thyroid cancer; liver cancer; intrahepatic bile duct cancer; hepatocellular carcinoma; adrenal cancer; gastric cancer; endometrial cancer; kidney cancer; renal pelvis cancer; bladder cancer; uterine corpus cancer; cervical cancer; vaginal cancer; ovarian cancer (including clear cell ovarian cancer); esophageal cancer; leukemia; acute myeloid leukemia; chronic myeloid leukemia; lymphocytic leukemia; myeloid leukemia; brain cancer; oropharyngeal cancer; laryngeal cancer; small bowel cancer; cervical cancer; and head cancer. In some embodiments, the cancer is selected from sarcoma; carcinoma; colon cancer; adenoma; colorectal adenoma; glioma; glioblastoma; melanoma; multiple myeloma; brain cancer; non-Hodgkin's lymphoma; villous colonic adenoma; neoplasm formation; epithelial characteristic neoplasm formation; lymphoma; breast cancer; basal cell carcinoma; squamous cell carcinoma; actinic keratosis; polycythemia vera; essential thrombocythemia; myelofibrosis with myeloid metaplasia; and Waldenström macroglobulinemia.

[0180] In some embodiments, the cancer is selected from lung cancer; bronchial cancer; prostate cancer; breast cancer (including sporadic breast cancer and Cowden's disease); pancreatic cancer; gastrointestinal cancer; colon cancer; rectal cancer; thyroid cancer; liver cancer; intrahepatic bile duct cancer; hepatocellular carcinoma; adrenal cancer; gastric cancer; endometrial cancer; kidney cancer; renal pelvis cancer; bladder cancer; uterine corpus cancer; cervical cancer; vaginal cancer; ovarian cancer (including clear cell ovarian cancer); esophageal cancer; brain cancer; oropharyngeal cancer; laryngeal cancer; small bowel cancer; cervical cancer; and head cancer. In some embodiments, the cancer is leukemia. In some embodiments, the cancer is acute myeloid leukemia; chronic myeloid leukemia; lymphocytic leukemia; or myeloid leukemia.

[0181] In some embodiments, the cancer is breast cancer (including sporadic breast cancer and Cowden's disease). In some embodiments, the cancer is breast cancer. In some embodiments, the cancer is ER+ / HER2- breast cancer. In some embodiments, the cancer is ER+ / HER2- breast cancer, and the subject is intolerant to or unsuitable for apracoxib treatment. In some embodiments, the cancer is sporadic breast cancer. In some embodiments, the cancer is Cowden's disease.

[0182] In some embodiments, the cancer is ovarian cancer. In some embodiments, the ovarian cancer is clear cell ovarian cancer.

[0183] In some embodiments, the cancer is squamous cell carcinoma. In some embodiments, the cancer is squamous cell carcinoma of the head and neck.

[0184] In some embodiments, the cancer is cervical cancer.

[0185] In some embodiments, proliferative diseases have mutated PI3Kα. In some embodiments, cancers have mutated PI3Kα. In some embodiments, breast cancer has mutated PI3Kα. In some embodiments, ovarian cancer has mutated PI3Kα. In some embodiments, clear cell ovarian cancer has mutated PI3Kα.

[0186] In some embodiments, proliferative disorders have PI3Kα containing at least one of the following mutations: H1047R, E542K, and E545K. In some embodiments, proliferative disorders have PI3Kα containing at least one of the following mutations: E542K, E542Q, E545A, E545G, E545K, E545Q, Q546E, Q546K, Q546L, Q546P, Q546R, H1047R, H1047L, H1047Y, G1049R, and G1049S. In some embodiments, cancer has PI3Kα containing at least one of the following mutations: H1047R, E542K, and E545K. In some embodiments, the cancer has a PI3Kα mutation containing at least one of the following: E542K, E542Q, E545A, E545G, E545K, E545Q, Q546E, Q546K, Q546L, Q546P, Q546R, H1047R, H1047L, H1047Y, G1049R, and G1049S. In some embodiments, the breast cancer has a PI3Kα mutation containing at least one of the following: H1047R, E542K, and E545K. In some embodiments, breast cancer has a PI3Kα mutation containing at least one of the following: E542K, E542Q, E545A, E545G, E545K, E545Q, Q546E, Q546K, Q546L, Q546P, Q546R, H1047R, H1047L, H1047Y, G1049R, and G1049S. In some embodiments, ovarian cancer has a PI3Kα mutation containing at least one of the following: H1047R, E542K, and E545K. In some embodiments, ovarian cancer has a PI3Kα mutation containing at least one of the following: E542K, E542Q, E545A, E545G, E545K, E545Q, Q546E, Q546K, Q546L, Q546P, Q546R, H1047R, H1047L, H1047Y, G1049R, and G1049S. In some embodiments, clear cell ovarian cancer has a PI3Kα mutation containing at least one of the following: H1047R, E542K, and E545K. In some embodiments, clear cell ovarian cancer has PI3Kα containing at least one of the following mutations: E542K, E542Q, E545A, E545G, E545K, E545Q, Q546E, Q546K, Q546L, Q546P, Q546R, H1047R, H1047L, H1047Y, G1049R, and G1049S.

[0187] In some embodiments, the cancer is an adenoma; carcinoma; sarcoma; glioma; glioblastoma; melanoma; multiple myeloma; or lymphoma. In some embodiments, the cancer is a colorectal adenoma or villous colonic adenoma. In some embodiments, the cancer is colon cancer; brain cancer; breast cancer; basal cell carcinoma; or squamous cell carcinoma. In some embodiments, the cancer is vesicle formation or epithelial-characteristic vesicle formation. In some embodiments, the cancer is non-Hodgkin's lymphoma. In some embodiments, the cancer is actinic keratosis; polycythemia vera; essential thrombocythemia; myelofibrosis with myeloid metaplasia; or Waldenström macroglobulinemia.

[0188] In some embodiments, proliferative disorders exhibit overexpression or amplification of PI3Kα, somatic mutations in PIK3CA, germline or somatic mutations in PTEN, or mutations and translocations of p85α in the p85-p110 complex as described above. In some embodiments, proliferative disorders exhibit overexpression or amplification of PI3Kα. In some embodiments, proliferative disorders exhibit somatic mutations in PIK3CA. In some embodiments, proliferative disorders exhibit germline or somatic mutations in PTEN. In some embodiments, proliferative disorders exhibit mutations and translocations of p85α in the p85-p110 complex as described above.

[0189] Other diseases

[0190] In some embodiments, PI3Kα-mediated conditions are selected from the group consisting of: polycythemia vera, essential thrombocythemia, myelofibrosis with myeloid metaplasia, asthma, COPD, ARDS, PRO (PI3K-associated overgrowth syndrome), venous malformations, Loffler's syndrome, eosinophilic pneumonia, parasitic (especially metazoan) infections (including tropical eosinophilia), bronchopulmonary aspergillosis, and polyarteritis nodosa (including Churg-Strauss syndrome). Eosinophilic granulomatosis, eosinophilic-associated diseases affecting the respiratory tract due to drug reactions, psoriasis, contact dermatitis, atopic dermatitis, alopecia areata, erythema multiforme, herpetic dermatitis, scleroderma, vitiligo, allergic vasculitis, urticaria, bullous pemphigoid, lupus erythematosus, pemphigus, acquired epidermolysis bullosa, autoimmune blood disorders (e.g., hemolytic anemia, aplastic anemia, pure red cell aplasia, and idiopathic thrombocytopenic purpura), systemic lupus erythematosus, polychondritis, Wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, Stevens-Johnson syndrome, idiopathic diarrhea, autoimmune inflammatory bowel diseases (e.g., ulcerative colitis and Crohn's disease). Diseases including endocrine ophthalmopathy, Graves' disease, sarcoidosis, alveolitis, chronic allergic pneumonia, multiple sclerosis, primary biliary cirrhosis, uveitis (anterior and posterior), pulmonary interstitial fibrosis, psoriatic arthritis, glomerulonephritis, cardiovascular diseases, atherosclerosis, hypertension, deep vein thrombosis, stroke, myocardial infarction, unstable angina, thromboembolism, pulmonary embolism, thrombolytic diseases, acute arterial ischemia, peripheral thrombotic occlusion and coronary artery disease, reperfusion injury, and retinopathy (e.g., diabetic retinopathy or hyperbaric oxygen-induced retinopathy and conditions characterized by increased intraocular pressure or aqueous humor secretion, such as glaucoma).

[0191] In some embodiments, PI3Kα-mediated conditions include polycythemia vera, essential thrombocythemia, or myelofibrosis with myeloid metaplasia. In some embodiments, PI3Kα-mediated conditions include asthma, COPD, ARDS, PROS (PI3K-associated overgrowth syndrome), venous malformations, Löffler's syndrome, eosinophilic pneumonia, parasitic (especially metazoan) infections (including tropical eosinophilia), or bronchopulmonary aspergillosis. In some embodiments, PI3Kα-mediated conditions include polyarteritis nodosa (including Chag-Strauss syndrome), eosinophilic granuloma, eosinophilic-associated conditions affecting the respiratory tract due to drug reactions, psoriasis, contact dermatitis, atopic dermatitis, alopecia areata, erythema multiforme, dermatitis herpetiformis, or scleroderma. In some embodiments, PI3Kα-mediated conditions include vitiligo, allergic vasculitis, urticaria, bullous pemphigoid, lupus erythematosus, pemphigus, acquired epidermolysis bullosa, or autoimmune blood disorders (e.g., hemolytic anemia, aplastic anemia, pure red cell aplasia, and idiopathic thrombocytopenic purpura). In some embodiments, PI3Kα-mediated conditions include systemic lupus erythematosus, polychondritis, scleroderma, Wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, Stephen Johnson syndrome, idiopathic diarrhea, or autoimmune inflammatory bowel disease (e.g., ulcerative colitis and Crohn's disease).

[0192] In some embodiments, PI3Kα-mediated conditions include endocrine ophthalmopathy, Graves' disease, sarcoidosis, alveolitis, chronic allergic pneumonia, multiple sclerosis, primary biliary cirrhosis, uveitis (anterior and posterior), pulmonary interstitial fibrosis, or psoriatic arthritis. In some embodiments, PI3Kα-mediated conditions include glomerulonephritis, cardiovascular disease, atherosclerosis, hypertension, deep vein thrombosis, stroke, myocardial infarction, unstable angina, thromboembolism, pulmonary embolism, thrombolytic disease, acute arterial ischemia, peripheral thrombotic occlusion, and coronary artery disease or reperfusion injury. In some embodiments, PI3Kα-mediated conditions include retinopathy, such as diabetic retinopathy or hyperbaric oxygen-induced retinopathy, and conditions characterized by elevated intraocular pressure or aqueous humor secretion (e.g., glaucoma).

[0193] Application route and dosage form

[0194] According to the methods of this disclosure, the compounds and compositions can be administered in any amount and via any route of administration to effectively treat a condition (e.g., proliferative diseases) or reduce its severity. The precise amount required will vary from subject to subject, depending on the subject's species, age and general condition, severity of infection, specific agent, mode of administration, etc. For ease of administration and dosage uniformity, the compounds of this disclosure are preferably formulated into unit dosage forms. As used herein, the term "unit dosage form" refers to a physically discrete unit of the agent suitable for the patient to be treated. However, it should be understood that the total daily dosage of the compounds and compositions of this disclosure will be determined by the attending physician within the bounds of reasonable medical judgment. The specific effective dose level for any particular patient or organism will depend on a variety of factors, including the condition being treated and its severity; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health, sex, and diet; the timing, route of administration, and elimination rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors well known in the medical field.

[0195] The pharmaceutically acceptable compositions of this disclosure can be administered to humans and other animals via oral, rectal, parenteral, intracerebrospinal, vaginal, intraperitoneal, topical (e.g., by powder, ointment, or drops), buccal, oral, or nasal spray form. In some embodiments, the compounds of this disclosure may be administered once or more daily at a dose level of about 0.01 mg to about 50 mg per kilogram of subject body weight per day, and preferably at a dose level of about 1 mg to about 25 mg per kilogram of subject body weight per day, via oral or parenteral administration to obtain the desired therapeutic effect.

[0196] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, liquid dosage forms may contain inert diluents commonly used in the field, such as water or other solvents; solubilizers and emulsifiers, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, methyl benzoate, propylene glycol, 1,3-butanediol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitol; and mixtures thereof. In addition to inert diluents, oral compositions may also include adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and aromatizers.

[0197] Injectable formulations can be formulated using suitable dispersants or wetting agents and suspending agents according to known techniques, such as sterile injectable aqueous or oily suspensions. Sterile injectable formulations can also be sterile injectable solutions, suspensions, or emulsions in non-toxic, parenteral-acceptable diluents or solvents, such as solutions in 1,3-butanediol. Among acceptable mediators and solvents, water, Ringer's solution, USP, and isotonic sodium chloride solution can be used. Additionally, sterile non-volatile oils are routinely used as solvents or suspension media. For this purpose, any mild, non-volatile oil can be used, including synthetic monoglycerides or diglycerides. Furthermore, fatty acids, such as oleic acid, are used in the preparation of injectable formulations.

[0198] Injectable formulations can be sterilized, for example, by filtration through a bacterial trapping filter or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable media before use.

[0199] To prolong the effects of the disclosed compounds, it is generally necessary to slow down the absorption of the compounds from subcutaneous or intramuscular injection. This can be achieved by using liquid suspensions of crystalline or amorphous materials with poor water solubility. The absorption rate of the compound then depends on its solubility, which in turn depends on the crystal size and crystal form. Alternatively, delayed absorption of the parenteral compound form can be achieved by dissolving or suspending the compound in an oil-based medium. Injectable storage forms are prepared by forming microcapsule matrices of the compound in a biodegradable polymer, such as polylactide-polyglycolic acid. The release rate of the compound can be controlled depending on the ratio of compound to polymer and the properties of the specific polymer used. Examples of other biodegradable polymers include poly(orthoester) and poly(anhydride). Reservoir-type injectable formulations are also prepared by encapsulating the compound in tissue-compatible lipid particles or microemulsions.

[0200] Compositions for rectal or vaginal administration are preferably suppositories, which can be prepared by mixing the disclosed compounds with a suitable non-irritating excipient or carrier (e.g., cocoa butter, polyethylene glycol, or suppository wax) that is solid at ambient temperature but liquid at body temperature and thus melts in the rectal or vaginal cavity and releases the active compound.

[0201] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one pharmaceutically acceptable inert excipient or carrier (e.g., sodium citrate or dicalcium phosphate) and / or the following: a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and silica; b) binders, such as carboxymethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic; c) humectants, such as glycerin; d) disintegrants, such as agar-agar, calcium carbonate, potato or cassava starch, alginate, certain silicates, and sodium carbonate; e) solvent inhibitors, such as paraffin; f) absorption accelerators, such as quaternary ammonium compounds; g) humectants, such as cetyl alcohol and glyceryl monostearate; h) absorbents, such as kaolin and bentonite; and i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also contain a buffer.

[0202] Similar solid compositions can also be used as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose / milk sugar and high molecular weight polyethylene glycol. Solid dosage forms of tablets, sugar-coated pills, capsules, pellets, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings well known in pharmaceutical formulation techniques. They may optionally contain emulsifiers and may also have compositions that release the active ingredient only or preferentially in a portion of the intestine or optionally in a delayed manner. Examples of usable encapsulation compositions include polymeric substances and waxes. Similar solid compositions can also be used as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose / milk sugar and high molecular weight polyethylene glycol.

[0203] The active compound may also be present in microencapsulation form with one or more of the excipients mentioned above. Solid dosage forms of tablets, sugar-coated pills, capsules, pellets, and granules may be prepared with coatings and shells, such as enteric coatings, release-controlled coatings, and other coatings well known in pharmaceutical formulation techniques. In said solid dosage forms, the active compound may be mixed with at least one inert diluent, such as sucrose, lactose, or starch. Conventionally, such dosage forms may also contain additional substances besides inert diluents, such as tablet-making lubricants and other tablet-making aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pellets, the dosage form may also contain a buffer. It may optionally contain an emulsifier and may also have a composition that releases or optionally releases the active ingredient only or preferentially in a portion of the intestine. Examples of encapsulation compositions that can be used include polymers and waxes.

[0204] Dosage forms for topical or transdermal application of the compounds of this disclosure include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalers, or patches. The active ingredient is blended under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or buffers. Ophthalmic formulations, ear drops, and eye drops are also contemplated within the scope of this disclosure. Additionally, the use of transdermal patches is contemplated, offering the added advantage of providing controlled delivery of the compound to the body. Such dosage forms can be prepared by dissolving or dispensing the compound in a suitable medium. Absorption enhancers may also be used to increase the transdermal amount of the compound. The rate can be controlled by providing a rate-controlled membrane or by dispersing the compound in a polymer matrix or gel.

[0205] Dosage and regimen

[0206] According to the method of this disclosure, a therapeutically effective amount of the disclosed compound is administered to a subject, for example, to alleviate or improve the subject's symptoms. This amount can be readily determined by those skilled in the art based on known procedures, including analysis of titration curves established in vivo and the methods and determinations disclosed herein.

[0207] In some embodiments, the method comprises administering a therapeutically effective dose of a compound of the present disclosure. In some embodiments, the therapeutically effective dose is at least about 0.0001 mg / kg body weight, at least about 0.001 mg / kg body weight, at least about 0.01 mg / kg body weight, at least about 0.05 mg / kg body weight, at least about 0.1 mg / kg body weight, at least about 0.25 mg / kg body weight, at least about 0.3 mg / kg body weight, at least about 0.5 mg / kg body weight, at least about 0.75 mg / kg body weight, at least about 1 mg / kg body weight, at least about 2 mg / kg body weight, at least about 3 mg / kg body weight, at least about 4 mg / kg body weight, at least about 5 mg / kg body weight, at least about 6 mg / kg body weight, at least about 7 mg / kg body weight, at least about 8 mg / kg body weight, at least about 9 mg / kg body weight, at least about 10 mg / kg body weight, at least about 15 mg / kg body weight, at least about 20 mg / kg body weight, at least about 25 mg / kg body weight, at least about 30 mg / kg body weight, at least about 40 mg / kg body weight, at least about 50 mg / kg body weight, at least about 75 mg / kg body weight. The dosages listed herein may be at least approximately 100 mg / kg body weight, at least approximately 200 mg / kg body weight, at least approximately 250 mg / kg body weight, at least approximately 300 mg / kg body weight, at least approximately 350 mg / kg body weight, at least approximately 400 mg / kg body weight, at least approximately 450 mg / kg body weight, at least approximately 500 mg / kg body weight, at least approximately 550 mg / kg body weight, at least approximately 600 mg / kg body weight, at least approximately 650 mg / kg body weight, at least approximately 700 mg / kg body weight, at least approximately 750 mg / kg body weight, at least approximately 800 mg / kg body weight, at least approximately 900 mg / kg body weight, or at least approximately 1000 mg / kg body weight. It should be understood that any of the dosages listed herein may constitute an upper or lower limit dose range and may be combined with any other dosage to constitute a dose range that includes both the upper and lower limits.

[0208] In some embodiments, the effective therapeutic dose ranges from about 0.1 mg to about 10 mg per kilogram of body weight, from about 0.1 mg to about 6 mg per kilogram of body weight, from about 0.1 mg to about 4 mg per kilogram of body weight, or from about 0.1 mg to about 2 mg per kilogram of body weight.

[0209] In some embodiments, the range of therapeutically effective doses is about 1 to 500 mg, about 2 to 150 mg, about 2 to 120 mg, about 2 to 80 mg, about 2 to 40 mg, about 5 to 150 mg, about 5 to 120 mg, about 5 to 80 mg, about 10 to 150 mg, about 10 to 120 mg, about 10 to 80 mg, about 10 to 40 mg, about 20 to 150 mg, about 20 to 120 mg, about 20 to 80 mg, about 20 to 40 mg, about 40 to 150 mg, about 40 to 120 mg, or about 40 to 80 mg.

[0210] In some embodiments, the method comprises a single dose or administration (e.g., in the form of a single injection or deposition). Alternatively, in some embodiments, the method comprises administering the drug to a subject in need once daily, twice daily, three times daily, or four times daily for a period of about 2 to about 28 days, or about 7 to about 10 days, or about 7 to about 15 days or longer. In some embodiments, the method comprises long-term administration. In yet other embodiments, the method comprises administration over a course of weeks, months, years, or decades. In other embodiments, the method comprises administration over a course of weeks. In other embodiments, the method comprises administration over a course of months. In other embodiments, the method comprises administration over a course of years. In other embodiments, the method comprises administration over a course of decades.

[0211] The dosage may vary depending on known factors, such as the pharmacodynamic characteristics of the active ingredient and its administration modality and route; the timing of administration of the active ingredient; the recipient's age, sex, health status, and weight; the nature and severity of symptoms; the type, frequency, and desired effect of concurrent treatments; and the rate of secretion. All of these factors are readily determined and can be used by those skilled in the art to adjust or customize the dosage and / or dosing regimen.

[0212] Inhibition of protein kinases

[0213] According to one embodiment, this disclosure relates to a method for inhibiting the activity of a protein kinase in a biological sample, comprising the step of contacting the biological sample with a compound of the present disclosure or a composition containing the compound. According to another embodiment, this disclosure relates to a method for inhibiting the activity of PI3K or a mutant thereof in a biological sample, comprising the step of contacting the biological sample with a compound of the present disclosure or a composition containing the compound. According to another embodiment, this disclosure relates to a method for inhibiting the activity of PI3Kα or a mutant thereof in a biological sample, comprising the step of contacting the biological sample with a compound of the present disclosure or a composition containing the compound. In some embodiments, PI3Kα is a mutant PI3Kα. In some embodiments, PI3Kα contains at least one of the following mutations: H1047R, E542K, and E545K.

[0214] In another embodiment, this disclosure provides a method for selectively inhibiting PI3Kα relative to one or both of PI3Kδ and PI3Kγ. In some embodiments, the compounds of this disclosure have selectivity greater than 5-fold relative to PI3Kδ and PI3Kγ. In some embodiments, the compounds of this disclosure have selectivity greater than 10-fold relative to PI3Kδ and PI3Kγ. In some embodiments, the compounds of this disclosure have selectivity greater than 50-fold relative to PI3Kδ and PI3Kγ. In some embodiments, the compounds of this disclosure have selectivity greater than 100-fold relative to PI3Kδ and PI3Kγ. In some embodiments, the compounds of this disclosure have selectivity greater than 200-fold relative to PI3Kδ and PI3Kγ. In some embodiments, PI3Kα is a mutant PI3Kα. In some embodiments, PI3Kα contains at least one of the following mutations: H1047R, E542K, and E545K.

[0215] In another embodiment, this disclosure provides a method for selectively inhibiting mutant PI3Kα relative to wild-type PI3Kα. In some embodiments, the compounds of this disclosure exhibit more than 5-fold selectivity for mutant PI3Kα relative to wild-type PI3Kα. In some embodiments, the compounds of this disclosure exhibit more than 10-fold selectivity for mutant PI3Kα relative to wild-type PI3Kα. In some embodiments, the compounds of this disclosure exhibit more than 50-fold selectivity for mutant PI3Kα relative to wild-type PI3Kα. In some embodiments, the compounds of this disclosure exhibit more than 100-fold selectivity for mutant PI3Kα relative to wild-type PI3Kα. In some embodiments, the compounds of this disclosure exhibit more than 200-fold selectivity for mutant PI3Kα relative to wild-type PI3Kα. In some embodiments, the mutant PI3Kα contains at least one of the following mutations: H1047R, E542K, and E545K.

[0216] As used herein, the term “biological sample” includes (but is not limited to) cell cultures or extracts thereof; biopsy material obtained from mammals or extracts thereof; and blood, saliva, urine, feces, semen, tears or other bodily fluids or extracts thereof.

[0217] Inhibiting the activity of PI3K (e.g., PI3Kα, or mutants thereof) in biological samples is suitable for a variety of purposes known to those skilled in the art. Examples of such purposes include, but are not limited to, bioassays for blood transfusions, organ transplantation, and biological specimen storage.

[0218] Another embodiment of this disclosure relates to a method for inhibiting protein kinase activity in a patient, comprising the step of administering a compound of this disclosure or a composition comprising the compound to the patient.

[0219] According to another embodiment, this disclosure relates to a method for inhibiting the activity of PI3K or a mutant thereof in a patient, comprising the step of applying a compound of the present disclosure or a composition comprising the compound to the patient. In some embodiments, this disclosure relates to a method for inhibiting the activity of PI3Kα or a mutant thereof in a patient, comprising the step of applying a compound of the present disclosure or a composition comprising the compound to the patient. In some embodiments, PI3Kα is mutant PI3Kα. In some embodiments, PI3Kα contains at least one of the following mutations: H1047R, E542K, and E545K.

[0220] According to another embodiment, this disclosure provides a method for treating a patient in need of a condition mediated by PI3K or a mutant thereof, the method comprising the step of administering to the patient a compound according to this disclosure or a pharmaceutically acceptable composition thereof. In some embodiments, this disclosure provides a method for treating a patient in need of a condition mediated by PI3Kα or a mutant thereof, comprising the step of administering to the patient a compound according to this disclosure or a pharmaceutically acceptable composition thereof. In some embodiments, PI3Kα is a mutant PI3Kα. In some embodiments, PI3Kα contains at least one of the following mutations: H1047R, E542K, and E545K.

[0221] According to another embodiment, this disclosure provides a method for inhibiting the signal transduction activity of PI3Kα or a mutant thereof in a subject, comprising administering to a subject in need a therapeutically effective amount of a compound according to this disclosure or a pharmaceutically acceptable composition thereof. In some embodiments, this disclosure provides a method for inhibiting the signal transduction activity of PI3Kα in a subject, comprising administering to a subject in need a therapeutically effective amount of a compound according to this disclosure or a pharmaceutically acceptable composition thereof. In some embodiments, PI3Kα is a mutant PI3Kα. In some embodiments, PI3Kα contains at least one of the following mutations: H1047R, E542K, and E545K. In some embodiments, the subject has a mutant PI3Kα. In some embodiments, the subject has PI3Kα containing at least one of the following mutations: H1047R, E542K, and E545K.

[0222] The compounds described herein can also inhibit PI3Kα function by incorporation into agents that catalyze PI3Kα destruction. For example, the compounds can be incorporated into protein hydrolysis-targeting chimeras (PROTACs). PROTACs are bifunctional molecules, with one part binding to an E3 ubiquitin ligase and the other part binding to a target protein intended for degradation via cellular protein biomechanical control mechanisms. The target protein is recruited to a specific E3 ligase, which then labels it for destruction (i.e., ubiquitination) and subsequently degrades it by the proteasome. Any E3 ligase can be used. The portion of the PROTAC that binds to the E3 ligase is linked to the portion of the PROTAC that binds to the target protein via a linker composed of a variable atomic chain. Therefore, recruiting PI3Kα to an E3 ligase will result in the destruction of the PI3Kα protein. The variable atomic chain may include, for example, rings, heteroatoms, and / or repeating polymer units. It can be rigid or flexible. It can be linked to the two parts using standard techniques in the field of organic synthesis.

[0223] combination therapy

[0224] Depending on the specific condition, symptom, or disease to be treated, additional therapeutic agents typically administered for treating said condition may be combined with the compounds and compositions disclosed herein. As used herein, additional therapeutic agents typically administered for treating a specific disease or symptom are referred to as “the disease or symptom to be treated”.

[0225] In addition, PI3K acts as a second messenger node that integrates parallel signaling pathways, and there is evidence that combinations of PI3K inhibitors with inhibitors of other pathways may be suitable for the treatment of cancer and proliferative diseases.

[0226] Therefore, in some embodiments, the treatment method comprises administering a compound or composition of the present disclosure in combination with one or more other therapeutic agents. In some other embodiments, the treatment method comprises administering a compound or composition of the present disclosure as the sole therapeutic agent.

[0227] Approximately 20-30% of human breast cancers overexpress Her-2 / neu-ErbB2, which is the target of the drug trastuzumab. Although trastuzumab has shown durable responses in some patients expressing Her2 / neu-ErbB2, only a fraction of these patients respond. Recent studies have indicated that this limited response rate can be significantly improved by combining trastuzumab with PI3K or PI13K / AKT pathway inhibitors (Chan et al., Breast Cancer Research and Treatment 91:187 (2005), Woods Ignatoski et al., British Journal of Cancer 82:666 (2000), Nagata et al., Cancer Cell 6:117 (2004)). Therefore, in some embodiments, the treatment method comprises administration of a combination of the compounds or compositions of this disclosure and trastuzumab. In some embodiments, the cancer is human breast cancer that overexpresses Her-2 / neu-ErbB2.

[0228] Many human malignancies express activating mutations or increased levels of Her1 / EGFR, and various antibodies and small molecule inhibitors targeting this receptor tyrosine kinase have been developed, including tarceva, gefitinib, and erbitux. However, although EGFR inhibitors have shown antitumor activity in some human tumors (e.g., NSCLC), they have failed to increase overall patient survival in all patients with EGFR-expressing tumors. This can be justified by the fact that many downstream targets of Her1 / EGFR are frequently mutated or dysregulated in a variety of malignancies, including the PI3K / Akt pathway.

[0229] For example, gefitinib inhibits the growth of adenocarcinoma cell lines in in vitro assays. However, subclones of these cell lines resistant to gefitinib can be selected, exhibiting increased activation of the PI3 / Akt pathway. Downregulation or inhibition of this pathway makes the resistant subclones sensitive to gefitinib (Kokubo et al., *British Journal of Cancer* 92:1711 (2005)). Furthermore, a synergistic effect is observed in in vitro models of breast cancer with cell lines carrying PTEN mutations and overexpressing both the PI3K / Akt pathway and EGFR-inhibited cells (She et al., *Cancer Cells* 8:287-297 (2005)). These results suggest that the combination of gefitinib with PI3K / Akt pathway inhibitors could be an attractive cancer treatment strategy.

[0230] Therefore, in some embodiments, the treatment method comprises administering a combination of the compounds or compositions of this disclosure with a Her1 / EGFR inhibitor. In some embodiments, the treatment method comprises administering a combination of the compounds or compositions of this disclosure with one or more of the following: Dextromethorphan, gefitinib, and erbitrol. Therefore, in some embodiments, the treatment method comprises administering a combination of the compounds or compositions of this disclosure with gefitinib. In some embodiments, the cancer expresses activating mutations or has increased levels of Her1 / EGFR.

[0231] The combination of AEE778 (an inhibitor of Her-2 / neu / ErbB2, VEGFR and EGFR) and RAD001 (an inhibitor of mTOR, a downstream target of Akt) produced greater combined efficacy in glioblastoma xenograft models than either agent alone (Goudar et al., Molecular Cancer Ther. 4:101-112 (2005)).

[0232] Anti-estrogens, such as tamoxifen, inhibit breast cancer growth by inducing cell cycle arrest, which requires the action of the cell cycle inhibitor p27Kip. Recently, activation of the Ras-Raf-MAP kinase pathway has been shown to alter the phosphorylation state of p27Kip, weakening its cell cycle-inhibiting activity and thus contributing to anti-estrogenic resistance (Donovan et al., *Journal of Biol. Chem.* 276:40888, (2001)). According to Donovan et al., treatment with a MEK inhibitor to inhibit MAPK signaling reverses the phosphorylation state of p27 in hormone-refractory breast cancer cell lines, thereby restoring hormone sensitivity. Similarly, phosphorylation of p27Kip via Aid also eliminates its cell cycle-inhibiting effect (Viglietto et al., *Nature Medicine* 8:1145 (2002)).

[0233] Therefore, in some embodiments, the treatment method comprises administering a combination of the compounds or compositions of this disclosure with the treatment of hormone-dependent cancer. In some embodiments, the treatment method comprises administering a combination of the compounds or compositions of this disclosure with tamoxifen. In some embodiments, the cancer is a hormone-dependent cancer, such as breast cancer and prostate cancer. This use is intended to reverse hormone tolerance common in these cancers using conventional anticancer agents.

[0234] In hematologic malignancies, such as chronic myeloid leukemia (CML), chromosomal translocations lead to constitutive activation of the BCR-Abl tyrosine kinase. Patients with this disease respond to the small-molecule tyrosine kinase inhibitor imatinib due to inhibition of Abl kinase activity. However, many patients with advanced disease who initially respond to imatinib subsequently relapse due to resistance-conferred mutations in the Abl kinase domain. In vitro studies have shown that BCR-Ab1 activates its action via the Ras-Raf kinase pathway. Furthermore, inhibition of more than one kinase in the same pathway provides additional protection against resistance-conferred mutations.

[0235] Therefore, in another aspect, the compounds and compositions of this disclosure are combined with at least one additional agent selected from the group of kinase inhibitors (e.g., imatinib) to treat hematologic cancers, such as chronic myeloid leukemia (CML). This use is intended to reverse or prevent resistance to said at least one additional agent.

[0236] Since activation of the PI3K / Akt pathway drives cell survival, pathway inhibition in combination with therapies that drive cancer cell apoptosis (including radiotherapy and chemotherapy) will result in improved responses (Ghobrial et al., CA Cancer J. Clin 55:178-194 (2005)). For example, the combination of PI3 kinase inhibitors with carboplatin has shown synergistic effects in in vitro proliferation and apoptosis assays and in vivo tumor efficacy in ovarian cancer xenograft models (Westfall and Skinner, Molecular Cancer Ther. 4:1764-1771 (2005)).

[0237] In some embodiments, one or more additional therapeutic agents are selected from antibodies, antibody-drug conjugates, kinase inhibitors, immunomodulators, and histone deacetylase inhibitors. Synergistic combinations with PIK3CA inhibitors and other therapeutic agents are described, for example, in Castel et al., *Molecular and Cellular Oncology* (2014) 1(3)e963447.

[0238] In some embodiments, one or more additional therapeutic agents are selected from the following agents or pharmaceutically acceptable salts thereof: BCR-ABL inhibitors (see, for example, Ultimo et al., Oncotarget (2017) 8(14) 23213-23227): such as imatinib, inilotinib, nilotinib, dasatinib, bosutinib, ponatinib, bafetinib, danusertib, saracatinib, PF03814735; ALK inhibitors (see Yang et al., Tumor Biology (2014) 35(10)). 9759-67): For example, crizotinib, NVP-TAE684, ceritinib, alectinib, brigatinib, entrecinib, lorlatinib; BRAF inhibitors (see, for example, Silva et al., *Molecular Cancer Research* (2014) 12, 447-463): For example, vemurafenib, dabrafenib; FGFR inhibitors (see, for example, *Molecular Cancer Therapeutics* (2017) 16(4)). 637-648): For example, infigratinib, dovitinib, erdafitinib, TAS-120, pemigatinib, BLU-554, AZD4547; FLT3 inhibitors: For example, sunitinib, midostaurin, tanutinib, sorafenib, lestaurtinib, quizartinib, and crenolanib; MEK inhibitors (see, for example, Jokinen et al., Adv. Med. Oncol.).(2015) 7(3) 170-180): for example, trametinib, cobimetinib, binimetinib, selumetinib; ERK inhibitors: for example, ulixertinib, MK 8353, LY 3214996; KRAS inhibitors: for example, AMG-510, MRTX849, ARS-3248; tyrosine kinase inhibitors (see, for example, Makhov et al., Molecular Cancer Therapeutics (2012) 11(7)1510-1517): for example, erlotinib, linifanib, sunitinib, pazopanib; epidermal growth factor receptor (EGFR) inhibitors (see, for example, She et al., BMC Cancer (2016) 16, 587): Gefitinib, osimertinib, cetuximab, panitumumab; HER2 receptor inhibitors (see, for example, Lopez et al., Molecular Cancer Therapeutics (2015) 14(11) 2519-2526): for example, trastuzumab, pertuzumab, neratinib, lapatinib, lapatinib; MET inhibitors (see, for example, Hervieu et al., Frontiers in Molecular Biosciences).(2018) 5,86): e.g., crizotinib, cabozantinib; CD20 antibodies: e.g., rituximab, tositumomab, ofatumumab; DNA synthesis inhibitors: e.g., capecitabine, gemcitabine, nelarabine, hydroxycarbamide; anti-hypertrophic agents (see e.g., Wang et al., Cell Death & Disease (2018) 9, 739): For example, oxaliplatin, carboplatin, cisplatin; immunomodulators: for example, afutuzumab, lenalidomide, thalidomide, pomalidomide; CD40 inhibitors: for example, dacetuzumab; pro-apoptotic receptor agonists (PARA): for example, dulanermin; heat shock protein (HSP) inhibitors (see, for example, Chen et al., Tumor Targets (2014) 5 (9). 2372-2389): For example, tanespimycin; hedgehog protein antagonists (see, for example, Chaturvedi et al., Tumor Targets (2018) 9 (24)). 16619-16633): e.g., vismodegib; proteasome inhibitors (see, e.g., Lin et al., *International Journal of Oncology* (2014) 44(2), 557-562): e.g., bortezomib; PI3K inhibitors: e.g., pictilisib, dactolisib, apiride, bupalisib, tasilisib, idelalisib, duvelisib, umbralisib; SHP2 inhibitors (see, e.g., Sun et al., *American Journal of Cancer Research*).(2019) 9 (1), 149-159: e.g., SHP099, RMC-4550, RMC-4630); BCL-2 inhibitors (see e.g., Bojarczuk et al., Blood (2018) 133 (1), 70-80): e.g., venetoclax; aromatase inhibitors (see e.g., Mayer et al., Clin. CancerRes. (2019) 25 (10), 2975-2987): exemestane, letrozole, anastrozole, fulvestrant, tamoxifen; mTOR inhibitors (see e.g., Woo et al., Oncogenesis (2017) 6, e385): for example, temsirolimus, ridaforolimus, everolimus, sirolimus; CTLA-4 inhibitors (see, for example, O'Donnell et al., (2018) 48, 91-103): for example, tremelimumab, ipilimumab; PD1 inhibitors (see, O'Donnell, ibid.): for example, nivolumab, pembrolizumab; immunoadhesins; other immune checkpoint inhibitors (see, for example, Zappasodi et al., Cancer Cell (2018) 33, 581-598), where the term "immune checkpoint" refers to a group of molecules on the cell surface of CD4 and CD8 T cells. Immune checkpoint molecules include, but are not limited to, planned death protein 1 (PD-1), cytotoxic T-lymphocyte antigen 4 (CTLA-4), B7H1, B7H4, OX-40, CD137, CD40, and LAG3. Immunotherapy agents that can act as immune checkpoint inhibitors suitable for the methods of this disclosure include, but are not limited to, inhibitors of PD-L1, PD-L2, CTLA4, TIM3, LAG3, VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4, and / or TGFR β: e.g., pidilizumab, AMP-224; PDL1 inhibitors (see, for example, otonic, above): e.g., MSB0010718C; YW243.55.S70, MPDL3280A; MEDI-4736, MSB-0010718C or MDX-1105; histone deacetylase inhibitors (HDI, see, for example, Rahmani et al., Clinical Cancer Research (2014) 20(18), 4849-4860): for example, vorinostat; androgen receptor inhibitors (see, for example, Thomas et al., Molecular Cancer Therapeutics (2013) 12(11), 2342-2355): for example, enzalutamide, abiraterone acetate. Acetate), orteronel, galeterone, seviteronel, bicalutamide, flutamide; androgens: such as fluoxymesterone; CDK4 / 6 inhibitors (see, for example, Gul et al., *American Journal of Cancer Research* (2018) 8(12), 2359-2376): such as alvocidib, palbociclib, ribociclib, trilaciclib, abemaciclib.

[0239] In some embodiments, one or more additional therapeutic agents are selected from the following: anti-FGFR antibodies; FGFR inhibitors; cytotoxic agents; estrogen receptor-targeted or other endocrine therapies; immune checkpoint inhibitors; CDK inhibitors; receptor tyrosine kinase inhibitors; BRAF inhibitors; MEK inhibitors; other PI3K inhibitors; SHP2 inhibitors; and SRC inhibitors. (See Katoh, *Nature Reviews Clin. Oncol.* (2019), 16:105-122; Chae et al., *Targets for Tumors* (2017), 8:16052-16074; Formisano et al., *Nature Communications* (2019), 10:1373-1386; and references cited therein).

[0240] In some embodiments, estrogen receptor-targeted therapy is a selective estrogen receptor degrader (SERD, such as fulvestrant, elacestrant, or giredestrant). In some embodiments, estrogen receptor-targeted therapy is a PROTAC that degrades estrogen receptors (e.g., ARV-471). In some embodiments, endocrine therapy is an aromatase inhibitor (e.g., anastrozole, letrozole, or exemestane).

[0241] In some embodiments, one or more additional therapeutic agents are inhibitors of one or more of the CDK2, CDK4, and CDK6 enzymes. In some embodiments, the CDK inhibitor is a CDK2 inhibitor (e.g., PF-07104091). In some embodiments, the CDK inhibitor is a CDK4 inhibitor (e.g., PF-07220060, AU2-94). In some embodiments, the CDK inhibitor is a dual CDK4 / 6 inhibitor (e.g., peroxizob, abemaxib, ribociclib, triazobicil). In some embodiments, the CDK inhibitor is a CDK2 / 4 / 6 inhibitor.

[0242] In some embodiments, more than one CDK inhibitor is administered together with a compound of this disclosure. In some embodiments, an additional therapeutic agent comprises one or more CDK inhibitors and an estrogen receptor-targeting therapy. In some embodiments, an additional therapeutic agent comprises a selective estrogen receptor degrader and one or more CDK inhibitors.

[0243] The structures of active compounds identified by code number, generic name, or trademark name can be obtained from the current version of the standard compilation "The Merck Index" or from databases such as Patents International (e.g., IMS World Publications).

[0244] The compounds disclosed herein can also be used in combination with known treatment methods, such as hormone administration or radiation. In some embodiments, the provided compounds are used as radiosensitizers, particularly for treating tumors that exhibit poor sensitivity to radiation therapy.

[0245] The compounds disclosed herein may be administered alone or in combination with one or more other therapeutic compounds. Possible combination therapies may involve the administration of the compounds disclosed herein and one or more other therapeutic compounds in a fixed combination or alternately or independently, or a fixed combination combined with one or more other therapeutic compounds. Alternatively or additionally, the compounds disclosed herein may be administered, particularly in combination with chemotherapy, radiotherapy, immunotherapy, phototherapy, surgical intervention, or combinations thereof, for cancer therapy. As described above, long-term therapy is also possible, similar to adjuvant therapy in the context of other treatment strategies. Other possible treatments include therapies to maintain the patient's condition after tumor regression, or even chemopreventive therapy (e.g., for patients at risk).

[0246] These additional agents may be administered separately from the composition containing the compounds of the present invention as part of a multiple-dose regimen. Alternatively, the agents may be mixed with the compounds of the present disclosure in a single composition as part of a single dosage form. If administered as part of a multiple-dose regimen, the two active agents may be provided simultaneously, sequentially, or at intervals between each other (typically within five hours).

[0247] As used herein, the terms “combination,” “combined,” and related terms refer to the simultaneous or sequential administration of a therapeutic agent according to this disclosure. For example, a compound of this disclosure may be administered simultaneously or sequentially with another therapeutic agent in an individual unit dosage form or together in a single unit dosage form. Thus, this disclosure provides a single unit dosage form comprising a compound of this disclosure, an additional therapeutic agent, and a pharmaceutically acceptable carrier, adjuvant, or mediator.

[0248] The amounts of the compounds of the present invention that can be combined with carrier materials to produce a single dosage form and additional therapeutic agents (in those compositions comprising the additional therapeutic agents described above) will vary depending on the host being treated and the specific administration method. Preferably, the compositions of the present disclosure are formulated such that the compounds of the present invention can be administered at doses between 0.01 and 100 mg / kg body weight / day.

[0249] In compositions containing an additional therapeutic agent, the additional therapeutic agent may act synergistically with the compounds of this disclosure. Therefore, the amount of the additional therapeutic agent in such compositions will be less than that required in a monotherapy using only the therapeutic agent. In such compositions, the additional therapeutic agent can be administered at a dose between 0.01 and 1,000 micrograms per kilogram of body weight per day.

[0250] The amount of additional therapeutic agent present in the compositions disclosed herein will not exceed the amount typically applied in a composition comprising that therapeutic agent as the sole active agent. Preferably, the amount of additional therapeutic agent in the currently disclosed compositions will be in the range of about 50% to 100% of the amount typically present in a composition comprising said pharmaceutical agent as the sole active agent.

[0251] The compounds of this disclosure or pharmaceutical compositions thereof may also be incorporated into compositions for coating implantable medical devices, such as prostheses, artificial valves, vascular grafts, stents, and catheters. Intravascular stents have been used, for example, to overcome restenosis (restriction of the vessel wall after injury). However, patients using stents or other implantable devices are at risk of clot formation or platelet activation. These undesirable effects can be prevented or mitigated by pre-coating the device with a pharmaceutically acceptable composition containing a kinase inhibitor. Implantable devices coated with the compounds of this disclosure are another embodiment of this disclosure.

[0252] Any of the compounds and / or compositions disclosed herein may be provided in a kit comprising said compounds and / or compositions. Therefore, in some embodiments, the compounds and / or compositions disclosed herein are provided in a kit.

[0253] This disclosure is further described by the following non-limiting examples.

[0254] Example

[0255] This document provides examples to facilitate a more thorough understanding of this disclosure. The following examples are exemplary ways of making and practicing the subject matter of this disclosure. However, the scope of this disclosure should not be construed as limited to the specific embodiments disclosed in these examples, which are merely illustrative.

[0256] As described in the examples below, in some exemplary embodiments, the compounds are prepared according to the following general procedure. It should be understood that although the general method describes the synthesis of some compounds of this disclosure, the following general method and other methods known to those skilled in the art can also be applied to other classes and subclasses and species of each of these compounds described herein. Other compounds of this disclosure are prepared by methods substantially similar to those described in the examples herein and methods known to those skilled in the art.

[0257] In the following description of the synthetic methods, unless otherwise stated, it should be understood that all reaction conditions (e.g., reaction solvent, atmosphere, temperature, duration, and procedure) are selected from the standard conditions of the described reaction, unless otherwise indicated. The starting materials in the examples are commercially available or readily prepared from known materials by standard methods.

[0258] The compounds described herein can be prepared in a variety of ways based on the teachings contained herein and synthetic procedures known in the art. The following non-limiting examples illustrate the disclosure herein.

[0259] Instruments and methods

[0260] X-ray powder diffraction (XRPD)

[0261] Methods. Instrumentation: PANalytical Empyrean. XRPD diffraction patterns were collected on a PANalytical Empyrean diffractometer using Cu Kα radiation (45 kV, 40 mA) transmission geometry. A 0.5° slit, a 4 mm mask, and a 0.04 radian Soller slit, along with a focusing lens, were used for the incident beam. A PIXcel was placed on the diffraction beam. 3D The detector is equipped with a receiving slit and a 0.04 radian Soler slit. The software used for data collection is the X'Pert Data Collector with the X'Pert operator interface. HighScore Plus is used for data analysis and presentation. Samples are prepared and analyzed in transmission mode in either a metal or Millipore 96-well plate. An X-ray transparent membrane is used between metal sheets on the metal plate, and powder (approximately 1–2 mg) is used as is. With the Millipore plate, solids from the suspension are separated and analyzed by adding a small amount of suspension directly to the plate and then filtering under low vacuum. The scanning mode for the metal plate uses an angular scanning axis, while the Millipore plate utilizes a 2θ scan. The parameters for the standard screening data collection method are:

[0262] Angular range: 2.5 to 32.0° 2θ

[0263] Step size: 0.0130° 2θ

[0264] Collection time: 12.75 seconds / step (total collection time: 2.07 min)

[0265] Differential scanning calorimetry (DSC)

[0266] Method 1. Instrument: TA Instruments Discovery DSC. DSC data were collected on a TA Instruments Discovery DSC equipped with a 50-position autosampler. Typically, 0.5–3 mg samples in a pinhole aluminum dish were heated from 25°C to 300°C at 10°C / min. Dry nitrogen was maintained on the samples at a rate of 50 ml / min. The instrument control software was TRIOS, and the data were analyzed using TRIOS or Universal Analysis.

[0267] Method 2. Instrument: TA Instruments DSC2500. DSC data were collected on a TA Instruments DSC2500 equipped with a 54-bit autosampler. Typically, 0.5–3 mg samples in a pinhole aluminum pan were heated from 25°C to 300°C at 10°C / min. Dry nitrogen purging at 50 ml / min was maintained on the samples. Temperature-controlled DSC (MDSC) was performed using a basic heating rate of 2°C / min and temperature adjustment parameters of ±0.64°C per 60-second intervals. The instrument control software was TRIOS, and the data were analyzed using TRIOS or Universal Analysis.

[0268] Thermogravimetric analysis (TGA)

[0269] Instrument Discovery 5500 or Q5000

[0270] Sample tray, aluminum, open type

[0271] Initial temperature, environmental conditions (below 35℃)

[0272] Final temperature, 300°C, or if weight < 80% (w / w), then discontinue the next section.

[0273] (The weight loss of the compound shall not exceed 20% (w / w))

[0274] Heating rate, 10℃ / min

[0275] Nitrogen flow equilibration, 10 mL / min; sample chamber, 25 mL / min

[0276] Sample weight, approximately 2-10 mg

[0277] Dynamic gas phase adsorption (DVS)

[0278] Method 1

[0279] Instruments, Intrinsic, Advantage, or Adventure

[0280] Total airflow, 200 sccm

[0281] Oven temperature: 25℃

[0282] Solvent, water

[0283] Method loop: 40-0-95-0-40% RH

[0284] Phase step size: 10%

[0285] Equilibrium: 0.002 dm / dt (% / min)

[0286] Minimum dm / dt stability duration: 60 min

[0287] Maximum dm / dt stage time: 360 min

[0288] Method 2

[0289] Instruments, Intrinsic, Advantage, or Adventure

[0290] Total airflow, 200 sccm

[0291] Oven temperature: 25℃

[0292] Solvent, water

[0293] Method cycle: 40-95-0-95-40% RH

[0294] Phase step size: 10%

[0295] Equilibrium: 0.002 dm / dt (% / min)

[0296] Minimum dm / dt stability duration: 60 min

[0297] Maximum dm / dt stage time: 360min

[0298] Nuclear magnetic resonance (NMR)

[0299] Instrument: Bruker Avance-AV 400M (for 1H-NMR, 19F-NMR, and 31P-NMR)

[0300] Bruker Avance-III, 400M (for 13C-NMR)

[0301] Frequency, 400MHz

[0302] Probe, 5 mm, PABBO BB / 19F-1H / D Z-GRD Z108618 / 0406 (for 1H-NMR, 19F-NMR and 31P-NMR)

[0303] 5 mm PABBO BB-1H / D Z-GRD Z108618 / 0229 (for 13C NMR)

[0304] Number of scans, 8

[0305] Temperature, 297.6K

[0306] Relaxation delay, 1 second

[0307] High-performance liquid chromatography (HPLC)

[0308] Instruments: Agilent 1260, SHIMADZU CBM-40

[0309] Chiral purity wavelength: 220 nm

[0310] Column: Daicel OD-RH (4.6×150 mm×5 µm)

[0311] Detectors: DAD, PDA

[0312] Column temperature: 40℃

[0313] Flow rate: 1 mL / min

[0314] Mobile phase A: Water containing 10 mM NH4OAc

[0315] Mobile phase B: ACN

[0316] Diluent: ACN

[0317] Injection volume: 5 µL

[0318] Sample preparation: 2 mg / mL

[0319] Solution for washing needles: ACN:H2O = 90:10 (v / v)

[0320] Gradient: Isocratic elution

[0321]

[0322] FeSSIF (Feeding State Simulated Intestinal Fluid)

[0323] Acetate-Alkaline Buffer Solution (FeSSIF), pH 5.0: Dissolve sodium hydroxide (4.061 g), acetic acid (8.25 ml), and sodium chloride (11.88 g) in approximately 800 ml of deionized water in a 1000 ml volumetric flask. Mix this solution thoroughly and then adjust the pH to 5.0 ± 0.05 using 1 M hydrochloric acid / sodium hydroxide. Then bring the solution to volume with deionized water and record the pH (pH 5.00).

[0324] FeSSIF medium: Dissolve 0.56 g of Phares SIF (simulated intestinal fluid) powder in approximately 40 ml of alkaline buffer in a 50 ml volumetric flask. Mix this solution thoroughly and then bring it to volume with the same alkaline buffer. Record the final pH (pH 4.93).

[0325] Fasting intestinal fluid simulation (FaSSIF)

[0326] Phosphate-based alkaline buffer (FaSSIF), pH 6.5: Dissolve sodium hydroxide (0.403 g), sodium dihydrogen phosphate (3.95 g), and sodium chloride (6.50 g) in approximately 800 ml of deionized water in a 1000 ml volumetric flask. Mix this solution thoroughly and then adjust the pH to 6.5 ± 0.05 using 1 M hydrochloric acid / sodium hydroxide. Then bring the solution to volume with deionized water and record the pH (pH 6.47).

[0327] FaSSIF medium: Dissolve 0.112 g of Phares SIF (simulated intestinal fluid) powder in approximately 40 ml of alkaline buffer in a 50 ml volumetric flask. Mix this solution thoroughly and then bring it to volume with the same alkaline buffer. Record the final pH (pH 6.46).

[0328] Fasting gastric juice simulation (FaSSGF)

[0329] Alkaline buffer (FaSSGF), pH 1.6: Dissolve 1.99 g of sodium chloride in approximately 800 ml of deionized water in a 1000 ml volumetric flask. Mix this solution thoroughly, and then adjust the pH to 1.6 ± 0.05 with concentrated hydrochloric acid. Next, bring the solution to volume with deionized water and record the pH (pH 1.60).

[0330] FaSSGF medium: Dissolve 0.03 g of Phares SIF (simulated intestinal fluid) powder in approximately 40 ml of alkaline buffer in a 50 ml volumetric flask. Mix this solution thoroughly and then bring it to volume with the same alkaline buffer. Record the final pH (pH 1.63).

[0331]

[0332] Example 1

[0333] Preparation of synthetic intermediates

[0334] 1.1. Preparation of intermediate SM1

[0335]

[0336] Step 1. Preparation of methyl (1R,4S)-4-((tert-butoxycarbonyl)amino)cyclopent-2-ene-1-carboxylate (1)

[0337]

[0338] A solution of (1R,4S)-4-aminocyclopentan-2-ene-1-carboxylate methyl hydrochloride (5000.00 g, 28.149 mol, 1 eq) in DCM (30 L) was treated with triethylamine (2.848 kg, 28.149 mol, 1.0 eq) at 0 °C. The temperature was controlled at 0–5 °C. Di-tert-butyl dicarbonate (6.143 kg, 28.149 mol, 1.0 eq) was added dropwise to the resulting mixture at 0–5 °C, while maintaining the internal temperature of the mixture below 10 °C. The mixture was then heated to 22 °C and stirred for 16 hours.

[0339] The reaction mixture was diluted with water (8000 mL) and the organic layer was separated. The aqueous layer was extracted with DCM (5000.0 mL), and the DCM layer was washed with brine (5.0 L), dried over anhydrous MgSO4, filtered, and concentrated. Methyl (1R,4S)-4-((tert-butoxycarbonyl)amino)cyclopent-2-ene-1-carboxylate (6.20 kg, 91.3% yield) was obtained as a white solid.

[0340] Step 2. Preparation of methyl (3aR,5R,6R,6aR)-6-bromo-2-oxohexahydro-2H-cyclopentane[d]oxazol-5-carboxylate (2)

[0341]

[0342] A solution of (1R,4S)-4-((tert-butoxycarbonyl)amino)cyclopent-2-ene-1-carboxylate (5500.00 g, 22.795 mol, 1 eq) in THF (22 L) and H₂O (2.2 L) was cooled to 0 °C. Then, N-bromosuccinimide (4.463 kg, 25.074 mol, 1.1 eq) was added in portions. The mixture was heated to room temperature and stirred for 16 hours. The solution was concentrated to dryness. The residue was slurried in DCM (15 L) and washed with 1 M HCl (5 L), saturated Na₂SO₃ (6 L), and brine (6.0 L), dried over anhydrous MgSO₄, filtered, and concentrated. The residue was slurried in n-heptane (4 L) for 30 min, filtered and dried to obtain methyl (3aR,5R,6R,6aR)-6-bromo-2-oxohexahydro-2H-cyclopentane[d]oxazol-5-carboxylate (4.033 kg, 67.0%) as a white solid.

[0343] Step 3. Preparation of (3R,4S)-4-((tert-butoxycarbonyl)amino)-3-hydroxycyclopent-1-ene-1-carboxylic acid (3)

[0344]

[0345] A solution of methyl (3aR,5R,6R,6aR)-6-bromo-2-oxohexahydro-2H-cyclopentane[d]oxazol-5-carboxylate (2000.00 g, 7.574 mol, 1 eq) in MeOH (10 L) and H₂O (10 L) was cooled to 0 °C. KOH (1.700 kg, 30.295 mol, 4.0 eq) was then added in portions. The mixture was heated at 80 °C for 16 hours. The mixture was cooled to 20 °C and concentrated under vacuum. THF (4.0 L) containing di-tert-butyl dicarbonate (1.653 kg, 7.574 mol, 1 eq) was added dropwise to the residue. The mixture was heated to room temperature and stirred for 16 hours. The mixture was redissolved in EA (3 L) and H2O (3 L), and 1 M HCl was added dropwise to the mixture to adjust the pH to 3–4. Extraction was performed with EA (5 L × 3). The organic layer was separated, washed with brine (5 L), dried over anhydrous MgSO4, filtered, and concentrated to dryness. (3R,4S)-4-((tert-butoxycarbonyl)amino)-3-hydroxycyclopent-1-ene-1-carboxylic acid (1.48 kg, 80.0% yield) was obtained as a white solid.

[0346] Step 4. Preparation of methyl (3R,4S)-4-((tert-butoxycarbonyl)amino)-3-hydroxycyclopent-1-ene-1-carboxylate (4)

[0347]

[0348] A solution of (3R,4S)-4-((tert-butoxycarbonyl)amino)-3-hydroxycyclopent-1-ene-1-carboxylic acid (3,2900.00 g, 11.922 mol, 1 eq) in DMF (14.5 L) was cooled to 0 °C, followed by the addition of K₂CO₃ (2.471 kg, 17.882 mol, 1.5 eq) in portions. The mixture was heated to room temperature and stirred for 1 hour. Iodomethane (3.384 kg, 23.843 mol, 2.0 eq) was added dropwise to the resulting mixture at 0–5 °C, and the mixture was heated to room temperature and stirred for 16 hours. The mixture was quenched with H₂O (40.0 L) and subsequently extracted with EA (10 L × 2). The organic layer was separated, washed with brine (5 L), dried over anhydrous MgSO₄, filtered, and concentrated to dryness. The residue was slurried in (PE:EA=3:1 (2 L)) for 30 min, filtered and dried to obtain methyl (3R,4S)-4-((tert-butoxycarbonyl)amino)-3-hydroxycyclopent-1-ene-1-carboxylate (2.200 kg, 71.7% yield) as a white solid.

[0349] Step 5. Preparation of methyl (1S,3S,4R)-3-((tert-butoxycarbonyl)amino)-4-hydroxycyclopentane-1-carboxylate (SM1)

[0350]

[0351] Glucose (1800.00 g) was dissolved in 14.4 L of 100 mM potassium phosphate buffer at pH 7. Subsequently, 800.00 g of wet whole cells of *E. coli* BL21(DE3) overexpressing Ene reductase 10 (PE10) were added to the potassium phosphate buffer and allowed to fully suspend to obtain a homogeneous suspension. Next, 8.00 g of glucose dehydrogenase powder and nicotinamide adenine dinucleotide phosphate were added to the potassium phosphate buffer. 800.00 g of DMSO containing the substrate ((3R,4S)-4-((tert-butoxycarbonyl)amino)-3-hydroxycyclopent-1-ene-1-carboxylate) was added as a co-solvent. The mixture was stirred at 37 °C. Since gluconic acid was generated during the reaction, the pH was adjusted to 7.0 using 3 M K₂CO₃. The mixture was stirred for 16 hours.

[0352] The reaction mixture was filtered through diatomaceous earth. The filtrate was extracted with EA (3 L × 3), and the filter cake was slurried with 8 L EA. The combined organic layers were washed with brine (5 L), dried over anhydrous MgSO4, filtered, and concentrated to dryness. The residue was slurried in n-heptane (1 L) for 30 min, filtered, and dried to give methyl (1S,3S,4R)-3-((tert-butoxycarbonyl)amino)-4-hydroxycyclopentane-1-carboxylate (0.685 kg, 85.0% yield) as a white solid. 1 ¹H NMR (400MHz, CDCl₃) δ 4.81 (s, ¹H), 4.28 (s, ¹H), 3.67 (s, ³H), 3.08–3.15 (m, ¹H), 2.07–2.27 (m, ¹H), 2.02–2.05 (m, ²H), 1.84–1.86 (m, ²H), 1.44 (s, ⁹H). HPLC purity = 98.80%. ee% = 100%.

[0353] 1.2. Preparation of intermediate A1

[0354]

[0355] Step 1. Preparation of ethyl 4-methylenecyclohexane-1-carboxylate

[0356] Under N2, THF (72.0 L, 8.0 V) containing Ph3MeBr (28.35 kg, 1.5 eq) was added to the reaction vessel. At approximately 0 °C, t-BuOK (8.91 kg, 1.5 eq) was added to the vessel. THF (18.0 L, 2.0 V) containing ethyl 4-oxocyclohexane-1-carboxylate (9.00 kg, 1.00 eq) was added dropwise to the mixture, and the mixture was stirred at approximately 0 °C for 10 min. Subsequently, a 30% aqueous citric acid solution (1 V) was added to the reaction vessel to adjust the pH to approximately 4.5. The mixture was stirred at 20–25 °C for 30 min. The aqueous phase was extracted with heptane (2 × 4 V), and the organic phases were combined, concentrated to approximately 1 V, and loaded into the vessel with heptane (2 V), and stirred for 30 min. The mixture was filtered, dried over MgSO4 and concentrated to give ethyl 4-methylenecyclohexane-1-carboxylate (6.93 kg, 78% yield), which was a pale yellow liquid.

[0357] Step 2. Preparation of ethyl 4-(bromomethyl)-4-fluorocyclohexane-1-carboxylate

[0358] DCM (93.3 L, 10 V) and ethyl 4-methylenecyclohexane-1-carboxylate (9.33 kg, 1.0 eq) were charged into a reaction vessel. Triethylamine trifluoride (22.39 kg, 2.5 eq) was added to the mixture over 10 min at approximately 0 °C. N-bromosuccinimide (14.74 kg, 1.5 eq) was then added to the mixture, and the mixture was stirred at 20–25 °C for 10 min. Distilled water (140.0 kg, 15.0 V) was then added, and the mixture was stirred for 30 min. The mixture was then washed with distilled water (93.0 kg, 10.0 V), and the organic layers were combined, dried over MgSO4, filtered, and concentrated to obtain crude ethyl 4-(bromomethyl)-4-fluorocyclohexane-1-carboxylate (10.81 kg crude product) as a brown liquid.

[0359] Step 3. Preparation of ethyl 4-fluorobicyclo[2.2.1]heptane-1-carboxylate

[0360] THF (20 L, 10 V), MTBE (6.0 L, 3 V), and crude ethyl 4-(bromomethyl)-4-fluorocyclohexane-1-carboxylate (2.0 kg, 1.0 eq) were charged into a reaction vessel. Potassium bis(trimethylsilyl)amidide (13.1 L, 1.75 eq) was added to the solution over 60 min at 30–40 °C. Subsequently, a 10% aqueous solution of ammonium chloride (5 V) was added dropwise to the mixture, and it was stirred at approximately 0 °C for 30 min. The aqueous phase was extracted twice with MTBE (4 V), and the combined organic layers were concentrated into a crude pale yellow liquid. Under N2 conditions, N-(2-hydroxyethyl)-2-pyrrolidone (5.0 V), activated carbon (10%), and the crude pale yellow concentrate were added to a separate vessel. The mixture was stirred, filtered, and concentrated to give ethyl 4-fluorobicyclo[2.2.1]heptane-1-carboxylate (2.51 kg, 58.7% yield) as a brown liquid.

[0361] Step 4. Preparation of 4-fluoronorbornene-1-carboxaldehyde

[0362] Preparation of SDBBA reducing agent: DIBAL-H (2.115 kg, 14.98 mol, 9.99 L, 1.860 eq) was added dropwise to a solution of t-BuONa (1.500 kg, 15.608 mol, 1.938 eq) in THF (20.0 L) at -10 to 0 °C. The reaction mixture was stirred at 25 °C for 2 hours to obtain the reducing agent SDBBA.

[0363] Freshly prepared SDBBA was added to a solution of ethyl 4-fluorobicyclo[2.2.1]heptane-1-carboxylate (1500.0 g, 8.055 mol, 1.0 eq) in THF (20.0 L) at -10 to 0 °C, and the mixture was stirred at -10 to 0 °C for 16 hours. The reactants were then quenched dropwise by adding 1 N HCl (30.0 L) and stirred for 2 hours. The mixture was extracted with MTBE (10.0 L × 3), the combined organic layers were dried over MgSO4, filtered, and concentrated at 10–20 °C to obtain the residue. The residue was redissolved in MTBE (1.0 L), a saturated solution of NaHSO3 (6 L) was added, and the mixture was stirred at 25 °C for 16 hours. The mixture was extracted with MTBE (5.0 L × 4), and the aqueous layer was collected. The pH of the aqueous layer was adjusted to pH 9–10 by adding saturated Na2CO3 (10 L), and the mixture was stirred at 25 °C for another 16 hours. The mixture was extracted with MTBE (5.0 L × 5), the organic phase was combined and dried with MgSO4, filtered and concentrated at 15-20 °C to obtain 4-fluorobicyclo[2.2.1]heptane-1-carboxaldehyde (640.6 g, 74.488% yield) as a colorless oil.

[0364] Step 5. Preparation of (R,Z)-N-[(4-fluoronorbornen-1-yl)methylene]-2-methyl-propane-2-sulfinamide (A1)

[0365] At 25°C, (R)-2-methylpropane-2-sulfinamide (545.60 g, 4.502 mol, 1.0 eq) and tetraisopropyl titanate (TIPT, 2.561 kg, 9.003 mol, 2.0 eq) were added to a solution of 4-fluorobicyclo[2.2.1]heptane-1-carboxaldehyde (640.0 g, 4.502 mol, 1.0 eq) in THF (15.0 L). The reaction mixture was stirred at 70–75°C for 3 hours. The reaction mixture was allowed to cool naturally to 25°C and diluted with EA (10.0 L), then poured into water (10.0 L) at 10–20°C. The mixture was extracted with EA (10.0 L × 3), the combined organic layers were washed with saturated NaCl solution (4.5 L), dried over MgSO4, filtered, and concentrated at 35–45°C to obtain the residue. The residue was ground with (n-heptane:EA=10:1) (1.5L) at 0-10℃ for 2 hours, and then filtered to obtain (R,Z)-N-[(4-fluoronorbornen-1-yl)methylene]-2-methyl-propane-2-sulfinamide (521.02 g, 47.172%) as a white solid.

[0366] Example 2

[0367] Synthesis of Compound I

[0368]

[0369]

[0370] Step 1. Preparation of (1S,3S,4R)-3-((tert-butoxycarbonyl)amino)-4-hydroxycyclopentane-1-carboxylic acid (S1)

[0371] At 0–5 °C, 1 M LiOH aqueous solution (3.5 L, 5 V) was added to a solution of (1R,3R,4S)-3-((tert-butoxycarbonyl)amino)-4-hydroxycyclopentane-1-carboxylate (700 g, 1.00 eq) in THF (1.4 L, 2 V) and MeOH (1.4 L, 2 V), and the mixture was stirred at 0–5 °C for 1 h. The reaction was monitored by HPLC and LCMS. At 0–5 °C, the reaction mixture was neutralized to pH 6–7 with 1 N HCl. The reaction mixture was concentrated below 40 °C to remove the organic solvent. At 20–25 °C, the solution was acidified to pH 2–3 with 1 N HCl and extracted with 2-MeTHF (10 V × 3). The combined organic layers were washed with brine (10 V) and dried over sodium sulfate. After filtration and concentration at 40°C, the product was dried under vacuum at 50°C to give (1S,3S,4R)-3-((tert-butoxycarbonyl)amino)-4-hydroxycyclopentane-1-carboxylic acid (S1) as a white solid (191 g, 92.9% yield).

[0372] Step 2. Preparation of (R)-N-((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)-2-methylpropane-2-sulfinamide

[0373] In a nitrogen atmosphere at -65°C, n-BuLi (2.50 M, 1585 mL, 1.30 eq) was added to a solution of 1-chloro-2,4-difluorobenzene (590.5 g, 1.30 eq) in THF (5.25 L, 7 V). The mixture was stirred at -65°C for 0.5 h, followed by dropwise addition of (R,Z)-N-((4-fluorobicyclo[2.2.1]hept-1-yl)methylene)-2-methylpropane-2-sulfinamide (750 g, 1.0 eq) in THF (3.75 L, 5 V) at -65°C. The mixture was stirred at -65°C for 2 h. The reaction was monitored by HPLC and LCMS. The reaction mixture was added to a saturated NH4Cl solution (11.25 L, 15 V) at -10-0°C. The aqueous phase was extracted with ethyl acetate (10 V × 2). The combined organic phases were washed with saturated brine (15 V × 2), dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum to give a crude product. The crude product was dissolved in ethanol (750 mL, 1 V). Water (3.75, 5 V) was added, and the mixture was stirred for 2 h. The resulting solid was filtered. The solid was slurried with heptane / EA (10:1, 5 V) for 5 h, filtered, and dried in air at 50 °C to give a pure (R)-N-((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)-2-methylpropane-2-sulfinamide (995 g, 79% yield) as a white solid.

[0374] Step 3. Preparation of (S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methylamine

[0375] At 25 °C, HCl / MeOH (3980 mL, 4 V) was added to a solution of (R)-N-((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)-2-methylpropane-2-sulfinamide (995 g, 1.00 eq) in MeOH (2985 mL, 3 V), and the mixture was stirred at 25 °C for 1 h. The reaction was monitored by HPLC and LCMS. The reaction mixture was concentrated under vacuum to give a crude product. The residue was ground with ethyl acetate / n-heptane (1:10, 5 V) at 25 °C for 2 h, filtered, and the filter cake was dried under vacuum at 50 °C. The compound (S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methylamine (738 g, 90.5% yield) was given as a white solid.

[0376] Step 4. Preparation of ((1S,2R,4S)-4-(((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)carbamoyl)-2-hydroxycyclopentyl)tert-butyl carbamate

[0377] At 0–5 °C, DIEA (4.0 eq) and HOBt (1.3 eq) were added to a solution of (1S,3S,4R)-3-((tert-butoxycarbonyl)amino)-4-hydroxycyclopentane-1-carboxylic acid (490 g, 1.0 eq) in DCM (7 V), followed by the addition of (S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methylamine (619 g, 0.95 eq) under N2 atmosphere. EDCI (497.8 g, 1.3 eq) was added to the mixture. The resulting reaction mixture was stirred at 0–5 °C under N2 for 24 h and monitored by HPLC and LCMS. The reaction mixture was washed with semi-saturated K2CO3 (15 V × 2), 1 N HCl (15 V × 2), and then with brine (15 V). The organic phase was separated and concentrated under reduced pressure at 40 °C. The crude product was crystallized using EtOAc / n-heptane (1.5 V: 6 V). The solid was filtered and the filter cake was dried under vacuum at 50 °C to give tert-butyl ((1S,2R,4S)-4-(((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)carbamoyl)-2-hydroxycyclopentyl)carbamate (852 g, 83% yield) as a white solid.

[0378] Step 5. Preparation of (1S,3S,4R)-3-amino-N-((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)-4-hydroxycyclopentane-1-carboxamide hydrochloride

[0379] At 25°C, concentrated HCl (425.5 mL, 3 eq) was added to a solution of ((1S,2R,4S)-4-(((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)carbamoyl)-2-hydroxycyclopentyl)carbamate tert-butyl (880 g, 1.00 eq) in DCM (2640 mL, 3 V), and the mixture was stirred at 25°C for 3 hours. The reaction was monitored by HPLC and LCMS. The reaction mixture was concentrated under vacuum to give a semi-oily crude product (770 g net weight of crude material), which was used in the next step without further purification.

[0380] Step 6. Preparation of N-((1S,2R,4S)-4-(((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)carbamoyl)-2-hydroxycyclopentyl)pyrimidine-5-carboxamide (Compound I)

[0381] Under a nitrogen atmosphere, N-methylmorpholine (542 g, 3.0 eq) was added to a solution of pyrimidine-5-carboxylic acid (232.8 g, 1.05 eq) in DCM (5670 mL, 7 V). This was followed by the addition of DCM (2430 mL, 3 V) containing (1S,3S,4R)-3-amino-N-((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)-4-hydroxycyclopentane-1-carboxamide hydrochloride (810 g net weight, 1.0 eq) at 0–5 °C. Then, HATU (747.4 g, 1.1 eq) was finally added to the mixture under nitrogen atmosphere at 0–5 °C. The resulting reaction mixture was stirred under nitrogen atmosphere at 0–5 °C for 2 h and monitored by HPLC and LCMS. N-methylmorpholine (361 g, 2.0 eq) was added to the reaction mixture at 0–5 °C, followed by washing with saturated Na₂CO₃ (12.15 L, 15 V). The organic phase was concentrated under reduced pressure at 40 °C and dissolved in EA (1.0 L, 10 V). The resulting solution was washed with 0.5 N HCl (15 V × 3) followed by brine (15 V). The organic phase was separated and concentrated under reduced pressure at 40 °C to give the crude product. It was purified by silica gel column chromatography (eluting with DCM containing 0-10% MeOH) and dried under vacuum at 40 °C to give N-((1S,2R,4S)-4-(((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)carbamoyl)-2-hydroxycyclopentyl)pyrimidine-5-carboxamide (compound I) (733 g, 82% yield) as a grayish-white solid.

[0382] Example 3

[0383] Synthesis of Compound II

[0384]

[0385] Step 1. Synthesis of (1S,3S,4R)-3-acetamido-N-((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)-4-hydroxycyclopentane-1-carboxamide

[0386] To a round-bottom flask, add (1S,3S,4R)-3-amino-N-((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)-4-hydroxycyclopentane-1-carboxamide (1.45 g, 3.48 mmol), acetic acid (209 mg, 3.48 mmol), NaHCO3 (1.46 g, 17.4 mmol), HATU (2.64 g, 6.96 mmol), and a stir bar. Add DMF (15 mL) and stir the solution at room temperature for 1 hour. The crude material was purified by preparative HPLC (column: LuxCellulose-34.6×100 mm, 3 µm; mobile phase A: water, mobile phase B: MeOH (MeOH containing 0.5% 2 M NH3); flow rate: 4 mL / min; gradient: 20% B isocratic) to obtain (1S,3S,4R)-3-acetamido-N-((S)-(3-chloro-2,6-difluorophenyl)(4-fluorobicyclo[2.2.1]hept-1-yl)methyl)-4-hydroxycyclopentane-1-carboxamide (1.34 g, 2.93 mmol), which was a grayish-white amorphous solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.22 (d, J = 8.2 Hz, 1H), 7.61-7.48(m, 2H), 7.19-7.09 (m, 1H), 5.26 (d, J = 8.1 Hz, 1H), 4.78 (d, J = 3.3 Hz,1H), 3.96-3.85 (m, 2H), 3.15-3.03 (m, 1H), 1.82 (d, J = 5.0 Hz, 2H), 1.81 (s,3H), 1.75 (ddd, J = 21.1, 11.5, 8.1 Hz, 8H), 1.59 (d, J = 8.8 Hz, 2H), 1.45(d, J = 9.6 Hz, 2H). [M+H] + 459.05.

[0387] Example 4

[0388] Preparation and characterization of crystal form

[0389] Compound I form A

[0390] A sample (30 mg) of compound I was weighed into an HPLC vial and 5 volumes of ethyl acetate were added. The sample was aged in an aging chamber at room temperature and 50°C for 24 hours (approximately 4 hours for each 1°C increase). Subsequently, 500 µl of heptane was added to the vial, and the sample was aged for another 24 hours. Under vacuum, the suspension was filtered through an XRPD Millipore plate for 10 minutes to obtain a crystalline solid. The observed XRPD chromatogram was designated as crystalline form A of compound I (see [reference]). Figure 1 ).

[0391] Form A appears to be substantially anhydrous, as only trace amounts of ethyl acetate (0.09 eq) were detected in the NMR spectrum, and the total weight loss observed from TGA analysis also totaled 0.09 eq of ethyl acetate. A broad endothermic peak was observed at the beginning of the thermal trace (solvent loss), followed by a melting endothermic peak at 174 °C. After storage at 40 °C and 75% RH, the sample remained in form A as monitored by X-ray powder diffraction.

[0392] Table 2. Characteristic description of compound I, form A

[0393]

[0394] Compound I form B

[0395] Weigh 30 mg of compound I into an HPLC vial and add 10 volumes of isopropyl acetate. Cherish the sample in an aging chamber at 24 hours between room temperature and 50°C (approximately 4 hours for each 1°C increase). Cherish the sample again for 24 hours. Filter the suspension through an XRPD Millipore plate under vacuum for 10 minutes to obtain a crystalline solid. Designate the observed XRPD chromatogram as crystalline form B of compound I (see [link to XRPD analysis]). Figure 2 ).

[0396] Characterization of form B indicates that, based on the small amount of isopropyl acetate (0.26 eq) present in the sample, it can also be in an anhydrous form. The DSC trace of form B shows a broad endothermic peak at the beginning of the trace (possibly due to solvent loss), followed by another endothermic peak at 141 °C, and finally a melting endothermic peak at 172 °C. After accelerated storage conditions at 40 °C and 75% RH, conversion of form B to form A was also observed.

[0397] Table 3. Characteristic description of compound I, form B

[0398]

[0399] Compound I form C

[0400] A sample (30 mg) of compound I was weighed into an HPLC vial and 5 volumes of ethanol were added. The sample was aged in an aging chamber at room temperature and 50°C for 24 hours (approximately 4 hours for each 1°C increase). Subsequently, 500 µl of heptane was added to the sample, and aging was continued for another 24 hours. Under vacuum, the suspension was filtered through an XRPD Millipore plate for 10 minutes to obtain a crystalline solid. The observed XRPD chromatogram was designated as crystalline form C of compound I (see [reference]). Figure 3 ).

[0401] Analysis of form C indicated it to be an ethanol solvate. NMR analysis showed approximately 0.82 eq of ethanol in the sample, and TGA analysis also showed a weight loss of approximately 0.9 eq of ethanol. The sample transformed into an amorphous material after storage under accelerated storage conditions.

[0402] Table 4. Characteristic description of compound I, form C

[0403]

[0404] Compound II form A

[0405] Weigh 30 mg of compound II into an HPLC vial and add 20 times its volume of water. Cherish the sample in an aging chamber at 24 hours between room temperature and 50°C (approximately 4 hours for each 1°C increase). Cherish the sample again for 24 hours. Filter the suspension through an XRPD Millipore plate under vacuum for 10 minutes to obtain a crystalline solid. Designate the observed XRPD chromatogram as crystalline form A of compound II (see [link to XRPD analysis]). Figure 4 ).

[0406] Form A TGA analysis showed a similar weight loss of water (1.79 eq water) to that seen in KF (1.31 eq). The difference in water volume between the TGA and KF analyses is likely due to surface-bound water evaporating prior to the KF analysis. DSC showed a broad endothermic peak starting at 46.8 °C, likely due to water loss from the sample, followed by several endothermic events and an exothermic peak at 129.6 °C, with the melting peak likely at 225.1 °C.

[0407] GVS analysis showed a reversible weight loss of 1.8% w / w (0.47 eq water) between 20% and 90% RH, followed by a significant loss / adsorption of 4.46% w / w (1.19 eq water) between 20% and 0% RH, indicating that this is a hydrated form. The sample retained form A after GVS and static storage conditions, and XRPD measurements showed minimal change in sample purity. Upon heating to 160°C at 10°C / min and cooling to room temperature, the sample converted to form B.

[0408] Table 5. Characteristic description of compound form II A

[0409]

[0410] Compound II form B

[0411] Weigh 30 mg of compound II into an HPLC vial and add 20 volumes of ethyl acetate. Aging the sample in an aging chamber between room temperature and 50°C for 24 hours (approximately 4 hours for each 1°C increase). Aging the sample again for 24 hours. Under vacuum, filter the suspension through an XRPD Millipore plate for 10 minutes to obtain a crystalline solid. The observed XRPD chromatogram is designated as crystalline form B of compound II (see [link to XRPD analysis]). Figure 5 ).

[0412] Based on the trace amount of residual organic solvent (ethyl acetate, 0.02 eq) observed in the NMR trace and the lack of weight loss observed in the TGA thermal trace, form B appears to be anhydrous. The only endothermic peak observed for form B is the melting endothermic peak at an initial temperature of 229 °C. The sample then begins to degrade above 250 °C. After static storage at 40 °C and 75% RH, the sample remains in form B.

[0413] Table 6. Characteristic description of compound II form B

[0414]

[0415] Example 5

[0416] Solubility assessment of crystalline form in media

[0417] Compound I form A

[0418] Form A maintained high solubility, especially at 2.3 mg / ml in FeSSIF medium. Solubility in FaSSIF was 0.51 mg / ml (slightly less than the amorphous form), and in FaSSGF it was 0.34 mg / ml, and in deionized water it was 0.45 mg / ml. XRPD analysis of the residues showed that the sample remained in form A.

[0419]

[0420] Compound I form B

[0421] Solubility assessment of form B showed extremely high solubility in the selected medium. However, XRPD analysis of the residue following solubility analysis confirmed that the crystal structure of form B collapsed, leaving mostly amorphous material. This indicates that the high solubility of the sample is attributable to the transformation of the sample into an amorphous material and is unrelated to the actual solubility of form B.

[0422]

[0423]

[0424] Compound II form A

[0425] Solubility assessment of form A showed a maximum solubility of 0.3 mg / ml in FeSSIF, and approximately 0.1 mg / ml in the three other media. XRPD analysis of the residues following the solubility assessment confirmed that the sample remained in form A.

[0426]

[0427] Compound II form B

[0428] Form B has a slightly lower solubility than form A (but is mostly equivalent), and the residue collected after solubility analysis showed that the solid was still form B.

[0429]

[0430] Example 6

[0431] Competitive slurry experiment

[0432] Compound I (form A and form B)

[0433] Compound I form A (100 mg) and compound I form B (100 mg) were weighed into 4 ml vials and mixed for 2 hours. The samples were recovered and analyzed by XRPD for reference. Saturated solutions were prepared simultaneously by pipetting 500 µl of the selected solvent system into HPLC vials and stirring at 5°C, 25°C, and 50°C (see table below). Compound I was added to each sample vial until a suspension was formed. After 1 hour, all samples were observed to still be in suspension, and the mixture was stirred for another 24 hours.

[0434] All samples remained suspensions. Each suspension was pipetted into a syringe and filtered through a nylon filter cartridge, with the solution collected in a new vial. 15 mg of reference material (1:1 form A / form B) was added to each solution, and the vials were stirred at their respective temperatures for 24 hours. Observations in the vials were recorded, and the first aliquot was pipetted into a metal XRPD plate and analyzed by XRPD. The samples were stirred for another 5 days, after which the second aliquot was obtained and analyzed by XRPD.

[0435] Competitive slurries of compound I after 24 hours showed that all samples remained suspensions after slurrying at 50°C and 5°C. XRPD analysis of these samples showed that some samples were completely converted to form A, while others remained a mixture of forms A and B.

[0436] Five days later (out of a total of six days), all samples remained suspensions, and XRPD analysis of these samples showed that more samples had completely converted to pure form A. The samples still in mixed form were predominantly in heptane, but two of the three samples had largely formed form A, accompanied by some smaller peaks associated with form B. This may be attributed to the lower solubility in heptane, which slowed the conversion to form A. This suggests that given sufficient time, complete conversion to form A is possible.

[0437]

[0438]

[0439] Compound II (Form A and Form B)

[0440] Compound II form A (100 mg) and compound II form B (100 mg) were weighed into 4 ml vials and mixed for 2 hours. The samples were recovered and analyzed by XRPD for reference. Saturated solutions were prepared simultaneously by pipetting 500 µl of the selected solvent system into HPLC vials and stirring at 5 °C, 25 °C, and 50 °C (see table below). Compound II was added to each sample vial until a suspension was formed. After 1 hour, all samples were observed to still be in suspension, and the mixture was stirred for another 24 hours.

[0441] All samples remained suspensions. Each suspension was pipetted into a syringe and filtered through a nylon filter cartridge, with the solution collected in a new vial. 15 mg of reference material (1:1 form A / form B) was added to each solution, and the vials were stirred at their respective temperatures for 24 hours. Observations in the vials were recorded, and the first aliquot was pipetted into a metal XRPD plate and analyzed by XRPD. The samples were stirred for another 5 days, after which the second aliquot was obtained and analyzed by XRPD.

[0442] All samples remained in suspension, and XRPD analysis of these samples showed complete conversion to form B after 24 hours. Five days later, the samples were still in suspension, and all samples remained in form B. This suggests that form B may be a more thermodynamically stable form compared to form A.

[0443]

[0444] References included

[0445] All disclosures and patents mentioned herein are incorporated herein by reference in their entirety for all purposes, just as individual disclosures or patents are specifically and individually incorporated by reference. In case of conflict, this application (including any definitions herein) shall prevail.

[0446] equivalent

[0447] Although specific embodiments of this disclosure have been discussed, the above description is illustrative and not restrictive. Many variations of this disclosure will become apparent to those skilled in the art upon review of this specification. The full scope of this disclosure, as well as the full scope of its equivalents, the specification, and such variations, should be determined with reference to the claims.

[0448] Unless otherwise indicated, all figures used in this specification and claims representing quantities of ingredients, reaction conditions, etc., should in all cases be understood to be modified by the term "about". Therefore, unless indicated to the contrary, the numerical parameters set forth in this specification and appended claims are approximate values ​​that may vary depending on the desired characteristics sought to be obtained by this disclosure.

Claims

1. A compound in solid form, wherein the compound is compound I: ， Or its solvates.

2. The compound according to claim 1, wherein the compound is amorphous.

3. The compound according to claim 1, wherein the compound is crystalline.

4. The compound according to claim 1 or 3, wherein the solid form is crystalline form A, form B or form C.

5. The compound according to any one of claims 1, 3 and 4, wherein the solid form is form A.

6. The compound of claim 5, wherein the solid form has an X-ray powder diffraction pattern comprising at least two peaks, each peak selected from the group consisting of about 17.4 2θ, about 13.3 2θ, and about 16.5 2θ.

7. The compound of claim 5, wherein the solid form has an X-ray powder diffraction pattern comprising at least two peaks, each peak selected from the group consisting of: about 17.4 2θ, about 13.3 2θ, about 16.5 2θ, about 24.2 2θ, about 19.8 2θ, about 14.5 2θ, and about 18.2 2θ.

8. The compound of claim 5, wherein the solid form has an X-ray powder diffraction pattern comprising at least three peaks, each peak selected from the group consisting of: about 17.4 2θ, about 13.3 2θ, about 16.5 2θ, about 24.2 2θ, about 19.8 2θ, about 14.5 2θ, and about 18.2 2θ.

9. The compound of claim 5, wherein the solid form has an X-ray powder diffraction pattern comprising at least four peaks, each peak selected from the group consisting of: about 17.4 2θ, about 13.3 2θ, about 16.5 2θ, about 24.2 2θ, about 19.8 2θ, about 14.5 2θ, and about 18.2 2θ.

10. The compound of claim 5, wherein the solid form has an X-ray powder diffraction pattern comprising at least five peaks, each peak selected from the group consisting of: about 17.4 2θ, about 13.3 2θ, about 16.5 2θ, about 24.2 2θ, about 19.8 2θ, about 14.5 2θ, and about 18.2 2θ.

11. The compound of claim 5, wherein the solid form has an X-ray powder diffraction pattern comprising at least six peaks, each peak selected from the group consisting of: about 17.4 2θ, about 13.3 2θ, about 16.5 2θ, about 24.2 2θ, about 19.8 2θ, about 14.5 2θ, and about 18.2 2θ.

12. The compound of claim 5, wherein the solid form has an X-ray powder diffraction pattern containing peaks at about 17.4 2θ, about 13.3 2θ, about 16.5 2θ, about 24.2 2θ, about 19.8 2θ, about 14.5 2θ and about 18.2 2θ.

13. The compound of claim 5, wherein the solid form has an X-ray powder diffraction pattern comprising at least three peaks, the at least three peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

1.

14. The compound of claim 5, wherein the solid form has an X-ray powder diffraction pattern comprising at least four peaks, the at least four peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

1.

15. The compound of claim 5, wherein the solid form has an X-ray powder diffraction pattern comprising at least five peaks, the at least five peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

1.

16. The compound of claim 5, wherein the solid form has an X-ray powder diffraction pattern comprising at least six peaks, the at least six peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

1.

17. The compound of claim 5, wherein the solid form has an X-ray powder diffraction pattern comprising at least seven peaks, the at least seven peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

1.

18. The compound according to claim 5, wherein the solid form has an X-ray powder diffraction pattern substantially similar to that depicted in Figure 1.

19. The compound according to any one of claims 1, 3 and 4, wherein the solid form is form B.

20. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern comprising at least two peaks, each selected from the group consisting of about 18.5 2θ, about 11.4 2θ, and about 17.5 2θ.

21. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern comprising at least two peaks, each peak being selected from the group consisting of: about 18.5 2θ, about 11.4 2θ, about 17.5 2θ, about 5.7 2θ, about 13.6 2θ, about 17.3 2θ, and about 10.7 2θ.

22. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern comprising at least three peaks, each peak selected from the group consisting of: about 18.5 2θ, about 11.4 2θ, about 17.5 2θ, about 5.7 2θ, about 13.6 2θ, about 17.3 2θ, and about 10.7 2θ.

23. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern comprising at least four peaks, each peak selected from the group consisting of: about 18.5 2θ, about 11.4 2θ, about 17.5 2θ, about 5.7 2θ, about 13.6 2θ, about 17.3 2θ, and about 10.7 2θ.

24. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern comprising at least five peaks, each peak selected from the group consisting of: about 18.5 2θ, about 11.4 2θ, about 17.5 2θ, about 5.7 2θ, about 13.6 2θ, about 17.3 2θ, and about 10.7 2θ.

25. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern comprising at least six peaks, each peak selected from the group consisting of: about 18.5 2θ, about 11.4 2θ, about 17.5 2θ, about 5.7 2θ, about 13.6 2θ, about 17.3 2θ, and about 10.7 2θ.

26. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern containing peaks at about 18.5 2θ, about 11.4 2θ, about 17.5 2θ, about 5.7 2θ, about 13.6 2θ, about 17.3 2θ and about 10.7 2θ.

27. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern comprising at least three peaks, the at least three peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

2.

28. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern comprising at least four peaks, the at least four peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

2.

29. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern comprising at least five peaks, the at least five peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

2.

30. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern comprising at least six peaks, the at least six peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

2.

31. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern comprising at least seven peaks, the at least seven peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

2.

32. The compound of claim 19, wherein the solid form has an X-ray powder diffraction pattern substantially similar to that depicted in Figure 2.

33. The compound according to any one of claims 1, 3 and 4, wherein the solid form is form C.

34. The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern comprising at least two peaks, each peak being selected from the group consisting of about 19.3 2θ, about 19.9 2θ, and about 20.0 2θ.

35. The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern comprising at least two peaks, each peak selected from the group consisting of: about 19.3 2θ, about 19.9 2θ, about 20.0 2θ, about 10.1 2θ, about 19.6 2θ, about 15.2 2θ, and about 22.0 2θ.

36. The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern comprising at least three peaks, each peak selected from the group consisting of: about 19.3 2θ, about 19.9 2θ, about 20.0 2θ, about 10.1 2θ, about 19.6 2θ, about 15.2 2θ, and about 22.0 2θ.

37. The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern comprising at least four peaks, each peak selected from the group consisting of: about 19.3 2θ, about 19.9 2θ, about 20.0 2θ, about 10.1 2θ, about 19.6 2θ, about 15.2 2θ, and about 22.0 2θ.

38. The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern comprising at least five peaks, each peak selected from the group consisting of: about 19.3 2θ, about 19.9 2θ, about 20.0 2θ, about 10.1 2θ, about 19.6 2θ, about 15.2 2θ, and about 22.0 2θ.

39. The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern comprising at least six peaks, each peak selected from the group consisting of: about 19.3 2θ, about 19.9 2θ, about 20.0 2θ, about 10.1 2θ, about 19.6 2θ, about 15.2 2θ, and about 22.0 2θ.

40. The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern containing peaks at about 19.3 2θ, about 19.9 2θ, about 20.0 2θ, about 10.1 2θ, about 19.6 2θ, about 15.2 2θ and about 22.0 2θ.

41. The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern comprising at least three peaks, the at least three peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

3.

42. The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern comprising at least four peaks, the at least four peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

3.

43. The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern comprising at least five peaks, the at least five peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

3.

44. The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern comprising at least six peaks, the at least six peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

3.

45. The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern comprising at least seven peaks, the at least seven peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

3.

46. ​​The compound of claim 33, wherein the solid form has an X-ray powder diffraction pattern substantially similar to that depicted in FIG3.

47. A compound in solid form, wherein the compound is compound II: ， Or its solvates.

48. The compound according to claim 47, wherein the compound is amorphous.

49. The compound according to claim 47, wherein the compound is crystalline.

50. The compound according to claim 47 or claim 49, wherein the solid form is form A or form B.

51. The compound according to any one of claims 47, 49 and 50, wherein the solid form is form A.

52. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern comprising at least two peaks, each peak being selected from the group consisting of about 15.7 2θ, about 14.7 2θ, and about 20.0 2θ.

53. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern comprising at least two peaks, each peak being selected from the group consisting of: about 15.7 2θ, about 14.7 2θ, about 20.0 2θ, about 21.0 2θ, about 23.2 2θ, about 16.5 2θ, about 17.5 2θ, and about 13.3 2θ.

54. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern comprising at least three peaks, each peak selected from the group consisting of: about 15.7 2θ, about 14.7 2θ, about 20.0 2θ, about 21.0 2θ, about 23.2 2θ, about 16.5 2θ, about 17.5 2θ, and about 13.3 2θ.

55. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern comprising at least four peaks, each peak selected from the group consisting of: about 15.7 2θ, about 14.7 2θ, about 20.0 2θ, about 21.0 2θ, about 23.2 2θ, about 16.5 2θ, about 17.5 2θ, and about 13.3 2θ.

56. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern comprising at least five peaks, each peak selected from the group consisting of: about 15.7 2θ, about 14.7 2θ, about 20.0 2θ, about 21.0 2θ, about 23.2 2θ, about 16.5 2θ, about 17.5 2θ, and about 13.3 2θ.

57. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern comprising at least six peaks, each peak selected from the group consisting of: about 15.7 2θ, about 14.7 2θ, about 20.0 2θ, about 21.0 2θ, about 23.2 2θ, about 16.5 2θ, about 17.5 2θ, and about 13.3 2θ.

58. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern containing peaks at about 15.7 2θ, about 14.7 2θ, about 20.0 2θ, about 21.0 2θ, about 23.2 2θ, about 16.5 2θ, about 17.5 2θ and about 13.3 2θ.

59. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern comprising at least three peaks, the at least three peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

4.

60. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern comprising at least four peaks, the at least four peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

4.

61. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern comprising at least five peaks, the at least five peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

4.

62. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern comprising at least six peaks, the at least six peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

4.

63. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern comprising at least seven peaks, the at least seven peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

4.

64. The compound of claim 51, wherein the solid form has an X-ray powder diffraction pattern substantially similar to that depicted in FIG4.

65. The compound according to any one of claims 47, 49 and 50, wherein the solid form is form B.

66. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern comprising at least two peaks, each peak being selected from the group consisting of about 23.6 2θ, about 10.9 2θ, and about 16.9 2θ.

67. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern comprising at least two peaks, each peak being selected from the group consisting of: about 23.6 2θ, about 10.9 2θ, about 16.9 2θ, about 15.1 2θ, about 26.1 2θ, about 12.9 2θ, and about 19.4 2θ.

68. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern comprising at least three peaks, each peak selected from the group consisting of: about 23.6 2θ, about 10.9 2θ, about 16.9 2θ, about 15.1 2θ, about 26.1 2θ, about 12.9 2θ, and about 19.4 2θ.

69. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern comprising at least four peaks, each peak selected from the group consisting of: about 23.6 2θ, about 10.9 2θ, about 16.9 2θ, about 15.1 2θ, about 26.1 2θ, about 12.9 2θ, and about 19.4 2θ.

70. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern comprising at least five peaks, each peak selected from the group consisting of: about 23.6 2θ, about 10.9 2θ, about 16.9 2θ, about 15.1 2θ, about 26.1 2θ, about 12.9 2θ, and about 19.4 2θ.

71. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern comprising at least six peaks, each peak selected from the group consisting of: about 23.6 2θ, about 10.9 2θ, about 16.9 2θ, about 15.1 2θ, about 26.1 2θ, about 12.9 2θ, and about 19.4 2θ.

72. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern containing peaks at about 23.6 2θ, about 10.9 2θ, about 16.9 2θ, about 15.1 2θ, about 26.1 2θ, about 12.9 2θ, and about 19.4 2θ.

73. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern comprising at least three peaks, the at least three peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

5.

74. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern comprising at least four peaks, the at least four peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

5.

75. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern comprising at least five peaks, the at least five peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

5.

76. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern comprising at least six peaks, the at least six peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

5.

77. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern comprising at least seven peaks, the at least seven peaks being in units of ° 2θ and selected from the group of peaks listed in Table 1.

5.

78. The compound of claim 65, wherein the solid form has an X-ray powder diffraction pattern substantially similar to that depicted in FIG5.

79. A pharmaceutical composition comprising a compound according to any one of claims 1 to 78 and a pharmaceutically acceptable carrier.

80. A method for inhibiting PI3Kα activity in a subject in need, comprising: The subject is given a therapeutically effective amount of the compound according to any one of claims 1 to 78 or the pharmaceutical composition according to claim 79.

81. A method of treating cancer in a subject in need, comprising administering to the subject a therapeutically effective amount of a compound according to any one of claims 1 to 78 or a pharmaceutical composition according to claim 79.

82. The method according to claim 80 or 81, further comprising administering a therapeutically effective amount of an antibody, antibody-drug conjugate, kinase inhibitor, immunomodulator, or histone deacetase inhibitor.

83. A kit comprising the compound according to any one of claims 1 to 78.

84. The kit according to claim 83, further comprising a written instruction describing the preparation of a pharmaceutical composition suitable for administration to a patient from the solid form or the compound.

85. The kit according to claim 83 or 84, further comprising a written instruction manual describing how to administer the obtained composition to the patient.

86. The kit according to any one of claims 83 to 85, further comprising a pharmaceutically acceptable excipient.

87. A method for preparing a crystalline form of compound I, comprising: (a) preparing a solution of compound I; (b) adjusting the temperature to precipitate a solid crystalline form of compound I from the solution; and (c) separating the solid crystalline form.

88. A method for preparing a crystalline form of compound I, comprising: (a) preparing a solution of compound I in a solvent; (b) adjusting the temperature; (c) adding heptane to the solution; and (d) separating the solid crystalline form.

89. A method for preparing a crystalline form of compound II, comprising: (a) preparing a solution of compound II; (b) adjusting the temperature such that a solid crystalline form of compound II precipitates out of the solution; and (c) separating the solid crystalline form.

90. A method for preparing a crystalline form of compound II, comprising: (a) preparing a solution of compound II in a solvent; (b) adjusting the temperature; (c) adding heptane to the solution; and (d) separating the solid crystalline form.

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