Form and manufacturing method of G1T38

Abstract

The present invention provides an unexpectedly stable, highly crystalline form of the di-HCl salt of 2'-((5-(4-isopropylpiperazin-1-yl)pyridin-2-yl)amino)-7',8'-dihydro-6'H-spiro[cyclohexane-1,9'-pyrazino[1',2':1,5]pyrrolo[2,3-d]pyrimidin]-6'-one for suitable therapeutic pharmaceutical efficacy and dosage form stability.

Description

Technical Field

[0001] [Cross - reference to Related Applications] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 526,937, filed Jun. 29, 2017. The entire disclosure of this application is incorporated herein by reference in its entirety for all purposes.

[0002] The present invention provides a preferred isolated morphic form of the dihydrochloride salt of G1T38, which is (2’ - ((5 - (4 - isopropylpiperazin - 1 - yl)pyridin - 2 - yl)amino)-7’,8’ - dihydro - 6’H - spiro[cyclohexane - 1,9’ - pyrazino[1’,2’:1,5]pyrrolo[2,3 - d]pyrimidin] - 6’ - one).

Background Art

[0003] Patent Document 1, Patent Document 2, Patent Document 3, Patent Document 4 assigned to G1 Therapeutics, Inc. and corresponding Patent Document 5 describe N - (heteroaryl) - pyrrolo[3,2 - d]pyrimidine - 2 - amine cyclin - dependent kinase inhibitors of the type containing 2’ - ((5 - (4 - isopropylpiperazin - 1 - yl)pyridin - 2 - yl)amino)-7’,8’ - dihydro - 6’H - spiro[cyclohexane - 1,9’ - pyrazino[1’,2’:1,5]pyrrolo[2,3 - d]pyrimidin] - 6’ - one (Compound 1) having the formula:

Chemical Formula

[0004] This compound is currently referred to as "G1T38." The diHCl salt of G1T38 (compound 2) is currently undergoing a Phase Ib / 2a human clinical trial in the United States by the U.S. Food and Drug Administration for the treatment of estrogen-positive, HER2-negative breast cancer after failure of endocrine therapy. G1T38 has also been favorably evaluated in a Phase Ia toxicity study in 75 women and has been found to be well-tolerated without significant adverse events. [ka]

[0005] G1T38 inhibits cell proliferation in various CDK4 / 6-dependent oncogenic cell lines, including breast cancer, melanoma, leukemia, and lymphoma cells, and inhibits RB phosphorylation in vitro and in vivo. Further favorable therapeutic properties of G1T38, including tumor selectivity across the entire plasma in mouse xenograft tumors, have attracted attention in a recently published paper in a peer-reviewed journal (Non-Patent Document 1). See also Patent Document 6.

[0006] Other publications describing this common type of compound include: Patent Document 7, filed by Strum et al. and assigned to G1 Therapeutics, describes compounds and methods for protecting normal cells during chemotherapy using pyrimidine CDK4 / 6 inhibitors. Patent Document 8, filed by Strum et al. and assigned to G1 Therapeutics, describes compounds and methods for protecting hematopoietic stem cells and progenitor cells against ionizing radiation using pyrimidine CDK4 / 6 inhibitors. Patent Document 9, filed by Strum et al. and assigned to G1 Therapeutics, describes HSPC-preserving therapy for abnormal cell proliferation using pyrimidine CDK4 / 6 inhibitors. Patent Document 10, filed by Strum et al. and assigned to G1 Therapeutics, describes highly active antineoplastic and antiproliferative pyrimidine CDK4 / 6 inhibitors. Patent document 11, filed by Strum et al. and assigned to G1 Therapeutics, describes a tricyclic pyrimidine CDK inhibitor used for radiation protection. Patent document 12, filed by Strum et al. and assigned to G1 Therapeutics, describes a similar tricyclic pyrimidine CDK inhibitor for cell protection during chemotherapy. Patent document 13, filed by Strum et al. and assigned to G1 Therapeutics, describes a similar tricyclic pyrimidine CDK inhibitor used for HSPC-preserving treatment of RB-positive abnormal cell proliferation. Patent document 14, filed by Strum et al. and assigned to G1 Therapeutics, describes a similar tricyclic pyrimidine CDK inhibitor used as an antineoplastic and antiproliferative agent. Patent document 15, filed by Strum et al. and assigned to G1 Therapeutics, describes the combination of a pyrimidine CDK4 / 6 inhibitor with other antineoplastic agents. Patent document 16, filed by Strum et al. and assigned to G1 Therapeutics, describes compounds and methods for treating certain Rb-negative cancers with CDK4 / 6 inhibitors and topoisomerase inhibitors.

[0007] Other biologically active condensed spirolactams and their synthesis are described in the following publications, for example: Non-Patent Document 2 describes a spirolactam for acetyl-CoA carboxylase inhibition, which is metabolically stable and has the lactam on the condensed ring. Patent Document 17, filed by Bell et al., describes an aliphatic spirolactam with the lactam on the spiro ring as a CGRP receptor antagonist. Patent Document 18, filed by Bell et al., describes an aryl spirolactam with the lactam on the spiro ring as a CGRP receptor antagonist. Patent Document 19, filed by Bell et al., describes a constrained spirolactam compound with the lactam on the spiro ring as a CGRP receptor antagonist. Patent Document 20, filed by Bell et al., describes a carboxamide spirolactam compound with the spirolactam on the spiro ring as a CGRP receptor antagonist. Patent documents 21, 22, and 23 filed by Bell et al. describe anilide spirolactam compounds in which the spirolactam is located on a spiro ring. Patent document 24 filed by Bhunai et al. describes spirolactam compounds in which the lactam is located on a spiro ring to which it may be further condensed.

[0008] Considering the therapeutic significance of G1T38 in patients suffering from proliferative disorders such as tumors or cancer, it is beneficial to provide a suitable delivery method that can enhance therapeutic activity and / or stability. [Prior art documents] [Patent Documents]

[0009] [Patent Document 1] U.S. Patent No. 8,822,683 [Patent Document 2] U.S. Patent No. 8,598,197 [Patent Document 3] U.S. Patent No. 8,829,102 [Patent Document 4] U.S. Patent No. 9,102,683 [Patent Document 5] International Publication No. 2012 / 061156 [Patent Document 6] U.S. Patent No. 9,527,857 [Patent Document 7] International Publication No. 2014 / 144326 [Patent Document 8] International Publication No. 2014 / 144596 [Patent Document 9] International Publication No. 2014 / 144847 [Patent Document 10] International Publication No. 2014 / 144740 [Patent Document 11] International Publication No. 2015 / 161285 [Patent Document 12] International Publication No. 2015 / 161287 [Patent Document 13] International Publication No. 2015 / 161283 [Patent Document 14] International Publication No. 2015 / 161288 [Patent Document 15] International Publication No. 2016 / 040858 [Patent Document 16] International Publication No. 2016 / 040848 [Patent Document 17] International Publication No. 2013 / 169574 [Patent Document 18] International Publication No. 2007 / 061677 [Patent Document 19] International Publication No. 2008 / 073251 [Patent Document 20] International Publication No. 2006 / 031606 [Patent Document 21] International Publication No. 2006 / 031610 [Patent Document 22] International Publication No. 2006 / 031491 [Patent Document 23] International Publication No. 2006 / 029153 [Patent Document 24] International Publication No. 2008 / 109464 [Non-Patent Document]

[0010] [Non-Patent Document 1] Bisi, et al., Preclinical development of G1T38: A novel, potent and selective inhibitor of cyclin dependent kinases 4 / 6 for use as an oral antineoplastic in patients with CDK 4 / 6 sensitive tumors", Oncotarget, March 15, 2017 [Non-Patent Document 2] Griffith, D. A., et al. (2013). "Spirolactam-Based Acetyl-CoA Carboxylase Inhibitors: Toward Improved Metabolic Stability of a Chromanone Lead Structure." Journal of Medicinal Chemistry 56(17): 7110-7119 [Summary of the Invention]

[0011] It has been discovered that compound 2, which is the dihydrochloride salt of G1T38 (2'-((5-(4-isopropylpiperazin-1-yl)pyridin-2-yl)amino)-7',8'-dihydro-6'H-spiro[cyclohexane-1,9'-pyrazino[1',2':1,5]pyrrolo[2,3-d]pyrimidin]-6'-one), can be prepared in a highly purified and suitable form referred to herein as Form B.

[0012] Form B of compound 2 is an unexpectedly highly stable, highly crystalline form of solid compound 2, which is beneficial for therapeutic efficacy and for the manufacture of pharmaceutical formulations. As discussed in Example 4, form B is stable for 7 days under thermal stress at 60°C. Furthermore, long-term stability studies at 25°C and 60% relative humidity have revealed that isolated form B of compound 2 is stable for at least one year (Example 7). In one embodiment, isolated form B of compound 2 is stable for at least about 6, 7, 8, 9, 10, 11, 12, 14, 16, 18, 20, 22, or 24 months. [ka]

[0013] Numerous crystallization and slurry experiments were conducted by varying the temperature, cooling procedure, and isolation procedure (Example 2, Tables 1-4). These experiments revealed 11 distinct forms of compound 2, but only forms A, B, and D were suitable for evaluation. The other forms resulted in weakly crystalline forms, solvates, unstable hydrates, or anhydrous forms. Of the three solid forms, form B was found to be an unexpectedly superior highly crystalline and stable material for therapeutic drug formulations. In dynamic vapor adsorption experiments, compound 2 remained in form B after exposure to 90% relative humidity (Example 3).

[0014] Form B possesses desirable properties for use as an active pharmaceutical ingredient in solid dosage forms and can enhance efficacy in such formulations. In one embodiment, Form B is produced by recrystallization from HCl and acetone, as described in more detail below. In one embodiment, Form B features an XRPD pattern substantially similar to the pattern shown in Figure 7. In one embodiment, Form B features an XRPD pattern including at least three 2-theta values ​​selected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.7±0.2°, and 22.4±0.2°. In one embodiment, Form B features an XRPD pattern including at least a 2-theta value of 9.5±0.2°. In some embodiments, Form B of isolated compound 2 is characterized by the absence of at least one peak at 4.6±0.2° in the 2-theta. In some embodiments, the isolated compound 2, form B, is characterized by the absence of a peak at 5.0 ± 0.2° in the 2-theta spectrum. In one embodiment, isolated form B is characterized by a 7.5% weight loss at 31°C to 120°C in thermogravimetric infrared (TG-IR) analysis. In one embodiment, isolated form B is characterized by an onset of endothermic activity in differential scanning calorimetry (DSC) at approximately 105 ± 20°C, approximately 220 ± 20°C, and approximately 350 ± 20°C, for example, 105°C, 220°C, and 350°C, or at 92°C, 219°C, and 341°C.

[0015] Accordingly, the present invention generally provides an isolated B form of compound 2, a pharmaceutical composition containing such form, a method for inhibiting or reducing the activity of CDK4 or CDK6 in a host using the isolated form described herein, a method for treating a host having pRb-positive cancer such as estrogen receptor-positive (ER+) breast cancer, non-small cell lung cancer (NSCLC), or prostate cancer, and a method for producing such form.

[0016] Form B of compound 2 can be produced, for example, by recrystallizing compound 1 in concentrated HCl and acetone. In one embodiment, compound 1 is dissolved in concentrated HCl and heated. After this, acetone is added, and the product is isolated by cooling and filtration.

[0017] In one embodiment, form B of compound 2 is produced by recrystallization of form D of compound 2. In an alternative embodiment, form B of compound 2 is produced by repeated recrystallization. In one embodiment, pure form B of compound 2 is purified from impure form B of compound 2 using a water:acetone (1:2) (v / v) slurry, and then vacuum dried.

[0018] Form A of compound 2 has lower stability than form B. Form A was produced when MeOH, EtOH, and 1-BuOH were used as solvents in single-solvent crystallization, and form A was also produced in binary solvent crystallization using water and MeOH as the main solvents. Similarly, form A was produced by slurry experiments using n-heptane and c-hexane.

[0019] Form D of compound 2 has lower stability than form B. In one embodiment, form D is produced by stirring a slurry of compound 2 in acetonitrile at room temperature. In another embodiment, form D is produced by dissolving compound 1 in concentrated HCl and then heating it. Subsequently, the solution is cooled, and acetone is added only after crystallization has begun to complete the precipitation. The precipitate is then isolated by filtration. In an alternative embodiment, form D is produced by dissolving compound 1 in concentrated HCl and then heating it. Subsequently, the solution is cooled, and acetone is added only after crystallization has occurred, and all solid matter is recovered by filtration.

[0020] In alternative embodiments, combinations of two or more forms of compound 2 are provided, for example, form B and form D, form B and form A, or form A and form D. In alternative embodiments, isolated combinations of three forms are provided, for example, form A, form B, and form D.

[0021] In one embodiment, a pharmaceutical composition is provided comprising the B form of isolated compound 2 and a pharmaceutically acceptable excipient. In another embodiment, the pharmaceutical composition further comprises one or more further therapeutic agents, which are, for example, but not limited to, anti-estrogens, anti-androgens, anti-cancer agents, aromatase inhibitors, Bruton's tyrosine kinase (BTK) inhibitors, CYP17 inhibitors, extracellular signal-regulated kinase (ERK) inhibitors, gonadotropin-releasing hormone superagonists (GnRH agonists), luteinizing hormone-releasing hormone (LH-RH) agonists, luteinizing hormone-releasing hormone (LH-RH) antagonists, and rapamycin. These include mTOR inhibitors, mitogenic factor-activated protein kinase (MEK) inhibitors, nucleoside or nucleotide analogs or prodrugs, phosphatidylinositol 3-kinase (PI3K) pathway inhibitors, rapidly progressive fibrosarcoma (RAF) kinase inhibitors, renin-angiotensin system (RAS) inhibitors, selective estrogen receptor degraders (SERDs), selective estrogen receptor modulators (SERMs), serine-threonine protein kinase B (Akt) inhibitors, or topoisomerase inhibitors. In one embodiment, one or more further therapeutic agents are selected from letrazole, anastrozole, fulvestrant, tamoxifen, etoposide, enzalutamide, pictilisib, exemestane, or combinations thereof.

[0022] In another embodiment, form B of compound 2 is used in combination with SERD as described in International Publication No. 2017 / 100712, International Publication No. 2017 / 100715, U.S. Patent Application Publication No. 2017 / 0166550, or U.S. Patent Application Publication No. 2017 / 0166551. In yet another embodiment, a pharmaceutical composition is provided comprising isolated form B of compound 2, a pharmaceutically acceptable excipient, and SERD as described in International Publication No. 2017 / 100712, International Publication No. 2017 / 100715, U.S. Patent Application Publication No. 2017 / 0166550, or U.S. Patent Application Publication No. 2017 / 0166551.

[0023] One aspect of the present invention provides a method for treating a CDK4 / 6-dependent cell proliferation disorder, comprising administering a therapeutically effective amount of an isolated form B of compound 2 to a host in need thereof.

[0024] Furthermore, the use of isolated B-forms in the manufacture of agents for treating pRb-positive cancers such as estrogen receptor-positive (ER+) breast cancer, non-small cell lung cancer (NSCLC), prostate cancer, or other abnormal cell proliferations in the host is provided. [Brief explanation of the drawing]

[0025] [Figure 1] This figure compares the XRPD patterns of morphology A, morphology B, and morphology C. These three morphologies were obtained from crystallization and slurry experiments, as described in Example 2 and shown in Tables 1 to 4. The x-axis represents 2 theta measured in degrees, and the y-axis represents intensity measured in counts. [Figure 2] This figure compares the XRPD patterns of morphology D, morphology E, and morphology F. These three morphologies were obtained from crystallization and slurry experiments, as described in Example 2 and shown in Tables 1 to 4. The x-axis represents 2 theta measured in degrees, and the y-axis represents intensity measured in counts. [Figure 3]This figure compares the XRPD patterns of morphology G and morphology H. These two morphologies were obtained from crystallization and slurry experiments, as described in Example 2 and shown in Tables 1 to 4. Morphology G is the anhydrous form, and morphology H is the n-PrOH solvate. The x-axis represents the 2-theta measured in degrees, and the y-axis represents the intensity measured in counts. [Figure 4A] This figure shows the results of a dynamic vapor adsorption analysis (Example 3) of a moisture adsorption experiment for morphology A. This material was found to be unstable, and XRPD analysis of the dried sample at the end of the experiment revealed a new morphology, morphology K. Morphology A adsorbed 14.9 wt% at 60% RH (relative humidity) and 15.8 wt% at 90% RH. The x-axis represents relative humidity measured as a percentage, and the y-axis represents the weight of water in the material measured as a percentage. [Figure 4B] This figure shows the results of a dynamic vapor adsorption analysis (Example 3) of the moisture adsorption experiment for morphology D. This material was found to be unstable, and XRPD analysis of the dried sample at the end of the experiment revealed a new morphology, morphology K. Morphology D adsorbed 4.4 wt% at 60% RH (relative humidity) and 4.4 wt% at 90% RH. The x-axis represents relative humidity measured as a percentage, and the y-axis represents the weight of water in the material measured as a percentage. [Figure 4C] This figure shows the results of a dynamic vapor adsorption analysis (Example 3) of the moisture adsorption experiment for Form B. The material was stable, and Form B was confirmed by XRPD analysis of the dried sample at the end of the experiment. Form B adsorbed 5.8 wt% at 60% RH (relative humidity) and 5.9 wt% at 90% RH. The x-axis represents relative humidity measured as a percentage, and the y-axis represents the weight of water in the material measured as a percentage. [Figure 5A] This figure compares the XRPD patterns of morphology A before (top) and after (bottom) the moisture adsorption experiment. After the moisture adsorption experiment, XRPD analysis revealed that morphology A was not stable and had been transformed into a new morphology, morphology K (Example 3). The x-axis represents 2-theta measured in degrees, and the y-axis represents intensity measured in counts. [Figure 5B]This figure compares the XRPD patterns of morphology D before (top) and after (bottom) the moisture adsorption experiment. After the moisture adsorption experiment, XRPD analysis revealed that morphology D was not stable and had been transformed into a new morphology, morphology K (Example 3). The x-axis represents 2-theta measured in degrees, and the y-axis represents intensity measured in counts. [Figure 6] This figure compares the XRPD patterns of morphology A, morphology B, and morphology C after stability studies (Example 4) with the XRPD patterns of reference morphology A, morphology B, and morphology C. The top three patterns are the reference morphologies of morphology A, morphology B, and morphology C. After a 7-day stability study, morphology A transformed into a new morphology (morphology A after study), but after 3 days of equilibrium at room temperature, the new morphology reverted to morphology A (morphology A after 3 days). Morphology B and morphology C remained unchanged during the stability study. The x-axis is the 2-theta measured in degrees, and the y-axis is the intensity measured in counts. [Figure 7] This is a diagram of the XRPD pattern for pure morphology B. The peaks indicated by bars are listed in Example 6. The x-axis is 2 theta measured in degrees, and the y-axis is intensity measured in counts. [Figure 8] This figure compares the XRPD pattern of a non-pure morphology B material with the XRPD pattern of a pure morphology B material, such as the one characterized in Example 6. The non-pure morphology B material has two peaks at approximately 4.0 and 5.6 degrees, which are not seen in the pure morphology B material. The x-axis is 2 theta measured in degrees, and the y-axis is the relative intensity as a means of comparing the two morphology B materials. [Figure 9]This figure compares impure form B, samples 1 and 4 from the slurry experiment described in Example 8, and pure form B. Pure form B is the form B characterized in Example 6. Numerous experiments, including slurry experiments using 0.1 M HCl:acetone at 1:1 (v / v) (sample 1) and 0.5 M HCl:acetone at 1:2 (v / v) (sample 4), were performed to convert impure form B to pure form B material. The acidic aqueous acetone mixture could not convert the impure material to the pure material. The XRPD patterns of samples 1 and 4 did not match the XRPD pattern of pure form B because a peak at approximately 4.0 degrees was still present. The x-axis represents relative humidity measured as a percentage, and the y-axis represents the weight of water in the material measured as a percentage. [Figure 10] This figure compares a non-pure sample of Form B, samples 3 and 5 from the slurry experiment described in Example 8, and pure Form B. Numerous experiments were conducted, including slurry experiments using 0.1 M HCl:acetone at 75:25 (v / v) (sample 3) and 0.5 M HCl:acetone at 50:50 (v / v) (sample 5), to convert non-pure Form B to pure Form B material. The acidic aqueous acetone mixture could not convert the non-pure material to the pure material. Pure Form B is Form B characterized in Example 6. Non-pure Form B is the material used as the starting material in the slurry experiment, and sample 2 of non-pure Form B is a second non-pure form used as a reference. The x-axis represents relative humidity measured as a percentage, and the y-axis represents the weight of water in the material measured as a percentage. [Figure 11]This figure compares impure form B, samples 6, 7, and 11 from the slurry experiment described in Example 8, and pure form B. Numerous experiments, including slurry experiments using a 1:2 (v / v) water:acetone mixture stirred at room temperature, were performed to convert impure form B to pure form B material. Samples 6, 7, and 11 varied with the concentration of impure form B and the length of time the samples were stirred (details are given in Table 12). Since the XRPD patterns of samples 6, 7, and 11 matched the XRPD pattern of pure form B, all three conditions converted impure form B to pure form B. Pure form B is the form B characterized in Example 6, while impure form B is the material used as the starting material in the slurry experiment. The x-axis represents relative humidity measured as a percentage, and the y-axis represents the weight of water in the material measured as a percentage. [Figure 12] This figure compares the impure form B, samples 12 and 14 from the slurry experiment described in Example 8, and pure form B. As part of efforts to improve the yield of the recrystallization process, slurry experiments were performed using 1:3 (v / v) water:acetone (sample 14) and 1:2 water:acetone, followed by further acetone (sample 12). The XRPD patterns of samples 12 and 14 did not match the XRPD pattern of form B, as a peak at approximately 4.0 degrees was still present. Pure form B is the form B characterized in Example 6, and impure form B is the material used as the starting material in the slurry experiment. The x-axis is relative humidity measured as a percentage, and the y-axis is the weight of water in the material measured as a percentage. [Figure 13] This graph shows the TG-IR experiment results for pure form B, sample 11 (Example 8). The TG data showed a weight loss of 6.4% between 33°C and 137°C. The x-axis represents temperature measured in degrees Celsius, and the y-axis represents the weight of the material measured as a percentage. [Figure 14]This is the IR data from the TG-IR experiment of pure form B, sample 11 (Example 8). The x-axis represents the wavelength measured at cm⁻¹ and the time measured in minutes. The y-axis represents the absorbance. [Figure 15] This figure compares the IR spectra of pure form B, sample 11, obtained at 2.691 min and 5.382 min in the TG-IR experiment, with those of water and hydrogen chloride. During the TG-IR experiment, only water was released as a volatile substance; hydrogen chloride was not released. The x-axis represents the wavelength measured at cm⁻¹, and the y-axis represents the absorbance. [Figure 16] This figure shows a comparison of sample 8, which was dried in a vacuum furnace at approximately 40°C for 15 hours (Example 8, Table 15). The XRPD after the vacuum procedure did not correlate with the XRPD pattern of pure morphology B. Dried sample 8 is a novel crystalline morphology. The x-axis is 2 theta measured in degrees, and the y-axis is intensity measured in counts. [Figure 17] The image shows the XRPD patterns of samples 11 and 23, both dried in a vacuum furnace under different conditions, compared to the XRPD pattern of pure morphology B (Example 8, Table 15). Both samples 11 and 23 showed the XRPD pattern of morphology B. The x-axis is 2 theta measured in degrees, and the y-axis is intensity measured in counts. [Figure 18] This figure compares the XRPD patterns of impure morphology B, pure morphology B, and a material converted from impure morphology B as described in Example 8. The XRPD pattern of the converted material matched that of the pure morphology B material. Pure morphology B is the morphology B characterized in Example 6, and impure morphology B is the material used as the starting material in the conversion procedure. The x-axis represents 2-theta measured in degrees, and the y-axis represents intensity measured in counts. [Figure 19]These are TGA data for batches converted from impure form B to pure form B material, as described in Example 8. The TGA data showed a weight loss of 7.6% between 31°C and 120°C, and approximately 20% between 120°C and 350°C. The x-axis represents temperature measured in degrees Celsius, and the y-axis represents the weight of the material measured as a percentage. [Figure 20] These are the XRPD patterns of morphology I and morphology J. The x-axis represents the 2-theta measured in degrees, and the y-axis represents the intensity measured in counts. [Figure 21] These are DSC data for a representative batch of morphology B material. The DSC data were collected by raising the temperature of the sample (3.9 mg) from 25°C to 400°C at a rate of 10°C / min. Endothermic reactions were observed at 113°C (1), 231°C (2), 262°C (3), and 348°C (4). Endothermic reaction 1 (integral = -237 mJ, normalized = -60 J / g) indicated a start at 113°C and an endset at 140°C. Endothermic reaction 2 (integral = -182 mJ, normalized = -46 J / g) indicated a start at 219°C and an endset at 239°C. Endothermic reaction 3 (integral = 177 mJ, normalized = 45 J / g) indicated a start at 250°C and an endset at 271°C. Endothermic test 4 (integral = -728 mJ, normalized = -186 J / g) shows a start at 341°C and an end at 350°C. The x-axis represents temperature measured in degrees Celsius, and the y-axis represents heat flow rate measured in milliwatts (mW). [Figure 22] These are DSC data from a representative batch of morphology A. The DSC data were collected by raising the temperature of the sample (4.4 mg) from 30°C to 350°C at a rate of 10°C / min. Endothermic reactions were observed at 110°C (1), 275°C (2), and 344°C (3). Endothermic reaction 1 (integral = -670 mJ, normalized = -151 J / g) indicated an onset at 84°C. Endothermic reaction 2 (integral = -480 mJ, normalized = -108 J / g) indicated an onset at 242°C. Endothermic reaction 3 indicated an onset at 344°C. The x-axis represents temperature measured in degrees Celsius, and the y-axis represents heat flow rate measured in milliwatts (mW). [Figure 23]These are DSC data from a representative batch of morphology B. The DSC data were collected by raising the temperature of the sample (2.6 mg) from 30°C to 350°C at a rate of 10°C / min. Endothermic reactions were observed at 95°C (1), 225°C (2), 254°C (3), and 348°C (4). Endothermic reaction 1 (integral = -256 mJ, normalized = -97 J / g) indicated the start at 75°C. Endothermic reaction 2 (integral = -265 mJ, normalized = -101 J / g) indicated the start at 199°C. Endothermic reaction 3 (integral = -140 mJ, normalized = -53 J / g) indicated the start at 239°C. Endothermic reaction 4 (integral = -94 mJ, normalized = -36 J / g) indicated the start at 344°C. The x-axis represents temperature measured in degrees Celsius, and the y-axis represents heat flow measured in milliwatts (mW). [Figure 24] These are DSC data from a representative batch of morphology C. The DSC data were collected by raising the temperature of the sample (2.5 mg) from 30°C to 350°C at a rate of 10°C / min. Endothermic reactions were observed at 95°C (1), 235°C (2), 257°C (3), and 344°C (4). Endothermic reaction 1 (integral = -88 mJ, normalized = -36 J / g) indicated an onset at 77°C. Endothermic reaction 2 (integral = -58 mJ, normalized = -23 J / g) indicated an onset at 216°C. Endothermic reaction 3 (integral = -31 mJ, normalized = -12 J / g) indicated an onset at 247°C. Endothermic reaction 4 (integral = -379 mJ, normalized = -154 J / g) indicated an onset at 338°C. The x-axis represents temperature measured in degrees Celsius, and the y-axis represents heat flow measured in milliwatts (mW). [Figure 25] These are DSC data from a representative batch of morphology D. The DSC data were collected by raising the temperature of the sample (2.5 mg) from 30°C to 350°C at a rate of 10°C / min. Endothermic reactions were observed at 103°C (1), 260°C (2), and 345°C (3). Endothermic reaction 1 (integral = -370 mJ, normalized = -149 J / g) indicated an onset at 73°C. Endothermic reaction 2 (integral = -271 mJ, normalized = -109 J / g) indicated an onset at 228°C. Endothermic reaction 3 (integral = -321 mJ, normalized = -129 J / g) indicated an onset at 340°C. The x-axis represents temperature measured in degrees Celsius, and the y-axis represents heat flow rate measured in milliwatts (mW). [Figure 26]These are DSC data from a representative batch of morphology E. DSC data were collected by raising the temperature of the sample (2.5 mg) from 30°C to 350°C at a rate of 10°C / min. Endothermic reactions were observed at 70°C (1), 219°C (2), 275°C (3), and 345°C (4). Endothermic reaction 1 (integral = -495 mJ, normalized = -194 J / g) indicated an onset at 38°C. Endothermic reaction 2 (integral = 25 mJ, normalized = 10 J / g) indicated an onset at 209°C. Endothermic reaction 3 (integral = -208 mJ, normalized = -81 J / g) indicated an onset at 242°C. Endothermic reaction 4 (integral = -339 mJ, normalized = -133 J / g) indicated an onset at 340°C. The x-axis represents temperature measured in degrees Celsius, and the y-axis represents heat flow measured in milliwatts (mW). [Figure 27] These are DSC data from a representative batch of morphology F. The DSC data were collected by raising the temperature of the sample (3.0 mg) from 30°C to 350°C at a rate of 10°C / min. Endothermic reactions were observed at 73°C (1), 214°C (2), 277°C (3), 303°C (4), and 329°C (5). Endothermic reaction 1 (integral = -991 mJ, normalized = -323 J / g) indicated an onset at 43°C. Endothermic reaction 2 (integral = -121 mJ, normalized = -39 J / g) indicated an onset at 205°C. Endothermic reaction 3 (integral = 98 mJ, normalized = 32 J / g) indicated an onset at 265°C. Endothermic reaction 4 (integral = -15 mJ, normalized = -5 J / g) indicated an onset at 297°C. Endothermic reaction 5 (integral = -283 mJ, normalized = -92 J / g) showed an initiation of 318°C. The x-axis represents temperature measured in degrees Celsius, and the y-axis represents heat flow rate measured in milliwatts (mW). [Figure 28]These are DSC data from a representative batch of morphology G. The DSC data were collected by raising the temperature of the sample (2.8 mg) from 30°C to 350°C at a rate of 10°C / min. Endothermic reactions were observed at 81°C (1), 120°C (2), 260°C (3), and 347°C (4). Endothermic reaction 1 (integral = -167 mJ, normalized = -59 J / g) indicated an onset at 56°C. Endothermic reaction 2 (integral = -183 mJ, normalized = -65 J / g) indicated an onset at 103°C. Endothermic reaction 3 (integral = -251 mJ, normalized = -89 J / g) indicated an onset at 235°C. Endothermic reaction 4 (integral = -164 mJ, normalized = -58 J / g) indicated an onset at 344°C. The x-axis represents temperature measured in degrees Celsius, and the y-axis represents heat flow measured in milliwatts (mW). [Figure 29] These are DSC data from a representative batch of morphology H. The DSC data were collected by raising the temperature of the sample (2.7 mg) from 30°C to 350°C at a rate of 10°C / min. Endothermic reactions were observed at 110°C (1), 225°C (2), 274°C (3), and 346°C (4). Endothermic reaction 1 (integral = -300 mJ, normalized = -110 J / g) indicated the start at 109°C. Endothermic reaction 2 (integral = -41 mJ, normalized = -15 J / g) indicated the start at 210°C. Endothermic reaction 3 (integral = -138 mJ, normalized = -50 J / g) indicated the start at 242°C. Endothermic reaction 4 (integral = -301 mJ, normalized = -110 J / g) indicated the start at 346°C. The x-axis represents temperature measured in degrees Celsius, and the y-axis represents heat flow measured in milliwatts (mW). [Figure 30]These are DSC data from a representative batch of morphology A. The DSC data were collected by raising the temperature of the sample (6.0 mg) from 30°C to 350°C at a rate of 10°C / min. Endothermic reactions were observed at 121°C (1), 242°C (2), 290°C (3), and 348°C (4). Endothermic reaction 1 (integral = -541 mJ, normalized = -90 J / g) indicated an onset at 93°C. Endothermic reaction 2 (integral = 133 mJ, normalized = 22 J / g) indicated an onset at 233°C. Endothermic reaction 3 (integral = -272 mJ, normalized = -45 J / g) indicated an onset at 268°C. Endothermic reaction 4 (integral = -1131 mJ, normalized = -198 J / g) indicated an onset at 344°C. The x-axis represents temperature measured in degrees Celsius, and the y-axis represents heat flow measured in milliwatts (mW). [Figure 31] This is a diagram of the XRPD patterns for morphology I and morphology J. The x-axis represents the 2-theta measured in degrees, and the y-axis represents the intensity measured in counts. [Modes for carrying out the invention]

[0026] It is impossible to predict in advance whether a compound exists in two or more solid forms, or, if one or more exist, what the various properties of any given solid form may be, or whether those properties are suitable for a therapeutic drug delivery. For example, the drug ritonavir is active in one polymorphic form and inactive in another, with the inactive form being more stable.

[0027] The solid form of a compound can be characterized by analytical methods such as X-ray powder diffraction (XRDP), thermogravimetric analysis (TGA), TGA with IR off-gas analysis, differential scanning calorimetry (DSC), melting point, FT-Raman spectroscopy, dynamic vapor adsorption (DVS), polarized light microscopy (PLM), or other techniques known in the art.

[0028] Eleven morphologies of compound 2 were discovered through slurry and crystallization experiments. Of these eleven morphologies, morphologies A, B, and D were found to possess properties suitable for further development. Water adsorption experiments revealed that morphology B is a stable solid with unexpectedly excellent crystalline properties.

[0029] Form B An isolated form B of compound 2 is provided in the present invention.

[0030] In one embodiment, form B features the XRPD pattern in Figure 7, or an XRPD pattern substantially similar to the pattern described in Figure 7. In one embodiment, form B features an XRPD pattern including at least three 2-theta values ​​selected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.7±0.2°, and 22.4±0.2°. In one embodiment, form B features an XRPD pattern including a peak having a 2-theta value of 9.5±0.4°.

[0031] In one embodiment, form B is characterized by a 7.5% weight loss at 31°C to 120°C in thermogravimetric infrared (TG-IR) analysis.

[0032] In one embodiment, form B of isolated compound 2 does not have a peak at either or both of 4.0±0.2° and 5.6±0.2° 2-theta, or the peaks at either or both of 4.0±0.2° and 5.6±0.2° 2-theta are present at 200 counts per second (CPS) or less, 150 CPS or less, 100 CPS or less, or 75 CPS or less.

[0033] Form B can be produced using selective crystallization. This method can be carried out by treating a solution containing a suitable solvent(s) and compound 2 in the presence of one or more species, optionally including form B, under conditions that provide crystallization of form B. Selective crystallization can be carried out in any suitable solvent. For example, selective crystallization can be carried out in a nonpolar solvent or a mixture thereof. Selective crystallization can be carried out at temperatures in the range of, for example, about 40°C to about 65°C. In another embodiment, selective crystallization can be carried out at temperatures in the range of, for example, about 45°C to about 60°C, or about 45°C to about 55°C.

[0034] In one embodiment, form B of compound 2 is produced by recrystallization in a hydrochloric acid solution. Compound 1 is dissolved in HCl water and heated to at least 55±10°C. The solution is stirred for approximately 45 minutes and filtered through an in-line filter. Acetone is gradually added to the hot solution to induce crystallization. Subsequently, the temperature of the solution is lowered to 25±5°C or below and stirred for at least 2 hours. The resulting solid is collected by filtration to obtain form B.

[0035] In an alternative embodiment, form B of compound 2 is recrystallized from form D of compound 2. Form D of compound 2 is first formed by dissolving compound 1 in HCl water and heating the solution to about 55±10°C. The solution is stirred for about 45 minutes, and the resulting solution is filtered through an in-line filter. Subsequently, the temperature of the solution is lowered to about 25±5°C, and the solution is stirred for at least 2 hours. After crystallization has begun, acetone is added over about 1 hour at a temperature of about 25±5°C to complete the crystallization. The solution is stirred for a further 2 hours, and the resulting solid is collected by filtration to obtain form D of compound 2. Next, form D is dissolved in concentrated HCl, and the solution is heated. After acetone is added to the hot solution, any solid is formed. After the solution is cooled, the solid is collected by filtration to obtain form B.

[0036] In one embodiment, the impure form B of compound 2 is converted to pure form B in a water:acetone (1:2) (v / v) slurry at 30°C. This is followed by slow filtration to produce a wet cake. The wet cake is dried at ambient temperature for approximately 3.5 hours, and then vacuum-dried at ambient temperature.

[0037] In a particular embodiment, form B features an XRPD pattern containing all of the following, or at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 2-theta values ​​selected from the following: a.6.5, 8.1, 9.4, 9.6, 10.2, 10.6, 11.2, 12.2, 12.9, 13.0, 13.3, 13.4, 14.0, 14.4, 14.6, 15.0, 15.9, 16.2, 16.4, 16.5, 16.8, 18.1, 18.4, 18.5, 18.6, 18.6, 18.9, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4, 20.6, 21.3, 21.4, 21.8, 22.0 , 22.2, 22.3, 22.4, 22.5, 22.8, 23.0, 23.1, 23.4, 23.8, 24.1, 24.2, 24.3, 24.4, 24.5, 24.6, 25.4, 25.6, 25.7, 25.9, 26.0, 26.1, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.2, 27.3, 27.5, 27.6, 27.7, 27.9, 28.3, 28.4, 28.5, 28.7, 28.9, 29.0, 29.1, 29 0.3, 29.4, 29.5, 29.6, 29.7, 29.8, 29.9, 30.0, 30.3, 30.4, 30.5, 30.6, 30.7, 30.9, 31.2, 31.5, 31.6, 31.7, 31.8, 31.9, 32.0, 32.2, 32.3, 32.4, 32.5, 32.6, 32.7, 32.8, 33.1, 33.2, 33.3, 33.6, 33.7, 33.8, 34.0, 34.1, 34.2, 34.3, 34.6, 34.7, 34.8, 35.0, 35.2, 35.3, 35.5, 35.6, 35.9, 36.0, 36.2, 36.5, 36.6, 36.7, 36.8, 36.9, 37.1, 37.2, 37.3, 37.4, 37.5, 37.6, 37.7, 37.8, 37.9, 38.2, 38.3, 38.4, 38.5, 38.6, 38.7, 38.8, 38.9, 39.0, 39.1, 39.2, 39.3, 39.4, 39.5, 39.6, 39.7, 39.8, 39.9, and 40.0°²θ, or, b.6.5, 9.4, 9.5, 9.6, 10.2, 10.6, 13.3, 13.4, 14.0, 14.4, 14.6, 15.0, 16.2, 16.4, 16.5, 16.8, 18.1, 18 .4, 18.5, 18.6, 18.9, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4, 22.3, 22.4, 22.5, 22. 8, 23.0, 23.1, 23.4, 23.8, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.2, 27.3, 27.5, 27.6, 27.7, 27.9, 28.3, 28.4, 28.5, 28.7, 28.9, 29.0, 29.1, 29.3, 29.4, 29.5, 29.6, 29.7, 29.8, 29.9, and 30.0, °2θ, or, c.6.5, 9.4, 9.5, 9.6, 10.2, 10.6, 13.3, 13.4, 14.0, 14.4, 14.6, 15.0, 16.2, 16.4, 16 .5, 16.8, 18.1, 18.4, 18.5, 18.6, 18.9, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19. 8, 19.9, 20.4, 22.3, 22.4, 22.5, 22.8, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, 27.9, 29.0, 29.1, 29.3, 29.4, 29.5, 29.6, 29.7, 29.8, 29.9, and 30.0, °2θ, or, d.6.5, 9.4, 9.5, 9.6, 10.2, 10.6, 13.3, 13.4, 14.0, 14.4, 14.6, 15.0, 16.2, 16.4, 16.5, 16.8, 18.1, 18.4, 18.5, 18.6, 18.9, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4, 22.3, 22.4, 22.5, 22.8, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, °2θ, or, e.6.5, 9.4, 9.5, 9.6, 10.2, 10.6, 14.0, 14.4, 14.6, 15.0, 16.2, 16.4, 16.5, 18.1, 18.4, 18.5, 18.6, 18.9, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4, 22.3, 22.4, 22.5, 22.8, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, °2θ, or, f.6.5, 9.5, 14.0, 14.4, 14.6, 18.1, 18.4, 18.5, 18.6, 18.9, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4, 22.3, 22.4, 22.5, 22.8, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, °2θ, or, g.9.5, 14.6, 18.1, 18.4, 18.5, 18.6, 18.9, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4, 22.3, 22.4, 22.5, 22.8, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, °2θ, or, h.9.5, 14.6, 18.1, 18.4, 18.5, 18.6, 18.9, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 20.4, 22.3, 22.4, 22.5, 22.8, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, °2θ, or, i.9.5, 14.6, 18.1, 18.4, 18.5, 18.6, 18.9, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 22.3, 22.4, 22.5, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, °2θ, or, j.9.5, 18.1, 18.4, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 22.3, 22.4, 22.5, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, °2θ, or, k.9.5, 18.1, 18.4, 19.3, 19.7, 22.3, 22.4, 22.5, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, °2θ, or, l.9.5, 18.1, 18.4, 19.3, 19.7, 22.4, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.7, 27.9, and 27.9, °2θ, or, m.9.5, 18.1, 18.4, 19.3, 19.7, 22.4, 26.6, 27.7, 27.9, and 27.9, °2θ, or, n.9.5, 18.1, 18.4, 19.3, 19.7, 22.4, 26.6, and 27.7, °2θ, or, 0.9.5, 18.1, 19.3, 19.7, 22.4, 26.6, and 27.7, °2θ, or, p.9.5, 18.1, 19.3, 22.4, 26.6, and 27.7, °2θ, or, Any of the above peaks where q.°2θ is ±0.1, or Any of the peaks in the above list where r.°2θ is ±0.2, Any of the above peaks where s.°2θ is ±0.3, Any of the above peaks where t.°2θ is ±0.4, Any of the peaks in the above list whose °2θ is ±0.4, located in u.9.5 The peak in v.9.5 has a °2θ of ±0.4, and any of the remaining peaks in the above peak list have a °2θ of ±0.1. Any of the peaks listed above in w.9.5, where the °2θ of the peak is ±0.4 and the remaining peaks are ±0.2°2θ.

[0038] In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak exceeding 200 CPS in the range of 4°2θ to 6°2θ. In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak exceeding 150 CPS in the range of 4°2θ to 6°2θ. In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak exceeding 100 CPS in the range of 4°2θ to 6°2θ. In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak exceeding 75 CPS in the range of 4°2θ to 6°2θ.

[0039] In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak at approximately 4.0°2θ above 150 CPS. In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak at approximately 4.0°2θ above 100 CPS. In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak at approximately 4.0°2θ above 75 CPS.

[0040] In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak at approximately 5.6°2θ with more than 150 CPS. In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak at approximately 5.6°2θ with more than 100 CPS. In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak at approximately 5.6°2θ with more than 75 CPS in the range of 4°2θ to 6°2θ.

[0041] In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak at approximately 5.3°2θ with more than 150 CPS. In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak at approximately 5.3°2θ with more than 100 CPS. In one embodiment, form B is characterized by the above-described XRPD pattern and is further characterized by not having a peak at approximately 5.3°2θ with more than 75 CPS in the range of 4°2θ to 6°2θ.

[0042] In a further embodiment, the CPS count is baseline-corrected.

[0043] The method used to produce form B is further described in Examples 2 and 8 below.

[0044] Form D In one embodiment, form D is characterized by the initiation of endothermic DSC at approximately 100±20°C, approximately 270±20°C, and approximately 347±20°C, for example, at 108.3°C, 266.1°C, and 347.0°C, or at 95°C, 257°C, and 344°C.

[0045] Form D can be prepared using selective crystallization. This method can be carried out by treating a solution containing a suitable solvent(s) and compound 2 in the presence of one or more species, optionally including form D, under conditions that provide crystallization of form D. Selective crystallization can be carried out in any suitable solvent. For example, selective crystallization can be carried out in a nonpolar solvent or a mixture thereof. In one embodiment, the solvent is acetonitrile. Selective crystallization can be carried out at temperatures, for example, in the range of about 5°C to about 55°C.

[0046] In one embodiment, form D of compound 2 is formed by dissolving compound 1 in 2M aqueous HCl (10 times volume) and heating the solution to 55±10°C. The solution is stirred for approximately 45 minutes, and the resulting solution is filtered through an in-line filter. Subsequently, the temperature of the solution is lowered to 25±5°C, and the solution is stirred for at least 2 hours. After crystallization begins, acetone (30 times volume) is added over 1 hour at a temperature of 25±5°C to complete the crystallization. The solution is stirred for a further 2 hours, and the resulting solid is collected by filtration to obtain form D of compound 2.

[0047] In an alternative embodiment, form D of compound 2 is formed by dissolving compound 1 in 2M aqueous HCl (10 times volume) and heating the solution to 55±10°C. The solution is stirred for 45 minutes, and the resulting solution is filtered through an in-line filter. The solution is cooled to 25±5°C, and the solution is stirred for at least 2 hours. The resulting solid is collected by filtration, and acetone is added to obtain form D of compound 2.

[0048] In one embodiment, form D is also recrystallized again to produce form B.

[0049] The method used to produce morphology D is further described in Example 2 below.

[0050] Form A In one embodiment, form A is characterized by XRPD peaks at two thetas of approximately 7.4±0.2 and 9.0±0.2. In a further embodiment, form A is characterized by the onset of endothermic DSC at approximately 110±20°C, approximately 275±20°C, and approximately 350±20°C, for example, at 110.3°C, 275.6°C, and 344.8°C, or at 103°C, 260°C, and 345°C.

[0051] Form A can be produced using selective crystallization. This method can be carried out by treating a solution containing a suitable solvent(s) and compound 2 in the presence of one or more species, optionally including Form A, under conditions that provide crystallization of Form D. Selective crystallization can be carried out in any suitable solvent. For example, selective crystallization can be carried out in a polar solvent or a mixture thereof. In one embodiment, the solvent is MeOH, EtOH, or 1-BuOH. Selective crystallization can be carried out at temperatures in the range of about 5°C to about 75°C, for example. In one embodiment, crystallization is carried out at a temperature of about 60°C.

[0052] The method used to produce morphology A is further described in Example 2 below.

[0053] Chemical descriptions and technical terms Compounds are described using their formal names. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which this invention pertains.

[0054] The terms "a" and "an" do not indicate a limit on quantity, but rather indicate the presence of at least one of the items mentioned. The term "or" means "and / or". Unless otherwise specified herein, the enumeration of value ranges is intended simply as a way to refer individually to each distinct value contained within that range, and each distinct value becomes part of this specification by being cited as if they were individually enumerated herein. All endpoints of a range are contained within that range and can be combined independently.

[0055] All methods described herein can be carried out in a preferred order unless otherwise specified herein or clearly rejected by the context. Any use of examples or illustrative language (e.g., "such as") is intended solely to better illustrate the invention and, unless otherwise asserted, does not imply any limitation of the scope of the invention.

[0056] "Activator" is a compound (including the compounds disclosed herein) that, when administered to a patient alone or in combination with another compound, element, or mixture, directly or indirectly confers a physiological effect to the patient. Indirect physiological effects may result from metabolites or other indirect mechanisms.

[0057] "Deuterated" and "deuterated" mean that hydrogen is replaced by deuterium so that deuterium is present in amounts exceeding its natural abundance and is therefore "concentrated." A 50% concentration means that the deuterium content is 50%, not hydrogen at the specified positions. For clarity, it is confirmed that the term "concentrated," as used herein, does not mean a percentage concentrated above its natural abundance. In other embodiments, a deuterium concentration of at least 80%, at least 90%, or at least 95% is observed at the specified deuterated position(s). In other embodiments, a deuterium concentration of at least 96%, at least 97%, at least 98%, or at least 99% is observed at the indicated specified deuterated position(s). Conversely, in the absence of the indication, the deuterium concentration at the specified positions of the compounds described herein is at least 90%.

[0058] "Dosage form" refers to the unit of administration of an active agent. Non-exclusive examples of dosage forms include tablets, capsules, injections, suspensions, liquids, intravenous solutions, emulsions, creams, ointments, suppositories, inhalable forms, and transdermal forms.

[0059] A “pharmaceutical composition” is a composition comprising at least one activator, such as one compound or salt of an active compound disclosed herein, and at least one other substance, such as a carrier. A pharmaceutical composition may optionally contain two or more activators. A “pharmaceutical combination” or “combination therapy” refers to the administration of at least two activators, which in one embodiment may be combined in a single-dosage form, or three, four or more activators which may be supplied together in separate dosage forms, along with instructions that the activators are used together to treat a disorder.

[0060] "Pharmacopoeia-acceptable salts" include derivatives of disclosed compounds in which the parent compounds are modified by producing appropriately non-toxic inorganic and organic acid or base addition salts thereof. Salts of these compounds can be synthesized from parent compounds containing basic or acidic moieties by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid form of these compounds with a stoichiometric amount of a suitable base (e.g., Na, Ca, Mg, or K hydroxide, carbonate, bicarbonate, etc.) or by reacting the free base form of these compounds with a stoichiometric amount of a suitable acid. Such reactions are usually carried out in water or an organic solvent, or in a mixture thereof. Pharmaceutically acceptable salts may exist in pure crystalline form or in monocrystalline form, or may be used in amorphous, non-crystalline, glassy, ​​or vitreous form, or in mixtures thereof. In alternative embodiments, the active compound may be supplied in solvate form.

[0061] Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid salts of basic residues such as amines, and alkali or organic salts of acidic residues such as carboxylic acids. Pharmaceutically acceptable salts include, for example, conventional non-toxic salts and quaternary ammonium salts of parent compounds formed from non-toxic inorganic or organic acids. For example, conventional non-toxic acid salts include those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, and nitric acid, as well as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, pamoic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, mesylic acid, ecylic acid, besylic acid, sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isethionic acid, and HOOC-(CH2) n Examples include salts prepared from organic acids such as -COOH (where n=0 to 4). A further list of suitable salts can be found, for example, in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., p. 1418 (1985).

[0062] The term "carrier" refers to a diluent, excipient, or vehicle provided with the active compound.

[0063] "Pharmacopoeia acceptable excipients" means excipients that are generally safe, sufficiently non-toxic, and useful in creating pharmaceutical compositions / combinations that are not biologically or otherwise undesirable. As used in this application, "pharmacopoeia acceptable excipients" encompasses both one such excipient and two or more such excipients.

[0064] "Patient" or "host" refers to a human or non-human animal, including but not limited to a monkey, bird, cat, dog, cattle, horse, or pig, that requires medical treatment. Medical treatment may include the treatment of an existing condition, such as a disease or disorder, or preventive or diagnostic treatment. In certain embodiments, the patient or host is a human patient. In alternative embodiments, the patient, such as a host, is treated or prevented from having a disorder or disease as described herein.

[0065] The term "isolated," as used herein, refers to a material in a substantially pure form. An isolated compound has no other components that substantially affect the properties of the compound. In certain embodiments, the isolated form is at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, or 99% pure.

[0066] Treatment method One embodiment provides a method for treating proliferative disorders in a host, including a human, comprising administering form B of an isolated compound 2 in an optionally pharmaceutically acceptable carrier. Non-limiting examples of disorders include tumors, cancers, disorders related to abnormal cell proliferation, inflammatory disorders, immune disorders, and autoimmune disorders.

[0067] Form B of compound 2, when administered in an effective dose to a host including humans, causes tumors, cancers (solid, non-solid, diffuse, hematological, etc.), abnormal cell proliferation, immune disorders, inflammatory disorders, hematological disorders, myeloproliferative or lymphoproliferative disorders such as B-cell or T-cell lymphoma, multiple myeloma, breast cancer, prostate cancer, AML, ALL, ACL, lung cancer, pancreatic cancer, colon cancer, skin cancer, melanoma, Waldenström macroglobulinemia, Wiscott-Aldrich syndrome, or post-transplant lymphoma. It is useful as a therapeutic agent in drug form for the treatment of proliferative disorders; autoimmune disorders, such as lupus, Crohn's disease, Addison's disease, celiac disease, dermatomyositis, Graves' disease, thyroiditis, multiple sclerosis, pernicious anemia, reactive arthritis, or morphological I diabetes mellitus; diseases of cardiac dysfunction, including hypercholesterolemia; infectious diseases, including viral and / or bacterial infections; and inflammatory conditions, including asthma, chronic peptic ulcers, tuberculosis, rheumatoid arthritis, periodontitis, ulcerative colitis, or hepatitis.

[0068] Examples of proliferative disorders include, but are not limited to, benign tumors, neoplasms, tumors, cancer (Rb-positive or Rb-negative), autoimmune disorders, inflammatory disorders, graft-versus-host rejection, and fibrous disorders.

[0069] Non-limiting examples of cancers that can be treated according to the present invention include acoustic neuroma, adenocarcinoma, adrenal carcinoma, anal cancer, angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma), appendiceal cancer, benign monoclonal gammaglobulinemia, biliary tract cancer (e.g., cholangiocarcinoma), bladder cancer, breast cancer (e.g., adenocarcinoma of the breast, papillary carcinoma of the breast, breast cancer, medullary carcinoma of the breast), and brain cancer (e.g., meningioma; glioma, e.g., astrocytoma, oligodendroglioma; medulloblastoma). ), bronchial cancer, carcinoid tumors, cervical cancer (e.g., cervical adenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma), epithelial carcinoma, ependymoma, endotheliosarcoma (e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma), endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophageal cancer (e.g., esophageal adenocarcinoma, Barrett's adenocarcinoma), Ewing's sarcoma, ocular cancer (e.g., intraocular melanoma, retinoblastoma), familial hypereosinophilia (familiar Leukemias such as hypereosinophilia, gallbladder cancer, gastric cancer (e.g., gastric adenocarcinoma), gastrointestinal stromal tumors (GIST), head and neck cancer (e.g., head and neck squamous cell carcinoma), oral cancer (e.g., oral squamous cell carcinoma (OSCC), pharyngeal cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer)), hematopoietic cancer (e.g., acute lymphoblastic leukemia (ALL), also known as acute lymphoblastic leukemia or acute lymphoblastic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myeloid leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myeloid leukemia (CML) (e.g., B-cell CML, T-cell CML), and chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL);Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non-Hodgkin lymphoma (NHL) (e.g., diffuse large cell lymphoma (DLCL) (e.g., B-cell NHL such as diffuse large B-cell lymphoma (DLBCL)), follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma (CLL / SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphoma (e.g., mucosa-associated lymphoid tissue (MALT) lymphoma, nodal marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (i.e., Lymphomas such as Waldenström's macroglobulinemia, hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B lymphoblastic lymphoma and primary central nervous system (CNS) lymphoma; and T-cell NHL such as precursor T lymphoblastic lymphoma / leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g., mycosis fungoides, Sézary syndrome), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplastic large cell lymphoma); and mixtures of one or more of the above leukemia / lymphomas;Also, multiple myeloma (MM), heavy chain disease (e.g., alpha chain disease, gamma chain disease, μ chain disease), hemangioblastoma, inflammatory myofibroblastic neoplasm, immunocytic amyloidosis, renal cancer (e.g., nephroblastoma, also known as Wilms' tumor, renal cell carcinoma), liver cancer (e.g., hepatocellular carcinoma (HCC), malignant hepatoma), lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung), leiomyosarcoma (LMS), mastocytosis (e.g., systemic mastocytosis), myelodysplastic syndromes Myeloproliferative disorders (MDS), mesothelioma, myeloproliferative disorders (MPD) (e.g., polycythemia vera (PV), essential thrombocythemia (ET), idiopathic myelometropia (AMM), also known as myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), eosinophilic syndrome (HES)), neuroblastoma, neurofibroma (e.g., neurofibromatosis type 1 or 2, schwannoma), neuroendocrine carcinoma (e.g., pancreatic gastrointestinal neuroendocrine tumor (GEP-N)) ET), carcinoid tumors, osteosarcoma, ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonic carcinoma, ovarian adenocarcinoma), papillary adenocarcinoma, pancreatic cancer (e.g., pancreatic adenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), islet cell tumor), penile cancer (e.g., Paget's disease of the penis and scrotum), pineal gland tumor, primitive neuroectodermal tumor (PNT), prostate cancer (e.g., prostatic adenocarcinoma), rectal cancer, rhabdomyosarcoma, salivary gland cancer, skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma) Examples of cancers that fall under this category include, but are not limited to, BCC (biocracytoma), small intestine cancer (e.g., appendiceal cancer), soft tissue sarcomas (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous gland cancer, sweat gland cancer, synoviomas, testicular cancer (e.g., seminomas, embryonal testicular cancer), thyroid cancer (e.g., papillary thyroid carcinoma, papillary thyroid carcinoma (PTC), medullary thyroid carcinoma), urethral cancer, vaginal cancer, and vulvar cancer (e.g., Paget's disease of the vulva).

[0070] In another embodiment, the disorder is myelodysplastic syndrome (MDS).

[0071] In certain embodiments, cancer is hematopoietic cancer. In certain embodiments, hematopoietic cancer is lymphoma. In certain embodiments, hematopoietic cancer is leukemia. In certain embodiments, leukemia is acute myeloid leukemia (AML).

[0072] In certain embodiments, the proliferative disorder is a myeloproliferative neoplasm. In certain embodiments, the myeloproliferative neoplasm (MPN) is primary myelofibrosis (PMF).

[0073] In certain embodiments, cancer is a solid tumor. As used herein, a solid tumor refers to an abnormal mass of tissue that does not typically contain cysts or fluid areas. Various types of solid tumors are named after the type of cells that form them. Examples of types of solid tumors include, but are not limited to, sarcomas, carcinomas, and lymphomas as described herein. Additional examples of solid tumors include, but are not limited to, squamous cell carcinoma, colon cancer, breast cancer, prostate cancer, lung cancer, liver cancer, pancreatic cancer, and melanoma.

[0074] In certain embodiments, the condition treated with form B of compound 2 is a disorder related to abnormal cell proliferation.

[0075] Abnormal cell proliferation, particularly hyperproliferation, can result from a wide range of factors, including gene mutations, infections, exposure to toxins, autoimmune disorders, and the induction of benign or malignant tumors.

[0076] Numerous skin disorders are associated with cell overgrowth. For example, psoriasis is a benign disease of human skin generally characterized by plaques covered with thickened scales. This disease is caused by an increase in the proliferation of epidermal cells of unknown origin. Chronic eczema is also associated with marked epidermal overgrowth. Other diseases caused by excessive proliferation of skin cells include atopic dermatitis, lichen planus, warts, pemphigus vulgaris, actinic keratosis, basal cell carcinoma, and squamous cell carcinoma.

[0077] Other hyperproliferative cell disorders include vascular disorders, fibrotic disorders, autoimmune disorders, graft-versus-host rejection, tumors, and cancers.

[0078] Vascular proliferative disorders include neovascularization and vascular disorders. The proliferation of smooth muscle cells during plaque formation in vascular tissue leads to conditions such as restenosis, retinopathy, and atherosclerosis. Both cell migration and cell proliferation play a role in the formation of atherosclerotic lesions.

[0079] Fibrotic disorders are often caused by abnormal formation of the extracellular matrix. Examples of fibrotic disorders include cirrhosis and mesangial proliferative cytotoxicity. Cirrhosis is characterized by an increase in extracellular matrix components that lead to the formation of liver scars. Cirrhosis can lead to diseases such as liver cirrhosis. The increase in extracellular matrix that leads to liver scars may also be due to viral infections such as hepatitis. Lipid cells appear to play an important role in liver cirrhosis.

[0080] Mesangial disorders are caused by the abnormal proliferation of mesangial cells. Mesangial hyperproliferative cytotoxicity includes various human kidney diseases such as glomerulonephritis, diabetic nephropathy, malignant nephrosclerosis, thrombotic microangiopathy syndrome, graft rejection, and glomerulopathy.

[0081] Another disease caused by proliferative components is rheumatoid arthritis. Rheumatoid arthritis is generally considered an autoimmune disease associated with the activity of autoreactive T cells and caused by autoantibodies produced against collagen and IgE.

[0082] Other disorders that may contain abnormal cell proliferation components include Behçet's syndrome, acute respiratory distress syndrome (ARDS), ischemic heart disease, post-dialysis syndrome, leukemia, acquired immunodeficiency syndrome, vasculitis, lipid histiocytosis, septic shock, and general inflammation.

[0083] In certain embodiments, the compounds of the present invention and their pharmaceutically acceptable derivatives, or pharmaceutically acceptable formulations containing these compounds, are also useful for the prevention and treatment of HBV infection, as well as anti-HBV antibody positivity and HBV-positive conditions, HBV-induced chronic hepatitis, cirrhosis, acute hepatitis, fulminant hepatitis, chronic persistent hepatitis, and other related conditions such as fatigue. These compounds or formulations can also be used prophylactically to prevent or slow the progression of clinical disease in individuals who are anti-HBV antibody or HBV antigen positive, or who have been exposed to HBV.

[0084] In certain embodiments, the pathological condition is associated with an immune response.

[0085] Skin contact hypersensitivity and asthma are just two examples of immune responses that may have a significant prevalence. Other examples include atopic dermatitis, eczema, Sjögren's syndrome including keratoconjunctivitis sicca secondary to Sjögren's syndrome, alopecia areata, allergic reactions to arthropod bites, Crohn's disease, aphthous ulcers, iritis, conjunctivitis, keratoconjunctivitis, ulcerative colitis, cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, and drug eruptions. These conditions may present with one or more of the following symptoms or signs: itching, swelling, redness, blistering, crusting, ulceration, pain, desquamation, cracking, hair loss, scarring, or exudation of fluid from the skin, eyes, or mucous membranes.

[0086] In atopic dermatitis and eczema, immune-mediated leukocyte infiltration into the skin (particularly mononuclear cells, lymphocytes, neutrophils, and eosinophils) generally plays a significant role in the development of these diseases. Chronic eczema is also associated with marked epidermal hyperplasia. Immune-mediated leukocyte infiltration can occur in areas other than the skin, such as the airways in asthma and the tear-producing glands in keratoconjunctivitis sicca.

[0087] In one non-limiting embodiment, the compounds of the present invention are used as topical agents for the treatment of contact dermatitis, atopic dermatitis, eczematous dermatitis, psoriasis, Sjögren's syndrome including keratoconjunctivitis sicca secondary to Sjögren's syndrome, alopecia areata, allergic reactions due to arthropod bites, Crohn's disease, aphthous ulcers, iritis, conjunctivitis, keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma, cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, and drug eruptions. This novel method may also be useful in reducing skin infiltration by malignant leukocytes in diseases such as mycosis fungoides. These compounds can also be used to treat patients suffering from tear-deficient dry eye conditions (such as immune-mediated keratoconjunctivitis) by topical administration of the compounds to the eyes.

[0088] The terms “neoplasia” or “cancer” are used throughout this specification to refer to the formation and growth of malignant or cancerous neoplasms, which are pathological processes resulting from the formation and growth of abnormal tissues (solid) or cells (non-solid) that often grow more rapidly than normal through cell proliferation and continue to grow after the stimulus that initiated new growth has ceased. Malignant neoplasms exhibit a partial or complete lack of structural organization and functional coordination with normal tissue, are prone to invading surrounding tissues, may metastasize to several sites, may recur after attempted removal, and may lead to patient death if not properly treated. As used herein, the term neoplasia is used to describe all cancerous disease conditions and includes or encompasses pathological processes associated with malignant hematological malignancies, ascites tumors, and solid tumors. Exemplary cancers that can be treated with the disclosed compounds alone or in combination with at least one additional anticancer agent include squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, hepatocellular carcinoma, and renal cell carcinoma; cancers of the bladder, intestine, breast, cervix, colon, esophagus, head, kidney, liver, lung, neck, ovaries, pancreas, prostate, and stomach; leukemia; benign and malignant lymphomas, particularly Burkitt lymphoma and non-Hodgkin lymphoma; benign and malignant melanoma; myeloproliferative disorders; Ewing's sarcoma, angiosarcoma, and capillary cancer. Sarcomas including disarcoma, liposarcoma, myosarcoma, peripheral neuroepithelioma, synovial sarcoma, glioma, astrocytoma, oligodendroglioma, ependymoma, glioblastoma, neuroblastoma, gangliocytoma, ganglioglioma, medulloblastoma, pineal cell tumor, meningioma, meningiosarcoma, neurofibroma, and Schwann cell tumor; intestinal cancer, breast cancer, prostate cancer, cervical cancer, uterine cancer, lung cancer, ovarian cancer, testicular cancer, thyroid cancer, astrocytoma, esophageal cancer, pancreatic cancer, gastric cancer, liver cancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease, Wilms' tumor, and teratoma. Additional cancers that can be treated with the compounds disclosed according to the present invention include, for example, acute granulocytic leukemia, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), adenocarcinoma, adenosarcoma, adrenal carcinoma, adrenocortical carcinoma, anal carcinoma, anaplastic astrocytoma, angiosarcoma, appendiceal carcinoma, astrocytoma, basal cell carcinoma, B-cell lymphoma, cholangiocarcinoma, bladder carcinoma, bone carcinoma, bone marrow carcinoma, intestinal carcinoma, brain carcinoma, brainstem glioma, breast carcinoma, triple (estrogen, progesterone, and HER-2) negative breast carcinoma, double negative breast carcinoma (estrogen,(Negative for both progesterone and HER-2), single-negative (negative for one of estrogen, progesterone, and HER-2), estrogen receptor positive, HER2-negative breast cancer, estrogen receptor-negative breast cancer, estrogen receptor-positive breast cancer, metastatic breast cancer, luminal A breast cancer, luminal B breast cancer, HER2-negative breast cancer, HER2-positive or negative breast cancer, progesterone receptor-negative breast cancer, progesterone receptor-positive breast cancer, recurrent breast cancer, carcinoid tumor, cervical cancer, cholangiocarcinoma, chondrosarcoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), colon cancer, colorectal cancer, craniopharyngioma, cutaneous lymphoma, cutaneous melanoma, diffuse astrocytoma, ductal carcinoma in situ (DCIS), endometrial cancer, ependymoma, epithelioid sarcoma, esophageal cancer, Ewing's sarcoma, extrahepatic cholangiocarcinoma, ocular cancer, Fallopius Ductal cancer, fibrosarcoma, gallbladder cancer, gastric cancer, gastrointestinal cancer, gastrointestinal carcinoid cancer, gastrointestinal stromal tumor (GIST), germ cell tumor, glioblastoma multiforme (GBM), glioma, hairy cell leukemia, head and neck cancer, vascular endothelium cancer, Hodgkin lymphoma, hypopharyngeal cancer, invasive ductal carcinoma (IDC), invasive lobular carcinoma (ILC), inflammatory breast cancer (IBC), intestinal cancer, intrahepatic cholangiocarcinoma, invasive / invasive breast cancer, islet cell carcinoma, jaw cancer, Kaposi's sarcoma, kidney Cancer, laryngeal cancer, leiomyosarcoma, leukemia, lip cancer, liposarcoma, liver cancer, lobular carcinoma in situ, low-grade astrocytoma, lung cancer, lymph node cancer, lymphoma, male breast cancer, medullary carcinoma, medulloblastoma, melanoma, meningioma, Merkel cell carcinoma, mesenchymal chondrosarcoma, mesenchymal mesothelioma, metastatic breast cancer, metastatic melanoma, metastatic squamous cell carcinoma of the neck, mixed glioma, monodermal teratoma, mouth cancer Cancer, mucinous carcinoma, mucosal melanoma, multiple myeloma, mycosis fungoides, myelodysplastic syndrome, nasal cavity cancer, nasopharyngeal cancer, cervical cancer, neuroblastoma, neuroendocrine tumor (NET), non-Hodgkin lymphoma, non-small cell lung cancer (NSCLC), ophthalmic cancer, intraocular melanoma, oligodendroglioma, oral cancer, oral cancer, oropharyngeal cancer, osteogenic sarcoma, osteosarcoma, ovarian cancer, epithelial ovarian cancer, ovarian germ cell tumor, primary peritoneal cancer of the ovary, ovarian cord-stromal tumor, Paget's disease, pancreatic cancer, papillary carcinoma, paranasal sinus cancer, parathyroid cancer, pelvic cancer, penile cancer, peripheral nerve cancer, peritoneal cancer, pharyngeal cancer, pheochromocytoma, pilocytic astrocytoma, pineal gland tumor, pineoblastoma, pituitary cancer, primary central nervous system (CNS) lymphoma, prostate cancer, rectal cancer, renal cell carcinoma,Renal pelvis cancer, rhabdomyosarcoma, salivary gland cancer, soft tissue sarcoma, bone sarcoma sarcoma), sarcoma, sinus cancer, skin cancer, small cell lung cancer (SCLC), small intestine cancer, spine cancer, spinal column cancer, spinal cord cancer, squamous cell carcinoma, gastric cancer, synovial sarcoma, T-cell lymphoma, testicular cancer, throat cancer, thymoma / thymic cancer, thyroid cancer, tongue cancer, tonsil cancer, transitional cell cancer, fallopian tube cancer, tubular cancer carcinoma), undiagnosed cancer, ureteral cancer, urethral cancer, uterine adenocarcinoma, uterine cancer, uterine sarcoma, vaginal cancer, vulvar cancer, T-cell acute lymphoblastic leukemia (T-ALL), T-cell lymphoblastic lymphoma (T-LL), peripheral T-cell lymphoma, adult T-cell leukemia, Pre-B ALL, Pre-B lymphoma, Large B-cell lymphoma, Burkitt lymphoma, B-cell ALL, Philadelphia chromosome-positive ALL, Philadelphia chromosome-positive CML, Juvenile myelomonocytic leukemia (JMML), Acute promyelocytic leukemia (a subtype of AML), Large granular lymphocytic leukemia, Adult T-cell chronic leukemia, Diffuse large B-cell lymphoma, Follicular lymphoma; Mucosa-associated lymphoid tissue lymphoma (MALT), Small cell lymphocytic lymphoma, Mediastinal large B-cell lymphoma, Nodal marginal zone B-cell lymphoma (NMZL); Splenic marginal zone lymphoma (SMZL); Intravascular large B-cell lymphoma; Primary exudative lymphoma; or Lymphomatoid granulomatosis; B-cell prelymphocytic leukemia; Unclassifiable splenic lymphoma / leukemia, Diffuse red pulp small B-cell lymphoma; Lymphoid Plasma cell lymphoma; heavy chain disease, e.g., α-heavy chain disease, γ-heavy chain disease, μ-heavy chain disease; plasma cell myeloma; solitary plasmacytoma of bone; extraskeletal plasmacytoma; primary cutaneous follicular lymphoma; T-cell / histiocyte-rich large B-cell lymphoma; DLBCL associated with chronic inflammation; Epstein-Barr virus (EBV) + DLBCL in the elderly; primary mediastinal (thymic) large B-cell lymphoma; primary cutaneous DLBCL lower extremity type; ALK + large B-cell lymphoma; plasmablastic lymphoma; large B-cell lymphoma arising in HHV8-associated multicentric Castleman disease; unclassifiable B-cell lymphoma with intermediate characteristics between diffuse large B-cell lymphoma and classical Hodgkin lymphoma; or unclassifiable B-cell lymphoma with intermediate characteristics between diffuse large B-cell lymphoma and classical Hodgkin lymphoma.

[0089] In another embodiment, a method is provided for increasing BIM expression (e.g., BCLC2L11 expression) to induce apoptosis in cells, comprising contacting cells with a compound of the present invention, or a pharmaceutically acceptable composition thereof. In certain embodiments, the method is an in vitro method. In certain embodiments, the method is an in vivo method. BCL2L11 expression is tightly regulated in cells. BCL2L11 encodes BIM, a pro-apoptotic protein. BCL2L11 is downregulated in many cancers, and BIM is inhibited in many cancers, including chronic myeloid leukemia (CML) and non-small cell lung cancer (NSCLC), and suppression of BCL2L11 expression confers resistance to tyrosine kinase inhibitors. See, for example, Ng et al., Nat. Med. (2012) 18:521-528.

[0090] In another embodiment, a method is provided for treating angiogenesis-related conditions such as diabetic conditions (e.g., diabetic retinopathy), inflammatory conditions (e.g., rheumatoid arthritis), macular degeneration, obesity, atherosclerosis, or proliferative disorders, comprising administering the compound of the present invention, or a pharmaceutically acceptable composition, salt, isotope analog, or prodrug thereof, to a subject in need of treatment.

[0091] In certain embodiments, the condition associated with angiogenesis is macular degeneration. In certain embodiments, a method for treating macular degeneration is provided, comprising administering the compound of the present invention, or a pharmaceutically acceptable composition, salt, isotope analog, or prodrug thereof, to a subject in need of treatment.

[0092] In certain embodiments, the condition associated with angiogenesis is obesity. As used herein, “obesity” and “obese” refer to Class I obesity, Class II obesity, Class III obesity, and pre-obesity (e.g., “overweight”) as defined by the World Health Organization. In certain embodiments, a method is provided for treating obesity, comprising administering a compound of the present invention, or a pharmaceutically acceptable composition, salt, isotope analog, or prodrug thereof, to a subject in need of treatment.

[0093] In certain embodiments, the condition associated with angiogenesis is atherosclerosis. In certain embodiments, a method is provided for treating atherosclerosis, comprising administering the compound of the present invention, or a pharmaceutically acceptable composition, salt, isotope analog, or prodrug thereof, to a subject in need of treatment.

[0094] In certain embodiments, the pathological condition associated with angiogenesis is a proliferative disorder. In certain embodiments, a method is provided for treating a proliferative disorder, comprising administering the compound of the present invention, or a pharmaceutically acceptable composition, salt, isotope analog, or prodrug thereof, to a subject in need of treatment.

[0095] In alternative embodiments, form A or form D of compound 2 is administered in an effective amount to treat the proliferative disorder.

[0096] In another alternative embodiment, form C, form E, form G, or form H of compound 2 is administered in an effective amount to treat the proliferative disorder.

[0097] Methods to reduce side effects associated with chemotherapy In certain embodiments, form B of isolated compound 2 reduces the toxic effects of chemotherapeutic agents (typically DNA damaging agents) on subjects exposed to, or having been exposed to, typically human hematopoietic stem cells and hematopoietic progenitor cells (both referred to as HSPCs), and / or CDK4 / 6 replication-dependent healthy cells such as renal epithelial cells.

[0098] In one embodiment, the subject is exposed to a chemotherapeutic agent, and the subject's CDK4 / 6 replication-dependent healthy cells are arrested in the G1 phase after exposure using, for example, form B of isolated compound 2 described herein, to mitigate DNA damage. In one embodiment, the compound is administered at least 1 / 2 hour, at least 1 hour, at least 2 hours, at least 3 hours, at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 10 hours, at least 12 hours, at least 14 hours, at least 16 hours, at least 18 hours, at least 20 hours or more after exposure to the chemotherapeutic agent.

[0099] In one embodiment, form B of isolated compound 2 can enable dose intensification (e.g., allowing more therapy to be administered over a period of time) in medical-related chemotherapy, which leads to better efficacy. Therefore, the method of the present disclosure can result in a less toxic and more effective chemotherapy regimen.

[0100] In some embodiments, the use of form B of isolated compound 2 described herein can reduce or substantially eliminate off-target effects related to the inhibition of kinases other than CDK4 and / or CDK6, such as CDK2. Furthermore, in certain embodiments, the use of form B of isolated compound 2 described herein does not induce cell cycle arrest in CDK4 / 6 replication-independent cells.

[0101] In some embodiments, the use of form B of isolated compound 2 described herein reduces the risk of undesirable off-target effects, including but not limited to long-term toxicity, antioxidant effects, and estrogen-like effects. Antioxidant effects can be determined by standard assays known in the art. For example, compounds with no significant antioxidant effect are those that do not significantly remove free radicals such as oxygen radicals. The antioxidant effect of a compound can be compared to known antioxidant compounds such as genistein. Therefore, compounds with no significant antioxidant activity may have antioxidant activity that is about 2, 3, 5, 10, 30, or 100 times less than genistein. Estrogen-like activity can also be determined by known assays. For example, non-estrogen compounds are those that do not significantly bind to or activate estrogen receptors. Compounds with substantially no estrogen-like effect may have estrogen-like activity that is about 2, 3, 5, 10, 20, or 100 times less than compounds with estrogen-like activity, such as genistein.

[0102] In alternative embodiments, form A or form D of compound 2 is administered in a dose effective in reducing the toxic effects of chemotherapeutic agents (typically DNA damaging agents) on healthy cells that are CDK4 / 6 replication-dependent, such as hematopoietic stem cells and hematopoietic progenitor cells (both referred to as HSPCs) and / or renal epithelial cells, in subjects, typically humans, who are scheduled to be exposed to, have been exposed to, or have been exposed to chemotherapeutic agents.

[0103] In alternative embodiments, form C, form E, form G, or form H of compound 2 is administered in a dose effective in reducing the toxic effects of chemotherapeutic agents (typically DNA damaging agents) on healthy cells that are CDK4 / 6 replication-dependent, such as hematopoietic stem cells and hematopoietic progenitor cells (both referred to as HSPCs) and / or renal epithelial cells, in subjects, typically humans, who are scheduled to be exposed to, have been exposed to, or have been exposed to chemotherapeutic agents.

[0104] Methods for treating abnormal proliferation of T cells, B cells, and / or NK cells. In certain embodiments, the present invention comprises the use of an effective amount of isolated compound 2 form B, or a pharmaceutically acceptable salt, prodrug, or isotopic variant thereof, in an optionally pharmaceutically acceptable amount in a pharmaceutical composition for the treatment of a host, typically human, having selected cancer, tumor, hyperproliferative condition, or inflammatory or immunodeficiency. Compound 2 form B is also highly active against T cell proliferation. Given the scarcity of drugs for T cell carcinoma and hyperproliferation, such use would represent a significant improvement in the pharmacotherapy of these diseases.

[0105] Abnormal proliferation of T cells, B cells, and / or NK cells can lead to a wide range of diseases, including cancer, proliferative disorders, and inflammatory / immune diseases. Treating a host suffering from any of these disorders, such as a human, with an effective amount of form B of isolated compound 2 can achieve symptom reduction (palliative agent) or reduction of the underlying disease (disease modifying agent).

[0106] Examples include T-cell or NK-cell lymphomas, such as, but not limited to, peripheral T-cell lymphoma; anaplastic large cell lymphomas, such as anaplastic lymphoma kinase (ALK)-positive, ALK-negative, or primary cutaneous anaplastic large cell lymphoma; angioimmunoblastic lymphoma; cutaneous T-cell lymphomas, such as mycosis fungoides, Sézary syndrome, primary cutaneous anaplastic large cell lymphoma, primary cutaneous CD30+ T-cell lymphoproliferative disorder; primary cutaneous progressive epidermotropic CD8+ cytotoxic T-cell lymphoma; primary cutaneous γ-δ T-cell lymphoma; primary cutaneous small / medium cell CD4+ T-cell lymphoma, and lymphomatoid papulosis; and adult T-cell leukemia. Disease / lymphoma (ATLL); blastic NK cell lymphoma; enteropathy-type T cell lymphoma; hepatosplenic γ-δ T cell lymphoma; lymphoblastic lymphoma; nasal NK / T cell lymphoma; treatment-related T cell lymphoma; for example, lymphoma occurring after solid organ or bone marrow transplantation; pre-T cell lymphocytic leukemia; T cell macrogranule lymphocytic leukemia; chronic lymphoproliferative disorder of NK cells; rapidly progressive NK cell leukemia; systemic EBV+ T cell lymphoproliferative disorder in children (associated with chronic active EBV infection); vaccinia-like varicella-like lymphoma; adult T cell leukemia / lymphoma; enteropathy-associated T cell lymphoma; hepatosplenic T cell lymphoma; or subcutaneous panniculitis-like T cell lymphoma.

[0107] In one embodiment, form B of isolated compound 2 disclosed herein, or its salt, prodrug, or isotopic variant, can be used in effective amounts to treat a host, such as a human, with lymphoma or lymphocytic or myeloid proliferative disorders or abnormalities. For example, form B of isolated compound 2 described herein can be administered to a host suffering from Hodgkin lymphoma or non-Hodgkin lymphoma. For example, the host may have, but is not limited to, AIDS-related lymphoma; anaplastic large cell lymphoma; angioimmunoblastic lymphoma; blastic NK cell lymphoma; Burkitt lymphoma; Burkitt-like lymphoma (small, non-incisional nuclear cell lymphoma); chronic lymphocytic leukemia / small lymphocytic lymphoma; cutaneous T-cell lymphoma; diffuse large B-cell lymphoma; enteropathy-type T-cell lymphoma; follicular lymphoma; hepatosplenic γ-δ T-cell lymphoma; lymphoblastic lymphoma; mantle cell lymphoma; marginal zone lymphoma; nasal T-cell lymphoma; pediatric lymphoma; peripheral T-cell lymphoma; primary central nervous system lymphoma; T-cell leukemia; transforming lymphoma; treatment-related T-cell lymphoma; or non-Hodgkin lymphoma such as Waldenström macroglobulinemia.

[0108] Alternatively, form B of isolated compound 2 disclosed herein, or its salts, prodrugs, or isotopic variants, may be used in effective amounts to treat hosts, such as humans, who have Hodgkin lymphomas including tuberous sclerosis classical Hodgkin lymphoma (CHL); mixed cell type CHL; lymphopenic CHL; lymphocyte-rich CHL; lymphocyte-dominant Hodgkin lymphoma; or nodular lymphocyte-dominant HL.

[0109] Alternatively, the isolated compound 2 form B disclosed herein, or its salts, prodrugs, or isotopic variants, may be used to treat, but are not limited to, multiple myeloma; diffuse large B-cell lymphoma; follicular lymphoma; mucosa-associated lymphoid tissue lymphoma (MALT); small cell lymphocytic lymphoma; mediastinal large B-cell lymphoma; nodal marginal zone B-cell lymphoma (NMZL); splenic marginal zone lymphoma (SMZL); intravascular large B-cell lymphoma; primary exudative lymphoma; or lymphomatoid granulomatosis; B-cell prelymphocytic leukemia; hairy cell leukemia; unclassifiable splenic lymphoma / leukemia; diffuse red pulp small B-cell lymphoma; hairy cell leukemia subtype; lymphoplasmacytic lymphoma; heavy chain disease, e.g., α-heavy chain disease, γ-heavy chain disease, μ-heavy chain disease; plasmacytic myeloma; and solitary phenotype of bone. It can be used in effective doses to treat specific B-cell lymphomas or proliferative disorders in hosts, such as humans, including cell tumors; extraskeletal plasmacytomas; primary cutaneous follicular lymphomas; T-cell / histiocyte-rich large B-cell lymphomas; DLBCL associated with chronic inflammation; Epstein-Barr virus (EBV) + DLBCL in the elderly; primary mediastinal (thymic) large B-cell lymphoma; primary cutaneous DLBCL lower extremity type; ALK + large B-cell lymphoma; plasmablastic lymphoma; large B-cell lymphoma occurring in HHV8-associated multicentric Castleman disease; unclassifiable B-cell lymphoma with intermediate characteristics between diffuse large B-cell lymphoma; or unclassifiable B-cell lymphoma with intermediate characteristics between diffuse large B-cell lymphoma and classical Hodgkin lymphoma.

[0110] In one embodiment, form B of isolated compound 2 disclosed herein, or its salt, prodrug, or isotopic variant, can be used in an effective amount to treat a host having leukemia, such as a human. For example, the host may have, but is not limited to, acute lymphoblastic leukemia (ALL); acute myeloid leukemia (AML); chronic lymphocytic leukemia (CLL); chronic myeloid leukemia (CML); juvenile myelomonocytic leukemia (JMML); hairy cell leukemia (HCL); acute promyelocytic leukemia (a subtype of AML); macrogranular lymphocytic leukemia; or acute or chronic leukemia of lymphocyte or myeloid origin, such as adult T-cell chronic leukemia. In one embodiment, the patient suffers from acute myeloid leukemia, such as undifferentiated AML (M0); myeloblastic leukemia (M1; with / without minimal cellular maturation); myeloblastic leukemia (M2; with cellular maturation); promyelocytic leukemia (M3 or subtype M3 (M3V)); myelomonocytic leukemia (M4 or subtype M4 with eosinophilia (M4E)); monocytic leukemia (M5); erythroleukemia (M6); or megakaryoblastic leukemia (M7).

[0111] In alternative embodiments, form A or form D of compound 2 is administered in a dose effective to treat a host, typically human, with selected cancer, tumor, hyperproliferative condition, or inflammatory or immunocompromised disorder. Given the shortage of drugs for T-cell cancer and hyperproliferative disorders, confirmation of such use would represent a substantial improvement in medical treatment for these diseases.

[0112] In alternative embodiments, form C, form E, form G, or form H of compound 2 is administered in a dose effective to treat a host, typically human, with selected cancer, tumor, hyperproliferative condition, or inflammatory or immunocompromised disorder. Given the shortage of drugs for T-cell cancer and hyperproliferative disorders, confirmation of such use represents a substantial improvement in medical treatment for these diseases.

[0113] Pharmaceutical compositions and dosage forms Form B of isolated compound 2 described herein, or alternative salts, isotopic analogs, or prodrugs, may be administered to a host in an effective dose to treat any of the disorders described herein using any preferred approach to achieve the desired therapeutic outcome. The amount and timing of administration of form B of isolated compound 2 will naturally depend on the host being treated, the instructions of the attending physician, the time course of exposure, the method of administration, the pharmacokinetic properties of the specific active compound, and the judgment of the prescribing physician. Therefore, the following dosages are guidelines due to inter-host variability, and the physician can determine the dosage of the compound and achieve the treatment that the physician deems appropriate for the host. In considering the desired degree of treatment, the physician can balance various factors such as the host's age and weight, the presence of pre-existing diseases, and the presence of other diseases.

[0114] Pharmaceutical compositions can be formulated in any pharmaceutically useful form, such as pills, capsules, tablets, transdermal patches, subcutaneous patches, dry powders, inhalation formulations in medical devices, suppositories, or oral or sublingual formulations. Some dosage forms, such as tablets and capsules, can be divided into appropriate-sized unit doses containing an appropriate amount of the active ingredient, for example, an effective amount to achieve the desired purpose.

[0115] The therapeutically effective dose of form B of isolated compound 2 described herein is determined by a healthcare professional according to the patient's condition, size and age, and route of delivery. In one non-limiting embodiment, doses ranging from about 0.1 mg / kg to about 200 mg / kg are therapeutically effective, with all weights calculated based on the weight of the active compound. In some embodiments, the dose may be the amount of form B of isolated compound 2 required to provide the active compound at serum concentrations of up to about 10 nM, 50 nM, 100 nM, 200 nM, 300 nM, 400 nM, 500 nM, 600 nM, 700 nM, 800 nM, 900 nM, 1 μM, 5 μM, 10 μM, 20 μM, 30 μM, or 40 μM.

[0116] In certain embodiments, the pharmaceutical composition is in a dosage form containing about 0.1 mg to about 2000 mg, about 10 mg to about 1000 mg, about 100 mg to about 800 mg, or about 200 mg to about 600 mg of the active compound in a unit dosage form, and optionally about 0.1 mg to about 2000 mg, about 10 mg to about 1000 mg, about 100 mg to about 800 mg, or about 200 mg to about 600 mg of form B of isolated compound 2, which is measured as either the active compound or a salt thereof. Examples of dosage forms include at least 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 50 mg, 100 mg, 200 mg, 250 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, or 750 mg of the active compound or a salt thereof. The pharmaceutical composition may also include form B of isolated compound 2 and an additional active agent in a molar ratio that achieves the desired result.

[0117] Form B of isolated compound 2 disclosed herein or used as described herein may be administered orally, topically, parenterally, by inhalation or spray, sublingually, by implant including an ocular implant, percutaneously, buccally, rectally, intramuscularly, by inhalation, intra-aortal, intracranial, subdermally, intraperitoneally, subcutaneously, nasally, sublingually or rectally, or by other means in a dosage unit formulation containing a conventional pharmaceutically acceptable carrier.

[0118] According to the method of this disclosure, oral administration may be in any desired form of isolated compound 2, form B, which is stable as a solid. In certain embodiments, isolated compound 2, form B, is delivered as solid particles or nanoparticles. When administered by inhalation, isolated compound 2, form B, may exist in the form of a plurality of solid particles or droplets of any desired particle size, for example, about 0.01 microns, 0.1 microns, or 0.5 microns to about 5 microns, 10 microns, 20 microns, or more, and optionally about 1 micron to about 2 microns. Isolated compound 2, form B, as disclosed in this invention, has good pharmacokinetic and pharmacokinetic properties when administered, for example, by an oral or intravenous route.

[0119] The pharmaceutical formulation may contain, in any pharmaceutically acceptable carrier, form B of isolated compound 2 as described herein or an alternative pharmaceutically acceptable salt thereof.

[0120] The carrier, containing excipients and diluents, must be sufficiently pure and sufficiently low in toxicity to be suitable for administration to the patient being treated. The carrier may be inert or may have its own pharmaceutically active properties. The amount of carrier used in combination with the compound is sufficient to provide a practical amount of material for administration per unit dose of the compound.

[0121] Examples of carriers include, but are not limited to, binders, buffers, colorants, diluents, disintegrants, emulsifiers, flavorings, lubricants, preservatives, stabilizers, surfactants, tableting agents, and wetting agents. Some carriers may be of two or more types; for example, vegetable oil can be used as a lubricant in some formulations and as a diluent in others. Exemplary pharmaceutically acceptable carriers include sugars, starches, cellulose, tragacanth powder, malt, gelatin; talc, and vegetable oils. Any active agent that does not substantially interfere with the activity of the compounds of the present invention may be included in the pharmaceutical composition.

[0122] Depending on the intended method of administration, the pharmaceutical composition may be in a solid or semi-solid dosage form in which form B of compound 2 isolated therein is stable, such as a tablet, suppository, pill, capsule, or powder, preferably a unit dosage form suitable for a precise single dose. The composition may contain an effective amount of the selected drug in combination with a pharmaceutically acceptable carrier, and may also contain other pharmaceuticals, adjuvants, diluents, buffers, etc.

[0123] Therefore, the compositions of this disclosure can be administered as pharmaceutical formulations suitable for oral (including buccal and sublingual), rectal, nasal, local, pulmonary, and vaginal administration, or in forms suitable for inhalation or inhalation. A preferred method of administration is oral administration using a simple daily dosing regimen that can be adjusted according to the degree of discomfort. Examples of conventional non-toxic solid carriers for the solid compositions include pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, and magnesium carbonate.

[0124] In another embodiment, permeation-promoting excipients are used that include polycations (chitosan and its quaternary ammonium derivatives, poly-L-arginine, amination gelatin); polyanions (N-carboxymethyl chitosan, poly-acrylic acid); and polymers such as thiolated polymers (carboxymethylcellulose-cysteine, polycarbophil-cysteine, chitosan-thiobutylamidine, chitosan-thioglycolic acid, chitosan-glutathione conjugate).

[0125] For oral administration, the compositions generally take the form of tablets or capsules. Tablets and capsules are preferred forms of oral administration. Tablets and capsules for oral use may contain one or more commonly used carriers, such as lactose and corn starch. Lubricants, such as magnesium stearate, are also typically added. Typically, the compositions of this disclosure can be combined with non-toxic, pharmaceutically acceptable, inert carriers for oral use, such as lactose, starch, sucrose, glucose, methylcellulose, magnesium stearate, dicalcium phosphate, calcium sulfate, mannitol, sorbitol, etc. Furthermore, suitable binders, lubricants, disintegrants, and colorants may be incorporated into the mixture as desired or required. Suitable binders include natural sugars such as starch, gelatin, glucose, or β-lactose, corn sweeteners, natural and synthetic gums such as gum arabic and tragacanth, or sodium alginate, carboxymethylcellulose, polyethylene glycol, wax, etc. Lubricants used in these drug formulations include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, and sodium chloride. Disintegrants, though not limited to these, include starch, methylcellulose, agar, bentonite, and xanthan gum.

[0126] In addition to the active compound or its salts, the pharmaceutical formulation may contain other additives such as pH adjusting additives. Particularly useful pH adjusting agents include acids such as hydrochloric acid, and bases or buffers such as sodium lactate, sodium acetate, sodium phosphate, sodium citrate, sodium borate, or sodium gluconate. Furthermore, the formulation may contain antimicrobial preservatives. Useful antimicrobial preservatives include methylparaben, propylparaben, and benzyl alcohol. Antimicrobial preservatives are typically used when the formulation is placed in a vial designed for multiple doses. The pharmaceutical formulations described herein can be freeze-dried using techniques known in the art.

[0127] For oral administration, pharmaceutical compositions can take the form of tablets, pills, capsules, powders, etc. Tablets containing various excipients such as sodium citrate, calcium carbonate, and calcium phosphate can be used in combination with binders such as polyvinylpyrrolidone, sucrose, gelatin, and gum arabic, along with various disintegrants such as starch (e.g., potato or tapioca starch) and certain complex silicates. Additionally, lubricants such as magnesium stearate, sodium lauryl sulfate, and talc are often extremely useful for tablet formation. Similar types of solid compositions can be used as fillers in soft and hard gelatin capsules.

[0128] For example, pharmaceutical formulations are also provided that result in the controlled release of the compounds described herein by the use of biodegradable polymers known in the art.

[0129] As used herein, the term “pharmaceutically acceptable salt” refers to a salt that is suitable for use in contact with a host (e.g., a human host) without excessive toxicity, irritation, allergic response, etc., within the bounds of sound medical judgment, that is effective for such use and that has a reasonable benefit-risk ratio, and, if possible, a zwitterionic form of the compound of the subject matter of this disclosure.

[0130] In an alternative embodiment, form B of compound 2 is not the HCl salt, but rather the salt described below.

[0131] In one embodiment, the further therapeutic agents described in the following combination section are administered as pharmaceutically acceptable salts, for example, the salts described below.

[0132] Therefore, the term “salt” refers to relatively non-toxic inorganic and organic acid addition salts of the compounds of this disclosure. These salts can be prepared during the final isolation and purification of the compounds, or by reacting the purified compound in its free base form separately with a suitable organic or inorganic acid and isolating the salt formed thereby. Basic compounds can form a wide range of salts with various inorganic and organic acids. Acid addition salts of basic compounds are prepared by conventional methods by contacting the free base form with a sufficient amount of the desired acid to produce a salt. The free base form can be regenerated by conventional methods by contacting the salt form with a base and isolating the free base. The free base form may differ from their respective salt forms in certain physical properties, such as solubility in polar solvents. Pharmaceutically acceptable base addition salts can be formed with metals or amines, such as alkali and alkaline earth metal hydroxides, or organic amines. Examples of metals used as cations include, but are not limited to, sodium, potassium, magnesium, and calcium. Suitable amines include, but are not limited to, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine, and procaine. Base addition salts of acidic compounds are prepared by conventional methods by contacting the free acid form with a sufficient amount of the desired base to produce a salt. The free acid form can be regenerated by contacting the salt form with an acid and isolating the free acid by conventional methods. The free acid forms may differ somewhat from their respective salt forms in certain physical properties, such as solubility in polar solvents.

[0133] Salts can be prepared from inorganic acids such as sulfate ions, pyrosulfate ions, bisulfate ions, sulfite ions, bisulfite ions, nitrate ions, phosphate ions, monohydrogen phosphate ions, dihydrogen phosphate ions, metaphosphate ions, pyrophosphate ions, chloride ions, bromide ions, and iodide ions, such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, and phosphorous acid. Typical salts include hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, laurylsulfonate, and isethionate. Salts can also be prepared from organic acids, such as aliphatic monocarboxylic acids and dicarboxylic acids, phenyl-substituted alkanes, hydroxyalkanoates, alkanedioates, aromatic acids, aliphatic and aromatic sulfonic acids, etc. Typical salts include acetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberinate, sebacinate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate, and methanesulfonate. Pharmaceutically acceptable salts include, but are not limited to, cations based on alkalis and alkaline earth metals such as sodium, lithium, potassium, calcium, and magnesium, as well as ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, and ethylamine, and may also include non-toxic ammonium, quaternary ammonium, and amine cations. Salts of amino acids such as arginates, glucons, and galacturons are also considered. See, for example, Berge et al., J. Pharm. Sci., 1977, 66, 1-19 (which is incorporated herein by reference).

[0134] Formulations suitable for rectal administration are typically provided as unit-dose suppositories. These can be prepared by mixing the disclosed active compound with one or more conventional solid carriers, such as cocoa butter, and then molding the resulting mixture.

[0135] Formulations suitable for topical application to the skin are preferably in the form of an ointment, cream, lotion, paste, gel, spray, aerosol, or oil that stably retains form B of the isolated compound 2. Suitable carriers include petrolatum, lanolin, polyethylene glycol, alcohol, transdermal enhancers, and combinations of two or more thereof.

[0136] Formulations suitable for transdermal administration can be provided as individual patches adapted to remain in close contact with the recipient's epidermis for extended periods. Formulations suitable for transdermal administration can also be delivered by iontophoresis (see, e.g., Pharmaceutical Research 3 (6):318 (1986)), typically in the form of an optionally buffered aqueous solution of the active compound. In one embodiment, a microneedle patch or device is provided for the delivery of drugs across or into biological tissues, particularly the skin. The microneedle patch or device enables drug delivery across or into the barriers of the skin or other tissues at clinically relevant rates with little or no tissue damage, pain, or irritation.

[0137] In alternative embodiments, form B of compound 2 is an HCl salt, for example, a monohydrochloride salt having about 1 hydrochloric acid unit per unit of compound 2, about 1.5 hydrochloric acid units per unit of compound 2, or about 2 hydrochloric acid units per unit of compound 2.

[0138] In one embodiment, "about" means ±1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or 10%. In one embodiment, form B of compound 2 has about 2 HCl ions per molecule of compound 2.

[0139] Formulations suitable for pulmonary administration can be delivered by various passive-respiratory and power-driven single / multi-dose dry powder inhalers (DPIs). The most commonly used devices for respiratory delivery include nebulizers, metered-dose inhalers, and dry powder inhalers. Several types of nebulizers are available, including jet nebulizers, ultrasonic nebulizers, and vibrating mesh nebulizers. The selection of an appropriate lung delivery device depends on parameters such as the properties of the drug and its formulation, the site of action, and the pathophysiology of the lung.

[0140] Combination therapy Form B of isolated compound 2 can be used alone or in combination with another compound of the present invention or another bioactive agent in an effective amount to treat a host such as a human having the disorder described herein.

[0141] Form B of isolated compound 2 described herein may be used alone or in combination with another compound of the present invention or another bioactive agent in an effective amount to treat a host such as a human having the disorder described herein.

[0142] The term "bioactive agent" is used to describe active substances other than the selected compounds of the present invention that can be used in combination with or alternately with the compounds of the present invention to achieve a desired therapeutic outcome. In one embodiment, the compounds of the present invention and the bioactive agent are administered such that they have overlapping periods during which they are active in vivo, for example, Cmax, Tmax, AUC, or other pharmacokinetic parameters. In another embodiment, isolated compound 2 form B and the bioactive agent, which do not have overlapping pharmacokinetic parameters but one has a therapeutic effect on the therapeutic efficacy of the other, are administered to a host in need.

[0143] In one embodiment of this invention, the bioactive agent is an immunomodulatory agent that includes, but is not limited to, PD-1 inhibitors, PD-L1 inhibitors, PD-L2 inhibitors, CTLA-4 inhibitors, LAG-3 inhibitors, TIM-3 inhibitors, checkpoint inhibitors including V-domain Ig suppressor of T-cell activation (VISTA) inhibitors, small molecules, peptides, nucleotides, or other inhibitors. In a particular embodiment, the immunomodulatory agent is an antibody such as a monoclonal antibody.

[0144] Examples of PD-1 inhibitors that block the interaction between PD-1 and PD-L1 by binding to the PD-1 receptor and thereby inhibit immunosuppression include nivolumab (Opdivo), pembrolizumab (Keytruda), pidilizumab, AMP-224 (AstraZeneca and MedImmune), PF-06801591 (Pfizer), MEDI0680 (AstraZeneca), PDR001 (Novartis), REGN2810 (Regeneron), SHR-12-1 (Jiangsu Hengrui Medicine Company and Incyte Corporation), TSR-042 (Tesaro), and the PD-L1 / VISTA inhibitor CA-170 (Curis Inc.). Examples of PD-L1 inhibitors that inhibit immunosuppression by blocking the interaction between PD-1 and PD-L1 by binding to the PD-L1 receptor include atezolizumab (Tecentriq), durvalumab (AstraZeneca and MedImmune), KN035 (Alphamab), and BMS-936559 (Bristol-Myers Squibb). Examples of CTLA-4 checkpoint inhibitors that inhibit immunosuppression by binding to CTLA-4 include, but are not limited to, ipilimumab, tremelimumab (AstraZeneca and MedImmune), AGEN1884, and AGEN2041 (Agenus). Examples of LAG-3 checkpoint inhibitors include, but are not limited to, BMS-986016 (Bristol-Myers Squibb), GSK2831781 (GlaxoSmithKline), IMP321 (Prima BioMed), LAG525 (Novartis), and MGD013 (MacroGenics), a dual inhibitor of PD-1 and LAG-3. An example of a TIM-3 inhibitor is TSR-022 (Tesaro).

[0145] In another embodiment, form B of isolated compound 2 may be administered in combination with, or alternately with, an effective dose of an estrogen inhibitor, including but not limited to, a SERM (selective estrogen receptor modulator), SERD (selective estrogen receptor degrader), a complete estrogen receptor degrader, or another form of partial or complete estrogen antagonist or agonist, for the treatment of abnormal tissues of the female reproductive system, such as breast cancer, ovarian cancer, endometrial cancer, or uterine cancer. Partial antiestrogens such as raloxifene and tamoxifen retain some estrogen-like effects, including estrogen-like stimulation of uterine growth and, in some cases, estrogen-like effects in progressing breast cancer that actually stimulate tumor growth. In contrast, fulvestrant, a complete antiestrogen, has no estrogen-like effects on the uterus and is effective in tamoxifen-resistant tumors. Non-limiting examples of anti-estrogen compounds are presented in International Publication 2014 / 19176, assigned to Astra Zeneca; International Publication 2013 / 090921, International Publication 2014 / 203129, International Publication 2014 / 203132, and U.S. Patent Application Publication 2013 / 0178445, assigned to Olema Pharmaceuticals; and U.S. Patents 9,078,871, 8,853,423, and 8,703,810, as well as U.S. Patent Application Publication 2015 / 0005286, International Publication 2014 / 205136, and International Publication 2014 / 205138.Additional, non-limiting examples of anti-estrogenic compounds include SERMs such as anoldrin, bazedoxifene, propalestrol, chlorotrianicene, clomiphene citrate, cyclophenyl, rasofoxifene, olmeroxifene, raloxifene, tamoxifene, toremifene, and fulvestrant; aromatase inhibitors such as aminoglutethimide, testolactone, anastrozole, exemestane, fadrozol, formestan, and letrozole; and antigonadotropins such as leuprorelin, cetrorelix, allylestrenol, chlormadinone acetate, cyproterone acetate, dermadinone acetate, dydrogesterone, medroxyprogesterone acetate, megestrol acetate, nomegestrol acetate, norethisterone acetate, progesterone, and spironolactone.Other estrogen ligands that can be used in accordance with the present invention include U.S. Patent Nos. 4,418,068; 5,478,847; 5,393,763; and 5,457,117, International Publication No. 2011 / 156518, U.S. Patent Nos. 8,455,534 and 8,299,112, U.S. Patent Nos. 9,078,871; 8,853,423; 8,703,810; U.S. Patent Application Publication No. 2015 / 0005286; and International Publication No. 2014 / 2 Patent No. 05138, U.S. Patent Application Publication No. 2016 / 0175289, U.S. Patent Application Publication No. 2015 / 0258080, International Publication No. 2014 / 191726, International Publication No. 2012 / 084711; International Publication No. 2002 / 013802; International Publication No. 2002 / 004418; International Publication No. 2002 / 003992; International Publication No. 2002 / 003991; International Publication No. 2002 / 003990; International Publication No. 2002 / 003989; International Publication No. 2002 / 003988 International Publication No. 2002 / 003986; International Publication No. 2002 / 003977; International Publication No. 2002 / 003976; International Publication No. 2002 / 003975; International Publication No. 2006 / 078834; US Patent No. 6821989; US Patent Application Publication No. 2002 / 0128276; US Patent No. 6777424; US Patent Application Publication No. 2002 / 0016340; US Patent No. 6326392; US Patent No. 6756401; US ​​Patent Application Publication No. 2002 / 0013327 It is described in U.S. Patent No. 6512002, U.S. Patent No. 6632834, U.S. Patent Application Publication No. 2001 / 0056099, U.S. Patent No. 6583170, U.S. Patent No. 6479535, International Publication No. 1999 / 024027, U.S. Patent No. 6005102, European Patent No. 0802184, U.S. Patent No. 5998402, U.S. Patent No. 5780497, U.S. Patent No. 5880137, International Publication No. 2012 / 048058 and International Publication No. 2007 / 087684.

[0146] In another embodiment, form B of isolated compound 2 may be administered in combination with or alternately in an effective amount of an androgen (e.g., testosterone) inhibitor, including but not limited to a selective androgen receptor modulator, selective androgen receptor degrader, complete androgen receptor degrader, or another form of partial or complete androgen antagonist, for the treatment of abnormal tissue of the male reproductive system, such as prostate cancer or testicular cancer. In one embodiment, prostate or testicular cancer is androgen resistant. Non-limiting examples of antiandrogen compounds are presented in International Publication No. 2011 / 156518 and U.S. Patents No. 8,455,534 and 8,299,112. Additional non-limiting examples of antiandrogen compounds include enzalutamide, apalutamide, cyproterone acetate, chlormadinone acetate, spironolactone, canrenone, drospirenone, ketoconazole, topirutamide, abiraterone acetate, and cimetidine.

[0147] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of abiraterone acetate (Zytiga) for the treatment of abnormal tissue in the male reproductive system.

[0148] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of abiraterone acetate (Zytiga) for the treatment of prostate cancer.

[0149] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of enzalutamide for the treatment of prostate cancer.

[0150] In one embodiment, the bioactive agent is an ALK inhibitor. Examples of ALK inhibitors include, but are not limited to, crizotinib, alectinib, ceritinib, TAE684 (NVP-TAE684), GSK1838705A, AZD3463, ASP3026, PF-06463922, entrectinib (RXDX-101), and AP26113. In one embodiment, the bioactive agent is an EGFR inhibitor. Examples of EGFR inhibitors include erlotinib (Tarceva), gefitinib (Iressa), afatinib (Gilotrif), rosiletinib (CO-1686), osimertinib (Tagrisso), olmutinib (Olita), nacotinib (ASP8273), nazartinib (EGF816), PF-06747775 (Pfizer), icotinib (BPI-2009), neratinib (HKI-272;PB272); abitinib (AC0010), EA Examples include I045, tarloxotinib (TH-4000;PR-610), PF-06459988 (Pfizer), tesevatinib (XL647;EXEL-7647;KD-019), transtinib, WZ-3146, WZ8040, CNX-2006, dacomitinib (PF-00299804;Pfizer), brigatinib (Alunbrig), lorlatinib, and PF-06747775.

[0151] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of afatinib dimaleate (Gilotrif) for the treatment of non-small cell lung cancer.

[0152] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of alectinib (Alecensa) for the treatment of non-small cell lung cancer.

[0153] In one embodiment, an effective amount of compound 2, form B, is administered in combination with a dose of ceritinib (Zykadia) effective for the treatment of non-small cell lung cancer.

[0154] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of crizotinib (Xalkori) for the treatment of non-small cell lung cancer.

[0155] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of osimertinib (Tagrisso) for the treatment of non-small cell lung cancer.

[0156] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of brigatinib (Alunbrig) for the treatment of non-small cell lung cancer.

[0157] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of lorlatinib for the treatment of non-small cell lung cancer.

[0158] In one embodiment, the bioactive agent is a HER-2 inhibitor. Examples of HER-2 inhibitors include trastuzumab, lapatinib, ado-trastuzumab emtansine, and pertuzumab.

[0159] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of lapatinib ditosylate for the treatment of breast cancer.

[0160] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of lapatinib ditosylate for the treatment of HER2+ breast cancer.

[0161] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of PF7775 for the treatment of non-small cell lung cancer.

[0162] In one embodiment, the bioactive agent is a CD20 inhibitor. Examples of CD20 inhibitors include obinutuzumab, rituximab, ofatumumab, ibritumomab, tocitumomab, and ocrelizumab.

[0163] In one embodiment, the bioactive agent is a JAK3 inhibitor. An example of a JAK3 inhibitor is tasocitinib.

[0164] In one embodiment, the bioactive agent is a BCL-2 inhibitor. Examples of BCL-2 inhibitors include venetoclax, ABT-199 (4-[4-[[2-(4-chlorophenyl)-4,4-dimethylcyclohexa-1-en-1-yl]methyl]piperazine-1-yl]-N-[[3-nitro-4-[[(tetrahydro-2H-pyran-4-yl)methyl]aminophenyl]sulfonyl]-2-[(1H-pyrrolo[2,3-b]pyridine-5-yl)oxy]benzamide), and ABT-737 (4-[4-[[2-(4-chlorophenyl)phenyl]methyl]pi [Perazin-1-yl]-N-[4-[[(2R)-4-(dimethylamino)-1-phenylsulfanylbutan-2-yl]amino]-3-nitrophenyl]sulfonylbenzamide)(navitoclax), ABT-263((R)-4-(4-((4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((4-((4-morpholino-1-(phenylthio)butan-2-yl)amino)-3((tri Fluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide), GX15-070 (Obatoclax mesylate, (2Z)-2-[(5Z)-5-[(3,5-dimethyl-1H-pyrrole-2-yl)methylidene]-4-methoxypyrrole-2-ylidene]indole; methanesulfonic acid), 2-methoxy-antimycin A3, YC137 (4-(4,9-dioxo-4,9-dihydronaphtho[2,3-d]thiazole-2-ylamino)-phenyl ester), pogosin, ethyl 2- Examples include amino-6-bromo-4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate, nilotinib-d3, TW-37 (N-[4-[[2-(1,1-dimethylethyl)phenyl]sulfonyl]phenyl]-2,3,4-trihydroxy-5-[[2-(1-methylethyl)phenyl]methyl]benzamide), apogossiporone (ApoG2), HA14-1, AT101, sabutoclax, gumbognate, or G3139 (oblimersene).

[0165] In one embodiment, a therapeutic regime is provided comprising the administration of form B of isolated compound 2 in combination with at least one additional chemotherapeutic agent. The combinations disclosed herein may be administered for beneficial additive or synergistic effects in the treatment of abnormal cell proliferation disorders.

[0166] In certain embodiments, the treatment regimen comprises administration of form B of isolated compound 2 in combination with at least one kinase inhibitor. In one embodiment, the at least one kinase inhibitor is selected from a phosphoinositide 3-kinase (PI3K) inhibitor, a Bruton's tyrosine kinase (BTK) inhibitor, a spleen tyrosine kinase (Syk) inhibitor, or a combination thereof.

[0167] The PI3k inhibitors that can be used in the present invention are known. Examples of PI3 kinase inhibitors include wartmannin, demethoxypyridine, perifosine, idelalisib, pictilisib, Palomid 529, ZSTK474, PWT33597, CUDC-907 and AEZS-136, duvelisib, GS-9820, BKM120, GDC-0032 (taselisib), (2-[4-[2-(2-isopropyl-5-methyl-1,2,4-triazole-3-yl)-5,6-dihydroimidazo[1,2-d][1,4]benzoxazepine-9- [Iyl]pyrazole-1-yl]-2-methylpropanamide), MLN-1117 ((2R)-1-phenoxy-2-butanyl hydrogen (S)-methylphosphonate; or methyl(oxo){[(2R)-1-phenoxy-2-butanyl]oxy}phosphonium)), BYL-719 ((2S)-N1-[4-methyl-5-[2-(2,2,2-trifluoro-1,1-dimethyl (Tyl)-4-pyridinyl]-2-thiazolyl]-1,2-pyrrolidinedicarboxamide), GSK2126458 (2,4-difluoro-N-{2-(methyloxy)-5-[4-(4-pyridazinyl)-6-quinolinyl]-3-pyridinyl}benzenesulfonamide) (omiparisib), TGX-221 ((±)-7-methyl-2-(morpholin-4-yl)-9-(1- Phenylaminoethyl)-pyrido[1,2-a]-pyrimidine-4-one), GSK2636771(2-methyl-1-(2-methyl-3-(trifluoromethyl)benzyl)-6-morpholino-1H-benzo[d]imidazole-4-carboxylate dihydrochloride), KIN-193((R)-2-((1-(7-methyl-2-morpholino-4-oxo-4H-pyrido[1,2-a]Pyrimidine-9-yl(ethyl)amino(benzoic acid), TGR-1202 / RP5264, GS-9820((S)-1-(4-((2-(2-aminopyrimidine-5-yl)-7-methyl-4-mohydroxypropane(mohydroxypropan)-1-one), GS-1101(5-fluoro-3-phenyl-2-([S)]-1-[9H-purine-6-ylamino]-propyl)-3H-quinazoline- 4-one), AMG-319, GSK-2269557, SAR245409 (N-(4-(N-(3-((3,5-dimethoxyphenyl)amino)quinoxaline-2-yl)sulfamoyl)phenyl)-3-methoxy-4 methylbenzamide), BAY80-6946 (2-amino-N-(7-methoxy-8-(3-morpholinopropoxy)-2,3-dihydroimidazo[1,2-c]quinaz), AS 252424 (5-[1-[5-(4-fluoro-2-hydroxyphenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidined-2,4-dione), CZ 24832 (5-(2-amino-8-fluoro-[1,2,4]triazolo[1,5-a]pyridin-6-yl)-N-tert-butylpyridine-3-sulfonamide), Buparlicib (5-[2,6-di(4-morpholinyl)-4-pyrimidinyl]-4-(trifluoromethyl)-2-pyridinamine), GDC-0941 (2-(1H-indazole-4-yl)-6-[[4-(methylsulfonyl)-1-piperazinyl]methyl]-4-(4-morpholinyl)thieno[3,2-d]pyrimidine), GDC-0980 ((S)-1-(4-( (2-(2-aminopyrimidine-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidine-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one (also known as RG7422)), SF1126((8S,14S,17S)-14-(carboxymethyl)-8-(3-guanidinopropyl)-17-(hydroxymethyl)-3,6,9,12,15-pentaoxo-1-(4-(4-oxo-8-phenyl-4H-chromen-2-yl)morpholino-4-ium)-2-oxa-7,10,13,16-Tetraazaoctadecane-18-Oate), PF-05212384 (N-[4-[[4-(dimethylamino)-1-piperidinyl]carbonyl]phenyl]-N'-[4-(4,6-di-4-morpholinyl-1,3,5-triazine-2-yl)phenyl]urea) (dactolisib), LY3023414, BEZ235 (2-methyl-2-{4-[3-methyl-2-oxo-8-(quinoline-3-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinoline-1-yl]phenyl}propannitrile) (dactolisib), XL-765 (N -(3-(N-(3-(3,5-dimethoxyphenylamino)quinoxaline-2-yl)sulfamoyl)phenyl)-3-methoxy-4-methylbenzamide) and GSK1059615(5-[[4-(4-pyridinyl)-6-quinolinyl]methylene]-2,4-thiazolidinedione), PX886([(3aR,6E,9S,9aR,10R,11aS)-6-[[bis(propa-2-enyl)amino]methylidene]-5-hydroxy-9-(methoxymethyl)-9a,11a-dimethyl-1,4,7-trioxo-2,3,3a,9,10,11-hexahydroindeno[4,5h]Isochromen-10-yl acetate (also known as sonolisib), LY294002, AZD8186, PF-4989216, pilaralisib, GNE-317, PI-3065, PI-103, NU7441 (KU-57788), HS 173, VS-5584 (SB2343), CZC24832, TG100-115, A66, YM201636, CAY10505, PIK-75, PIK-93, AS-605240, BGT226 (NVP-BGT226), AZD6482, voxtalisib, alpericib, IC-87114, TGI100713, CH5132799, PKI-402, copanlicib (BAY 80-6946), XL Examples include, but are not limited to, 147, PIK-90, PIK-293, PIK-294, 3-MA (3-methyladenine), AS-252424, AS-604850, apitricib (GDC-0980;RG7422), and the structures described in International Publication No. 2014 / 071109. In one embodiment, form B of isolated compound 2 is combined with a PIk3 inhibitor in a single-dose form.

[0168] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of alpelisib for the treatment of solid tumors.

[0169] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of alpelisib for the treatment of abnormal tissue in the female reproductive system.

[0170] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of alpelisib for the treatment of breast cancer.

[0171] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of copanlisib hydrochloride (Aliqopa) for the treatment of lymphoma.

[0172] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of copanlisib hydrochloride (Aliqopa) for the treatment of follicular lymphoma.

[0173] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of idelalisib (Zydelig) for the treatment of chronic lymphocytic leukemia.

[0174] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of idelalisib (Zydelig) for the treatment of non-Hodgkin lymphoma, including follicular B-cell non-Hodgkin lymphoma or small lymphocytic lymphoma.

[0175] The BTK inhibitors used in this invention are known. Examples of BTK inhibitors include ibrutinib (also known as PCI-32765) (Imbruvica®) (1-[(3R)-3-[4-amino-3-(4-phenoxyphenyl)pyrazolo[3,4-d]pyrimidine-1-yl]piperidine-1-yl]propa-2-en-1-one), dianilinopyrimidine inhibitors, such as AVL-101 and AVL-291 / 292 (N-(3-((5-fluoro-2-((4-(2-methoxyethoxy)phenyl)amino)pyrimidine-4-yl)amino)phenyl)acrylamide) (Avila Therapeutics) (see U.S. Patent Application Publication No. 2011 / 0117073, which in whole forms part of this specification), dasatinib (N-(2-chloro-6-methylphenyl)-2-(6-(4-(2-hydroxyethyl)piperazin-1-yl)-2-methylpyrimidine-4-ylamino)thiazole-5-carboxamide), LFM-A13 (α-cyano-β -Hydroxy-β-methyl-N-(2,5-dibromophenyl)propenamide), GDC-0834(RN-(3-(6-(4-(1,4-dimethyl-3-oxopiperazine-2-yl)phenylamino)-4-methyl-5-oxo-4,5-dihydropyrazine-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide), CGI-560 4-(tert-butyl)-N-(3-(8-(phenylamino)imidazo[1,2-a]pyrazine-6-yl)phenyl)benzamide, CGI-1746(4-(tert-butyl)-N-(2-methyl-3-(4-methyl-6-((4-(morpholine-4-carbonyl)phenyl)amino)-5-oxo-4,5-dihydropyrazine-2-yl)phenyl)benzamide), CNX-774(4-(4-((4-((3-acrylamidophenyl)amino)-5-fluoropyrimidine-2-yl)amino)phenoxy)-N-methylpicolinamide), CTA056(7-benzyl-1-(3-(piperidine-1-yl)propyl)-2-(4-(pyridine-4-yl)phenyl)-1H-imidazo[4,5-g]Quinoxaline-6(5H)-one), GDC-0834((R)-N-(3-(6-((4-(1,4-dimethyl-3-oxopiperazine-2-yl)phenyl)amino)-4-methyl-5-oxo-4,5-dihydropyrazine-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide), GDC- 0837((R)-N-(3-(6-((4-(1,4-dimethyl-3-oxopiperazine-2-yl)phenyl)amino)-4-methyl-5-oxo-4,5-dihydropyrazine-2-yl)-2-methylphenyl)-4,5,6,7-tetrahydrobenzo[b]thiophene-2-carboxamide), HM-71224, ACP-196, ONO-4059(Ono Pharmaceuticals), PRT062607 (4-((3-(2H-1,2,3-triazole-2-yl)phenyl)amino)-2-(((1R,2S)-2-aminocyclohexyl)amino)pyrimidine-5-carboxamide hydrochloride), QL-47 (1-(1-acryloylindorin-6-yl)-9-(1-methyl-1H-pyrazole-4-yl)benzo[h][1,6]naphthili Examples include dzin-2(1H)-one) and RN486(6-cyclopropyl-8-fluoro-2-(2-hydroxymethyl-3-{1-methyl-5-[5-(4-methyl-piperazin-1-yl)-pyridine-2-ylamino]-6-oxo-1,6-dihydropyridine-3-yl}phenyl)-2H-isoquinoline-1-one), as well as other molecules capable of inhibiting BTK activity, such as BTK inhibitors disclosed in Akinleye et al., Journal of Hematology & Oncology, 2013, 6:59 (which in whole constitutes part of this specification by reference). In one embodiment, an effective amount of isolated compound 2, form B, is combined with a BTK inhibitor in a single dosage form.

[0176] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of ibrutinib (Imbruvica) for the treatment of chronic lymphocytic leukemia.

[0177] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of ibrutinib (Imbruvica) for the treatment of lymphomas including small lymphocytic lymphoma, mantle cell lymphoma, marginal zone lymphoma, or Waldenström macroglobulinemia.

[0178] The Syk inhibitors used in the present invention are known, for example, cerdulatinib (4-(cyclopropylamino)-2-((4-(4-(ethylsulfonyl)piperazine-1-yl)phenyl)aminopyrimidine-5-carboxamide), entospletinib (6-(1H-indazole-6-yl)-N-(4-morpholinophenyl)imidazo[1,2-a]pyrazine-8-amine), fostamatinib ([6-({5-fluoro-2-[(3,4,5- Trimethoxyphenyl)amino]-4-pyrimidinyl}amino)-2,2-dimethyl-3-oxo-2,3-dihydro-4H-pyrido[3,2-b][1,4]oxazine-4-yl]methyl dihydrogen phosphate), fostamatinib disodium salt (sodium (6-((5-fluoro-2-((3,4,5-trimethoxyphenyl)amino)pyrimidin-4-yl)amino)-2,2-dimethyl-3-oxo-2H-pyrido[3,2-b][1,4]oxazine-4(3H)-yl)methyl phosphate), BAY 61-3606 (2-(7-(3,4-dimethoxyphenyl)-imidazo[1,2-c]pyrimidine-5-ylamino)-nicotinamide HCl), RO9021 (6-[(1R,2S)-2-amino-cyclohexylamino]-4-(5,6-dimethyl-pyridine-2-ylamino)-pyridazine-3-carboxylic acid amide), Imatinib (Gleevac; 4-[(4-methylpiperazine-1-yl)methyl]-N-(4-methyl-3-{[4-(pyridine-3-yl)pyrimidine-2-yl]amino}phenyl)benzamide), Staurosporine, GSK143 (2-(((3R,4R)-3-aminotetrahydro-2H-pyran-4-yl)amino)-4-(p-tolylamino)pyrimidine-5-carboxamide), PP2(1-(tert-butyl)-3-(4-chlorophenyl)-1H-pyrazolo[3,4-d]pyrimidine-4-amine), PRT-060318(2-(((1R,2S)-2-aminocyclohexyl)amino)-4-(m-tolylamino)pyrimidine-5-carboxamide), PRT-062607(4-((3-(2H-1,2,3-triazole-2-yl)phenyl)amino)-2-(((1R,R112(3,3'-((5-fluoropyrimidine-2,4-diyl)bis(azandiyl))diphenol), R348(3-ethyl-4-methylpyridine), R406(6-((5-fluoro-2-((3,4,5-trimethoxyphenyl)amino)pyrimidine-4-yl)amino)-2,2-dimethyl-2H-pyrido[3,2-b][1,4]oxazine-3(4H)-one), piceatannol(3-hydroxyresveratrol), YM193306(Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, See 3614-3643), 7-azaindole, piceatannol, ER-27319 (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643 (the entire text is incorporated herein by reference)), Compound D (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643 (the entire text is incorporated herein by reference)), PRT060318 (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, See 3614-3643 (the entirety of which constitutes part of this specification by reference), luteolin (Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55,See 3614-3643 (which in whole form constitutes part of this specification by reference), apigenin (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643 (which in whole form constitutes part of this specification by reference)), quercetin (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643 (which in whole form constitutes part of this specification by reference)), fisetin (see Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, Examples include 3614-3643 (see reference to the entire text which forms part of this specification), myricetin (see reference to Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643 (see reference to the entire text which forms part of this specification)), and morin (see reference to Singh et al. Discovery and Development of Spleen Tyrosine Kinase (SYK) Inhibitors, J. Med. Chem. 2012, 55, 3614-3643 (see reference to the entire text which forms part of this specification)). In one embodiment, an effective amount of isolated compound 2, form B, is combined with a Syk inhibitor in a single dosage form.

[0179] In one embodiment, at least one additional chemotherapeutic agent is a protein death-1 (PD-1) inhibitor. PD-1 inhibitors are known in the art and include, for example, nivolumab (BMS), pembrolizumab (Merck), pidilizumab (CureTech / Teva), AMP-244 (Amplimmune / GSK), BMS-936559 (BMS), and MEDI4736 (Roche / Genentech). In one embodiment, an effective amount of form B of isolated compound 2 is combined with a PIk3 inhibitor in a single-dose form.

[0180] In alternative embodiments, form A or form D of isolated compound 2 may be used alone or in combination with another compound or pharmacological agent of the present invention in an amount effective to treat a host such as a human having a disorder as described herein.

[0181] In one embodiment, at least one additional chemotherapeutic agent is a B-cell lymphoma 2 (Bcl-2) protein inhibitor. BCL-2 inhibitors are known in the art, for example, ABT-199 (4-[4-[[2-(4-chlorophenyl)-4,4-dimethylcyclohexa-1-en-1-yl]methyl]piperazine-1-yl]-N-[[3-nitro-4-[[(tetrahydro-2H-pyran-4-yl)methyl]aminophenyl]sulfonyl]-2-[(1H-pyrrolo[2,3-b]pyridine-5-yl)oxy]benzamide), ABT-737 (4-[4-[[2-(4-chlorophenyl (R)phenyl]methyl]piperazin-1-yl]-N-[4-[[(2R)-4-(dimethylamino)-1-phenylsulfanylbutan-2-yl]amino]-3-nitrophenyl]sulfonylbenzamide), ABT-263((R)-4-(4-((4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)piperazin-1-yl)-N-((4-((4-morpholino-1-(phenylthio)butan-2-yl )amino)-3((trifluoromethyl)sulfonyl)phenyl)sulfonyl)benzamide), GX15-070(Obatoclax mesylate, (2Z)-2-[(5Z)-5-[(3,5-dimethyl-1H-pyrrole-2-yl)methylidene]-4-methoxypyrrole-2-ylidene]indole;methanesulfonic acid), 2-methoxy-antimycin A3, YC137(4-(4,9-dioxo-4,9-dihydronaphtho[2,3-d]thiazole-2-ylamino)- Examples include phenyl esters), pogosine, ethyl 2-amino-6-bromo-4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate, nilotinib-d3, TW-37 (N-[4-[[2-(1,1-dimethylethyl)phenyl]sulfonyl]phenyl]-2,3,4-trihydroxy-5-[[2-(1-methylethyl)phenyl]methyl]benzamide), apogossiporone (ApoG2), or G3139 (oblimersen). In one embodiment, an effective amount of form B of isolated compound 2 is combined with a BCL-2 inhibitor in a single-dose form.

[0182] In one embodiment, the combination described herein can be further combined with additional therapeutic agents to treat cancer. The second therapy may be immunotherapy. As will be discussed in more detail below, an effective amount of form B of isolated compound 2 can be conjugated with an antibody, a radiopharmaceutical, or another targeting agent that directs the compound to diseased or abnormally proliferating cells. In another embodiment, the combination is used in combination with another pharmaceutical or biological agent (e.g., an antibody) to increase the efficacy of the treatment through a combination or synergistic approach. In one embodiment, the combination can be used with T-cell vaccination, which typically involves immunization with inactivated autoreactive T cells, to eliminate the cancer cell population described herein. In another embodiment, the combination is used in combination with a bispecific T-cell engager (BiTE), which is an antibody designed to simultaneously bind to endogenous T cells and specific antigens on cancer cells described herein, and to link these two types of cells.

[0183] In one embodiment, the bioactive agent is a MEK inhibitor. MEK inhibitors are known, for example, trametinib / GSK1120212 (N-(3-{3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidine-1(2H)-yl}phenyl)acetamide), selmetinib (6-(4-bromo-2-chloroanilino)-7-fluoro-N-(2-hydroxyethoxy)-3-methylbenzimidazole-5-carboxamide), and pimacertib / AS703026 / MSC 1935369((S)-N-(2,3-dihydroxypropyl)-3-((2-fluoro-4-iodophenyl)amino)isonicotinamide), XL-518 / GDC-0973(1-({3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]phenyl}carbonyl)-3-[(2S)-piperidine-2-yl]azetidine-3-ol), refametinib / BAY8697 66 / RDEA119(N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide), PD-0325901(N-[(2R)-2,3-dihydroxypropoxy]-3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-benzamide), TAK73 3((R)-3-(2,3-dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3-d]pyrimidine-4,7(3H,8H)-dione), MEK162 / ARRY438162(5-[(4-bromo-2-fluorophenyl)amino]-4-fluoro-N-(2-hydroxyethoxy)-1-methyl-1H-benzimidazole-6- Ruboxamide), R05126766 (3-[[3-fluoro-2-(methylsulfamoylamino)-4-pyridyl]methyl]-4-methyl-7-pyrimidine-2-yloxychromen-2-one), WX-554, R04987655 / CH4987655 (3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-5-((3-oxo-1,Examples include 2-oxazinan-2yl(methyl)benzamide) or AZD8330(2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carboxamide), U0126-EtOH, PD184352(CI-1040), GDC-0623, BI-847325, cobimetinib, PD98059, BIX02189, BIX02188, vinimetinib, SL-327, TAK-733, and PD318088.

[0184] In one embodiment, an effective amount of compound 2, form B, is administered in combination with a dose of binimetinib effective for the treatment of melanoma, including BRAF-mutated melanoma and NRAS-mutated melanoma.

[0185] In one embodiment, an effective amount of compound 2, form B, is administered in combination with cobimetinib (Cotellic) in an effective amount for the treatment of melanoma, including BRAF-mutated melanoma and NRAS-mutated melanoma.

[0186] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of binimetinib for the treatment of ovarian cancer.

[0187] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of selumetinib for the treatment of non-small cell lung cancer.

[0188] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of selumetinib for the treatment of thyroid cancer.

[0189] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of trametinib (Mekinist) for the treatment of thyroid cancer.

[0190] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of trametinib (Mekinist) for the treatment of melanoma.

[0191] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of trametinib (Mekinist) for the treatment of non-small cell lung cancer.

[0192] In one embodiment, the bioactive agent is a Raf inhibitor. Known Raf inhibitors include, for example, vemurafenib (N-[3-[[5-(4-chlorophenyl)-1H-pyrrolo[2,3-b]pyridine-3-yl]carbonyl]-2,4-difluorophenyl]-1-propanesulfonamide), sorafenib tosylate (4-[4-[[4-chloro-3-(trifluoromethyl)phenyl]carbamoylamino]phenoxy]-N-methylpyridine-2-carboxamide; 4-methylbenzenesulfonate), AZ62 8(3-(2-cyanopropan-2-yl)-N-(4-methyl-3-(3-methyl-4-oxo-3,4-dihydroquinazoline-6-ylamino)phenyl)benzamide), NVP-BHG712(4-methyl-3-(1-methyl-6-(pyridine-3-yl)-1H-pyrazolo[3,4-d]pyrimidine-4-ylamino)-N-(3-(trifluoromethyl)phenyl)benzamide), RAF-265(1-methyl-5-[2-[5-(trifluoro (Methyl)-1H-imidazole-2-yl]pyridine-4-yl]oxy-N-[4-(trifluoromethyl)phenyl]benzimidazole-2-amine), 2-bromoardicine (2-bromo-6,7-dihydro-1H,5H-pyrrolo[2,3-c]azepine-4,8-dione), Raf kinase inhibitor IV (2-chloro-5-(2-phenyl-5-(pyridine-4-yl)-1H-imidazole-4-yl)phenol), sorafenib N-oxide Examples include (4-[4-[[[[4-chloro-3(trifluoromethyl)phenyl]amino]carbonyl]amino]phenoxy]-N-methyl-2-pyridinecarboxamide 1-oxide), PLX4720, dabrafenib (GSK21184326), GDC-0879, RAF265, AZ628, SB590885, ZM336372, GW5074, TAK-632, CEP-32496, LY309120, and GX818 (encorafenib).

[0193] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of dabrafenib (Tafinlar) for the treatment of thyroid cancer.

[0194] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of dabrafenib (Tafinlar) for the treatment of melanoma.

[0195] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of dabrafenib (Tafinlar) for the treatment of non-small cell lung cancer.

[0196] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of encorafenib for the treatment of melanoma, including BRAF-mutated melanoma.

[0197] In one embodiment, the additional therapy is a monoclonal antibody (MAb). Some MAbs stimulate an immune response that destroys cancer cells. Similar to antibodies naturally produced by B cells, these MAbs "coat" the surface of cancer cells, inducing their destruction by the immune system. For example, bevacizumab is a protein secreted by tumor cells and other cells in the tumor microenvironment that targets vascular endothelial growth factor (VEGF), which promotes the development of tumor angiogenesis. When VEGF binds to bevacizumab, it is unable to interact with its cell receptor, thus interfering with the signaling that leads to the growth of new blood vessels. Similarly, cetuximab and panitumumab target the epidermal growth factor receptor (EGFR), and trastuzumab targets the human epidermal growth factor receptor 2 (HER-2). MAbs that bind to cell surface growth factor receptors prevent the target receptor from sending normal growth-promoting signals. They may also induce apoptosis and activate the immune system to destroy tumor cells.

[0198] Another group of cancer-treating MAbs are immunoconjugates. Sometimes called immunotoxins or antibody-drug conjugates, these MAbs consist of antibodies conjugated to cytotoxic substances such as plant or bacterial toxins, chemotherapeutic drugs, or radioactive molecules. The antibody strongly binds to its specific antigen on the surface of cancer cells, allowing the cytotoxic substance to be taken up by the cells. An FDA-approved conjugate MAb that works in this way includes ado-trastuzumab emtansine, which targets the HER-2 molecule to deliver the cell proliferation inhibitor DM1 to metastatic breast cancer cells expressing HER-2.

[0199] Immunotherapy using T cells modified to recognize cancer cells with bispecific antibodies (bsAbs) or chimeric antigen receptors (CARs) is an approach that has the potential to eliminate both proliferating and non-proliferating / slowly proliferating subpopulations of cancer cells.

[0200] Bispecific antibodies, which simultaneously recognize target antigens and activating receptors on the surface of immune effector cells, offer an opportunity to modify immune effector cells to kill cancer cells. Another approach is the creation of chimeric antigen receptors by fusing extracellular antibodies to intracellular signaling domains. Chimeric antigen receptor-modified T cells can specifically kill tumor cells independently of MHC.

[0201] In some embodiments, the above combinations can be administered to a subject in further combination with other chemotherapeutic agents. If convenient, the combinations described herein can be administered simultaneously with another chemotherapeutic agent to simplify the treatment regimen. In some embodiments, the combinations and other chemotherapeutic agents can be provided in a single formulation. In one embodiment, the use of the compounds described herein is combined in a treatment regime with other active agents. Such active agents include tamoxifen, midazolam, letrozole, bortezomib, anastrozole, goserelin, mTOR inhibitors, PI3 kinase inhibitors, dual mTOR-PI3K inhibitors, MEK inhibitors, RAS inhibitors, ALK inhibitors, HSP inhibitors (e.g., HSP70 and HSP90 inhibitors, or combinations thereof), BCL-2 inhibitors, apoptosis-inducing compounds, MK-2206, GSK690693, perifosine (KRX-0401), GDC-0068, trisirivine, AZD5363, honokiol, and PF-04. Examples include, but are not limited to, AKT inhibitors including 691502 and ipatasertib and miltefosine; PD-1 inhibitors including, but not limited to, nivolumab, CT-011, MK-3475, BMS936558 and AMP-514; or FLT-3 inhibitors including, but not limited to, P406, dovitinib, quizartinib (AC220), amvatinib (MP-470), tanzutinib (MLN518), ENMD-2076 and KW-2449; or combinations thereof.

[0202] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of ipatasertib for the treatment of breast cancer, including triple-negative breast cancer.

[0203] In one embodiment, the bioactive agent is an mTOR inhibitor. Examples of mTOR inhibitors include, but are not limited to, bisutucertib and rapamycin and their analogues, everolimus (Afinitor), temsirolimus, ridafololimus, sirolimus and defololimus. Examples of MEK inhibitors include trametinib / GSK1120212 (N-(3-{3-cyclopropyl-5-[(2-fluoro-4-iodophenyl)amino]-6,8-dimethyl-2,4,7-trioxo-3,4,6,7-tetrahydropyrido[4,3-d]pyrimidine-1(2H-yl}phenyl)acetamide) and selumetinib (6-(4-bromo-2-chloroanilino) -7-Fluoro-N-(2-hydroxyethoxy)-3-methylbenzimidazole-5-carboxamide), pimacertib / AS703026 / MSC1935369((S)-N-(2,3-dihydroxypropyl)-3-((2-fluoro-4-iodophenyl)amino)isonicotinamide), XL-518 / GDC-0973(1-({3,4-difluoro-2-[(2- Fluoro-4-iodophenyl)amino]phenyl}carbonyl)-3-[(2S)-piperidine-2-yl]azetidine-3-ol), refametinib / BAY869766 / RDEA119(N-(3,4-difluoro-2-(2-fluoro-4-iodophenylamino)-6-methoxyphenyl)-1-(2,3-dihydroxypropyl)cyclopropane-1-sulfonamide), PD-0325901(N-[(2R)-2,3-dihydroxypropoxy]-3,4-difluoro-2-[(2-fluoro-4-iodophenyl)amino]-benzamide), TAK733((R)-3-(2,3-dihydroxypropyl)-6-fluoro-5-(2-fluoro-4-iodophenylamino)-8-methylpyrido[2,3d]pyrimidine-4,7(3H,8H)-dione), MEK162 / ARRY438162(5-[(4-bromo-2-fluorophenyl)amino]-4-fluoro-N-(2-hydroxyethoxy)-1-methyl-1H-benzimidazole-6-carboxamide), R05126766(3-[[3-fluoro-2-(methylsulfamoylamino)-4-pyridyl]methyl]-4-methyl-7-pyrimidine-2-yloxychromen-2-one), WX-554, R04987655 / C Examples include, but are not limited to, H4987655 (3,4-difluoro-2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-5-((3-oxo-1,2-oxadinan-2-yl)methyl)benzamide) or AZD8330 (2-((2-fluoro-4-iodophenyl)amino)-N-(2-hydroxyethoxy)-1,5-dimethyl-6-oxo-1,6-dihydropyridine-3-carboxamide).

[0204] In one embodiment, the bioactive agent is a RAS inhibitor. Examples of RAS inhibitors include, but are not limited to, Reolysin and siG12D LODER.

[0205] In one embodiment, the bioactive agent is an ALK inhibitor. Examples of ALK inhibitors include, but are not limited to, crizotinib, AP26113, and LDK378.

[0206] In one embodiment, the bioactive agent is an HSP inhibitor. Examples of HSP inhibitors include, but are not limited to, geldanamycin or 17-N-allylamino-17-demethoxygeldanamycin (17AAG) and radicicol. In certain embodiments, the compounds described herein are administered in combination with letrozole and / or tamoxifen. Other chemotherapeutic agents that can be used in combination with the compounds described herein include, but are not limited to, chemotherapeutic agents that do not require cell cycle activity for their antitumor effect.

[0207] Additional bioactive compounds include, for example, everolimus, trabectedin, abraxane, TLK 286, AV-299, DN-101, pazopanib, GSK690693, RTA 744, ON 0910.Na, AZD 6244 (ARRY-142886), AMN-107, TKI-258, GSK461364, and AZD 1152, Enzastaurin, Vandetanib, ARQ-197, MK-0457, MLN8054, PHA-739358, R-763, AT-9263, FLT-3 inhibitor, VEGFR inhibitor, Aurora kinase inhibitor, PIK-1 modulator, HDAC inhibitor, c-MET inhibitor, PARP inhibitor, Cdk inhibitor, IGFR-TK inhibitor, Anti-HGF antibody, Focal adhesion plaque kinase inhibitor, Map kinase (mek) inhibitor, VEGF trap antibody, pemetrexed, panitumumab, amrubicin, olegobomab, Lep-etu, noratexide, azd2171, batabulin, ofatumumab, zanorimumab, edtecalin, tetrandrin, lubitecan, tesmilifene, oblimersen, tisilimmab, ipilimumab, gossypol, Bio 111, 131-I-TM-601, ALT-110, BIO 140, CC 8490, sirengitide, gimatecan, IL13-PE38QQR, INO 1001, IPdR1KRX-0402, rucanton, LY317615, neuradiab, vitespan, Rta 744, Sdx 102, Taranpanel, Atracentane, Xr 311, Romidepsin, ADS-100380, Sunitinib, 5-Fluorouracil, Vorinostat, Etoposide, Gemcitabine, Doxorubicin, Liposomal Doxorubicin, 5'-Deoxy-5-Fluorouridine, Vincristine, Temozolomide, ZK-304709, Sericiclib; PD0325901, AZD-6244, Capecitabine, L-Glutamic Acid, N-[4-[2-(2-amino-4,7-dihydro-4-oxo-1H-pyrrolo[2,3-d]pyrimidine-5-yl)ethyl]benzoyl]-, Disodium Salt Heptahydrate, Camptothecin, PEG-labeled Irinotecan, Tamoxifen, Toremifene Citrate, Anastrazole, Exemestane, Letrozole,DES (Diethylstilbestrol), Estradiol, Estrogen, Conjugated Estrogen, Bevacizumab, IMC-1C11, CHIR-258); 3-[5-(Methylsulfonylpiperazine methyl)-Indolyl-Quinolone, Batalanib, AG-013736, AVE-0005, Goserelin Acetate, Leuprolide Acetate, Triptorelin Pamoate, Medroxyprogesterone Acetate, Hydroxyprogesterone Caproate, Megestrol Acetate Raloxifene, bicalutamide, flutamide, nilutamide, megestrol acetate, CP-724714; TAK-165, HKI-272, erlotinib, lapatinib, canertinib, ABX-EGF antibody, Erbitux, EKB-569, PKI-166, GW-572016, ronafarnib, BMS-214662, tipifarnib; amifostin, NVP-LAQ824, suberoyl analide hydroxamic acid (acid), valproic acid, trichostatin A, FK-228, SU11248, sorafenib, KRN951, aminoglutethimide, amsacrin, anagrelide, L-asparaginase, Calmette-Guéran bacillus (BCG) vaccine, adriamycin, bleomycin, buserelin, busulfan, carboplatin, carmustine, chlorambucil, cisplatin, cladribine, clodronate, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, diethylstilbestrol, epirubicin, fludarabine, fludrocortisone, fluoxymesterone, flutamide, gleevec, gemcitabine, hidol Roxyurea, Idarubicin, Ifosfamide, Imatinib, Leuprolide, Levamisole, Lomustine, Mechloretamine, Melphalan, 6-Mercaptopurine, Mesna, Methotrexate, Mitomycin, Mitotane, Mitoxantrone, Niltamide, Octreotide, Oxaliplatin, Pamidronate, Pentostatin, Plicamycin, Porfimer, Procarbazine, Larcitrexed, Rituximab, Streptozocin, Teniposide, Testosterone, Thalidomide, Thioguanine, Thiotepa, Tretinoin, Vindesine, 13-Cis-Retinoic Acid, Phenylalanine Mustard, Uracil Mustard, Estramustine,Altretamine, phloxuridine, 5-deoxyuridine, cytosine arabinoside, 6-mercaptopurine, deoxycoformycin, calcitriol, barrubicin, mitramycin, vinblastine, vinorelbine, topotecan, razoxin, marimast, COL-3, neovastat, BMS-275291, squalamine, endostatin, SU5416, SU6668, EMD121974, interleukin-12, IM862, angiostatin, vitaxin, droloxifen, idoxyfen e) Spironolactone, finasteride, cimitidine, trastuzumab, denileukin difutitox, gefitinib, bortezomib, paclitaxel, paclitaxel without cremofol, docetaxel, epotilon B, BMS-247550, BMS-310705, droloxifen, 4-hydroxytamoxifen, pipendoxifen, ERA-923, alzoxifen, fulvestrant, acorbifen, rasofoxifen, idoxifen, TSE-424, HMR-3339, ZK186619, topotecan, PTK787 / ZK 222584, VX-745, PD 184352, Rapamycin, 40-O-(2-hydroxyethyl)-rapamycin, Temsirolimus, AP-23573, RAD001, ABT-578, BC-210, LY294002, LY292223, LY292696, LY293684, LY293646, Wartmannin, ZM336372, L-779,450, PEG-filgrastim, Darbepoetin, Erythropoetin, Granulocyte Colony-Stimulating Factor, Zoledronate, Prednisone, Cetuximab, Granulocyte Macrophage Colony Stimulation Stimulant factor, histrelin, pegylated interferon α-2a, interferon α-2a, pegylated interferon α-2b, interferon α-2b, azacitidine, PEG-L-asparaginase, lenalidomide, gemtuzumab, hydrocortisone, interleukin-11, dexrazoxane, alemtuzumab, all-trans retinoic acid, ketoconazole, interleukin-2, megestrol, immunoglobulin, nitrogen mustard, methylprednisolone, ibritumomab tiuxetan,Examples include androgens, decitabine, hexamethylmelamine, bexarotene, tositumomab, arsenic trioxide, cortisone, etidronate, mitotane, cyclosporine, liposomal daunorubicin, Edwina-asparaginase, strontium-89, casopitant, netupitant, NK-1 receptor antagonists, palonosetron, aprepitant, diphenhydramine, hydroxyzine, metoclopramide, lorazepam, alprazolam, haloperidol, droperidol, dronabinol, dexamethasone, methylprednisolone, prochlorperazine, granisetron, ondansetron, drasetron, tropisetron, pegfilgrastim, erythropoetin, platelet-derived growth factor receptor α (PDGFR-α), epoetin α, darbepoetin α, and mixtures thereof.

[0208] In one embodiment, an effective amount of form B of isolated compound 2 described herein may be used in combination with a PARP inhibitor selected from niraparib tosylate monohydrate (Zejula), olaparib (Lynparza), rucaparib cansylate (Rubraca), and talazoparib.

[0209] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of niraparib tosylate monohydrate (Zejula) for the treatment of abnormal tissues of the female reproductive system, including ovarian epithelial carcinoma or fallopian duct carcinoma.

[0210] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of niraparib tosylate monohydrate (Zejula) for the treatment of peritoneal cancer.

[0211] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of olaparib (Lynparza) for the treatment of abnormal tissues of the female reproductive system, including breast cancer, ovarian cancer, ovarian epithelial carcinoma, or fallopian duct carcinoma.

[0212] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of olaparib (Lynparza) for the treatment of BRAC1 or BRAC2 mutant breast cancer.

[0213] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of olaparib (Lynparza) for the treatment of HER2-type breast cancer.

[0214] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of olaparib (Lynparza) for the treatment of peritoneal cancer.

[0215] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of rucaparibucansylate (Rubraca) for the treatment of abnormal tissues of the female reproductive system, including breast cancer, ovarian cancer, ovarian epithelial carcinoma, or fallopian duct carcinoma.

[0216] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of rucaparibucansylate (Rubraca) for the treatment of peritoneal cancer.

[0217] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of talazoparib for the treatment of abnormal tissues of the female reproductive system, including breast cancer, ovarian cancer, ovarian epithelial carcinoma, or fallopian duct carcinoma.

[0218] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of talazoparib for the treatment of BRAC1 or BRAC2 mutant breast cancer.

[0219] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of olaratumab for the treatment of soft tissue sarcoma.

[0220] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of savolitinib for the treatment of adenocarcinoma.

[0221] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of savolitinib for the treatment of non-small cell lung cancer.

[0222] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of savolitinib for the treatment of renal cell carcinoma.

[0223] In one embodiment, an effective amount of compound 2 form B is administered in combination with a visucertib amount effective for the treatment of advanced breast cancer.

[0224] In one embodiment, an effective amount of compound 2 form B is administered in combination with a visucertib amount effective for the treatment of advanced breast cancer.

[0225] In one embodiment, form B of isolated compound 2 described herein is imatinib mesylate (Gleevac®), dasatinib (Sprycel®), nilotinib (Tasigna®), bosutinib (Bosulif®), trastuzumab (Herceptin®), pertuzumab (Perjeta®), lapatinib (Tykerb®), gefitinib (Iressa®), erlotinib (Tarceva®), cetuximab (Erbitux®), panitumumab (Vectibix®), vandetanib (Caprelsa®), vemurafenib (Zelboraf®), vorinostat (Zolinza®). It can be combined with chemotherapeutic agents selected from, but not limited to, romidepsin (Istodax®), bexarotene (Tagretin®), alitretinoin (Panretin®), tretinoin (Vesanoid®), carfilzomib (Kyprolis®), pralatrexate (Folotyn®), bevacizumab (Avastin®), Ziv-aflibercept (Zaltrap®), sorafenib (Nexavar®), sunitinib (Sutent®), pazopanib (Votrient®), regorafenib (Stivarga®), and cabozantinib (Cometriq®).

[0226] In one embodiment, an effective amount of form B of the isolated compound 2 described herein may be used in combination with a CD4 / 6 inhibitor, including abemaciclib (Versenio), palbociclib (Ibrance), or trilaciclib.

[0227] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of abemaciclib (Versenio) for the treatment of breast cancer.

[0228] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of abemaciclib (Versenio) for the treatment of HR+HER2- breast cancer.

[0229] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of palbociclib (Ibrance) for the treatment of breast cancer.

[0230] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of palbociclib (Ibrance) for the treatment of HR+HER2- breast cancer.

[0231] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of palbociclib (Ibrance) for the treatment of breast cancer.

[0232] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of palbociclib (Ibrance) for the treatment of metastatic triple-negative breast cancer.

[0233] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of palbociclib (Ibrance) for the treatment of small cell lung cancer.

[0234] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of cabozantinib S-malate (Cometriq®) for the treatment of thyroid cancer.

[0235] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of cabozantinib S-malate (Cometriq®) for the treatment of renal cell carcinoma.

[0236] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of dasatinib (Sprycel) for the treatment of leukemia, including acute lymphoblastic leukemia or chronic myeloid leukemia.

[0237] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of dasatinib (Sprycel) for the treatment of prostate cancer.

[0238] In one embodiment, an effective amount of Form B of Compound 2 is administered in combination with an effective amount of erlotinib (Tarceva™) for the treatment of prostate cancer.

[0239] In one embodiment, an effective amount of Form B of Compound 2 is administered in combination with an effective amount of gefitinib (Iressa™) for the treatment of prostate cancer.

[0240] In one embodiment, an effective amount of Form B of Compound 2 is administered in combination with an effective amount of imatinib mesylate (Gleevec) for the treatment of leukemia, including acute lymphoblastic leukemia, chronic eosinophilic leukemia, hypereosinophilic syndrome, or chronic myelogenous leukemia.

[0241] In one embodiment, an effective amount of Form B of Compound 2 is administered in combination with an effective amount of trastuzumab (Herceptin) for the treatment of adenocarcinoma.

[0242] In one embodiment, an effective amount of Form B of Compound 2 is administered in combination with an effective amount of trastuzumab (Herceptin) for the treatment of breast cancer, including HER2+ breast cancer.

[0243] In one embodiment, an effective amount of Form B of Compound 2 is administered in combination with an effective amount of imatinib mesylate (Gleevec) for the treatment of tumors including, but not limited to, dermatofibrosarcoma protuberans and gastrointestinal stromal tumors.

[0244] In one embodiment, an effective amount of Form B of Compound 2 is administered in combination with an effective amount of imatinib mesylate (Gleevec) for the treatment of myelodysplastic / myeloproliferative neoplasms.

[0245] In one embodiment, an effective amount of Form B of Compound 2 is administered in combination with an effective amount of imatinib mesylate (Gleevec) for the treatment of systemic mastocytosis.

[0246] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of nilotinib (Tasigna) for the treatment of chronic myeloid leukemia, including Philadelphia chromosome-positive chronic myeloid leukemia (Ph+CML).

[0247] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of pazopanib hydrochloride (Votrient) for the treatment of renal cell carcinoma.

[0248] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of pazopanib hydrochloride (Votrient) for the treatment of soft tissue sarcoma.

[0249] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of regorafenib (Stivarga) for the treatment of colorectal cancer.

[0250] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of regorafenib (Stivarga) for the treatment of gastrointestinal stromal tumors.

[0251] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of regorafenib (Stivarga) for the treatment of hepatocellular carcinoma.

[0252] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of sorafenib tosylate (Nexavar) for the treatment of carcinomas including hepatocellular carcinoma or renal cell carcinoma.

[0253] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of sunitinib malate (Sutent) for the treatment of gastrointestinal stromal tumors.

[0254] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of sunitinib malate (Sutent) for the treatment of pancreatic cancer.

[0255] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of sunitinib malate (Sutent) for the treatment of renal cell carcinoma.

[0256] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of vemurafenib (Zelboraf) for the treatment of Erdheim-Chester disease.

[0257] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of vemurafenib (Zelboraf) for the treatment of melanoma.

[0258] In certain embodiments, the additional therapeutic agent is an anti-inflammatory agent, a chemotherapeutic agent, a radiotherapeutic agent, an additional therapeutic agent, or an immunosuppressant.

[0259] Suitable chemotherapeutic agents include, but are not limited to, radioactive molecules, cytotoxins, or toxins also known as cytotoxic agents, as well as any active substance harmful to the viability of cells, and active substances containing chemotherapeutic compounds and liposomes or other vesicles. Common anticancer drugs include vincristine (Oncovin®) or liposomal vincristine (Marqibo®), daunorubicin (Daunomycin or Cerubidine®) or doxorubicin (Adriamycin®), cytarabine (cytosine arabinoside, ara-C or Cytosar®), L-asparaginase (Elspar®) or PEG-L-asparaginase (Pegasparagase or Oncaspar®), et Examples include poside (VP-16), teniposide (Vumon®), 6-mercaptopurine (6-MP or Purinethol®), methotrexate, cyclophosphamide (Cytoxan®), prednisone, dexamethasone (Decadron), imatinib (Gleevec®), dasatinib (Sprycel®), nilotinib (Tasigna®), bosutinib (Bosulif®), and ponatinib (Iclusig®). Examples of suitable additional chemotherapeutic agents include 1-dehydrotestosterone, 5-fluorouracil, dacarbazine, 6-mercaptopurine, 6-thioguanine, actinomycin D, adriamycin, aldesleukin, alkylating agents, allopurinol sodium, altoretamine, amiphostine, anastrozole, anthramycin (AMC), antimitotic agents, cis-dichlorodiamine platinum(II) (DDP) (cisplatin), diaminodichloroplatin, anthracyclines, antibiotics, antimetabolites, asparaginase, and BCG bacteria (BCG).(live) (in bladder), betamethasone sodium phosphate and betamethasone acetate, bicalutamide, bleomycin sulfate, busulfan, leucovorin calcium, calicheamicin, capecitabine, carboplatin, lomustine (CCNU), carmustine (BSNU), chlorambucil, cisplatin, cladribine, colchicine, conjugated estrogen, cyclophosphamide, cyclotosphamide, cytarabine, cytarabine, cytochalasin B, cytoxane, dacarbazine, dactinomycin, dactinomycin (formerly actinomycin), daunorubicin HCl, daunorubicin citrate, denileukin difutitox, dexrazoxane, dibromomannitol, dihydroxyanthracine dione dione), docetaxel, drasetron mesylate, doxorubicin HCl, dronabinol, Escherichia coli (E. coli)coli) L-asparaginase, emetine, epoetin-α, Erwinia L-asparaginase, esterified estrogen, estradiol, estramustine sodium phosphate, ethidium bromide, ethinylestradiol, etidronate, etoposide, citroborum factor, etoposide phosphate, filgrastim, floxuridine, fluconazole, fludarabine phosphate, fluorouracil, flutamide, folinic acid, gemcitabine HCl, glucocorticoid, goserelin acetate, gramicidin D, granisetron HCl, hydroxyurea, idarubicin HCl, ifosfamide, interferon α-2b, irinotecan HCl, letrozole, leucovorin calcium, leuprolide acetate, levamisole HCl, lidocaine, lomustine, maytansinoid, Mechloretamine HCl, medroxyprogesterone acetate, megestrol acetate, melphalan HCl, mercaptopurine, mesna, methotrexate, methyltestosterone, mitramycin, mitomycin C, mitotane, mitoxantrone, nilutamide, octreotide acetate, ondansetron HCl, paclitaxel, disodium pamidronate, pentostatin, pilocarpine HCl, primycin, polyfeprozan 20 carmustine implant, porfimer sodium, procaine, procarbazine HCl, propranolol, rituximab, salglamostim, streptozotocin, tamoxifen, taxol, teniposide, tenoposide, testactone, tetracaine, thioepachlorambucil Examples include, but are not limited to, chlorambucil, thioguanine, thiotepa, topotecan HCl, toremifene citrate, trastuzumab, tretinoin, barrubicin, vinblastine sulfate, vincristine sulfate, and vinorelbine tartrate.

[0260] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of bosutinib (Bosulif®) for the treatment of chronic myeloid leukemia (CML).

[0261] In one embodiment, an effective amount of Form B of Compound 2 is administered in combination with an effective amount of ponatinib hydrochloride (Iclusig) for the treatment of leukemia, including acute lymphoblastic leukemia and chronic myelogenous leukemia.

[0262] Additional therapeutic agents that can be administered in combination with the compounds disclosed herein include bevacizumab, sunitinib, sorafenib, 2-methoxyestradiol or 2ME2, finasunate, batatinib, vandetanib, aflibercept, brolucizumab, etaracizumab (MEDI-522), sirukumab, erlotinib, cetuximab, panitumumab, gefitinib, trastuzumab, dovitinib, figitumumab, atacicept, rituximab, alemtuzumab, aldesleukin, atorizumab, tocilizumab, temsirolimus, everolimus, lucatumumab, dacetuzumab, HLL1, huN901-DM1, atiprimod, natalizumab, bortezomib, carfilzomib, marizomib, tanespimycin, saquinavir mesylate, ritonavir, nelfinavir mesylate, indinavir sulfate, belinostat, panobinostat, mapatumumab, lexatumumab, dulanermin, ABT-737, oblimersen, plitidepsin, talmapimod, P276-00, enzastaurin, tipifarnib, perifosine, imatinib, dasatinib, lenalidomide, thalidomide, simvastatin, celecoxib, bazedoxifene, AZD4547, rilotumumab, oxaliplatin (Eloxatin), PD0332991, ribociclib (LEE011), abemaciclib (LY2835219), HDM201, fulvestrant (Faslodex), exemestane (Aromasin), PIM447, ruxolitinib (INC424), BGJ398, necitumumab, pemetrexed (Alimta), and ramucirumab (IMC-1121B).

[0263] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of everolimus (Afinitor) for the treatment of breast cancer.

[0264] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of everolimus (Afinitor) for the treatment of HR+, HER2- breast cancer.

[0265] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of everolimus (Afinitor) for the treatment of pancreatic cancer.

[0266] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of everolimus (Afinitor) for the treatment of gastrointestinal cancer.

[0267] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of everolimus (Afinitor) for the treatment of lung cancer.

[0268] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of everolimus (Afinitor) for the treatment of renal cell carcinoma.

[0269] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of everolimus (Afinitor) for the treatment of astrocytoma, including subependymal giant cell astrocytoma.

[0270] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of fulvestrant (Faslodex) for the treatment of breast cancer.

[0271] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of fulvestrant (Faslodex) for the treatment of HR+, HER2- breast cancer.

[0272] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of ramucirumab for the treatment of adenocarcinoma.

[0273] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of ramucirumab for the treatment of non-small cell lung cancer.

[0274] In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of ramucirumab for the treatment of colorectal cancer.

[0275] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of ribociclib (Kisqali) for the treatment of breast cancer.

[0276] In one embodiment, an effective amount of compound 2, form B, is administered in combination with an effective amount of ribociclib (Kisqali) for the treatment of HR+ and HER2- breast cancer.

[0277] In one embodiment of the present invention, the compounds described herein may be used in combination with at least one IDH1 or IDH2 inhibitor. In one embodiment, an effective amount of form B of compound 2 is administered in combination with an effective amount of enasidenib mesylate (Idhifa) for the treatment of acute myeloid leukemia.

[0278] In one embodiment of the present invention, the compounds described herein may be used in combination with at least one fibroblast growth factor receptor (FGFR) tyrosine kinase inhibitor. In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of erdafitinib for the treatment of urothelial carcinoma, including metastatic urothelial carcinoma.

[0279] In one embodiment of the present invention, the compounds described herein may be used in combination with at least one ERK inhibitor.

[0280] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of SCH772984 for the treatment of melanoma, including BRAF-mutated melanoma or NRAS-mutated melanoma.

[0281] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of urixertinib for the treatment of melanoma, including uveal melanoma.

[0282] In one embodiment, an effective amount of compound 2 form B is administered in combination with an effective amount of urixertinib for the treatment of pancreatic cancer.

[0283] In one aspect of the present invention, the compounds described herein can be combined with at least one immunosuppressant. The immunosuppressants include calcineurin inhibitors, such as cyclosporine or ascomycin, such as cyclosporine A (NEORAL®), FK506 (tacrolimus), and pimecrolimus; mTOR inhibitors, such as rapamycin or its derivatives, such as sirolimus (RAPAMUNE®), everolimus (Certican®), temsirolimus, zotarolimus, biolimus-7, biolimus-9; rapalog, such as ridafololimus; azathioprine; camphoglion; S1P receptor modulators, such as fingolimod or its analogues; anti-IL-8 antibodies; mycophenolic acid or its salts, such as sodium salt or its prodrugs, such as mycophenolate mofetil (CELLCEPT®), and OKT3 (ORTHOCLONE®). OKT3(trademark), prednisone, ATGAM(trademark), THYMOGLOBULIN(trademark), Brequinal sodium, OKT4, T10B9.A-3A, 33B3.1, 15-deoxysperguarine, tresperimus, leflunomide (ARAVA(trademark)), CTLAI-Ig, anti-CD25, anti-IL2R, basiliximab (SIMULECT(trademark)), daclizumab (ZENAPAX(trademark)), mizoribine, methotrexate, dexamethasone, ISAtx-247, SDZ ASM The following are selected from the group consisting of 981 (pimecrolimus, Elidel®), CTLA4Ig (abatacept), belatacept, LFA3Ig, etanercept (marketed as Enbrel® by Immunex), adalimumab (Humira®), infliximab (Remicade®), anti-LFA-1 antibody, natalizumab (Antegren®), enrimomab, gavilimomab, anti-thymocyte immunoglobulin, cyprizumab, alefacept, efalizumab, pentasa, mesalazine, asacol, codeine phosphate, benolilate, fenbufen, naprosin, diclofenac, etodolac, and indomethacin, aspirin, and ibuprofen.

[0284] In certain embodiments, the compounds described herein are administered to a subject before, during, or after treatment with another chemotherapeutic agent.

[0285] In some embodiments, an effective amount of isolated compound 2, form B, can be administered to a subject so that other chemotherapeutic agents can be administered at either higher doses (increased chemotherapeutic dose intensity) or more frequently (increased chemotherapeutic dose density). Dose-dense chemotherapy is a chemotherapy treatment plan in which drugs are administered at shorter intervals than in a standard chemotherapy treatment plan. Chemotherapy dose intensity represents the unit dose of a chemotherapeutic agent administered per unit time. Dose intensity can be increased or decreased by changing the dose, the time interval of administration, or both.

[0286] In one embodiment of the present invention, the compounds described herein may be administered in a co-administered regimen with another active agent, such as a non-DNA damaging targeted antitumor agent or a hematopoietic growth factor agent. It has recently been reported that mistiming of hematopoietic growth factors can lead to serious side effects. For example, the use of EPO family growth factors has been associated with arterial hypertension, cerebral convulsions, hypertensive encephalopathy, thromboembolism, iron deficiency, influenza-like syndrome, and venous thrombosis. G-CSF family growth factors have been associated with splenomegaly and rupture, respiratory distress syndrome, allergic reactions, and sickle cell complications.Thus, in one embodiment, the use of the compounds or methods described herein is as follows: granulocyte colony-stimulating factor (G-CSF, e.g., marketed as Neupogen (filgrastin), Neulasta (pegfilgrastim), or lenograstim), granulocyte-macrophage colony-stimulating factor (GM-CSF, e.g., marketed as morglamostim and salglamostim (Leukine)), M-CSF (macrophage colony-stimulating factor), thrombopoietin (megakaryocyte growth and differentiation factor (MGDF), e.g., marketed as romiplostim and eltrombopag), interleukin (IL)-12, interleukin-3, interleukin-11 (lipid-derived Combined with the use of hematopoietic growth factors, including but not limited to growth inhibitors (or oprelbequin), SCF (stem cell factor, steel factor, kit-ligand, or KL), and erytropoetin (EPO) and their derivatives (e.g., epoetin-α marketed as darbepoetin, Epocept, Nanokine, Epofit, Epogin, Eprex, and Procrit; epoetin-β marketed as NeoRecormon, Recormon, and Micera), epoetin-δ (e.g., marketed as Dynepo), epoetin-ω (e.g., marketed as Epomax), epoetin-ζ (e.g., marketed as Silapo and Reacrit), and hematopoietic growth factors (e.g., Epocept, EPOTrust, Erypro Safe, Repoeitin, Vintor, Epofit, Erykine, Wepox, Espogen, Relipoeitin, Shanpoietin, Zyrop, and EPIAO). In one embodiment, an effective amount of form B of isolated compound 2 is administered before the administration of hematopoietic growth factor. In one embodiment, the administration of hematopoietic growth factor is performed at the time when the effect of the compound on HSPC has disappeared. In one embodiment, the growth factor is administered at least 20 hours after the administration of the compound described herein.

[0287] If necessary, multiple doses of the compounds described herein may be administered to the subject. Alternatively, a single dose of the compounds described herein may be administered to the subject. In one aspect of the present invention, the compounds disclosed herein can be beneficially administered in combination with any therapeutic regimen involving radiotherapy, chemotherapy, or other therapeutic agents. In additional embodiments, the compounds disclosed herein can be beneficially administered in combination with therapeutic agents targeting autoimmune disorders.

[0288] In alternative embodiments, forms A, C, D, E, G, or H of compound 2 are administered in the above-described combinations to treat a selected host, typically human, with cancer, tumor, hyperproliferative condition, or inflammatory or immunodeficiency, instead of form B of compound 2. [Examples]

[0289] Example 1. Conversion of compound 1 to its HCl counterpart, compound 2. A typical synthesis of compound 2 is provided in Scheme 1. [ka]

[0290] Compound 1 (0.9 kg, 1.9 mol, 1 equivalent) was placed in a 22 L flask and dissolved in 3.78 L of 2 M hydrochloric acid aqueous solution. The solution was heated to 50 ± 5 °C and stirred for 30 minutes. The resulting mixture was filtered through Celite (or the solution may be filtered through a 0.45 micron in-line filter) to obtain Compound 2. The flask was rinsed with 0.1 M hydrochloric acid solution, and any further Compound 2 was collected. Next, Compound 2 was heated to 50 ± 5 °C while gradually adding acetone (6.44 L). The solution was stirred at 50 ± 5 °C for 30 minutes, the temperature was lowered to 20 ± 5 °C, and stirring was continued for 2 hours. The solid was collected by filtration, washed with acetone, and dried to obtain 820.90 g of Compound 2 (yield 82.1%). In one embodiment, ethanol is used instead of acetone.

[0291] Example 2. Form of Compound 2 Eleven unique XRPD patterns (morphologies A to K) of compound 2 were obtained from crystallization and slurry experiments using various solvents. The conditions for these crystallization experiments and the XRPD results are shown in Tables 1 to 4. Crystallization in a single solvent (Table 1) resulted in a weak crystalline form or morphology A. Binary solvent crystallization using water (Table 2) and MeOH (Table 3) as the main solvents resulted in a weak crystalline form as well as morphologies A, B, F, G, and H. Solids recovered from slurry experiments (Table 4) one and seven days after equilibration were analyzed by XRPD to determine the crystalline form, and morphologies A, B, C, D, and E were observed after seven days. Figure 1 shows the XRPD patterns of morphologies A, B, and C. Figure 2 shows the XRPD patterns of morphologies D, E, and F. Figure 3 shows the XRPD patterns of morphologies G and H.

[0292] [Table 1]

[0293] [Table 2]

[0294] [Table 3] TIFF0007850883000008.tif22168

[0295] [Table 4]

[0296] Example 2. Morphological characterization of compound 2 Table 5 provides a summary of the characterization data for all isolated forms of compound 2. Forms A, B, and D were evaluated as solid forms.

[0297] [Table 5]

[0298] In one embodiment, morphology A is characterized by at least one XRPD peak located at 7.4±0.2°, 9.0±0.2°, or 12.3±0.2° in theta. In one embodiment, morphology B is characterized by at least one XRPD peak located at 6.4±0.2° or 9.5±0.2° in theta. In one embodiment, morphology C is characterized by at least one XRPD peak located at 5.3±0.2° or 7.2±0.2° in theta. In one embodiment, morphology D is characterized by at least one XRPD peak located at 5.6±0.2° or 8.2±0.2° in theta. In one embodiment, morphology E is characterized by at least one XRPD peak located at 5.5±0.2° or 6.7±0.2° in theta. In one embodiment, morphology E is characterized by at least one XRPD peak located at 5.5±0.2° or 6.7±0.2° in theta. In one embodiment, morphology F is characterized by an XRPD peak located at 7.2 ± 0.2°² theta. In one embodiment, morphology G is characterized by an XRPD peak located at 6.7 ± 0.2°² theta. In one embodiment, morphology H is characterized by an XRPD peak located at 6.6 ± 0.2°² theta.

[0299] Example 3. Dynamic vapor adsorption experiment of forms A, B, and D Dynamic vapor adsorption experiments were conducted on morphologies A, B, and D. Table 6 shows the results of the DVS experiments.

[0300] [Table 6]

[0301] Morphology A was found to be unstable in the moisture adsorption test. The material adsorbed 14.9% by weight of moisture at 60% RH and 15.8% by weight at 90% RH. After the moisture adsorption experiment, the sample was dried at 60°C and 0% RH, and a new morphology (morphology K) was revealed by XRPD analysis of the dried sample. The DVS analysis of morphology A is shown in Figure 4A. Morphology D was also found to be unstable in the moisture adsorption experiment. The material adsorbed 4.4% by weight of moisture at 60% RH and 17.0% by weight at 90% RH. After the moisture adsorption experiment, the sample was dried at 60°C and 0% RH, and morphology K was revealed by XRPD analysis of the dried sample. The DVS analysis of morphology D is shown in Figure 4B.

[0302] Unlike morphologies A and D, morphology B was stable in the moisture adsorption experiment. The material adsorbed 5.8 wt% moisture at 60% RH and 5.9 wt% moisture at 90% RH. After drying at 60°C and 0% RH for 2 hours, the XRPD pattern remained unchanged as morphology B. The DVS analysis of morphology B is shown in Figure 4C.

[0303] Figure 5A compares the XRPD pattern of morphology A before DVS analysis with the new pattern (morphology K) that emerged from DVS. Figure 5B compares the XRPD pattern of morphology D before DVS with the pattern (morphology K) that emerged after DVS.

[0304] Example 4. Stability study of morphologies A, B, and D under thermal stress. Morphologies A, B, and D were stored in a furnace maintained at 60°C for 7 days. No changes in the XRPD patterns were observed for morphologies B or D. For morphology A, a new pattern was found at the end of the stability study, but after 3 days of equilibrium at room temperature, the new morphology's XRPD revealed that it had reverted to morphology A. Figure 6 compares the XRPD patterns of morphologies A, B, and D with those of the reference material. Figure 6 also shows the new pattern resulting from exposing morphology A to thermal stress, along with the morphology A pattern that developed after a further 3 days at room temperature.

[0305] Example 5. Recrystallization procedure to produce form B from compound 2. Recrystallization studies were conducted to define procedures for improving chromatographic purity. All recrystallization procedures in Table 7 involved dissolving compound 2 in concentrated HCl, followed by the addition of the anti-solvent acetone. While the process differences were minor, they were significant in terms of their results.

[0306] Recrystallization process 1: Compound 1 was placed in a flask or reactor of appropriate size, dissolved in an aqueous hydrochloric acid solution, and heated to at least 55±10°C. The solution was stirred for about 45 minutes, and the resulting mixture was filtered through an in-line filter. Acetone was added over 1 hour at 55±10°C, and the solution was stirred for a further 1 hour. The temperature was lowered to about 25±5°C, and the solution was stirred for at least 2 hours. The solid was collected by filtration, washed with acetone, and dried to obtain form B of compound 2.

[0307] Recrystallization process 2: Compound 1 was placed in a flask or reactor of appropriate size, dissolved in an aqueous hydrochloric acid solution, and heated to at least 55±10°C. The solution was stirred for about 45 minutes, and the resulting mixture was filtered through an in-line filter. The temperature was lowered to about 25±5°C, and the solution was stirred for at least 2 hours. Acetone was added over 1 hour at 25±5°C, and the solution was stirred for a further 2 hours. The solid was collected by filtration, washed with acetone, and dried to obtain form D of compound 2.

[0308] Recrystallization process 3: Compound 1 was placed in a flask or reactor of appropriate size, dissolved in an aqueous hydrochloric acid solution, and heated to at least 55±10°C. The solution was stirred for about 45 minutes, and the resulting mixture was filtered through an in-line filter. The temperature was lowered to about 25±5°C, and the solution was stirred for at least 2 hours. The solid was collected by filtration, washed with acetone, and dried to obtain form D of compound 2.

[0309] [Table 7]

[0310] Experiments shown in Table 7 revealed that not all recrystallization processes produce the preferred solid form, morphology B. Specifically, recrystallization processes 2 and 3 produced different solid forms (presumed morphology D), while recrystallization process 1 reliably provided morphology B. In one embodiment, compound 2 is converted to morphology D by recrystallization procedures 2 and 3, and morphology D is converted to morphology B by recrystallization process 1.

[0311] Example 6. XRPD analysis of form B of compound 2 XRPD patterns of morphology B were collected using a PANalytical X'Pert PRO MPD diffractometer with an incident beam of Cu radiation generated using an Optix long, high-precision focus source. Cu Kα X-rays were focused from the sample to the detector using an elliptically stepped multilayer mirror. Prior to analysis, a silicon sample (NIST SRM 640e) was analyzed to verify that the observed position of the Si 111 peak coincided with the NIST-certified position. The sample was sandwiched between 3 μm thick films and analyzed by transmission geometry. Background caused by air was minimized using a beam stop, a short spatter extension, and a spatter-proof knife edge. A solar slit on the incident diffraction beam was used to minimize axial divergence and spreading. Diffraction patterns were collected using a scanning position-sensitive detector (X'Celerator) positioned 240 mm from the sample and data collector software v.2.2b. The data collection parameters for each pattern are shown on the images in the data section of this report, including the divergent slit (DS) before the mirror.

[0312] The XRPD pattern of pure morphology B, along with the indexing solution, is shown in Figure 7. The XRPD pattern of pure morphology B shows sharp peaks, indicating that the sample is composed of crystalline material. Possible peak locations from the XRPD indexing solution are 6.5, 8.1, 9.4, 9.6, 10.2, 10.6, 11.2, 12.2, 12.9, 13.0, 13.3, 13.4, 14.0, 14.4, 14.6, 15.0, 15.9, 16.2, 16.4, 16.5, 16.8, 18.1, 18.4, 18.5, 18.6, 18.6, 18.9, 19.1, 19.2, 19.3, 19.4, 19.5, 19.6, 19.7, 19.8, 19.9, 20.4, and 20. 6, 21.3, 21.4, 21.8, 22.0, 22.2, 22.3, 22.4, 22.5, 22.8, 23.0, 23.1, 23.4, 23.8, 24.1, 24.2, 24.3, 24.4, 24.5, 24.6, 25.4, 25.6, 25.7, 25.9, 26.0, 26.1, 26.3, 26.4, 26.5, 26.6, 26.7, 26.8, 26.9, 27.2, 27.3, 27.5, 27.6, 27.7, 27.9, 28.3, 28.4, 28.5, 28.7, 28. 9, 29.0, 29.1, 29.3, 29.4, 29.5, 29.6, 29.7, 29.8, 29.9, 30.0, 30.3, 30.4, 30.5, 30.6, 30.7, 30.9, 31.2, 31.5, 31.6, 31.7, 31.8, 31.9, 32.0, 32.2, 32.3, 32.4, 32.5, 32.6, 32.7, 32.8, 33.1, 33.2, 33.3, 33.6, 33.7, 33.8, 34.0, 34.1, 34.2, 34.3, 34.6, 34.7, 34 The values ​​are 0.8, 35.0, 35.2, 35.3, 35.5, 35.6, 35.9, 36.0, 36.2, 36.5, 36.6, 36.7, 36.8, 36.9, 37.1, 37.2, 37.3, 37.4, 37.5, 37.6, 37.7, 37.8, 37.9, 38.2, 38.3, 38.4, 38.5, 38.6, 38.7, 38.8, 38.9, 39.0, 39.1, 39.2, 39.3, 39.4, 39.5, 39.6, 39.7, 39.8, 39.9 and 40.0°2θ.

[0313] For example, the XRPD values ​​for form B are as follows: 6.47, 8.08, 9.42, 9.59, 10.18, 10.62, 11.22, 12.17, 12.91, 12.97, 13.27, 13.37, 14.03, 14.37, 14.63, 15.02, 15.93, 16.20, 16.35, 16.43, 16.47, 16.81, 18.10, 18.35, 18.41, 18.50, 18.55, 18.6.0 18.91, 19.11, 19.15, 19.24, 19.34, 19.43, 19.51, 19.61, 19.65, 19.76, 19.85, 19.90, 20.44, 20.61, 21.34, 21.43, 21.84, 21.95, 22.17, 22.28, 22.30, 22.33, 22.44, 22.54, 22.76, 22.81, 22.97, 23.00, 23.11, 23.42, 23.80, 24.11, 24.22, 24.34 , 24.38, 24.40, 24.48, 24.56, 24.57, 25.40, 25.56, 25.57, 25.59, 25.72, 25.74, 25.94, 25.99, 26.11, 26.28, 26.29, 26.37, 26.51, 26.58, 26.61, 26.73, 26.81, 26.92, 27.15, 27.19, 27.23, 27.31, 27.49, 27.57, 27.61, 27.71, 27.88, 27.94, 28.2 7, 28.41, 28.53, 28.71, 28.74, 28.86, 28.94, 28.98, 29.03, 29.06, 29.08, 29.25, 29.30, 29.38, 29.51, 29.57, 29.61, 29.70, 29.73, 29.75, 29.90, 29.95, 30.31, 30.38, 30.42, 30.54, 30.55, 30.66, 30.73, 30.85, 30.87, 30.89, 31.23, 31.51, 31. 55, 31.61, 31.70, 31.76, 31.77, 31.80, 31.81, 31.82, 31.82, 31.90, 31.91, 31.95, 32.17, 32.21, 32.23, 32.25, 32.36, 32.37, 32.43, 32.53, 32.54, 32.56, 32.61, 32.73, 32.80, 32.82, 33.05, 33.13, 33.17, 33.22, 33.28, 33.30, 33.60, 33.65, 33.71, 33.76, 33.77, 33.99, 34.01, 34.01, 34.05, 34.10, 34.17, 34.29, 34.55, 34.60, 34.62, 34.63, 34.68, 34.75, 34.76, 35.03, 35.16, 35.19, 35.21, 35.25, 35.31, 35.46, 35.61, 35.63, 35.85, 3 5.86, 35.90, 35.97, 36.19, 36.45, 36.56, 36.58, 36.67, 36.68, 36.70, 36.71, 36.77, 36.85, 36.87, 36.90, 37.09, 37.19, 37.27, 37.28, 37.29, 37.32, 37.33, 37.37, 37.38, 37.48, 37.48, 37.50 , 37.51, 37.54, 37.61, 37.64, 37.65, 37.68, 37.69, 37.71, 37.74, 37.74, 37.76, 37.81, 37.83, 37.93, 37.94, 38.15, 38.19, 38.32, 38.36, 38.39, 38.46, 38.59, 38.63, 38.69, 38.76, 38.79, 38. The angles may also be labeled as 85, 38.87, 38.88, 38.96, 38.98, 39.02, 39.05, 39.19, 39.27, 39.33, 39.36, 39.39, 39.43, 39.44, 39.53, 39.53, 39.6, 39.61, 39.70, 39.71, 39.72, 39.82, 39.87, 39.9, and 39.98°2θ.

[0314] Observed peaks related to morphology B include 9.5±0.2, 18.1±0.2, 19.3±0.2, 22.4±0.2, 26.6±0.2, and 27.7±0.2°2θ.

[0315] The agreement between the possible peak positions indicated by the bars and the observed peaks demonstrated a consistent determination of the unit cell. Successful indexing of the pattern indicated that the sample consisted primarily of a single crystalline phase. The space group, unit cell parameters, and derived quantities, corresponding to the assigned annihilation symbols, are given in Table 8.

[0316] [Table 8]

[0317] In one embodiment, form B features an XRPD pattern including at least two 2-theta values ​​selected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.9±0.2°, and 22.4±0.2°. In one embodiment, form B features an XRPD pattern including at least three 2-theta values ​​selected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.9±0.2°, and 22.4±0.2°. In one embodiment, form B features an XRPD pattern including at least four 2-theta values ​​selected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.9±0.2°, and 22.4±0.2°. In one embodiment, form B features an XRPD pattern including at least five 2-theta values ​​selected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.9±0.2°, and 22.4±0.2°. In one embodiment, form B features an XRPD pattern including at least six 2-theta values ​​selected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.9±0.2°, and 22.4±0.2°. In one embodiment, form B features an XRPD pattern including 2-theta values ​​selected from 6.5±0.2°, 9.5±0.2°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.9±0.2°, and 22.4±0.2°. In one embodiment, form B features an XRPD pattern including at least a 2-theta value of 9.5±0.4°.

[0318] Example 7: Stability study of Form B at 6 and 12 months under 25°C / 60%RH and 40°C / 75%RH conditions. Form B was stored at 25°C / 60%RH for 12 months and at 40°C / 75%RH for 6 months.

[0319] Table 9 shows the results under 25°C / 60%RH storage conditions, and Table 10 shows the results under 40°C / 75%RH. Under both conditions, morphology B was 99.5% pure, and the XRPD spectrum matched the reference spectrum at the longest time point studied.

[0320] [Table 9]

[0321] [Table 10]

[0322] Example 8. Conversion of impure form B material to pure form B material Pure morphology B was isolated from impure morphology B, a material characterized as containing an unknown residual amount of morphology in addition to morphology B. Figure 8 shows the difference between the XRPD patterns of impure morphology B and pure morphology B (in the experiments described below, pure morphology B is morphology B as characterized in Example 6). The pattern of pure morphology B is visually similar to that of impure morphology B, except that there are no peaks at two theta angles of approximately 4.0° and 5.6°. As is common in XRPD analysis, there is also a difference in relative peak intensity, likely due to the effects of favorable orientation and / or particle statistics.

[0323] Solubility studies, small-scale scouting experiments, and experiments under dry conditions were first conducted to confirm the conditions suitable for conversion to pure form B. TG-IR characterization was performed on numerous isolated samples of form B. Once the conditions were confirmed, the conversion from impure material to pure material was carried out in a water:acetone 1:2 (v / v) slurry at a concentration of 125 mg / mL and 30°C for 43 hours, as described in more detail below.

[0324] Solubility estimation experiment for developing conditions suitable for recrystallization The solubility estimation of impure form B was attempted in various primarily HCl-acidic aqueous acetone solvent mixtures using an aliquot addition method including visual observation. Aliquots of various solvents or diluent / organic solvent mixtures were added to the measured amount of impure form B at ambient temperature with stirring (usually sonication) until complete solubility was achieved as determined by visual observation. Solubility was calculated based on the total solvent used to impart the solution. Actual solubility may be higher due to the volume of solvent portion used or slow dissolution. If dissolution did not occur as determined by visual evaluation, the value was reported as "<". If dissolution occurred with the first aliquot, the value was reported as ">". Due to shading of the obtained samples, effective solubility estimations were difficult to identify. In general, impure form B showed very limited solubility (3 mg / mL to 7 mg / mL) in the solvent mixtures tested (Table 11).

[0325] [Table 11]

[0326] Small-scale scouting experiments for developing conditions suitable for recrystallization Approximately 16 small-scale slurry experiments were performed by varying slurry concentration, temperature, HCl molar concentration, and their content in the aqueous acetone mixture, as well as the water content. Non-pure form B slurries were performed in a given solvent system at the target calculated concentration, at ambient temperature or high temperature, and for various time / durations. Solids were isolated by vacuum filtration and subjected to XRPD analysis. Specific experimental conditions are detailed in Table 12. In Table 12, the solvent system ratios are based on volume. Slurries in acidic aqueous acetone mixtures at ambient temperature (Sample 1, Sample 2, and Sample 4) could not convert non-pure form B to pure form B. Figure 9 compares the XRPD patterns of Sample 1 and Sample 4 with the XRPD patterns of non-pure form B, the starting material for the experiment. Figure 9 also compares Sample 1 and Sample 4 with the pure form B material previously characterized in Example 6.

[0327] Slurries in an acidic aqueous-acetone mixture at a high temperature of 50°C (Sample 3 and Sample 5) produced irregular material with two broad, low-angle peaks, suggesting a potential intermediate phase. Figure 10 compares the XRPD patterns of Sample 3 and Sample 5 with the impure morph B, the starting material of the experiment, and with pure morph B. For comparison purposes, the samples were also compared with a second impure sample of morph B (Impure Morph B Sample 2 in Figure 10). This second impure morph B contained a larger amount of the unknown morph than the impure morph B already described in Example 8. Increasing the molar concentration of HCl from 0.1 M (Sample 3) to 0.5 M (Sample 5) also increased the intensity of these two peaks.

[0328] Starting with the impure form B, several slurry experiments were conducted in a water:acetone solvent system, varying the water:acetone ratio, slurry concentration, and time. Based on the results of the initial slurry, experiments were conducted in a 1:2 (v / v) water:acetone mixture at ambient temperature using aliquots taken after 16 hours (sample 6) and aliquots taken after 20.5 hours (sample 7). The slurry in this solvent system was prepared at concentrations of 100 mg / mL to 125 mg / mL and under ambient air. The XRPD patterns of the obtained materials were consistent with pure form B (Figure 11). The use of a water:acetone (1:2) solvent system resulted in a low yield of 78% to 79%, which was calculated with respect to solids isolated by vacuum filtration without drying.

[0329] As part of efforts to improve yield, a water:acetone 1:3 (v / v) mixture was used at a concentration of 150 mg / mL (Samples 13 and 14), but the conversion was not completed even after 4 days (Figure 12). Therefore, one experiment was conducted using a water:acetone 1:2 (v / v) slurry for 18 hours, followed by the addition of acetone to reach a water:acetone ratio of 1:4 (v / v), and then slurrying for 4 hours (Sample 12). The XRPD pattern of the obtained material was consistent with morphology B, but one of the undesirable peaks that reappeared was shifted from 3.95°2θ to 4.2°2θ (Figure 12).

[0330] [Table 12] TIFF0007850883000018.tif54170

[0331] Slurry experiment with Lara Controlled Laboratory Reactor As part of an attempt to demonstrate applicable conditions for the conversion of impure form B to pure form B, several scale-up experiments were performed. The slurry conversion experiments were conducted using a 1 L round-bottom controlled laboratory reactor (Radleys Lara CLR) equipped with a Teflon anchor impeller, a Julabo temperature control unit, and a temperature probe for monitoring reactor temperature throughout the experiment. The Julabo FP50 temperature control unit contained Julabo Thermal C10 liquid, and the reactor temperature was measured using a morph KPTFE temperature probe. The experiments were performed using Lara Control software version 2.3.5.0. The software tracked the circulator temperature, vessel temperature, and stirring speed, recording readings every 0.1 seconds throughout the experiment.

[0332] Impure form B solids (58.86 g) were added to 471 mL of a water:acetone 1:2 (v / v) solvent system in a reaction vessel to achieve a slurry concentration of 125 mg / mL (Samples 20-23). ​​The resulting slurry was stirred at 30°C for a maximum of 43 hours at a stirring speed of 400 rpm. The slurry was cooled to 25°C over 30 minutes, removed from the reaction vessel, and immediately filtered gradually toward a dry place (drop by drop). A water:acetone 1:2 (v / v) washing solution was prepared in advance and used to wash the filter cake in one go.

[0333] Aspirates were taken over a typical 20-hour period, or later if necessary (Pulls were taken) (Table 13). Scale-up experiments showed that longer periods and slightly higher temperatures (30°C above ambient temperature) were required to convert impure morph B to pure morph B on a larger scale. Sample 22 was converted to pure morph B, while samples 21 and 23 were not analyzed. Sample 20 produced morph B, but a broad peak was also observed at 4.2°²θ.

[0334] [Table 13]

[0335] TG-IR characterization of compound 2, form B. TG analysis was performed using a TA Instrument Q5000 thermogravimetric analyzer. Temperature calibration was performed using nickel and Alumel. The sample was placed in a platinum dish and inserted into the TG furnace. The furnace was heated to 350°C at a rate of 10°C / min under nitrogen purging.

[0336] Thermogravimetric (TG-IR) analysis was performed using a TA Instruments Q5000 IR thermogravimetric (TG) analyzer coupled to a Magna-IR 560® Fourier transform infrared (FT-IR) spectrophotometer (Thermo Nicolet) equipped with an Ever-Glo mid / far IR source, potassium bromide (KBr) beam splitter, and cadmium mercury telluride (MCT-A) detector. FT-IR wavelength validation was performed using polystyrene, and the calibration standards for the TG were nickel and Alumel®. The sample was placed in a platinum sample dish, which was then inserted into the TG furnace. The TG instrument was started first, followed immediately by the FT-IR instrument. The TG instrument was operated under helium flow rates of 90 cc / min and 10 cc / min for purging and balancing, respectively. The furnace was heated to a final temperature of approximately 140°C at a rate of 20°C / min under helium. IR spectra were collected approximately every 32 seconds for approximately 7.5 minutes. Each IR spectrum was measured at 4 cm. -1This represents 32 co-added scans collected at the given spectral resolution. Volatile substances were identified from a search of the High Resolution Nicolet Vapor Phase spectral library.

[0337] As part of an attempt to study the stability of morph B at high temperatures by monitoring the potential release of hydrogen chloride, a TG-IR experiment was performed on pure morph B (sample 11 from a small-scale slurry experiment) for 20 hours at ambient temperature.

[0338] TG data showed a weight loss of 6.4% between 33°C and 137°C (Figure 13). The correlation between time and temperature is shown in Table 14. A series of IR spectra collected during the TG-IR experiment are shown in Figures 14 and 15. The spectra demonstrated that only water was detected as a volatile substance and that hydrogen chloride was not released.

[0339] [Table 14]

[0340] Drying experiment of compound 2, form B The weighed quantities of the impure and pure forms B samples from previous experiments (Sample 14, Sample 8, Sample 11, Sample 19, Sample 21, and Sample 23) were vacuum-dried at ambient temperature or high temperature using various vacuum levels ranging from approximately 14 to a maximum of 27-28 in Hg. After weighing the obtained materials, they were subjected to XRPD analysis.

[0341] The two samples (Sample 14 and Sample 8) were vacuum-dried at 40°C for 15 hours (approximately 29% of Hg), resulting in a weight loss of approximately 7.4%. One of the samples (Sample 8) was analyzed by XRPD, yielding a new crystalline XRPD pattern that did not correspond to morphology B (Figure 16).

[0342] Sample 11 showed a weight loss of 1.8% after vacuum drying at ambient temperature for 0.5 hours (approximately 14 in Hg) (calculated by weighing the sample before and after drying). The XRPD pattern of the obtained material was consistent with morphology B, but slight shifts were observed in the positions of some peaks in the XRPD pattern (Figure 17). A significant peak shift was observed in the XRPD pattern of sample 23 (Figure 17), which resulted in a weight loss of 4.7% after vacuum drying at ambient temperature for 1 hour (approximately 27-28 in Hg) (calculated by weighing the sample before and after drying).

[0343] [Table 15]

[0344] Transformation from impure form B to pure form B The conversion from impure form B to pure form B was carried out in a 125 mg / mL water:acetone 1:2 (v / v) slurry at 30°C for 43 hours. Very slow filtration was observed, and the wet cake was air-dried at ambient temperature for 3.5 hours, then vacuum-dried at ambient temperature and 15 in Hg for 0.5 hours, followed by 3.5 hours at approximately 27 in Hg to obtain 49.26 g (84%).

[0345] As shown in Table 16, XRPD patterns were obtained at various points in the conversion. After 42 hours of heating, XRPD analysis showed that the impure form B had been completely converted to pure form B. After filtering and drying the material, TG analysis was performed in addition to XRPD analysis.

[0346] The XRPD pattern shown by the converted batch after drying was consistent with the XRPD pattern of pure morphology B in Figure 7, and its peaks coincided with the acceptable peak positions from the pattern shown in Figure 7. Figure 18 compares the patterns of impure morphology B, pure morphology B as characterized in Example 6, and pure morphology B converted from impure morphology B as described in Example 8.

[0347] The TGA data for the converted batch of form B showed a weight loss of 7.5% at 31°C to 120°C (Figure 19).

[0348] [Table 16]

Claims

1. (i) comprising at least five two-theta values ​​selected from 6.5±0.2°, 9.5±0.4°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.3±0.2°, 19.9±0.2°, and 22.4±0.2°, or (ii) including at least three 2-theta values ​​selected from 6.5±0.2°, 9.5±0.4°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.3±0.2°, 19.9±0.2°, and 22.4±0.2° (except for further including a 2-theta value of 5.3±0.2° or 7.2±0.2°), or (iii) A structure characterized by an X-ray powder diffraction (XRPD) pattern including a two-theta value of 6.5±0.2° and at least two two-theta values ​​selected from 9.5±0.4°, 14.0±0.2°, 14.4±0.2°, 18.1±0.2°, 19.3±0.2°, 19.9±0.2°, and 22.4±0.2°: 【Chemistry 1】 Isolated crystal B of the diHCl salt.

2. The isolated crystal B according to claim 1, wherein the XRPD pattern includes at least two theta values ​​of 9.5 ± 0.4°.

3. The isolated crystal B according to claim 1 or 2, wherein the XRPD pattern includes at least two theta values ​​of 9.5 ± 0.2°.

4. The isolated crystal B according to any one of claims 1 to 3, wherein the XRPD pattern includes at least two theta values ​​of 19.3 ± 0.2°.

5. The isolated crystal B according to any one of claims 1 to 4, wherein the XRPD pattern includes at least two theta values ​​of 22.4 ± 0.2°.

6. An isolated crystal B according to any one of claims 1 to 5, having an endothermic start of differential scanning calorimetry (DSC) at 105 ± 20°C, 220 ± 20°C, and 350 ± 20°C.

7. An isolated crystal B according to any one of claims 1 to 5, having an endothermic start of differential scanning calorimetry (DSC) at 105 ± 10°C, 220 ± 10°C, and 350 ± 10°C.

8. A pharmaceutical composition comprising isolated crystal B according to any one of claims 1 to 7 in a pharmaceutically acceptable excipient for solid drug delivery.

9. The pharmaceutical composition according to claim 8, further comprising one or more further therapeutic agents.

10. The one or more further therapeutic agents include antiandrogens, anticancer agents, aromatase inhibitors, Bruton's tyrosine kinase (BTK) inhibitors, CYP17 inhibitors, extracellular signal-regulated kinase (ERK) inhibitors, gonadotropin-releasing hormone superagonists (GnRH agonists), luteinizing hormone-releasing hormone (LH-RH) agonists, luteinizing hormone-releasing hormone (LH-RH) antagonists, rapamycin target protein (mTOR) inhibitors, and mitogenic factor-activated protein kinase (M) inhibitors. The pharmaceutical composition according to claim 9, selected from an EK inhibitor, a nucleoside or nucleotide analog or prodrug, a phosphatidylinositol 3-kinase (PI3K) pathway inhibitor, a rapidly progressive fibrosarcoma (RAF) kinase inhibitor, a renin-angiotensin system (RAS) inhibitor, a selective estrogen receptor degrader (SERD), a selective estrogen receptor modulator (SERM), a serine-threonine protein kinase B (Akt) inhibitor, or a topoisomerase inhibitor.

11. A pharmaceutical composition for treating a disorder associated with abnormal cell proliferation in a host, comprising isolated crystal B according to any one of claims 1 to 7, or a pharmaceutical composition according to any one of claims 8 to 10.

12. The pharmaceutical composition according to claim 11, wherein the host is human.

13. A method for producing crystal B according to any one of claims 1 to 7, (i) A step of heating the free base of 2'-((5-(4-isopropylpiperazine-1-yl)pyridine-2-yl)amino)-7',8'-dihydro-6'H-spiro[cyclohexane-1,9'-pyrazino[1',2':1,5]pyrrolo[2,3-d]pyrimidine]-6'-one in HCl water to at least 55±10°C, (ii) A step of stirring the solution for at least 15 minutes and filtering the resulting solution, (iii) Adding acetone at a temperature of at least 55 ± 10°C and stirring the solution for at least 15 minutes, (iv) The step of lowering the temperature of the solution to 25±5℃ or below and stirring the solution for at least 30 minutes, (v) A step of filtering the solution and washing the solid recovered by filtration with acetone to obtain crystal B according to any one of claims 1 to 7, Methods that include...

14. The method according to claim 13, wherein the volume of acetone added in step (iii) is at least seven times the volume of the aqueous HCl solution.

15. The method according to claim 13 or 14, wherein acetone is added over a period of 1 hour in step (iii).

16. The method according to any one of claims 13 to 15, wherein the temperature is lowered at a rate of 20°C / hour in step (iv).

17. The method according to any one of claims 13 to 16, wherein the solution is stirred for 45 minutes in step (ii).

18. The method according to any one of claims 13 to 17, wherein the solution is heated to at least 50°C in step (i).

19. The method according to any one of claims 13 to 17, wherein the solution is heated to 55°C in step (i).

20. The method according to any one of claims 13 to 19, wherein the solvent is heated to at least 50°C in step (iii).

21. The method according to any one of claims 13 to 20, wherein the solution is stirred for at least one hour in step (iii).

22. The method according to any one of claims 13 to 21, wherein the solution is stirred for at least one hour in step (iv).

23. The method according to any one of claims 13 to 21, wherein the solution is stirred for at least two hours in step (iv).

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