Crystalline forms of 4-amino-n-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinoprop-1-yn-1-yl)-7h-pyrrolo[2,3-d]pyrimidine-5-carboxamide, methods of preparation and uses thereof

Abstract

Provided herein are various crystalline forms of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinoprop-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, methods of producing them, and methods of using them for RET inhibition.

Description

[0001] Related applications

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 116,191, filed November 20, 2020, the entire contents of which are incorporated herein by reference for any purpose. Technical Field

[0003] This invention discloses a selective RET inhibitor useful in cancer treatment, crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (also known as HM06 or TAS953). The invention also discloses crystalline forms of HM06, both in its free base and salt forms, and methods for producing them. Background Technology

[0004] Multiple protein kinases exist in the body and are known to participate in a wide range of functional regulation. RET is a receptor tyrosine kinase identified as one of the proto-oncogenes. RET binds to glial lineage-derived neurotrophic factor (GDNF) and GDNF receptors to form a complex that enables RET to perform physiological functions through intracellular phosphorylation signal transduction (Bavetsias et al., “Aurora Kinase Inhibitors: Current Status and Outlook”, Frontiers in Oncology, 2015, Vol. 5, Art. 278). Some studies have shown that in cancers such as lung cancer, thyroid cancer, breast cancer, pancreatic cancer, and prostate cancer, RET translocation, mutation, and overexpression enhance its activation, thereby contributing to cell growth, tumor formation, or tissue invasion.(Kohno et al., “KIF5B-RET fusions in lung adenocarcinoma,” Nature Med., 18(3): pp. 375-377, (2012); Santoro et al., “RET / PTC activation in papillary thyroid carcinoma: European Journal of Endocrinology Prize Lecture,” Eur J Endocrinol., 155: pp. 645-653, (2006); Yeganeh et al., “RET Prato Oncogene Mutation Detection and Medullary Thyroid Carcinoma Prevention,” Asian Pac J Cancer Prev., 16(6): pp. 2107-2117, (2015); Gattelli et al., “Ret inhibition decreases growth and metastatic potential of estrogen receptor positive breast cancer cells,” EMBO Mol Med., 5: pp. 1335-1350, (2013); Ito et al., “Expression of glial cell line-derived neurotrophic factor family members and their receptors in pancreatic cancers,” Surgery, 138: pp. 788-794, (2005); and Dawson et al., “Altered Expression of RET Proto-oncogene Product in Prostatic Intraepithelial Neoplasia and Prostate Cancer,” J Natl Cancer Inst., 90: pp. 519-523, (1998)).In addition, RET is known to be a poor prognostic factor for cancer, as indicated in some reports, where RET translocation and its enhanced activation level are negatively correlated with prognosis in cancer (Cai et al., “KIF5B-RET Fusions in Chinese Patients With Non-Small Cell Lung Cancer,” Cancer, 119: pp. 1486-1494, (2013); Elisei et al., “Prognostic Significance of SomaticRET Oncogene Mutations in Sporadic Medulllary Thyroid Cancer: A 10-Year Follow-Up Study,” J Clin Endocrinol Metab., 93(3): pp. 682-687, (2008); Gattelli et al., “Retinhibition decreases growth and metastatic potential of estrogen receptor-positive breast cancer cells,” EMBO Mol Med., 5: pp. 1335-1350, (2013); and Zeng et al., “The Relationship between “Over-expression of Glial Cell-derived Neurotrophic Factor and Its RET Receptor with Progression and Prognosis of Human Pancreatic Cancer,” J. Int. Med. Res., 36: pp. 656-664, (2008) . Therefore, inhibitors that can suppress RET activity are considered useful as therapeutic agents for diseases associated with abnormally enhanced RET signaling pathways, including cancer.

[0005] Furthermore, various cancers can lead to metastatic brain tumors. Symptomatic metastatic brain tumors have been reported in 8 to 10% of cancer patients, and there are also reports of brain metastases in lung cancer at a rate of 40 to 50% based on autopsy findings. (Qingbei Zeng, J Med Chem. 22; 58(20): 8200-15, (2015); Lakshmi Nayak, Curr Oncol Rep; 14(1): 48-54, (2012); Brunilde Gril, Eur J Cancer.; 46(7): 1204-10, (2010)). Therefore, it is hoped that effective treatments for cancer, including those for brain metastases, can be found.

[0006] Furthermore, it is desirable to administer treatment in a form that is easily absorbed by the body and also stores stably. The active pharmaceutical ingredient used to prepare the treatment should be as pure as possible, and its long-term storage stability under various environmental conditions should be ensured. These properties are useful in preventing the formation of unintentional degradation products in the pharmaceutical composition, which can be potentially toxic or simply reduce the potency of the composition.

[0007] A major challenge in the large-scale production of pharmaceutical compounds is that the active ingredient must possess a stable crystalline morphology to ensure consistent processing parameters and drug quality. If an unstable crystalline form is used, the morphology can change during production and / or storage, leading to quality control issues and inconsistencies in formulations. This variation can affect the reproducibility of the manufacturing process and thus result in the final formulation failing to meet the high quality and stringent requirements imposed on the pharmaceutical composition. In this regard, any change to a solid state of the pharmaceutical composition that can improve its physical and chemical stability should generally be considered to offer a significant advantage over different stable forms of the same drug.

[0008] When a compound crystallizes from a solution or slurry, it can crystallize in different spatial lattice arrangements; this property is called "polymorphism." Each of these crystalline forms is a "polymorph." Although polymorphs of a given substance have the same chemical composition, they can differ from each other with respect to one or more physical properties, such as solubility, dissociation, true density, dissolution, melting temperature, crystal form, compressive behavior, flow properties, and / or solid-state stability.

[0009] U.S. Patent No. 10,155,768 reports a RET inhibitory agent for 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (also known as HM06 or TAS0953). The molecular formula of HM06 / TAS0953 in its free base form is C0 26 H30 N6O3 has a molecular weight of 474.57, and the structural formula of its free base is:

[0010]

[0011] However, the crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide has not been disclosed to date. Invention Overview

[0013] Therefore, this paper discloses the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide in the form of a free base and an HCl salt, as well as methods for preparing said crystalline form and methods for using said form.

[0014] This invention discloses the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide. In one aspect of this invention, the crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is a free base. In one aspect of the present invention, the crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is an HCl salt, for example, a 1:1 or 1:2 HCl salt.

[0015] The present invention also relates to pharmaceutical compositions comprising at least one basic crystalline form as described herein and a pharmaceutically acceptable excipient.

[0016] The present invention also relates to a method of treating cancer in patients in need of it, comprising administering to the patient an effective amount of the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide.

[0017] Other objectives and advantages will be set forth in part in the following description, and will be apparent in part from the description and may be learned by practice. The objectives and advantages will be achieved and obtained by means of the elements and combinations specifically specified in the appended claims.

[0018] It should be understood that the foregoing general description and the following detailed description are merely exemplary and illustrative and are not intended to limit the scope of the claims.

[0019] The accompanying drawings, which are incorporated in and form part of this specification, illustrate one or more embodiments and, together with the description, serve to explain the principles described herein. Attached Figure Description

[0020] Figure 1A The XRPD pattern of HM06 crystals in free alkali form 1 obtained using CuKα radiation is shown.

[0021] Figure 1B The XRPD pattern of HM06 crystals in free alkali form 2, obtained using CuKα radiation, is shown.

[0022] Figure 1C The XRPD pattern of HM06 crystals in free alkali form 3, obtained using CuKα radiation, is shown.

[0023] Figure 1D The XRPD pattern of HM06 crystals in free alkali form 4, obtained using CuKα radiation, is shown.

[0024] Figure 1E The XRPD pattern of HM06 crystals in free alkali form 5, obtained using CuKα radiation, is shown.

[0025] Figure 1F The XRPD pattern of HM06 HCl crystal salt form A obtained using CuKα radiation is shown.

[0026] Figure 1G The XRPD patterns of five HM06 crystal free base forms and HM06 HCl crystal salt form A obtained using CuKα radiation are shown as overlapping.

[0027] Figure 2A The XRPD pattern of crystal HM06 1:1HCl form 1 obtained using CuKα radiation is shown.

[0028] Figure 2B The XRPD pattern of crystal HM06 1:1HCl form 1 obtained using CuKα radiation is shown.

[0029] Figure 2CThe XRPD pattern of crystal HM06 1:1HCl form 1 obtained using CuKα radiation is shown.

[0030] Figure 2D This is the DSC temperature spectrum of HM06 1:1HCl form 1.

[0031] Figure 2E TGA spectra of HM06 1:1HCl form 1 are provided.

[0032] Figure 3A The XRPD pattern of crystal HM06 1:2HCl form 1 obtained using CuKα radiation is shown.

[0033] Figure 3B TGA spectra of HM06 1:2HCl form 1 are provided.

[0034] Figure 3C This is the DSC temperature spectrum of HM06 1:2HCl crystal form 1.

[0035] Figure 3D The diagram shows twins formed by a space group transformation from orthorhombic to monoclinic. Macroscopic views of the crystal are shown from both bottom and top views. Twin monoclinic crystals are produced when molecules rearrange themselves in the same manner in A (viewed from the top) and B (viewed from the bottom). Monoclinic stacking is equivalent in both volumes, but independent of crystallographic symmetry; instead, the twin operator, rotating 180° around an axis in a plane perpendicular to the unique crystallographic b-axis of the monoclinic lattice, involves diffraction patterns in both volumes.

[0036] Figure 3E The ORTEP diagram of a single crystal of HM06 1:2HCl form 1 is provided.

[0037] Figure 4A The image shows a crystal packing view of HM06 1:2HCl form 1 along the "a-axis".

[0038] Figure 4B The image shows a crystal packing view of HM06 1:2HCl form 1 along the "b-axis".

[0039] Figure 5A The XRPD pattern of crystal HM06 1:2HCl form 1-bis obtained using CuKα radiation is shown.

[0040] Figure 5B The XRPD pattern of HM06 1:2HCl crystal form 1-bis obtained using CuKα radiation is shown.

[0041] Figure 5CIt is an overlap of the XRPD patterns of HM06 1:2HCl form 1-bis in vacuum (time = zero) and in air at 2 to 20 minutes in the range of 2θ = 25.5° to 27.5°.

[0042] Figure 5D The XRPD patterns of HM06 1:2HCl form 1-bis as the starting material are shown as an overlap of spectra under vacuum and after water absorption from air.

[0043] Figure 6A The overlay of XRPD patterns (top pattern) of HM06 1:2HCl Form 2 samples obtained from ethanol after a slurry test at 50°C is shown. For comparison, standard reference patterns (bottom two patterns) of Form 1 and Form 2 are reported.

[0044] Figure 6B The image shows the overlap of XRPD patterns (top two patterns) of HM06 1:2HCl Form 2 obtained from ethanol after a slurry experiment at 50°C. For comparison, standard reference patterns of Form 2 (bottom pattern) and Form 3 (second to last from the bottom pattern) are reported.

[0045] Figure 6C The XRPD pattern of HM06 1:2HCl form 2, obtained using CuKα radiation, was collected from ethanol and used as an STD reference after a 4-day slurry experiment at a scale of 100 mg.

[0046] Figure 6D This is the DSC temperature spectrum of HM06 1:2HCl form 2.

[0047] Figure 6E TGA spectra of HM06 1:2HCl form 2 are provided.

[0048] Figure 6F XRPD patterns of samples collected after small-scale scaling up of R01 (second from the top pattern) and R02 (top pattern) in Example 5 are provided. For comparison, standard reference XRPD patterns of Forms 2 and 3, provided as the bottom two patterns, are included.

[0049] Figure 6G XRPD patterns (top line) of samples collected after a small-scale amplification procedure using a concentration of 20 mg / mL are provided. For comparison, standard reference XRPD patterns of forms 2 and 3, provided as the bottom two lines, are included.

[0050] Figure 7AThe image shows the overlay of XRPD patterns of standard reference patterns of HM06 1:2HCl crystals obtained from acetonitrile after a slurry experiment at 50 °C using CuKα radiation. The patterns are: Form 3 (blue line), Form 1 (black line), Form 2 (green line), and Form 3 (pink line).

[0051] Figure 7B The XRPD pattern of HM06 1:2HCl form 3 collected from 1-propanol after rapid gradient precipitation at a scale of 100 mg obtained using CuKα radiation is shown.

[0052] Figure 7C This is the DSC temperature spectrum of HM06 1:2HCl form 3.

[0053] Figure 7D TGA spectra of HM06 1:2HCl form 3 are provided.

[0054] Figure 8A The XRPD pattern of HM06 1:2HCl form 4-bis obtained using CuKα radiation is shown (top pattern), and the XRPD pattern of HM06 1:2HCl form 1 as a reference is shown (bottom pattern).

[0055] Figure 8B The XRPD pattern of 4-bis in the form of HM06 1:2HCl is shown.

[0056] Figure 8C The XRPD patterns of HM06 1:2HCl form 4-bis (bottom pattern) and the same sample analyzed after 7 days of storage in a sealed vial (middle pattern) are shown as an overlap. For comparison, the XRPD pattern of HM06 1:2HCl form 1 (top pattern) is provided.

[0057] Figure 8D The XRPD pattern of HM06 1:2HCl form 4 obtained using CuKα radiation is shown.

[0058] Figure 8E The image shows an overlap of the XRPD pattern of HM06 1:2HCl form 4 (bottom pattern) and the pattern after overnight storage at 43% relative humidity (top pattern).

[0059] Figure 9A The XRPD pattern of HM06 1:2HCl form 5-bis (top pattern) is shown compared to form 5 (bottom pattern).

[0060] Figure 9B The XRPD pattern of HM06 1:2HCl form 5-bis obtained using CuKα radiation is shown.

[0061] Figure 9C The XRPD patterns of HM06 1:2HCl form 5-bis (blue top pattern) and the same sample analyzed after 18 hours of exposure (red bottom pattern) are shown to overlap.

[0062] Figure 9D The XRPD patterns of HM06 1:2HCl form 5-bis (blue top pattern) and the same sample analyzed after 7 days in a sealed vial (middle pattern) are shown as an overlay. For comparison, the XRPD pattern of HM06 1:2HCl form 1 (bottom pattern) is provided.

[0063] Figure 9E The XRPD pattern of HM06 1:2HCl form 5 obtained using CuKα radiation is shown.

[0064] Figure 9F This is the DSC temperature spectrum of HM06 1:2HCl form 5.

[0065] Figure 9G TGA spectra of HM06 in 1:2HCl form 5 are provided.

[0066] Figure 10 The XRPD pattern of HM06 1:2HCl form 6 is shown.

[0067] Figure 11 The XRPD patterns of the separated forms of crystal HM06 1:2HCl are shown as overlapping. Invention Details

[0069] As summarized above and described in detail below, this invention discloses the crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (also known as HM06 or TAS953). This invention also discloses a method for preparing the crystalline free base form and its HCl salt form, such as dichloride (or 1:2) HCl salt:

[0070]

[0071] This article also discloses methods for using the described crystal form for therapeutic treatments, such as for cancer.

[0072] Detailed information about the invention is set forth in the following description. While similar or equivalent methods and materials to those described herein may be used in the practice or testing of the invention, illustrative methods and materials are described hereafter. Other features, objectives, and advantages of the invention will be apparent from the description and the claims. Unless the context clearly requires, the singular form also includes the plural form in the specification and appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. All patents and patent disclosures referenced in this specification are incorporated herein by reference in their entirety.

[0073] This invention discloses the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide. In at least one aspect of this invention, the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is a free base.

[0074] In some embodiments disclosed in this invention, the basic crystalline form of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base is form 1. In at least one embodiment, form 1 of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base is characterized by its consistency with... Figure 1A The XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base form 1 is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 12.57°2θ, 13.36°2θ, 16.08°2θ, 18.86°2θ, 20.66°2θ, 21.73°2θ, 23.90°2θ, and 24.86°2θ.

[0075] In some embodiments disclosed in this invention, the basic crystalline form of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base is form 2. In at least one embodiment, the basic crystalline form of form 2 of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base is characterized by... Figure 1B The XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base form 2 is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 7.94°2θ, 10.47°2θ, 11.53°2θ, 15.75°2θ, 21.80°2θ, and 23.65°2θ.

[0076] In some embodiments disclosed in this invention, the basic crystalline form of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base is form 3. In at least one embodiment, the basic crystalline form of form 3 of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base is characterized by... Figure 1C The XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base form 3 is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 7.11°2θ, 7.83°2θ, 14.12°2θ, 16.15°2θ, 20.61°2θ, 21.19°2θ, 26.37°2θ, and 28.59°2θ.

[0077] In some embodiments disclosed in this invention, the basic crystalline form of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base is form 4. In at least one embodiment, the basic crystalline form of form 4 of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base is characterized by... Figure 1D The XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base form 4 is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 7.77°2θ, 9.48°2θ, 11.54°2θ, 16.34°2θ, 20.21°2θ, 23.24°2θ, and 24.77°2θ.

[0078] In some embodiments disclosed in this invention, the basic crystalline form of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base is form 5. In at least one embodiment, the basic crystalline form of form 5 of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base is characterized by... Figure 1E The XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of the 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide free base form 5 is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 9.51°2θ, 13.52°2θ, 18.71°2θ, 21.26°2θ, 21.49°2θ, 28.60°2θ, and 29.05°2θ.

[0079] In some embodiments disclosed in this invention, the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is a mixture in the form of a free base.

[0080] In some embodiments, the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is HCl salt form A. In at least one embodiment, the basic crystalline form of HCl salt form A of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is characterized by its similarity to... Figure 1F The XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of the HCl salt form A of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 5.67°2θ, 7.19°2θ, 7.32°2θ, 10.90°2θ, 14.31°2θ, 14.59°2θ, 20.08°2θ, and 21.24°2θ.

[0081] In some embodiments disclosed in this invention, the basic crystalline form is a 1:1 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide. In at least one embodiment, the basic crystalline form of the 1:1 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is form 1. In at least one embodiment, the basic crystalline form of the 1:1 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is characterized by its... Figure 2A , 2BOr an XRPD pattern substantially the same as 2C. In at least one embodiment, the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide in 1:1 HCl form 1 is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 6.53°2θ, 7.37°2θ, 9.07°2θ, 14.60°2θ, 16.35°2θ, 21.26°2θ, and 26.12°2θ. In at least one embodiment, the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide in 1:1 HCl form 1 has at least one characteristic selected from the following: Figure 2D Essentially the same DSC thermogram, and with Figure 2E The TGA spectra are basically the same.

[0082] In some embodiments disclosed in this invention, the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is a mixture of HCl form A and 1:1 HCl form 1.

[0083] In some embodiments, the basic crystalline form is 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide 1:2HCl salt.

[0084] In some embodiments, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is form 1. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, form 1, is characterized by its similarity to... Figure 3AThe XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide 1:2HCl is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 6.59°2θ, 7.40°2θ, 9.12°2θ, 14.57°2θ, 16.39°2θ, 26.06°2θ, 26.57°2θ, and 27.07°2θ. In at least one embodiment, the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide in 1:2HCl form 1 has at least one characteristic selected from the following: Figure 3C Essentially the same DSC thermogram, and with Figure 3B The TGA spectra are basically the same.

[0085] In some embodiments, the basic crystalline form is 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide 1:2HCl salt in form 1-bis. In at least one embodiment, the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide 1:2HCl salt form 1-bis is characterized by its similarity to... Figure 5A or Figure 5B The XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of 1-bis, a 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 6.66°2θ, 7.63°2θ, 9.31°2θ, 10.74°2θ, 13.09°2θ, 16.45°2θ, 21.36°2θ, 26.70°2θ, and 29.01°2θ.

[0086] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is a mixture of form 1 and form 1-bis.

[0087] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is form 2. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, form 2, is characterized by its similarity to... Figure 6C The XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 7.13°2θ, 12.21°2θ, 14.22°2θ, 15.50°2θ, 17.18°2θ, 21.60°2θ, 22.23°2θ, 23.26°2θ, 26.72°2θ, and 27.69°2θ. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide has at least one characteristic selected from the following: Figure 6D Essentially the same DSC thermogram, and with Figure 6E The TGA spectra are basically the same.

[0088] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is a mixture of form 1 and form 2.

[0089] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is form 3. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, form 3, is characterized by its similarity to... Figure 7B The XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 5.44°2θ, 9.94°2θ, 14.85°2θ, 22.39°2θ, 22.84°2θ, and 27.96°2θ. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide has at least one characteristic selected from the following: Figure 7C Essentially the same DSC thermogram, and with Figure 7D The TGA spectra are basically the same.

[0090] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is a mixture of forms 2 and 3.

[0091] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is form 4-bis. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, form 4-bis, is characterized by... Figure 8BThe XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of 4-bis, a 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 4.35°2θ, 5.98°2θ, 6.20°2θ, 8.54°2θ, 17.39°2θ, 21.28°2θ, 21.58, and 21.89°2θ.

[0092] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is form 4. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, form 4, is characterized by... Figure 8D The XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 4.38°2θ, 6.15°2θ, 8.60°2θ, 9.62°2θ, 21.46°2θ, 21.90°2θ, and 26.14°2θ.

[0093] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is a mixture of form 4-bis(4-bis) and form 4.

[0094] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is a mixture of form 1 and form 4.

[0095] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is form 5-bis. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, form 5-bis, is characterized by... Figure 9B The XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of 5-bis, a 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 6.08°2θ, 6.82°2θ, 7.11°2θ, 7.51°2θ, 8.92°2θ, 9.35°2θ, 11.34°2θ, 17.29°2θ, 20.02°2θ, 21.21°2θ, 22.36°2θ, and 23.15°2θ.

[0096] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is form 5. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, form 5, is characterized by... Figure 9EThe XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of 5-bis, a 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 6.74°2θ, 7.11°2θ, 8.10°2θ, 13.10°2θ, 17.16°2θ, 23.28°2θ, 24.22°2θ, 25.15°2θ, and 26.24°2θ. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide has at least one characteristic selected from the following: Figure 9F Essentially the same DSC thermogram, and with Figure 9G The TGA spectra are basically the same.

[0097] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is a mixture of form 5-bis and form 5.

[0098] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is form 6. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide, form 6, is characterized by... Figure 10The XRPD patterns are substantially the same. In at least one embodiment, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is characterized by an XRPD pattern containing one or more peaks selected from the following: peaks located at about 5.88°2θ, 7.01°2θ, 8.81°2θ, 11.51°2θ, 13.12°2θ, 18.36°2θ, 21.4°2θ, and 22.92°2θ.

[0099] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is a mixture of forms 1, 2 and 3.

[0100] In some embodiments disclosed in this invention, the basic crystalline form of the 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide is a mixture of at least one of the forms selected from form 1, form 1-bis, form 2, form 3, form 4-bis, form 4, form 5-bis, form 5, and form 6.

[0101] The basic crystal form disclosed herein can be at least 50% crystal form, such as at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% crystal.

[0102] This invention also relates to pharmaceutical compositions comprising at least one basic crystalline form as disclosed herein and a pharmaceutically acceptable excipient. For example, in some embodiments, the pharmaceutical composition may comprise the basic crystalline form of a 1:2 HCl salt of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide.

[0103] The present invention also relates to a method of treating cancer in patients in need of treatment, comprising administering to the patient an effective amount of the basic crystalline form of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide. In at least one embodiment, the basic crystalline form is the free base form 1 of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide. In at least one embodiment, the basic crystalline form is 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide 1:1HCl form 1. In at least one embodiment, the basic crystalline form is 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide 1:2HCl form 1.

[0104] Table 1. Abbreviations and names used in the following embodiments and elsewhere herein include:

[0105]

[0106] Example

[0107] Detailed information on experiments and instruments

[0108] Unless otherwise stated, the following instruments and parameters are used for the physical characterization of the crystal forms disclosed herein.

[0109] X-ray powder diffraction (XRPD) analysis

[0110] Instrument type: Rigaku MiniFlex600

[0111] Application of Switch: Miniflex Guidance

[0112] Measurement details

[0113] Measurement type: Single scan

[0114] Sampling method: reflection

[0115] scanning

[0116] Scan range: 3,000–40,000° (2θ)

[0117] Step size: 0.01°(2θ)

[0118] Speed: 10.0° / min(2θ)

[0119] Scanning method: Continuous

[0120] The wavelength used

[0121] Expected wavelength type: Kα1

[0122] Kα1:

[0123] Kα2:

[0124] Kα2 / Kα1 intensity ratio: 0.50

[0125] Kα:

[0126] Kα:

[0127] Instrument Details

[0128] X-ray generator

[0129] Tube output voltage: 40kV

[0130] Tube output: 15mA

[0131] High voltage generation method: High-frequency Cockcroft-Walton method

[0132] Stability: For a ±10% input power variation, both tube voltage and tube current remain within ±0.05%.

[0133] X-ray tube

[0134] Name: Toshiba Analix A-26L

[0135] Anode material: Cu

[0136] Maximum output: 0.60kW

[0137] Focal point size: 1×10mm

[0138] Kβ filter

[0139] Name: Ni-Filter

[0140] Thickness (mm): 0.015

[0141] Material: Ni

[0142] Goniometer (angle measuring device)

[0143] Type: Vertical θ / 2θ

[0144] Goniometer radius: 150mm

[0145] Scan axis: θ / 2θ connection

[0146] 2θ scan range: +2° to +140°

[0147] Minimum step angle along the θ / 2θ axis: 0.005°(2θ)

[0148] Position velocity: 500° / min(2θ)

[0149] Scanning speed: 0.01 to 100° / min

[0150] Data angle: 2θ = 10°

[0151] X-ray deviation angle: 6° (fixed)

[0152] gap

[0153] DS: 1.25°

[0154] IHS: 10.0mm

[0155] SS: None (On)

[0156] RS: None (On)

[0157] Soler gap on the incident side: 2.5°

[0158] Receiving side Soler gap: 2.5°

[0159] detector

[0160] Name: D / teX Ultra High-speed 1D Detector

[0161] Detection element: 1D semiconductor element

[0162] Window material: Be

[0163] Effective window dimensions: 13mm (H) × 20mm (W)

[0164] Size: 80mm (L)

[0165] Thermal analysis

[0166] DSC analysis was performed using the DSC Mettler Toledo DSC1.

[0167] Weigh the sample in an aluminum pan with an airtight lid. Analyze the sample by heating it from 25°C to 320°C at a rate of 10 K / min.

[0168] TG analysis was performed using a Mettler Toledo TGA / DSC1.

[0169] The sample was weighed in an aluminum pan with an airtight cap pierced by an aluminum puncture. The sample was then heated from 25°C to 320°C at a rate of 10 K / min to perform the analysis.

[0170] Example 1. Preparation and characterization of crystalline form 1 of the free base of 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (HMO6)

[0171] Example 1A: Preparation of (HM06 free base form 1)

[0172] HM06 in its free base form 1 was prepared using a Sonogashira cross-coupling reaction mediated by Pd(PPh3)2Cl2 and CuI in ACN. More specifically, 8.2 kg of 4-amino-6-bromo-N-(4-(methoxymethyl)phenyl)-7-(1-methylcyclopropyl)-6,7-dihydro-5H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (which can be prepared according to known methods, such as Example 55 of U.S. Patent No. 10,155,768) was added to the reaction vessel at room temperature, along with the following reagents: CuI (0.108 kg), Pd(PPh3)2Cl2 (0.402 kg), and CH3CN (82.5 L). 6.683 kg of 4-(prop-2-yn-1-yl)morpholine was added to the mixture. 7.9 L of TEA was then added. The material was inertized by evacuation and nitrogen purging (500 mbar / 1030 mbar) five times. The material was heated to Tj 60°C (Ti: 56°C) under N2 and stirred at Tj 60°C for 14 hours to obtain a solution, which was then cooled to Ti 18-22°C. THF (82.2 L) was added, and the mixture was heated to Ti 40°C. The heated mixture was filtered through an 8-10 μm PTFE filter under pressure (at least 2 bar). The filtered mixture was cooled to Ti 20-25°C. The mixture was then passed through a resin plug (…). Si-thiols; by application rate: 220 L / h, Ti 25-25 °C mixture). Clean the reaction vessel, filter, and resin cake with THF (13 L). Discard the wet resin. Distill the solvent under vacuum at Tj: 45 °C until Vmax is applied (if necessary, by stopping stirring). Add MeTHF (164.5 L) to the residue and stir at room temperature to obtain a homogeneous suspension.

[0173] Add a 0.1 M solution of N-acetylcysteine ​​(2.6 kg) in water (161.6 L) to the organic suspension. Heat the mixture to Tj: 50 °C (Ti: 45-48 °C) and stir for 3 hours. Then stop stirring and allow phase separation for at least 20 minutes. After phase separation, back-extract the aqueous layer with MeTHF (81.9 L) at Ti: 45-48 °C. Stir the mixture for 30 minutes, then stop stirring and allow the layers to separate at Ti: 45-48 °C for at least 20 minutes. Then, separate the layers and drain the aqueous layer. Combine the two organic layers and add 20% NaCl (41 L) at Ti: 45-48 °C. Stir the mixture for 30 minutes, then stop stirring and allow phase separation for at least 20 minutes. Remove and drain the aqueous layer. Add water (66.4 L) to the organic layer at Ti: 45-48 °C. Stir the mixture at the same temperature for 30 minutes, then stop stirring and allow phase separation for at least 20 minutes. Remove and discard the water layer again.

[0174] Under vacuum, at Tj: 45°C, the organic layer was distilled down to the residue until Vmax was applied. The residue was then stripped overnight without stirring at Tj: 45°C and Vmax. Acetone was added (17 L) to the residue at Tj: 45°C, and then heated to Ti: 48°C (Tj: 52°C). The mixture was stirred for at least 1 hour to obtain a homogeneous suspension. The suspension was cooled to Ti: -10°C (Tj: -15°C) over at least 3 hours. The product was then separated at Tj: -10°C by filtration through a 20 μm filter under vacuum and pressure (at least 2 bar). The filter cake was washed with pre-cooled acetone (3.6 L; Ti: -10°C) under pressure (at least 2 bar) and vacuum until deliquoring was no longer observed. The solids were dried at Tj: 60°C for at least 24 hours to obtain the final product (6.8 kg). The product was stored at Tj: 2–8°C.

[0175] Figure 1A The XRPD pattern of HM06 crystals in free alkali form 1 obtained using CuKα radiation is shown. Figure 1A The peaks identified include those described in Table 2:

[0176] Table 2

[0177]

[0178] Example 1B: Preparation and characterization of HM06 free alkali crystals 2-5 and HCl salt crystals A

[0179] Five (5) new crystalline phases were observed in the form of free base and HCl crystalline salt after recrystallization experiments using the solvents listed in Table 3.

[0180] Table 3

[0181]

[0182]

[0183] Figure 1B The XRPD pattern of HM06 crystals in free alkali form 2, obtained using CuKα radiation, is shown. Figure 1B The peaks identified include those listed in Table 4:

[0184] Table 4

[0185]

[0186] Figure 1C The XRPD pattern of HM06 crystals in free alkali form 3, obtained using CuKα radiation, is shown. Figure 1C The peaks identified include those described in Table 5:

[0187] Table 5

[0188]

[0189] Figure 1D The XRPD pattern of HM06 crystals in free alkali form 4, obtained using CuKα radiation, is shown. Figure 1D The peaks identified include those described in Table 6:

[0190] Table 6

[0191]

[0192] Figure 1E The XRPD pattern of HM06 crystals in free alkali form 5, obtained using CuKα radiation, is shown. Figure 1E The peaks identified include those listed in Table 7:

[0193] Table 7

[0194]

[0195]

[0196] Figure 1F The XRPD pattern of HM06 HCl salt form A obtained using CuKα radiation is shown. Figure 1F The peaks identified include those described in Table 8:

[0197] Table 8

[0198]

[0199] Figure 1G The XRPD patterns of five HM06 crystals in the free alkali form and HM06 HCl salt form A, obtained using CuKα radiation, are shown to overlap.

[0200] Example 2. Synthesis and characterization of crystalline 4-amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide (HMO6) 1:1 HCl salt form 1

[0201] In a 100 ml single-necked round-bottom flask, add 1.16 ml (1.1 eq.) of 2 M HCl solution in water to 1 g of free HM06 base. Add 40 ml of tetrahydrofuran and stir the suspension at room temperature (25 °C) (800 rpm) for 2 days.

[0202] Samples were collected, filtered, and analyzed by XRPD. The remaining suspension was recovered by aspiration and dried under vacuum (50 mbar) at 25°C for 1 day. The dried samples were then analyzed by XRPD.

[0203] Figure 2A The XRPD pattern of crystal HM06 1:1HCl form 1 obtained using CuKα radiation is shown. Figure 2B and 2C The XRPD pattern of crystal HM06 1:1HCl form 1 obtained using CuKα radiation is also shown. Figure 2A-2C The peaks identified for crystal HM061:1HCl include those listed in Table 9.

[0204] Table 9

[0205]

[0206] Thermal analysis

[0207] The DSC spectrum of HM06 1:1HCl form 1 recorded in the sealed disk shows that the events after 200 °C can be attributed to sample melting and degradation. Figure 2DThis is the DSC temperature spectrum of HM06 1:1HCl form 1. The signal loss and baseline changes in the DSC spectrum related to solvent release may be due to the influence of solvent release on the sealing disk used.

[0208] The TGA spectrum of HM06 1:1HCl form 1 shows a 4.8% weight loss in the range of 40°C–170°C, consistent with water evaporation as recorded by EGA. Degradation occurs after 200°C. Figure 2E TGA spectra of HM06 1:1HCl form 1 are provided. The heat flow recorded in the TGA shows extensive and large events occurring within the water loss range, as well as signals of degradation attributable to melting after 200 °C.

[0209] Example 3. Preparation of HM06 1:2HCl

[0210]

[0211] A. Synthesis of 1g of HMO6 1:2HCl using an aqueous HCl solution

[0212] Weigh 1 g of free HM06 base and transfer it to a 250 mL reactor equipped with a magnetic stir bar. Then add 50 mL of ethanol and heat the resulting mixture until the solid is completely dissolved (T = 80 °C). When no solid material is observed, cool the solution to 50 °C. Slowly add 532 μL (3 equivalents) of 37% HCl to the reactor. Solid formation is immediately observed. Cool the mixture to 25 °C over 25 minutes, then stir for another hour. After this, separate the formed solid by vacuum filtration, wash with ethanol, and dry at 40 °C and 30 mbar for 24 hours. 1.08 g of the product as a white solid was recovered in near-quantitative yield.

[0213] B. Synthesis of HM06 1:2 HCl using 5g of HCl aqueous solution

[0214] Weigh 5 g of free HM06 base and transfer it to a 250 mL reactor equipped with a magnetic stir bar. Then add 70 mL of ethanol and heat the resulting mixture until the solid is completely dissolved (T = 80 °C). When no solid material is observed, cool the solution at 50 °C. A small amount of solid precipitate is observed, so the solution is heated again until completely dissolved, and then cooled to 60 °C. At this temperature, no precipitate formation is observed. Then, dissolve 2.5 mL (3 equivalents) of 37% HCl in 10 mL of ethanol and slowly add the resulting solution to the reactor. Solid formation is immediately observed. Cool the mixture at 25 °C for 35 minutes, and then stir for another hour. After this, separate the formed solid by vacuum filtration, wash with ethanol, and dry at 40 °C and 30 mbar for 24 hours. 5.64 g of the product as a white solid is recovered in near-quantitative yield.

[0215] C. Synthesis of HMO6 1:2 HCl using 2.5 g of anhydrous HCl

[0216] Weigh 2.5 g of free HM06 base and transfer it to a 250 mL reactor equipped with a magnetic stir bar. Then add 35 mL of ethanol and heat the resulting mixture until the solid is completely dissolved (T = 80 °C). When no solid material is observed, cool the solution at 50 °C. Mix 4.8 mL (3 equivalents) of anhydrous HCl 3.3 M in ethanol with 5 mL of ethanol and slowly add the resulting solution to the reactor. Solid formation is immediately observed. Cool the mixture to 25 °C over 35 minutes and then stir for another hour. After this, separate the formed solid by vacuum filtration, wash with another 10 mL of ethanol, collect and dry at 25 °C and 0.1 mbar for 24 hours. Further dry the solid at 100 °C and 30 mbar for another 72 hours. TG / EG analysis confirmed the recovery of the anhydrous compound. 2.69 g of the product as a white solid was recovered in near-quantitative yield.

[0217] D. Determination by stoichiometry

[0218] To determine the stoichiometry of the salt, chloride analysis was performed by ion chromatography. A solution of HM06 1:2 HCl was prepared by dissolving 149.2 mg of powder in a volumetric flask (10 mL) containing water (HPLC grade). Based on a TGA analysis of the batch prior to chloride determination, the amount of anhydrous salt was considered to be 140.4 mg (94.1%) (weight reduction related to a 5.9% water content).

[0219] Using a stoichiometric ratio of (HMO6:HCl) of 1:2, the molecular weight of the anhydrous salt is 547.5 g / mol, corresponding to a concentration of 0.0256 mmol / mL of HMO6 in the preparation solution. The chloride concentration, determined by ion chromatography, was found to be 0.05056 mmol / mL, corresponding to a chloride / HMO6 molar ratio of 1.98, thus confirming the 1:2 stoichiometric ratio of HMO6:HCl.

[0220] Example 4. Preparation and characterization of polymorphs 1 and 1-bis of HM06 1:2HCl

[0221] Example 4A: Synthesis of HM061:2HCl Form 1

[0222] Add 562.0 g of free HM06 base to hot ethanol (7885.5 mL) and heat at Ti: 75 °C with stirring until the solid is completely dissolved. Then, subject the solution to a fine filtration through a 1 μm column (PP or PTFE). Over at least 1 hour, add a mixture of 33% HCl (283.4 mL) and EtOH (283.5 mL) to the pre-filtered solution with stirring while maintaining Ti: 65–75 °C (target 70 °C). Then, cool the mixture to Ti: 20–25 °C over at least 30 minutes, and then stir at Ti: 20–25 °C for at least 1 hour. Then, separate the mixture by filtration through a 20 μm screen under pressure (at least 2 bar) and vacuum until no further liquid removal is observed. Wash the filter cake twice with EtOH (1070.8 mL × 2) under pressure (at least 2 bar) and vacuum until no further liquid removal is observed. Dry the wet product at Ti: 40 °C for at least 12 hours. The product HM06 in 1:2HCl form 1 (626 g) was obtained. The product was stored at Tj: 2-8℃.

[0223] Figure 3A The XRPD pattern of crystal HM06 1:2HCl form 1 obtained using CuKα radiation is shown. Figure 3A The peaks identified include those described in Table 10:

[0224] Table 10

[0225]

[0226] Thermal analysis was performed after storing the product in a sealed container at low temperature (4-10°C) for 2 months. Figure 3BTGA spectra of HM061:2HCl form 1 are provided. A 4.5% weight loss attributable to water release, as confirmed by EGA, was observed in the range of 30–160 °C. Degradation occurred above approximately 200 °C. TGA showed a further weight loss attributable to methanol release, as confirmed by EGA. Figure 3C This is the DSC temperature spectrum of HM06 1:2HCl form 1. As shown in the figure, the DSC reveals a broad endothermic event attributable to the release of water, starting at 25 °C and continuing up to approximately 160 °C. The endothermic peak below 195.13 °C (starting at 188.11 °C) is associated with sample melting.

[0227] HM06 1:2HCl form 1 single crystal

[0228] Crystals of HM06 in form 1:2HCl were obtained by slow evaporation. These crystals exhibited sufficiently large diffraction for single crystals, but were both affected by non-single-plane twinning, meaning that two crystals were grown together to form identical macroscopic samples. Separating the two crystals was impossible, and the collected data clearly showed the presence of two reciprocal lattices (see [link to data]). Figure 3D Solution and structural refinement are affected by this situation.

[0229] Two sets of data were collected for two different crystals. Both cases involved twins, with the second lattice obtained by rotating 180° along the b* axis of the first lattice. In the first case, the crystal consisted of two nearly identical components, and the non-short-facet twins significantly affected the data and did not allow for fine-tuning the structure to obtain favorable R values. The second data collection was characterized by a principal component and a second weak component. In this case, fine-tuning was acceptable.

[0230] HM06 1:2HCl form 1 crystallized as a monoclinic crystal in space group P21\c and the parameters β=94.163(7)° and V=2821.4(4) 3 The asymmetric unit consists of a diprotonated HMO6, two chloride ions, and 1.35 water molecules located at two positions (see [link to relevant documentation]). Figure 3E The chloride-labeled Cl2 is disordered at three positions, with occupancy rates of 0.35, 0.36, and 0.29 for Cl2A, Cl2B, and Cl2C, respectively. Possibly, the position of Cl2 depends on the number of water molecules in the unit cell, since Cl- and oxygen molecules in water molecules can repel each other.

[0231] The HM06 molecule forms pillars along the b-axis, and its existence can be attributed to the short contacts of π-stacking interactions. (molecular distance). The column has a cross-section that may prevent the structure from collapsing during water molecule removal (see [reference]). Figure 4A and 4B ).

[0232] Table 11 –HM06 1:2HCl Form 1 Crystal data and structural refinement

[0233]

[0234]

[0235] Example 4B: HM06 1:2HCl form 1-bis

[0236] To determine the hydration / anhydrous properties of HM06 1:2HCl form 1 and to limit the exact amount of water present in the lattice, a set of basic dehydration / drying experiments were performed, which led to the discovery of form 1-bis.

[0237] Figure 5A The XRPD pattern of crystal HM06 1:2HCl form 1-bis obtained using CuKα radiation is shown. Figure 5A The peaks identified include those described in Table 12:

[0238] Table 12

[0239]

[0240] VP-XRPD measurements were performed on a Panalytical X'pert equipped with an Anton Paar TTK450 chamber, which allows for in-situ measurement of powders under controlled temperatures and / or under vacuum.

[0241] The first measurement was collected at RT and atmospheric pressure. The sample was then held under vacuum (0.07 mbar) for 15 minutes. Figure 5B The XRPD pattern of HM06 1:2HCl crystal form 1-bis obtained using CuKα radiation is shown. Figure 5B As shown, the second pattern, labeled Form 1-bis, is different from the pattern of the starting material (Form 1) because the vacuum causes the sample to dehydrate.

[0242] It can be observed that some peaks do not change their positions, while others clearly migrate to higher θ values; it is possible that the release of water molecules affects some crystal planes without significantly altering the structure. Based on the structure determined by SC-XRD, form 1-bis is considered a highly unstable anhydrous form, not significantly different from form 1. This behavior allows for the easy absorption of water molecules over a short period of time.

[0243] The absorption of water by form 1-bis was observed via XRPD by tracking differences in patterns within the 2θ range of 25.5°–27.5° (see [link to XRPD]). Figure 5C Under vacuum, the highest peak is at 2θ = 26.7°, while after the sample is exposed to air for 2 minutes, the highest peak is at 2θ = 26.4°. The peak at 2θ = 26.4° shifts to 2θ = 26.2° within 20 minutes. Once the powder is exposed to air, the HM06 1:2HCl crystalline form 1-bis absorbs water from the atmosphere, and it reaches the diffraction pattern of the starting material within 20 minutes (see [reference]). Figure 5D The experiment was conducted in an environment with a room RH of approximately 80%.

[0244] Example 5. Synthesis and characterization of crystalline HM06 in form 1:2HCl 2

[0245] HM06 1:2HCl Form 2 was observed in a mixture with Form 1 from a high-temperature (50°C) slurry experiment using ethanol. 15 mg of HM06 1:2HCl Form 1 was suspended in 1.5 mL of ethanol and stirred at 50°C for three days. After this, the suspension was vacuum filtered at approximately 45-50% RH and analyzed by XRPD. The XRPD pattern of Form 1 was compared with that of the standard pattern of Form 2 (two lines at the bottom). Figure 6A The diffraction pattern was reported as the top pattern in the diagram.

[0246] Repeat and small-scale scaling procedures

[0247] 1. Repeat the procedure

[0248] The crystallization procedure was repeated twice. Repetition R01 resulted in HM061:2HCl form 2, affected by trace amounts of form 3, while pure form 2 was recovered from repetition R02. For both experiments, filtration and plate preparation for XRPD analysis were performed at 7% RH. XRPD measurements were performed using Kapton membranes. A summary of the results obtained and relative XRPD patterns are reported in Table 13.

[0249] Table 13 Example 5: Repeat the procedure results.

[0250] Crystallization process repeat result Experiments with 3-day slurry derived from ethanol at 50°C R00 A mixture of form 1 and form 2 <![CDATA[3-day slurry experiment from ethanol at 50 °C 1 > R01 Form 2 + tiny trace amounts of Form 3 <![CDATA[3-day slurry experiment from ethanol at 50 °C 1 > R02 Form 2

[0251] 1 Powder separation by vacuum filtration and preparation of sample plates covered with Kapton film were carried out at 7% RH.

[0252] exist Figure 6B The image shows the XRPD patterns of the results for R01 and R02 compared to the standard reference patterns of Forms 2 and 3.

[0253] Small-scale scaling procedure

[0254] Different small-scale scale-up procedures were explored to obtain sufficient powder for further testing and feasibility studies. The first experiment was conducted on 100 mg of HM06 1:2HCl form 1. The powder was suspended in 10 mL of ethanol (10 mg / mL) and stirred at 50 °C for 4 days. Following this, the suspension was vacuum filtered at 5% RH, and XRPD plates covered with a Kapton membrane were prepared at the same % RH. HM06 1:2HCl form 2 was isolated, and the collected XRPD pattern was used as a standard (STD) reference pattern for form 2.

[0255] The procedure was repeated twice, and the separation step was performed 5 days later under 5% RH conditions. The first repetition (R01) resulted in the production of HM06 1:2HCl form 3 affected by a certain trace amount of form 2, while the second repetition (R02) resulted in the production of form 3 with a signal at 7.2°2θ, which can be attributed to form 2.

[0256] Considering this data, try other procedures. Suspend 100 mg of HM06 1:2 HCl form 1 in 5 mL of ethanol (20 mg / mL) and keep stirring at 50 °C for 10 days. Following this procedure, form 2, with a small signal at 5.5°2θ, which is characteristic of form 3, was isolated.

[0257] The results are summarized in Table 14.

[0258] Table 14 Example 5: Results of small-scale scaling-up procedure.

[0259]

[0260]

[0261] 1 The experiment was conducted using a concentration of 20 mg / ml.

[0262] Figure 6C The XRPD pattern of HM06 1:2HCl form 2, obtained at a scale of 100 mg after 4-day slurry experiments using CuKα radiation and collected from ethanol, is shown and used as an STD reference. Figure 6C The peaks identified include those described in Table 15:

[0263] Table 15

[0264]

[0265] Thermal analysis

[0266] Figure 6D The image shows the DSC temperature spectrum of HM06 1:1HCl form 2, which reveals an endothermic event at 219.8 °C (starting at 205.5 °C) consistent with sample melting. Degradation occurs above approximately 200 °C.

[0267] Figure 6E TGA spectra of HM06 1:1HCl form 2 are provided, showing no weight loss. The sample can be considered anhydrous. Methanol and HCl release were detected during degradation.

[0268] Figure 6F and 6G The image shows a small-scale amplified relative XRPD pattern obtained using CuKα radiation from Example 5.

[0269] Example 6. Synthesis and characterization of form 3 of HM06 1:2HCl

[0270] 15 mg of HM06 in 1:2 HCl form 1 was suspended in 1.5 mL of acetonitrile and stirred at 50 °C for three days. Afterward, the suspension was vacuum filtered at approximately 45–50% RH and analyzed by XRPD. Figure 7A The image shows an overlay of XRPD patterns of HM06 1:2HCl crystals obtained from acetonitrile after an HT (50 °C) slurry experiment using CuKα radiation, in Form 3 (top line) and standard reference patterns in Form 1 (black line), Form 2 (bottom line), and Form 3 (pink line).

[0271] Repeat and small-scale scaling procedures

[0272] Repeat procedure

[0273] The crystallization process was repeated twice, extending the time to up to 9 days, and the recovered powder was treated under 7% RH conditions. Kapton films were used to prepare XRPD plates. Repetition of R01 resulted in a stage where some trace amounts of form 2 were observed. Repetition of R02 resulted in the production of form 3 and some peaks that were not further allocated.

[0274] Since pure Form 3 was collected from the rapid precipitation experiment using 1-propanol and used as a standard reference, other replicates were attempted. All three rapid gradient experiments were prepared as follows: 1.5 mL of 1-propanol was added to 15 mg HM06 1:2 HCl Form 1. The suspension was heated to up to the solvent boiling point for several minutes. A clear solution was observed immediately. It was then rapidly cooled to 10°C using an ice bath. Precipitation occurred immediately. The powder was recovered by vacuum filtration and XRPD plates were prepared using a Kapton membrane. All these experiments were treated under controlled 4–5% RH conditions.

[0275] The results are summarized in Table 16.

[0276] Table 16 Example 6: Repeat the procedure results.

[0277]

[0278] 1 Powder separation by vacuum filtration and preparation of sample plates covered with Kapton film were carried out at 7% RH.

[0279] 2 Compared to the standard reference pattern of form 3, the signal strength at 5.5°2θ is lower.

[0280] 3 Powder separation by vacuum filtration and preparation of sample plates covered with Kapton film were carried out at 4-5% RH.

[0281] Small-scale scaling procedure

[0282] Different small-scale scale-up procedures were explored to obtain sufficient powder for further testing and investigation of process feasibility. All experiments were conducted under controlled %RH conditions between 5 and 7%, for both the separation step and the preparation of XRPD sample plates using Kapton films.

[0283] A first test was performed on 100 mg HM06 1:2HCl form 1. The powder was suspended in 10 mL of acetonitrile (10 mg / mL) and stirred at 50 °C for 4 days. After this, the suspension was vacuum filtered and analyzed by XRPD. A mixture of forms 2 and 3 was recovered. In an attempt to achieve pure form 3, a first replicate R01 was planned, which extended the slurry time to 12 days. In parallel, the same experiment was also prepared for a concentration test at 20 mg / mL. From replicate R01, form 2 aggregated, while the other tests resulted in a mixture of forms 2 and 3, with an undistributed peak observed at 6.3°2θ.

[0284] Since pure form 3 was achieved from a rapid gradient precipitation of 15 mg from 1-propanol, and although repetition did not yield form 3, the small-scale scale-up procedure continued. 100 mg of HM06 1:2 HCl form 1 was suspended in 10 mL of 1-propanol. It was heated to the solvent boiling point. After several minutes, the resulting clear solution was rapidly cooled at 10 °C and held with magnetic stirring for 5 minutes. The powder was then separated by filtration and analyzed by XRPD. Pure form 3 was achieved, and its XRPD pattern was used as a standard reference. A very small signal was observed at 7.2°2θ, which could likely be attributed to form 2. The filtration step and the preparation of XRPD sample plates covered with a Kapton membrane were performed at 4% RH.

[0285] R01 was repeated and the drying process was applied according to the procedure mentioned in Table 17. Sampling was analyzed by XRPD. Form 3 was obtained by treating the entire wet filter cake at 40°C / 50 mbar for 3 hours, and then measuring again. Form 3 was achieved, although a small signal at 7.2°2θ attributable to Form 2 was detected.

[0286] Three additional replicates (R02 and R03) were performed according to the same procedure. Form 3 was collected, although a small signal at 7.2°2θ, attributable to form 2, was detected. No further drying steps were applied to R03. Instead, replicate R02 underwent two drying steps followed by TGA-EGA analysis. The low crystallinity shown by R02 was due to the smaller amount used for XRPD analysis.

[0287] Based on these results, rapid gradient precipitation from 1-propanol can be considered a suitable procedure for obtaining form 3.

[0288] The results are summarized in Table 17.

[0289] Table 17 Example 6: Results of small-scale scaling-up procedure.

[0290]

[0291] 1 The experiment was conducted using a concentration of 20 mg / mL.

[0292] 2 Powder separation by vacuum filtration and preparation of sample plates covered with Kapton film were carried out between 3-7% RH.

[0293] Figure 7B The XRPD pattern of HM06 1:2HCl form 3 collected from 1-propanol after rapid gradient precipitation at a scale of 100 mg obtained using CuKα radiation is shown. Figure 7BThe peaks identified in the study from HM06 1:2HCl form 3 include those listed in Table 17:

[0294] Table 17

[0295]

[0296] Figure 7C The image shows the DSC temperature spectrum of HM06 1:2HCl form 3, which reveals an endothermic event at 214 °C at which the sample melts (starting at 202 °C). Degradation occurs above approximately 200 °C.

[0297] Figure 7D TGA spectra of HM06 1:2HCl form 3 are provided, showing a very slight weight loss of 0.7% up to 180 °C. During degradation, methanol and HCl were detected by EGA.

[0298] Example 7. Synthesis and characterization of form 4 and form 4-bis of HM06 1:2HCl

[0299] Crystallization procedures for Form 04 and Form 04-bis

[0300] Following the procedure below: at 25°C and under low pressure, 4-bis form of methanol was collected from methanol after an evaporation experiment.

[0301] A saturated solution of approximately 50 mg / mL HM06 1:2HCl form 1 in methanol was prepared and stirred overnight (18 hours) at room temperature. Afterward, the solution was filtered and evaporated at 25 °C / 700 mbar. Sample plates sealed with a Kapton membrane were prepared at 40–45% RH. Figure 8A The report presents collected XRPD patterns compared to Form 1. Some very small signals exist in Form 1.

[0302] Repeat procedure

[0303] The experiment was repeated four times following the procedure reported above (repeated R01-R04). XRPD sample plates sealed with a Kapton film were prepared at 40-45% RH. Form 1 was recovered in all experiments. The procedure was attempted four more times, but this time the samples were separated at a controlled % RH with values ​​between 7-8% RH. A new pattern labeled Form 4, influenced by the signal of Form 1, was observed in all analyzed samples. From a qualitative point of view, repeat R05 showed the lowest amount of Form 1. Table 18 describes the repeat procedure and results.

[0304] Table 18 Example 7: Repeat procedure results.

[0305] Crystallization process repeat result Low-pressure room temperature evaporation experiment from a methanol saturated solution R01 Form 1 Low-pressure room temperature evaporation experiment from a methanol saturated solution R02 Form 1 Low-pressure room temperature evaporation experiment from a methanol saturated solution R03 Form 1 Low-pressure room temperature evaporation experiment from a methanol saturated solution R04 Form 1 <![CDATA[Low-pressure room-temperature evaporation experiment from a methanol saturated solution 1 > R05 Form 4 + Form 1 <![CDATA[Low-pressure room-temperature evaporation experiment from a methanol saturated solution 1 > R06 Form 4 + Form 1 <![CDATA[Low-pressure room-temperature evaporation experiment from a methanol saturated solution 1 > R07 Form 4 + Form 1 <![CDATA[Low-pressure room-temperature evaporation experiment from a methanol saturated solution 1 > R08 Form 4 + Form 1

[0306] 1 Powder separation by vacuum filtration and preparation of sample plates covered with Kapton film were carried out at 7-8% RH.

[0307] Figure 8B The XRPD pattern of 4-bis in the form of HM06 1:2HCl is shown. Figure 8B The peaks identified for the 4-bis form of HM061:2HCl include those listed in Table 19:

[0308] Table 19

[0309]

[0310] Stability evaluation

[0311] The stability of form 4-bis was evaluated after 7 days of storage in sealed vials. Figure 8C As shown, it demonstrates the complete transformation to form 1.

[0312] Form 4 Crystallization Procedure

[0313] As described in the previous section, HM06 1:2HCl form 4 was isolated from the mixture with form 1 in an attempt to achieve form 4-bis from the repeated experiments performed.

[0314] Repeat procedure

[0315] As described in the previous section and Table 18, form 4 was recovered from all four replicates (R05-R08) performed. From a qualitative point of view, replicate R05 appears to have the lowest amount of form 1.

[0316] Figure 8D The XRPD pattern of HM06 1:2HCl form 4 obtained using CuKα radiation is shown. Figure 8D The peaks of HM06 1:2HCl form 4 identified in the study include those listed in Table 20:

[0317] Table 20

[0318]

[0319] The HM06 1:2HCl form 4 exposed powder was stored overnight at room temperature under 43% RH. Figure 8E As shown, no significant changes were observed.

[0320] Example 8. Synthesis and characterization of form 5 and form 5-bis of HM06 1:2HCl

[0321] Form 5 and Form 5-bis

[0322] Form 5-bis

[0323] 1. Crystallization process

[0324] Evaporation experiments of HM06 1:2HCl form 1 mixed with 50 / 50 water / dimethylformamide at 60 °C resulted in the separation of new XRPD patterns, in which some signals from form 5 were observed. Due to similarity, this is labeled form 5-bis. Sample processing was performed under 40–45% RH conditions.

[0325] Figure 9A The XRPD pattern of HM06 1:2HCl form 5-bis is shown compared to form 5.

[0326] 2. Repeat the procedure

[0327] The evaporation experiment was repeated twice, and orange powder was collected. Both samples were analyzed by XRPD, revealing diffraction patterns of form 5. Some subtle signals, possibly related to form 5-bis, were not further assigned, specifically a peak with discontinuous intensity at 18°2θ. Based on these results, form 5-bis was considered non-reproducible.

[0328] Figure 9B The XRPD pattern of 5-bis in HM06 1:2HCl form is shown. Figure 9B The peaks identified for the 5-bis form of HM061:2HCl include those listed in Table 21:

[0329] Table 21

[0330]

[0331]

[0332] 3. Stability test

[0333] The stability of the HM06 1:2HCl 5-bis sample was evaluated after 18 hours of exposure to air and after one week in a sealed vial (both at room temperature).

[0334] After exposing the powder to room temperature for 18 hours, the form 5-bis was measured. %RH was approximately 45%. Its XRPD pattern showed some changes: for example, a lack of signal at 6° and 12° 2θ and an increase in the peak at 6.4° 2θ.

[0335] These changes may not be related to the transformation to one of the other isolated polymorphs observed during the study: therefore, the sample is considered unstable. Figure 9C The XRPD pattern of form 5-bis (blue top pattern) and the XRPD pattern of the same sample analyzed after 18 hours of exposure (red bottom pattern) are shown.

[0336] After 7 days in a sealed vial at room temperature, the HM06 1:2HCl form 5-bis was measured. Figure 9D As shown, the sample begins to transform into form 1.

[0337] Form 5 Crystallization Procedure / Repetitive Procedure

[0338] Evaporation experiments of HM06 1:2HCl form 1 at 60 °C in dimethyl sulfoxide yielded form 1 with signals that were not further distributed. To better understand the nature of these few new signals, the recrystallization procedure was repeated twice: R01 and R02. Both samples showed a dark brown color, while R02 was completely glassy, ​​and a small amount of powder could be recovered from R01. The powder from R01 was analyzed by XRPD and showed a diffraction pattern with low crystallinity, which was labeled as form 5.

[0339] Figure 9E The XRPD pattern of HM06 1:2HCl form 5 obtained using CuKα radiation is shown. Figure 9E The peaks identified for HM061:2HCl form 5 include those listed in Table 22:

[0340] Table 22

[0341]

[0342] Figure 9F The DSC temperature spectrum of HM06 1:2HCl form 5 shows a wide range of endothermic events at 30℃-90℃ attributable to solvent release, as observed in TGA-EGA. Two consecutive endothermic events were also observed at 158.7℃ (occurring at 144.6℃) and 164.8℃ (occurring at 158.3℃).

[0343] Figure 9G TGA spectra of HM06 1:2HCl form 5 are provided, showing a weight loss of water up to 130 °C. It is impossible to definitively attribute the water escape to dehydration or the release of adsorbed water. Degradation occurs above 200 °C. EGA did not detect HCl escape, as observed for other separated forms. The formation of salts with lower HCl content may not be ruled out; the stoichiometry of the salts in form 5 is not definitively determined.

[0344] Example 9. Synthesis and characterization of form 6 of HM06 1:2HCl

[0345] Form 6 was collected after evaporation of HM06 1:2 HCl form 1 in a 1:1 acetonitrile / water solution at low pressure and at room temperature. Conversion to form 1 was observed after 6 days of storage under these conditions. No further analysis was performed due to the batch exhibiting an orange color and its phase instability.

[0346] Figure 10 The XRPD pattern of HM06 1:2HCl form 6 using CuKα radiation is shown. Figure 10 The peaks identified for HM06 1:2HCl form 6 include those listed in Table 23:

[0347] Table 23

[0348]

[0349] Figure 11 Overlapping of XRPD patterns for all isolated forms of HM06 1:2HCl is reported.

[0350] Example 10: Storage stability test of HM06 1:2HCl form 1

[0351] The sample of HM06 1:2HCl form 1 was subjected to accelerated storage conditions and long-term storage. The results confirmed that HM06 1:2HCl form 1 remained stable and crystalline.

[0352] 1. Accelerate storage conditions

[0353] Samples from the same batch of HM06 1:2HCl form 1 were stored at 40°C and 75% relative humidity for 24 months and tested periodically. Table 24 describes the sample analysis at 0, 1 month, 3 months, and 6 months.

[0354] Table 24: Acceleration Conditions – 40℃ / 75% Relative Humidity

[0355]

[0356]

[0357] *Report all impurities >0.05% (A%). **RRT (Relative Retention Time) 0.92-0.94

[0358] 2.24 months of storage

[0359] Samples from the same batch of HM06 1:2HCl form 1 were stored at 25°C and 60% relative humidity for 6 months and tested periodically. Table 25 describes the sample analyses at 0, 3, 6, 9, 12, 18, and 24 months.

[0360] Table 25: Long-term conditions – 25°C and 60% relative humidity

[0361]

[0362] *Report all impurities >0.05% (A%); **RRT 0.92-0.94

[0363] The foregoing written description is considered sufficient to enable those skilled in the art to practice the embodiments. The foregoing description and examples detail certain embodiments and describe the best mode contemplated by the inventors. However, it will be understood that no matter how detailed the foregoing may be in the text, the embodiments can be practiced in various ways and should be understood in accordance with the appended claims and any equivalents thereof.

[0364] As used herein, whether explicitly stated or not, the term "approximately" refers to numerical values, including, for example, integers, fractions, and percentages. The term "approximately" generally indicates a range of numerical values ​​(e.g., + / - 5% to 10% of the listed numerical values) that a person skilled in the art would consider equal to (e.g., having the same function or result). When a term such as "at least" or "approximately" precedes a list of numerical values ​​or ranges, the term modifies all values ​​or ranges provided in that list. In some cases, the term "approximately" may include numerical values ​​rounded to the nearest significant figure.

Claims

1,4-Amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide 1:2HCl salt, in crystalline form 1, characterized by an XRPD plot containing the following peaks: at 6.59°2θ, 7.40°2θ, 9.12°2θ, 14.57°2θ, 16.39°2θ, 26.06°2θ, 26.57°2θ, and 27.07°2θ. 2,4-Amino-N-[4-(methoxymethyl)phenyl]-7-(1-methylcyclopropyl)-6-(3-morpholinopropyl-1-yn-1-yl)-7H-pyrrolo[2,3-d]pyrimidine-5-carboxamide 1:2HCl salt, in crystalline form 1, characterized by an XRPD plot containing the following peaks: peaks at 6.59°2θ, 7.40°2θ, 9.12°2θ, 14.57°2θ, 16.39°2θ, 26.06°2θ, 26.57°2θ, and 27.07°2θ, the values ​​of which include the listed values ​​within ±5%.

3. The product as claimed in claim 1 or 2, characterized by the same XRPD plot as FIG3A.

4. The product as claimed in claim 1 or 2, wherein the XRPD pattern in crystalline form is obtained using CuKα radiation.

5. The product as claimed in claim 1 or 2, characterized by at least one of the same DSC thermogram as in FIG3C and the same TGA spectrum as in FIG3B.

6. The product of claim 4, characterized by at least one of the same DSC thermogram as in FIG3C and the same TGA spectrum as in FIG3B.

7. The product as claimed in claim 1 or 2, wherein it is in crystalline form at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100%.

8. The product of claim 4, wherein it is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% crystalline.

9. The product of claim 5, wherein it is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% crystalline.

10. The product of claim 6, wherein it is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% crystalline.

11. A pharmaceutical composition comprising the crystalline form 1 as described in any one of claims 1 to 10 and a pharmaceutically acceptable excipient.

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