Biological activities of the crystal, phosphate salt, and enantiomers thereof of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol

Abstract

The disclosure of this specification relates to the biological activities and the preparation of a stable crystalline form (Compound A-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A), its tablets (Compound A-Xln tablets), its phosphate (Compound A-P) and a phosphate stable crystalline form (Compound A-P-Xln), as well as its enantiomers (Compound R-A, Compound S-A). Furthermore, their anti-cancer activities, stability, chirality, solubility, and therapeutic indications have also been investigated.

Description

Technical Field

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 351,205, filed on June 10, 2022. This application claims the benefit of U.S. Patent Application Publication No. 2023 / 399313, filed on June 6, 2023.

[0002] The present disclosure relates to the biological activity and preparation of stable crystalline forms of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A) (Compound A-Xln), tablets thereof (Compound A-Xln tablets), its phosphate (Compound A-P) and phosphate stable crystalline form (Compound A-P-Xln), and its enantiomers (Compound R-A, Compound S-A).

Background Art

[0003] Protein tyrosine kinases (PTKs) are a series of enzymes that catalyze the phosphorylation process of transferring a phosphate group from a nucleoside triphosphate (often ATP) to an amino acid residue of a protein. Through this phosphorylation process, phosphorylated proteins can be activated. Therefore, PTKs play a role as a "switch" that regulates many cellular functions by controlling signal cascades that extend from outside the cell, through the membrane, into the cytoplasm, and further into the nucleus.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Patent Document 2

Patent Document 3

Summary of the Invention

[0005] Some embodiments disclosed herein relate to PTK inhibitors with improved stability. These inhibitors can be used for the treatment of PTK-mediated disorders and diseases. For example, depending on the position on different extracellular domains, PTKs can be classified into epidermal growth factor receptor (EGFR), platelet-derived growth factor receptor (PDGFR), vascular endothelial growth factor receptor (VEGFR), fibroblast growth factor receptor (FGFR), etc. Many studies have demonstrated that in normal cells, PTK activity is usually low or not active at all, while in many tumor cells, especially gliomas and cancers, overexpression of PTK is characteristic. Apparently, abnormal overactivity of PTK is closely correlated with the growth and angiogenesis processes of tumor cells.

[0006] Interrupting or blocking PTK activity may be able to dramatically suppress the growth of tumor cells by inhibiting cell signaling. Therefore, targeted therapy that suppresses PTK expression via PTK inhibitors has become a widely accepted treatment method.

[0007] The novel PTK inhibitor 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A) is characterized by the potential to inhibit the activities of many protein tyrosine kinases (PTKs) such as, but not limited to, VEGFr, EGFr, c-kit, PDGF, FGF, SRC, Aurora B. The structure and synthetic route of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A) have already been disclosed in International Publication No. WO2010 / 021918.

[0008] Some embodiments relate to a stable crystalline form (Compound A-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol.

[0009] In some embodiments, the stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol is produced by a method that includes recrystallizing the amorphous form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol.

[0010] In some embodiments, the stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol is produced by a method that includes two recrystallization steps.

[0011] In some embodiments, the stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol is produced by a method that includes a recrystallization process carried out using a high-boiling solvent or a mixture of solvents having a high boiling point together.

[0012] In some embodiments, the stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol is produced by a method that includes a recrystallization process carried out using a low-boiling solvent or a mixture of solvents having a low boiling point together.

[0013] In some embodiments, a stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol is produced by a method comprising a first recrystallization process and a second recrystallization process.

[0014] In some embodiments, a stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol is produced by a method comprising a first recrystallization process using a high-boiling solvent or a mixture of solvents having a combined high boiling point, and a second recrystallization process using a low-boiling solvent or a mixture of solvents having a combined low boiling point. In some embodiments, the high-boiling solvent is DMF and the low-boiling solvent is EtOH. Some embodiments relate to a crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol exhibiting at least one of the following properties: DSC melting point range (Endo): 240 - 260 °C, peak temperature range: 244 - 254 °C, more specifically DSC melting point range (Endo): 247 - 253 °C, peak temperature = 249 °C, pattern shown in Figure 2. Or a TGA thermogram showing a non-solvated substance having a weight loss at 250 °C or higher, pattern shown in Figure 3.

[0015] Some embodiments relate to a crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol characterized by the XRPD pattern shown in Figure 4.

[0016] Some embodiments relate to a crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol, characterized in that the XRPD results include 14 characteristic peaks with an intensity (%) exceeding 10%, represented by d-values and angles as follows:

[0017] [Table 1]

[0018] Some embodiments relate to a crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol, characterized by an XRPD pattern as shown in Figure 11.

[0019] Some embodiments relate to a crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol, characterized in that the XRPD results include 43 characteristic peaks with all intensities (%), represented by d-values and angles as follows:

[0020] [Table 2]

[0021] In some embodiments, the crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol has improved stability and / or solubility compared to its amorphous form.

[0022] In some embodiments, the stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol is a pharmaceutically acceptable salt. In some embodiments, the stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol is a phosphate salt.

[0023] In some embodiments, the phosphate salt is prepared by a method comprising the step of neutralizing a solution of phosphoric acid in a solvent or mixture of solvents and a recrystallization process.

[0024] In some embodiments, the solvent in which the neutralization and / or recrystallization is carried out is EtOH.

[0025] In some embodiments, the phosphate salt of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol is characterized by one or more of the following properties: DSC melting point range (Endo): 221 - 235 °C, peak temperature = 229 °C, and this pattern is shown in Figure 9. In TGA, a slight weight loss is shown at about 30 - 60 °C, and a significant weight loss is shown above 210 °C, and this pattern is shown in Figure 10.

[0026] In some embodiments, the phosphate salt of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol comprises the XRPD pattern shown in Figure 11. The XRPD results include all characteristic peaks. All intensity % are represented by d-value and angle as follows:

[0027]

Table 3

[0028] Some embodiments relate to a compound having a structure represented by 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol, its stable crystalline forms, its salt forms, and / or its stable crystalline salt forms for use in the inhibition of protein tyrosine kinase (PTK). In some embodiments, the PTK is selected from, but not limited to, FGFRl(h), FGFR2(h), FGFR3(h), Fltl(h) (VEGFrl), Flt4(h) (VEGFr3), KDR(h) (VEGFr2), Aurora-B PDGFRα(h), PDGFRα(h), and PDGFRβ(h). In some embodiments, the salt form is a phosphate form.

[0029] Some embodiments relate to a compound having a structure represented by 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol, its stable crystalline forms, its salt forms, and / or its stable crystalline salt forms, which compound inhibits common cancer cell lines including, but not limited to, PANC-1, NCI-H157, MDA-MB-231, Hela, PC-3, BEL7404, MKN45, Ishikawa, Saos-2, SKOV3, SW579, and HCT116. In some embodiments, the salt form is a phosphate form.

[0030] Some embodiments relate to a pharmaceutical composition comprising an active ingredient that is a compound described above or elsewhere in this specification, and a pharmaceutically acceptable carrier and / or a pharmaceutically acceptable excipient.

[0031] Some embodiments relate to pharmaceutical compositions that include a compound described above or elsewhere in this specification as an active ingredient, and a pharmaceutically acceptable carrier and / or a pharmaceutically acceptable excipient, thereby forming tablets.

[0032] Some embodiments relate to pharmaceutical compositions that include an active ingredient selected from the stable crystalline free base form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol or the stable crystalline form of the phosphate, and a pharmaceutically acceptable carrier.

[0033] Some embodiments relate to methods of treating neoplastic diseases, the methods comprising administering to a subject in need thereof a pharmaceutical composition comprising a compound as defined above or elsewhere in this specification, or such a compound and a pharmaceutically acceptable excipient. In some embodiments, the methods comprise administering an additional agent, the additional agent being a chemotherapeutic compound and / or an immunotherapeutic agent. In some embodiments, the neoplastic disease is a solid tumor selected from lung cancer, renal cancer, colorectal cancer, gastric cancer, melanoma, head and neck cancer, thyroid cancer, pancreatic cancer, liver cancer, prostate cancer, bladder cancer, brain cancer, sarcoma, breast cancer, ovarian cancer, cervical cancer, and endometrial cancer, and a hematologic cancer selected from ALL, CLL, AML, CML, and multiple myeloma, in particular for human clinical trials for treating small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), and ovarian cancer. Some embodiments relate to methods of treatment, wherein the combination chemotherapeutic agent is selected from platinum-based or taxane-based or topoisomerase 1 inhibitors or alkaloids or alkylating agents. In some embodiments, the chemotherapeutic agent is selected from cisplatin, carboplatin, paclitaxel, or cisplatin / paclitaxel, carboplatin / paclitaxel, or carboplatin / etoposide, or topotecan, irinotecan, or lomustine. In some embodiments, the immunotherapeutic agent is selected from PD-1 or PD-L1 antibodies including, but not limited to, nivolumab, pembrolizumab, ipilimumab, blinatumomab, elotuzumab, daratumumab, semipilimab, avelumab, durvalumab, atezolizumab, tripalimumab, sintilimab, camrelizumab, tislelizumab, AK104, pemprinumab, KN035, CS1001, talimogene laherparepvec.

Brief Description of the Drawings

[0034]

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Modes for Carrying Out the Invention

[0035] Some embodiments relate to the compound 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A), its stable crystalline form (Compound A-Xln) and amorphous form (Compound A-Amp), its stabilized crystalline form (Compound A-P-Xln) crystallized from the phosphate (Compound A-P), and the enantiomers (two chiral isomers) of any of the aforementioned phosphates or crystals (e.g., Compound R-A, Compound S-A, Compound R-A-Xln, Compound S-A-Xln, Compound R-A-P-Xln, Compound S-A-P-Xln). Compound A is represented by the following structure:

[0036] [Chemical formula]

[0037] The following description, while indicating circumstances and examples, should not be construed as limiting the scope of the invention covered by the claims following this specification or any other application claiming priority herefrom. A single component (including method steps) or a collection of components (e.g., multiple steps) is not essential or indispensable. Any feature, structure, component, material, step, or method described and / or illustrated in any embodiment of this specification can be used with, or in place of, any feature, structure, component, material, step, or method described and / or illustrated in any other embodiment of this specification.

[0038] The terms and phrases used in this application, and their variations, especially the terms and phrases used in the appended claims, should be construed as open-ended and not limiting unless explicitly stated otherwise. The term "including" in the above example means "including, without limitation", "including but not limited to", etc. The term "comprising" as used herein is synonymous with "including", "containing", or "characterized by", is inclusive or open-ended, and does not exclude additional, unrecited elements or method steps.

[0039] The term "having" should be interpreted as "having at least". The term "include" should be interpreted as "including but not limited to these". The term "example" is used to present exemplary examples of the item being discussed and is not an exhaustive or limiting list thereof. Also, the use of terms such as "preferably", "preferred", "desired", "desirable" and terms of similar meaning does not mean that a particular function is critical, essential or important to the structure or function of the present invention, but is only intended to highlight alternative or additional features that may or may not be used in a particular embodiment of the present invention and should be understood as such. Further, the term "comprising" is interpreted synonymously with the phrase "having at least" or "including at least". When used in the context of a process, the term "comprising" means that the process includes at least the steps described, but may include additional steps. When used in the context of a compound, composition or device, the term "comprising" means that the compound, composition or device includes at least the features or components described, but may also include additional features or components. Similarly, a group of items connected by the conjunction "and" should not be read as requiring that each and every one of those items be present within the group, but should be read as "and / or" unless explicitly stated otherwise. Similarly, a group of items connected by the conjunction "or" should not be read as requiring mutual exclusivity within the group, but should be read as "and / or" unless explicitly stated otherwise.

[0040] Furthermore, the phrase "consisting essentially of" is understood to include the specifically recited elements and additional elements that do not substantially affect the basic and novel characteristics of the claimed technology. The phrase "consisting of" does not include any elements not specified.

[0041] The section headings used in this specification are for organizational purposes only and should not be construed as limiting the subject matter described. Features disclosed under one heading (such as compositions) can be used in combination with features disclosed under another heading (such as methods of treatment). Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art.

[0042] It should be noted that when a particular term is used to describe a particular feature or aspect of the present disclosure, this does not mean that the term is redefined herein in such a way as to be restricted to include any particular features of the feature or aspect of the present disclosure to which the term relates.

[0043] When referring to various features, the term "including the foregoing values, and / or ranges spanning the foregoing values" may be used. These terms (and their variations) mean including any of the foregoing values, or any range spanning the foregoing values. For example, with respect to temperature, the temperature may be expressed as "equal to or at least approximately the following values": 40°C, 50°C, 60°C, 70°C, or ranges including the foregoing values and / or ranges spanning the foregoing values. This language includes not only the specific temperatures provided and ranges above those temperatures (e.g., about 40°C or at least about 40°C, about 50°C or at least about 50°C, about 60°C or at least about 60°C, and about 70°C or at least about 70°C), but also temperature ranges spanning those values (e.g., 40°C to 50°C, 40°C to 60°C, 40°C to 70°C, 50°C to 60°C, 50°C to 70°C, or 60°C to 70°C). Similarly, with respect to temperature, it may be expressed as "less than or equal to the following values": 40°C, 50°C, 60°C, 70°C, or ranges including the foregoing values and / or ranges spanning the foregoing values. This language includes not only the specific temperatures provided and ranges above those temperatures (e.g., about 40°C or less, about 50°C or less, about 60°C or less, and about 70°C or less), but also temperature ranges spanning those values (e.g., 40°C to 50°C, 40°C to 60°C, 40°C to 70°C, 50°C to 60°C, 50°C to 70°C, or 60°C to 70°C).

[0044] The terms "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" include any solvent, dispersion medium, coating, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for pharmaceutically active substances is well known in the art. Any conventional media or agents are contemplated for use in therapeutic compositions, except when they are incompatible with the active ingredient. Additionally, it may include various adjuvants as are commonly used in the art. Considerations regarding the inclusion of various components in pharmaceutical compositions are described, for example, in Gilman et al. (Eds.) (1990); Goodman and Gilman’s: The Pharmacological Basis of Therapeutics, 8th Ed., Pergamon Press, which is hereby incorporated by reference in its entirety.

[0045] As disclosed herein, the "patient" or "subject" being treated is, in some embodiments, a human patient; however, it should be understood that the principles of the subject matter disclosed herein indicate that the subject matter disclosed herein is effective against all vertebrate species, including the mammals intended to be covered by the terms "subject" and "patient". Suitable subjects are generally mammalian subjects. The subject matter described herein has been found to be useful not only in research but also in veterinary and medical applications. The term "mammal" as used herein includes, but is not limited to, humans, non-human primates, cows, sheep, goats, pigs, horses, cats, dogs, rabbits, rodents (e.g., rats or mice), monkeys, etc. Subjects include neonatal, infant, juvenile, adult, and geriatric subjects.

[0046] As used herein, the term "therapeutically effective amount" refers to an amount of a therapeutic agent that confers a modulating effect, such as may be a beneficial effect, on a subject afflicted with a disorder, disease or illness, including improvement of the subject's symptoms (e.g., modulation of one or more symptoms), delay or reduction in the progression of symptoms, prevention or delay in the onset of a disorder, and / or changes in clinical parameters, disease or illness. For example, in some embodiments, an effective amount can refer to an amount of a composition, compound, or agent that improves the subject's condition by at least 5%, such as at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100%. The actual dosage levels of the active ingredients and agents in the disclosed subject matter can be varied so as to administer an amount of the active agent(s) that is effective to achieve the desired response for a particular subject and / or use. The selected dosage level will vary depending upon a variety of factors including, but not limited to, the activity of the composition, formulation, route of administration, combination with other drugs or treatments, the severity of the symptoms being treated, and the physical condition and history of the subject being treated. Determination and adjustment of effective dosages, as well as evaluation of when and how such adjustments are made, are contemplated herein. The term "therapeutically effective amount" can mean a therapeutically sufficient amount to prevent the spread of cancer or reverse cancer.

[0047] Compound Some embodiments relate to therapeutic compounds (e.g., therapeutics). Some embodiments relate to 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A), its crystalline forms (Compound A-Xln), stable crystalline forms of its phosphate (Compound A-P) (Compound A-P-Xln), and any enantiomers of the foregoing (e.g., Compound R-A, Compound S-A, Compound R-A-Xln, Compound S-A-Xln, Compound R-A-P-Xln, Compound S-A-P-Xln), each of which is a therapeutic compound. In some embodiments, the S in the name of Compound A indicates the S configuration, and the R in the name of Compound A indicates the R configuration. The two enantiomers of Compound A, Compound R-A and Compound S-A, are represented by the following structures, respectively:

[0048] [Chemical formula]

[0049] [Chemical formula]

[0050] In some embodiments, a crystalline form of Compound A is provided. In some embodiments, the crystalline form of Compound A is in the free base form. In some embodiments, the crystalline form is represented as Compound A-Xln. In some embodiments, the crystalline form is characterized by one or more peaks in the XRPD pattern. In some embodiments, the crystalline form is characterized by XRPD peaks at angles selected from 8.899, 9.927, 11.586, 13.145, 14.938, 15.374, 16.104, 16.558, 17.327, 17.820, 18.492, 18.809, 19.912, 20.445, 21.511, 22.004, 22.499, 23.188, 23.859, 24.314, 24.590, 25.171, 25.952, 26.464, 26.841, 27.217, 27.648, 28.457, 29.310, and 29.856, or one or more of any combination of the foregoing. In some embodiments, the crystalline form is characterized by XRPD peaks having an intensity of greater than 10% at angles selected from 8.899, 9.927, 14.938, 15.374, 16.104, 16.558, 18.492, 19.912, 20.445, 21.511, 23.859, 24.314, 24.590, and 25.952, or one or more of any combination of the foregoing. In some embodiments, the crystalline form is characterized by XRPD peaks having an intensity of greater than 20% at angles selected from 9.927, 14.938, 16.558, 20.445, and 21.511, or one or more of any combination of the foregoing.

[0051] In some embodiments, the DSC melting point range of Compound A-Xln is characterized by a melting point range of about 245 °C, 250 °C, 255 °C or less, or a range including and / or spanning the foregoing values. In some embodiments, the DSC melting point range of Compound A-Xln is characterized by a melting point of 249 °C.

[0052] In some embodiments, the TGA peak weight loss range of compound A-Xln is about 245 °C, 250 °C, 255 °C or below, or a range including and / or spanning the aforementioned values. In some embodiments, the TGA peak weight loss of compound A-Xln occurs at a temperature of 249 °C.

[0053] In some embodiments, a crystalline form of a salt of compound A is provided. In some embodiments, the crystalline form is represented as compound A-P-Xln. In some embodiments, the crystalline form is characterized by one or more peaks in the XRPD pattern. In some embodiments, the crystalline form is characterized by XRPD peaks at angles selected from one or more of 5.268, 7.139, 9.805, 10.455, 11.799, 12.417, 12.669, 13.672, 14.307, 15.546, 16.064, 16.719, 17.495, 18.035, 18.802, 19.530, 21.095, 22.535, 23.426, 25.916, and 26.577, or any combination thereof. In some embodiments, the crystalline form is characterized by XRPD peaks having an intensity of more than 50% at angles selected from one or more of 7.139, 12.417, 12.669, 13.672, 16.064, 16.719, 19.530, 21.095, and 23.426, or any combination thereof. In some embodiments, the crystalline form is characterized by XRPD peaks having an intensity of more than 80% at angles selected from one or more of 7.139, 13.672, 19.530, and 23.426, or any combination thereof.

[0054] In some embodiments, the DSC melting point range of compound A-P-Xln is characterized by a melting point range of about 220 °C, 225 °C, 230 °C, 235 °C, 240 °C, 245 °C or below, or a range including and / or spanning the aforementioned values. In some embodiments, the DSC melting point range of compound A-P-Xln is characterized by a melting point of 229 °C.

[0055] In some embodiments, the TGA peak weight loss range of compound A-P-Xln is about 205°C, 210°C, 215°C, 220°C, 225°C, 230°C or below, or a range including and / or spanning the aforementioned values. In some embodiments, the TGA peak weight loss of compound A-P-Xln occurs at a temperature of 210°C.

[0056] Method for preparing crystalline form Some embodiments relate to a method for preparing a stable crystalline form of compound A (compound A-Amp) from an amorphous form of compound A, for example, compound A-Xln, compound S-A-XIn, compound R-A-Xln, compound A-P-Xln, compound S-A-P-Xln, or compound R-A-P-Xln, etc., such as 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro-[2.4]-heptan-7-ol ((compound A). In some embodiments, the method for preparing a stable crystalline form of compound A includes one or more crystallization and / or recrystallization steps.

[0057] In some embodiments, preparing a stable crystalline form of compound A (e.g., compound A-Xln, compound S-A-Xln, compound R-A-Xln, compound A-P-Xln, compound S-A-P-Xln, or compound R-A-P-Xln) includes at least one recrystallization step. In some embodiments, the method for preparing compound A-Xln includes multiple recrystallization steps (e.g., 2 steps, 3 steps, 4 steps, etc.). In some embodiments, preparing a stable crystalline form of compound A (e.g., compound A-Xln, compound S-A-Xln, compound R-A-Xln, compound A-P-Xln, compound S-A-P-Xln, or compound R-A-P-Xln) includes at least two recrystallization processes (e.g., two recrystallization steps). In other embodiments, the preparation includes only one recrystallization step.

[0058] In some embodiments, the preparation of the stable crystalline forms of Compound A disclosed herein (e.g., Compound A-Xln, Compound A-P-Xln) may include a recrystallization process in a high-boiling solvent. In some embodiments, the high-boiling solvent is a solvent (or a mixture of solvents) having a boiling point equal to or at least approximately the next boiling point of: 100 °C, 110 °C, 120 °C, 130 °C, 140 °C, 150 °C, 160 °C, 170 °C, 180 °C, 190 °C, 200 °C, 250 °C, or a range including and / or spanning the aforementioned values. In some embodiments, after this recrystallization step, one or more stable forms of Compound A-Xln are provided. In some embodiments, additional recrystallization steps are performed to provide stable forms of Compound A-P-Xln, Compound S-A-P-Xln, or Compound R-A-P-Xln.

[0059] In some embodiments, the preparation of the stable crystalline forms of Compound A disclosed herein (e.g., Compound A-Xln, Compound S-A-Xln, Compound R-A-Xln, Compound A-P-Xln, Compound S-A-P-Xln, or Compound R-A-P-Xln) may include a recrystallization process in a low-boiling solvent (or a selected mixture of solvents). In some embodiments, the low-boiling solvent is a solvent (or a mixture of solvents) having a boiling point below the following: 30 °C, 40 °C, 50 °C, 60 °C, 70 °C, 80 °C, 90 °C, <100 °C, or a range including and / or spanning the aforementioned values.

[0060] In some embodiments, when at least two crystallization processes are used to prepare the stable crystalline form of Compound A, one process may include a recrystallization process in a high-boiling solvent (e.g., a solvent having a high boiling point or a mixture of solvents having a high boiling point together), and another process may include a recrystallization process in a low-boiling solvent (a solvent having a low boiling point or a mixture of solvents having a low boiling point together). In some embodiments, when multiple recrystallization processes are used, the first recrystallization process may be carried out using a high-boiling solvent (or a selected mixture of solvents including a high-boiling solvent and another solvent that may or may not have a high boiling point). In some embodiments, when two or more recrystallization processes are used, the second recrystallization process may be carried out using a low-boiling solvent (or a selected mixture of solvents including a low-boiling solvent and another solvent that may or may not have a low boiling point).

[0061] Alternatively, recrystallization in a low-boiling solvent may precede recrystallization in a high-boiling solvent.

[0062] As disclosed elsewhere herein, when a mixture of solvents is used, if the mixture itself has a high boiling point, the mix of components may also be regarded here as a "high-boiling solvent". Similarly, when a mixture of solvents is used, if the mixture itself has a low boiling point, the mix of components may also be regarded here as a "low-boiling solvent".

[0063] In some embodiments, for the preparation of a stable crystalline form (Compound A-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A), the high-boiling solvent (e.g., for the first recrystallization process) is selected from, but not limited to, DMF, DMA, NMP, a mixture of any of the aforementioned selected solvents, or others. In some embodiments, for the preparation of a stable crystalline form (Compound A-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A), the low-boiling solvent (e.g., for the second recrystallization process) is selected from, but not limited to, MeOH, EtOH, IPA, a mixture of any of the aforementioned selected solvents, or others.

[0064] In some embodiments, for the preparation of a stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A), the high-boiling solvent for the first recrystallization process is DMF, and the low-boiling solvent for the continuous recrystallization process (the second recrystallization process) is EtOH.

[0065] Different crystalline forms of a drug are characterized by different physical and chemical properties (such as stability, solubility, dissolution rate, bioavailability, etc.), which can thereby result in differences in the efficacy, safety, or quality of the drug. In some embodiments, one or more stability tests and / or dissolution tests can be performed. The crystalline form of Compound A-Xln has been found to have better stability and dissolution rate compared to the amorphous form of Compound A-Amp.

[0066] As disclosed elsewhere in this specification, in some embodiments, the present disclosure relates to methods of preparing salt forms of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A), pharmaceutically acceptable salts of Compound A, and the like. The concept of "pharmaceutically acceptable salts" includes, but is not limited to, acid addition salts formed from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid; or 1-hydroxy-2-naphthoic acid, 2,2-dichloroacetic acid, 2-hydroxyethanesulfonic acid, 2-oxoglutaric acid, 4-acetamidobenzoic acid, 4-aminosalicylic acid, acetic acid, adipic acid, ascorbic acid (L), aspartic acid (L), benzenesulfonic acid, benzoic acid, borneol acid (+), borneol-10-sulfonic acid (+), capric acid (decanoic acid), caproic acid (hexanoic acid), caprylic acid (octanoic acid), carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecyl sulfate, ethane-1,2-disulfonic acid, ethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid (D), gluconic acid (D), glucuronic acid (D), glutamic acid, glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid (DL), lactobionic acid, lauric acid, maleic acid, malic acid (-L), malonic acid, mandelic acid (DL), methanesulfonic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, nicotinic acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, phosphoric acid, propionic acid, pyroglutamic acid (-L), salicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid (+L), thiocyanic acid, toluenesulfonic acid (para-), undecylenic acid, and other organic acids.

[0067] Some embodiments relate to the preparation of salts and / or stable crystalline salt forms of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol. Some embodiments relate to the preparation of the phosphate salt (Compound A-P) and / or stable crystalline phosphate salt form (Compound A-P-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A).

[0068] In some embodiments, it is for preparing the stable crystalline phosphate salt form (Compound A-P-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A), and this method includes a neutralization process of a solution of a solvent (or a mixture of solvents) containing Compound A and phosphoric acid. In some embodiments, the recrystallization of Compound A-P is carried out in a solvent (or a mixture of solvents).

[0069] In some embodiments, for preparing the stable crystalline phosphate salt form (Compound A-P-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A), the solvents for the neutralization process and / or the recrystallization process include solvents selected from, but not limited to, MeOH, EtOH, IPA, any mixture of the foregoing, or other solvents.

[0070] In some embodiments, for preparing the stable crystalline phosphate form (Compound A-P-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A), the solvent for the neutralization process and / or the recrystallization process is EtOH.

[0071] In some embodiments, as disclosed elsewhere herein, Compound A-Xln, Compound S-A-Xln, or Compound R-A-Xln can be prepared using one recrystallization step (e.g., in a high-boiling solvent or a mixture of solvents). In some embodiments, Compound A is heated in a high-boiling solvent. In some embodiments, a portion of the high-boiling solvent is removed (e.g., under vacuum). In some embodiments, the solution is allowed to stand (e.g., cooled), at which point crystals form. In some embodiments, the crystals are collected.

[0072] In some embodiments, Compound A-P-Xln, Compound S-A-P-Xln, or Compound R-A-P-Xln can be prepared using a subsequent recrystallization process. In some embodiments, for example, Compound A-P-Xln, Compound S-A-P-Xln, or Compound R-A-P-Xln is prepared by introducing phosphoric acid (e.g., excess phosphoric acid) into a second recrystallization solvent (e.g., a low-boiling solvent). In some embodiments, Compound A is heated in a solvent (e.g., a low-boiling solvent). In some embodiments, a portion of the solvent is removed (e.g., under vacuum). In some embodiments, the solution is allowed to stand (e.g., cooled), at which point crystals form. In some embodiments, the crystals are collected. In some embodiments, when excess phosphoric acid is used, a portion of the phosphoric acid is neutralized.

[0073] Pharmaceutical composition In some embodiments, a pharmaceutical composition is provided. In some embodiments, the pharmaceutical composition comprises a stable crystalline form of compound A or a stable crystalline form of a salt of compound A, as disclosed elsewhere herein, and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition comprises a stable crystalline form of compound A or a stable crystalline form of a salt of compound A, as disclosed elsewhere herein, and a pharmaceutically acceptable excipient.

[0074] In some embodiments, provided is a method for preparing a pharmaceutical composition comprising a stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A-P-Xln), or a phosphate salt (Compound A-P) or a stable crystalline phosphate form (Compound A-P-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol, and a pharmaceutically acceptable carrier or excipient. In some embodiments, the excipient or carrier (or both) is mixed with Compound A-Xln or Compound A-P-Xln.

[0075] Treatment methods In some embodiments, a treatment method is provided. In some embodiments, the method comprises obtaining a pharmaceutical composition comprising a stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A-P-Xln), or a phosphate salt (Compound A-P) or a stable crystalline phosphate form (Compound A-P-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol.

[0076] In some embodiments, the method comprises administering to a patient suffering from a disease a compound A-Xln, a compound A-P, or a compound A-P-Xln. In some embodiments, the disease is a neoplastic disease.

[0077] In some embodiments, the compound A-Xln, the compound A-P, or the compound A-P-Xln comprises one chiral isomer of the compound A-Xln, the compound A-P, or the compound A-P-Xln. In some embodiments, a method for preparing the chiral isomer is implemented. In some embodiments, preparative HPLC with a chiral column is used to separate the enantiomers (two chiral isomers, compound R-A and compound S-A) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (compound A) from each other. In some embodiments, the chiral isomers can be recrystallized (and / or crystallized) as disclosed elsewhere herein.

[0078] Some embodiments relate to the compound A-Xln in a stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol, or a phosphate or stable crystalline phosphate form, or two chiral isomers (Compound R-A, Compound S-A) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol for use in the manufacture of a medicament for a method of treating neoplastic diseases. Some embodiments relate to a method of treating a disease selected from the group consisting of lung cancer, renal cancer, colorectal cancer, gastric cancer, melanoma, head and neck cancer, thyroid cancer, pancreatic cancer, liver cancer, prostate cancer, bladder cancer, brain cancer, sarcoma, breast cancer, ovarian cancer, cervical cancer, endometrial cancer, particularly small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC) and ovarian cancer, tumors caused by one or more of the foregoing, blood cancers (selected from ALL, CLL, AML, CML and multiple myeloma), and combinations thereof. In some embodiments, the method of treatment comprises administering the compound A-Xln in a stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol, or a phosphate or stable crystalline phosphate form, or two chiral isomers (Compound R-A, Compound S-A) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol.

[0079] Some embodiments relate to stable crystalline forms or salts or stable crystalline salt forms or two chiral isomers (Compound R-A, Compound S-A) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol for use in methods, either as monotherapy or in combination with additional chemotherapeutic agents. In some embodiments, the additional chemotherapeutic agent is selected from platinum-based agents or taxane-based agents.

[0080] In some embodiments, the method of treatment comprises selecting a patient suffering from a selected disease including a solid tumor. In some embodiments, the method of treatment comprises selecting a patient suffering from a disease selected from the group consisting of lung cancer, renal cancer, colorectal cancer, gastric cancer, melanoma, head and neck cancer, thyroid cancer, pancreatic cancer, liver cancer, prostate cancer, bladder cancer, brain cancer, sarcoma, breast cancer, ovarian cancer, cervical cancer, endometrial cancer, particularly small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC) and ovarian cancer, tumors caused by one or more of the foregoing, blood cancers (selected from ALL, CLL, AML, CML and multiple myeloma), and combinations thereof. In some embodiments, the method comprises administering to the patient a therapeutically effective amount of a stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol, or a phosphate or stable crystalline phosphate form, or two chiral isomers (Compound R-A, Compound S-A) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol.

[0081] In some embodiments, an effective amount of Compound A-Xln, Compound A-P, and / or Compound A-P-Xln is administered to a patient in need of treatment (e.g., a subject suffering from a disease disclosed herein). In some embodiments, an effective amount of Compound A-Xln, Compound A-P, and / or Compound A-P-Xln is administered to a patient in need of treatment (e.g., a subject suffering from a disease disclosed herein).

[0082] Some embodiments relate to a stable crystalline form or salt or stable crystalline salt form or two chiral isomers (Compound R-A, Compound S-A) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol for use in a method, either alone or in combination with an immunotherapeutic agent. In some embodiments, the immunotherapeutic agent is selected from PD-1, PD-L1, oncolytic virus therapy, bispecific T cell engagers (BiTE), and chimeric antigen receptor (CAR) T cell therapy-based reagents, including, but not limited to, nivolumab, pembrolizumab, ipilimumab, blinatumomab, elotuzumab, daratumumab, semipilimab, avelumab, durvalumab, atezolizumab, toripalimab, sintilimab, camrelizumab, tislelizumab, AK104, pemprinumab, KN035, CS1001, talimogene laherparepvec. In some embodiments, this combination is used to treat a disease or disorder as disclosed elsewhere herein. For example, it is used in the treatment of solid tumors selected from lung cancer, renal cancer, colorectal cancer, gastric cancer, melanoma, head and neck cancer, thyroid cancer, pancreatic cancer, liver cancer, prostate cancer, bladder cancer, brain cancer, sarcoma, breast cancer, ovarian cancer, cervical cancer, and endometrial cancer, and blood cancers selected from ALL, CLL, AML, CML, and multiple myeloma.

[0083] To test the biological activity, the PTK inhibitory activity of Compound A or Compound A-Xln was compared with the commercial drug sunitinib. In vitro tyrosine kinase inhibitory activity can be measured via commercial resources. Some of these tests can also be contracted with Eurofin or Reaction Biology for screening.

[0084] In the kinase inhibition test, the receptor tyrosine kinases are selected from FGFRl(h), FGFR2(h), FGFR3(h), Fltl(h)(VEGFrl), Flt4(h)(VEGFr3), KDR(h)(VEGFr2), Aurora-B, PDGFRα(h), and PDGFRβ(h). Compound A shows inhibition against all targets, and the IC50 values range from less than nanomolar to the micromolar scale. Compared with the commercial drug sunitinib, Compound A shows a higher inhibitory effect.

[0085] The inhibition of Compound A-Xln, Compound A-P-Xln, and two chiral isomers (Compound R-A, Compound S-A) against cancer cell lines was tested by in vitro MTT assay. The cancer cell lines are selected from PANC-1, NCI-H157, MDA-MB-231, Hela, PC-3, BEL7404, MKN45, Ishikawa, Saos-2, SKOV3, SW579, and HCT116 cell lines. The IC50 values are in micromolar units.

[0086] The following examples further illustrate the invention but should not be construed as limiting its scope in any way.

Example

[0087] Abbreviations and Definitions For easy reference, the following abbreviations are used and have the following meanings. EtOH: Ethanol, MeOH: Methanol, IPA: Isopropanol, EtOAc: Ethyl acetate, DCM: Dichloromethane, DMF: N,N-Dimethylformamide, DMA: N,N-Dimethylacetamide, NMP: N-Methyl-2-pyrrolidone, RT: Room temperature, Temp: Temperature, eq: Equivalent, g: Gram, mg: Milligram, ml: Milliliter, min: Minute. API: Active pharmaceutical ingredient. DSC: Differential scanning calorimetry, TGA: Thermogravimetric analysis, XRPD: X-ray powder diffraction, Exo: Exothermic, Endo: Endothermic. ALL: Acute lymphoblastic leukemia or lymphoblastic leukemia, CLL: Chronic lymphocytic leukemia or lymphoblastic leukemia, AML: Acute myeloid leukemia, CML: Chronic myeloid leukemia, NSCLC: Non-small cell lung cancer, SCLC: Small cell lung cancer, ULMS: Uterine leiomyosarcoma (sarcoma), OC: Ovarian cancer, BT: Brain tumor, SD: Stable, PR: Partial response, PFS: Progression-free survival, IC50: Half maximal inhibitory concentration. This is the concentration of the inhibitor in cell culture medium required to inhibit the transport activity of the protein by 50%. MTT: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. Example 1: Preparation of crystalline form 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A-Xln)

[0088] The crude compound A (18 g) prepared in the same manner according to International Publication No. 2010 / 021918 was obtained as an amorphous solid (Compound A-Amp) having the main peak of the DSC of Compound A at 245.99 °C. DMF (180 ml) was added thereto, and the mixture was stirred at 120 °C until the solid dissolved. The heated solution was filtered, and the filtrate was concentrated under vacuum to about half the volume and further cooled to room temperature. The precipitate was filtered, and the filter cake was washed with a small amount of DMF and then with a small amount of water. Next, the filter cake was mixed with ethanol (4 l) and heated to dissolve. The solution was concentrated under vacuum to about half the volume and further cooled to room temperature. The precipitate was filtered and dried in an oven to obtain a stable crystal (Compound A-Xln) as the desired product having only one peak in the DSC at 248.79 °C.

[0089] The DSC graph of the amorphous form of Compound A (Compound A-Amp) is shown in the drawing section of Figure 1.

[0090] The DSC, TGA, XRPD and 1 1H-NMR graphs of the crystalline form of Compound A (Compound A-Xln) are shown in the drawing section as Figure 2, Figure 3, Figure 4 and Figure 5 respectively. MS: (M+H + ) / z 478; 1 1H-NMR (DMSO-d6) δ ppm: 0.37 - 0.39 (m, 1H), 0.50 - 0.57 (m, 2H), 0.82 - 0.85 (m, 1H) 2.43 (s, 3H), 2.54 - 2.59 (d, 2H), 3.11 - 3.13 (d, 1H), 3.26 (s, 2H), 3.42 - 3.46 (m, 1H), 3.82 - 3.84 (t, 1H), 3.98 (s, 3H), 4.39 (t, 2H), 6.28 (s, 2H), 6.35 - 6.36 (d, 1H), 6.98 - 7.02 (t, 1H), 7.22 - 7.24 (d, 1H), 7.48 (s, 1H), 7.62 (s, 1H), 8.43 - 8.46 (d, 1H), 11.42 (s, 1H). DSC melting point range (Endo): 247 - 253 °C, peak temperature = 248.79 °C. In TGA, it is shown to be an unsolvated substance with weight loss above 250 °C. XRPD with 14 characteristic peaks where the intensity % exceeds 10%, or a pattern containing 43 characteristic peaks where all intensity % are represented by d values and angles as follows:

[0091] [Table 4] [Examples]

[0092] Based on the inventor's research experience regarding the preparation of the crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A) in Example 1, the preparation of the following crystalline forms of Compound A is expected. The preparation of the crystalline form of Compound A (Compound A-Xln) shall include at least one recrystallization process, preferably two recrystallization processes. Preferably, for the first recrystallization, relatively high-boiling solvents (or solvent mixtures) such as DMF, DMA, and NMP, but not limited thereto, are required. For the subsequent recrystallization process, certain solvents (or solvent mixtures) such as MeOH, EtOH, and IPA, but not limited thereto, are required. [Examples]

[0093] Comparison of the stability of the amorphous (Compound A-Amp) and crystalline form (Compound A-Xln) APIs of (2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro-[2.4]-heptan-7-ol (Compound A).

[0094] Samples of the amorphous and crystalline form APIs of Compound A were placed in clean containers, and the environmental temperature was controlled at 60 °C. The purity of the amorphous (Compound A-Amp) and crystalline form (Compound A-Xln) of Compound A was tested on day 0, day 10, and day 30. The amorphous and crystalline form API samples were measured, an appropriate amount of diluent was added, and they were dissolved by sonication. The concentration was controlled at 0.2 mg / ml, and purification was determined by HPLC equipped with a UV detector at 230 nm. The results are shown in the following table, and the fitting curve is shown in the drawing section of Figure 6. A-Xln has better stability than A-Amp.

[0095]

Table 5

Example

[0096] (2-(4-(4-Fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]-heptan-7-ol (Compound A) tablets of the amorphous form (Compound A-Amp) and crystalline form (Compound A-Xln) were compared for stability.

[0097] Samples of the amorphous and crystalline form tablets (10 mg or 30 mg) of Compound A (named Compound A-Amp or A-Xln; Compound A-Amp tablets or Compound A-Xln tablets) were prepared by the following process with the tablet compositions shown below:

[0098] (1) Sieving process: Compound A (A-Amp or A-Xln), lactose T80, microcrystalline cellulose (PH101), and sodium carboxymethyl starch were each sieved through an 80-mesh screen.

[0099] (2) Granulation process: The sieved compound A (A-Amp or A-Xln), lactose T80, microcrystalline cellulose, sodium carboxymethyl starch, and silicon dioxide were charged into a V-shaped blender and blended as Blend 1. After Blend 1, this material was fed into a roller compactor for dry granulation. The dried granules were sieved through a 24-mesh screen, and the obtained powder was sieved through a 50-mesh screen to obtain the desired fine powder accounting for 10% - 13% of the total particle weight. The sieved granules were blended with pre-sieved magnesium stearate for Blend 2, sampled for the content of the final blend, and tested for blend assay (8.9% - 10.3%).

[0100] (3) Tableting process: After setting the tablet press machine to compress at 10 mg and 30 mg, the final blend was compressed into tablets. During the process for weight variation and hardness, in-process control (IPC) tests were carried out (weight variation was ±6.0%, hardness was 7 - 13 kg / mm2).

[0101] (4) Coating process: A coating solution was prepared according to the ratio of gastric-soluble film coating premix: purified water = 15:85 (w:w), and stirring was continued for 45 minutes. The increase in coating weight was about 2% - 3%.

[0102] (5) Packaging process: The recovered tablets were processed by weight sorting. The qualified tablets were recovered and packaged in an aluminum-plastic blister at 7 tablets / board.

[0103]

Table 6

[0104]

Table 7

[0105] The prepared tablets were tested using the same protocol as described in Example 3, and the results are shown in Table 4. The fitting curve is shown in the drawing section of Figure 7.

[0106]

Table 8

[0107]

Table 9

[0108]

Table 10

[0109]

Table 11

[0110]

Table 12

[0111]

Table 13

[0112]

Table 14

[0113]

Table 15

[0114]

Table 16

Example

[0115] Comparison of in vitro dissolution curves of Compound A tablets Depending on the crystalline form of the compound, physical and chemical properties such as dissolution rate that can affect the efficacy of the drug may change. The measurement of the dissolution rate can be carried out in a dissolution test. Samples of amorphous (Compound A - Amp) tablets and crystalline (Compound A - Xln) tablets were dissolved in a suitable solvent (0.1 mol hydrochloric acid solution) and stirred at an appropriate speed (ZRC - 8D intelligent dissolution apparatus, speed 50 rpm) to promote the dissolution process. The dissolution rate was measured by HPLC with the wavelength of the UV detector set at 230 nm. Samples were taken at 5 minutes, 10 minutes, 20 minutes, 30 minutes, and 45 minutes. The experimental results are shown in the table below, and the dissolution curve can be described in the drawing section as Figure 8. After comparing the experimental data, it can be observed that the crystalline tablets dissolve faster than the amorphous tablets.

[0116]

Table 17

[0117]

Table 18

Example

[0118] Preparation of phosphate and crystalline form (Compound A - P - Xln) of 5-(2-(4-(4 - fluoro - 2 - methyl - 1H - indol - 5 - yloxy)-6 - methoxyquinolin - 7 - yloxy)ethyl)-5 - azaspiro[2.4] - heptan - 7 - ol Compound A (250 mg) was dissolved in EtOH (110 ml) under reflux, and EtOH containing 1 M H3PO4 (0.53 ml) was added to the resulting solution. The reaction mixture was refluxed for 1 hour and cooled to room temperature with slow stirring overnight. The solid was filtered, washed with EtOH by rinsing, and further dried in an oven at 50 °C for 10 hours to obtain a white solid as the phosphate salt of Compound A with a stable crystalline form (Compound A-P-Xln).

[0119] DSC, TGA, XRPD of the phosphate crystal (Compound A-P-Xln) and 1 The 1H-NMR graphs are shown in FIGS. 9, 10, 11 and 12 respectively. 1 1H NMR (DMSO-d6) δ ppm: 0.37 - 0.39 (m, 1H), 0.50 - 0.57 (m, 2H), 0.82 - 0.85 (m, 1H) 2.43 (s, 3H), 2.51 - 2.56 (m, IH), 2.58 - 2.60 (d, IH), 2.72 - 2.74 (d, 1H), 2.90 (t, 2H), 3.14 - 3.18 (m, IH), 3.77 - 3.82 (m, IH), 3.97 (s, 3H), 4.24 (t, 2H), 4.60 (s, IH), 6.28 (s, IH), 6.33 - 6.34 (d, IH), 7.00 (t, IH), 7.22 - 7.24 (d, IH), 7.42 (s, IH), 7.60 (s, IH), 8.42 - 8.43 (d, IH), 11.42 (s, IH). DSC melting point range (Endo): 221 - 235 °C, peak temperature = 229 °C. In TGA, a weight loss of 1.84% was shown at about 30 - 60 °C, and a significant weight loss was shown above 210 °C. In XRPD with a pattern containing 21 characteristic peaks, all intensity % are represented by d values and angles as follows:

[0120]

Table 19

Example

[0121] Based on the inventors' experience in the research on the preparation of the stable crystalline phosphate form (Compound A-P-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A), the preparation of the following stable crystalline phosphate forms of Compound A is expected. The preparation of the stable crystalline phosphate compound (Compound A-P-Xln) shall include a neutralization process of Compound A with a phosphoric acid solution in a certain solvent (or a mixture of solvents) and a recrystallization process in a certain solvent (or a mixture of solvents). Preferably, this stable crystalline phosphate form (Compound A-P-Xln) can be obtained from the recrystallization of a solvent (or a mixture of solvents) including but not limited to MeOH, EtOH, and IPA.

Example

[0122] Preparation of Enantiomers (Chiral Isomers) Compound A (10 mg) was dissolved in IPA (10 ml), and 2 ml of this solution was injected into a chiral HPLC apparatus equipped with a chiral separation column CHIRALCEL OD 500x50 mm at a flow rate of 5 ml / min and UV 240 nm using the mobile phase hexane:IPA:diethylamine (85:15:0.1).

[0123] A total of 5 injections were repeated, and the collected fractions were combined and further evaporated to obtain Compound R-A (specific rotation: -23.49, 1.3 mg) and Compound S-A (specific rotation: +22.51, 1.6 mg) as two enantiomers. R-A: 11H-NMR (DMSO-d6) δ ppm: 0.34 - 0.40 (m, 1H), 0.48 - 0.60 (m, 2H), 0.80 - 0.87 (m, 1H), 2.42 (s, 3H), 2.59 - 2.62 (d, 1H), 2.73 - 2.75 (d, 1H), 2.87 - 2.92 (m, 1H), 2.89 (t, 2H), 3.14 - 3.19 (m, 1H), 3.74 - 3.79 (m, 1H), 3.96 (s, 3H), 4.24 (t, 2H), 6.28 (s, 1H), 6.32 - 6.33 (d, 1H), 6.99 (t, 1H), 7.20 - 7.23 (d, 1H), 7.41 (s, 1H), 7.59 (s, 1H), 8.41 - 8.43 (d, 2H), 11.42 (s, 1H). S-A: 1 1H-NMR (DMSO-d6) δ ppm: 0.34 - 0.41 (m, 1H), 0.49 - 0.59 (m, 2H), 0.80 - 0.85 (m, 1H), 2.42 (s, 3H), 2.59 - 2.62 (d, 1H), 2.73 - 2.76 (d, 1H), 2.87 - 2.92 (m, 1H), 2.89 (t, 2H), 3.14 - 3.19 (m, 1H), 3.74 - 3.79 (m, 1H), 3.96 (s, 3H), 4.24 (t, 2H), 6.28 (s, 1H), 6.32 - 6.33 (d, 1H), 6.99 (t, 1H), 7.20 - 7.23 (d, 1H), 7.41 (s, 1H), 7.59 (s, 1H), 8.41 - 8.43 (d, 2H), 11.42 (s, 1H).

Example

[0124] Kinase inhibition To test the biological activity of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A-Xln), the inhibition of kinases FGFR1(h), FGFR2(h), FGFR3(h), Flt1(h) (VEGFr1), Flt4(h) (VEGFr3), KDR(h) (VEGFr2), Aurora-B, PDGFRα(h), PDGFRα(h), PDGFRβ(h) was tested, and the IC50 values are shown in the table below. Furthermore, the kinase inhibition of the commercially available drug sunitinib was also tested, and the results were compared with those of Compound A-Xln.

[0125]

Table 20

Example

[0126] The results of ion chromatography analysis and water solubility of the phosphate salt of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A-P) are shown in the table below.

[0127]

Table 21

Example

[0128] An in vitro MTT (proliferation) assay was performed using the compounds of the above examples, and the following inhibition results were obtained.

[0129]

Table 22

[0130]

Table 23

Example

[0131] Efficacy of compound A-Xln against tumor cell lines by combination with chemotherapy or immunotherapy in vitro and in vivo, and in vivo xenograft models against various tumor cell lines

[0132] The results of the MTT assay for the combined therapy of compound A-Xln were investigated. 100 μL of a cell suspension at a concentration of 5x104 cells / ml was added to each well of a 96-well plate and placed in an incubator at 37°C and 5% CO2. After 24 hours, the compound A-Xln sample solution was added to duplicate wells at a rate of 10 μl / well according to Table 13. After incubation at 37°C and 5% CO2 for 72 hours, 20 μl of a 5 mg / ml MTT solution was added to each well, and after 4 hours, 100 μl / well of a dissolution solution was added. The samples were placed in the incubator, and after dissolution, a full-wavelength multifunctional enzyme labeler was used. The OD value at 570 nm was measured. Figures 13 and 14 show the growth inhibition of breast cancer cell lines MCF7 and BT474 by compound A-Xln, the anti-breast cancer agents anastrozole and letrozole, the combination of compound A-Xln and anastrozole, and the combination of compound A-Xln and letrozole, respectively.

[0133]

Table 24

[0134] The samples in Table 14 were also tested for growth inhibitors of three colon cancer cell lines, HT29, HCT116, and colo205, by compound A-Xln, the anti-colon cancer agent 5FU / O, and the combination of compound A-Xln and 5FU / O. The results are shown in Figures 15 to 17.

[0135]

Table 25

[0136] The efficacy of immunotherapies such as the combined therapy of compound A-Xln and anti-mouse PD-1 antibody was also tested.

[0137] Nude mice were inoculated with mouse colon CT26 cell line for 7 days, randomly divided into groups of 6 each, and then treated with vehicle or compound A-Xln once a day in two dosing groups, anti-mouse PD-1 (PD-1), 200 μg / mouse once every 3 days in one group, or a combination of compound A-Xln and anti-PD-1 in two dosing groups. Table 15 shows the changes in tumor size after administration of each compound. Figure 18 shows the comparison of the average tumor volume changes after administration of compound A-Xln and anti-PD1 antibody agent, and after combination therapy of compound A-Xln and anti-PD1 antibody agent.

[0138]

Table 26

[0139] The in vivo antitumor efficacy of compound A-Xln against various tumor cell lines in a xenograft model was tested.

[0140] The antitumor activities of animals in an in vivo xenograft model using various tumor cell lines (SCLCNCI-H1436, NSCLC95-D, ovarian SKVO3, renal 786-O, liver Bel-7420, brain U87) were carried out as follows: Tumor tissues with well-grown tumor cell lines were cut into 3-mm fragments, and one fragment per nude mouse was subcutaneously inoculated into the right flank. Animals were divided into groups and administered at pre-designed doses. Treatment was started when the tumor size exceeded 100 mm 3 after 10 - 14 days. According to the tumor size, animals with tumors that were too large or too small were excluded, and animals with similar average tumor volumes were divided into groups. Then, the animals were orally administered daily continuously for 14 - 21 days. Thrice a week for 13 days after inoculation, the major diameter a (mm) and minor diameter b (mm) were measured with calipers. Each tumor volume was calculated by the following formula: TV = ab 2 / 2, and the relative tumor volume was calculated as follows: RTV = V t / V0, where V0 represents the tumor volume on the first day of treatment, and V trepresents the tumor volume on each measurement date. The animals were sacrificed 20 - 30 days after inoculation, and the tumors were excised by dissection. The individual body weights and tumor weights were measured and calculated. The tumor inhibition rate was calculated as [l - ((TW t ) / (TW c ))] × 100%, where TW t represents the average tumor weight of the treatment group on the last day of the experiment, and TW c represents the average tumor weight of the control group on the last day of the experiment. The experimental dosage and efficacy results are shown in Figures 19 - 24. The range of the best tumor inhibition rate is over 70% - 100%.

[0141] The conclusion is as follows: It has been demonstrated that by combining compound A - Xln with anti - breast cancer agents such as the aromatase inhibitors anastrozole and letrozole, the inhibition of the growth of human cancer cells such as breast cancer cell lines MCF7 and BT474 is enhanced.

[0142] It has been demonstrated that by combining compound A - Xln with chemotherapeutic agents such as anti - colon cancer 5FU / O, the inhibition of the growth of human cancer cells such as colon cancer cell lines HT29, HCT116, and colo205 is enhanced. The inhibitory activity is higher in the regimen where the chemotherapeutic agent is administered first and then compound A is administered, and there is also a slight difference shown in continuous administration.

[0143] The mouse xenograft model shows a combined inhibitory effect on cancer cell growth, such as the growth of mouse colon cancer cell line CT26. The combination treatment of an anti - PD1 antibody agent and compound A - Xln against cancer cell growth provides strong evidence showing improved efficacy in an in vivo animal model.

[0144] Combination therapy with compound A-Xln and an immunotherapeutic agent such as a PD-1 or PD-L1 antibody, for example, but not limited to, nivolumab, pembrolizumab, ipilimumab, blinatumomab, elotuzumab, daratumumab, semiprimab, avelumab, durvalumab, atezolizumab, toripalimab, sintilimab, camrelizumab, tislelizumab, AK104, AK105, pemprimumab, KN035, CS1001, talimogene laherparepvec, etc., can produce a clinical synergistic effect.

[0145] In various human cancer cell line xenograft models, particularly in SCLC, NSCLC, ovarian cancer, renal cancer, liver cancer, and brain cancer, significant tumor inhibitory activity by compound A-Xln has been demonstrated.

[0146] Example 13: Compound A-Xln tablets in human clinical trials for solid tumors such as NSCLC, SCLC, ovarian cancer, endometrial cancer, cervical cancer, uterine leiomyosarcoma, and peritoneal mesothelioma cancer.

[0147] A: Overview Compound A-Xln tablets were orally administered once daily at 90 mg, 80 mg, 70 mg, 60 mg, or 40 mg for 28 days as one cycle to various cancer patients until intolerance or disease progression (PD) was observed according to the RECISIT 1.1 evaluation. If no intolerance was observed, the dose could be decreased from 90 mg once daily to 80 mg, 70 mg, 60 mg, 50 mg, 40 mg, 30 mg, or 20 mg at 10 mg intervals. The treatment results of several representative patients showing remarkable unexpected antitumor activity are shown below (Cutoff, March 2022). In the treatment after the third treatment (after secondary treatment experience) of SCLC, a very good objective response rate (ORR) exceeding 20% was observed, and in the treatment after the third treatment (after secondary treatment experience) of platinum-resistant ovarian cancer, approximately 20% was observed. After the end of the primary standard-of-care (SOC) cycle, there was a certain degree of efficacy in SCLC patients who received maintenance therapy with Compound A-Xln tablets. As an option for second-line treatment (with primary treatment experience), there was a certain degree of efficacy in SCLC patients treated with Compound A-Xln tablets. Efficacy was also observed in sarcoma and brain tumors.

[0148]

Table 27

[0149] Furthermore, in a completed SCLC clinical trial involving 30 patients with primary treatment experience (Group B) and 30 patients with secondary or later treatment experience (Group C), positive clinical efficacy was demonstrated for the following progression-free survival (PFS). B: 3.68 months and C: 3.62 months, overall survival (OS) of B: 10.39 months, and C: >15 months. These are far superior to the available treatment options currently reported for primary treatment experience or treatment after secondary treatment. The Kaplan-Meier curves are shown in Figures 25 and 26.

[0150] B: Representative subject cases (using 10 mg tablets each)

[0151] (1): Representative subject S01013 in Phase 1b For the NSCLC subject S01013 (treatment experience of three or more times), compound A-Xln tablets (60 mg, 10 mg x 6) were orally administered once a day for a 28-day cycle. In the second cycle, the dosage was reduced to 40 mg and 30 mg, and the same treatment regimen as described above was used in subsequent cycles. The best response of the subject to the treatment was PR (a 54.70% decrease compared to the baseline target lesions).

[0152] (2): Representative subject S01014 in Phase 1b For the NSCLC subject S01014 (treatment experience = 3), compound A-Xln tablets (60 mg) were orally administered once a day for a 28-day cycle. In the third cycle, the dosage was reduced to 40 mg, and the same treatment regimen as described above was used in subsequent cycles. The best response of the subject to the treatment was PR (a 56.95% decrease compared to the baseline target lesions).

[0153] (3): Representative subject S05002 in Phase 1b For the NSCLC subject S05002 (treatment experience = 2), compound A-Xln tablets (60 mg) were orally administered once a day for a 28-day cycle. The same treatment regimen as described above was also used in subsequent cycles. The best response of the subject to the treatment was PR (a 38.42% decrease compared to the baseline target lesions).

[0154] (4): Representative subject 020207 in Phase 1 For the subject 020207 with peritoneal mesothelioma (treatment experience = 3), compound A-Xln tablets (40 mg) were orally administered once a day for a 28-day cycle in combination with tripalimab injection (240 mg, once every 3 weeks). The same treatment regimen as described above was also used in subsequent cycles. The best response of the subject to the treatment was PR (a 36.4% decrease compared to the baseline target lesions).

[0155] (5): Representative subject 020202 in Phase 1 Compound A-Xln tablets (40 mg) were orally administered once daily for a 28-day cycle in combination with toripalimab injection (240 mg, once every 3 weeks) to Subject 020202 with small cell lung cancer (treatment experience = 1). The same treatment regimen as described above was used in subsequent cycles. The best response of the subject to the treatment was PR (a 38.22% decrease compared to the baseline target lesions).

Claims

1. A stable crystalline form (Compound A-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A), which is produced by a method comprising subjecting the amorphous form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol to recrystallization, which is selected from using a high-boiling solvent or a mixture of solvents having a high boiling point together, and using a low-boiling solvent or a mixture of solvents having a low boiling point together, not limited to these. Here, the high-boiling solvent is DMF and the low-boiling solvent is EtOH, a stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A).

2. The stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A) according to Claim 1, exhibiting at least one of the following characteristics: DSC melting point range (Endo): 240 - 260 °C, peak temperature range: 244 - 254 °C; And a specific DSC melting point range (Endo): 247 - 253 °C, peak temperature 249 °C. A TGA thermogram indicating that it is a non-solvated substance having a weight loss at 250 °C or higher.

3. The stable crystalline form (Compound A-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]heptan-7-ol (Compound A) according to Claim 1, characterized by an XRD pattern including 14 characteristic peaks with an intensity % exceeding 10% at 2θ of 8.9, 9.9, 14.9, 15.4, 16.1, 16.6, 18.5, 19.9, 20.4, 21.5, 23.9, 24.3, 24.6, and 26.0 degrees.

4. Characterized by an XRPD pattern comprising characteristic peaks of 43, and all intensity % being represented by d-value and angle as follows, the stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]-heptan-7-ol (Compound A) according to claim 1 (Compound A-Xln). 【Table 1】

5. The stable crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]-heptan-7-ol (Compound A) according to claim 1 (Compound A-Xln), characterized substantially by the XRPD pattern shown in Figure 4.

6. The stable crystalline form of the phosphate of the crystalline form of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]-heptan-7-ol (Compound A) according to claim 1 (Compound A-Xln) (Compound A-P-Xln), prepared by a method comprising the steps of neutralizing a solution of a solvent or a mixture of solvents containing phosphoric acid and a recrystallization process.

7. The compound according to claim 6, wherein the solvent in which the neutralization and / or recrystallization is carried out is EtOH.

8. The phosphate of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]-heptan-7-ol, characterized by one or more of the following properties, the compound according to claim 6: DSC melting point range (Endo): 221 - 235 °C, peak temperature 229 °C; TGA showing a slight weight loss at about 30 - 60 °C and a significant weight loss above 210 °C.

9. The phosphate of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]-heptan-7-ol, characterized by an XRPD pattern comprising all characteristic peaks, and all intensity % being represented by d-value and angle as follows, the compound according to claim 6. 【Table 2】

10. A compound having the structure represented by 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]-heptan-7-ol according to claim 1, for use in the inhibition of protein tyrosine kinase (PTK), its stable crystalline form, its salt form, and / or its stable crystalline salt form, and / or its enantiomer.

11. The compound according to claim 10, wherein the PTK is selected from, but not limited to, FGFRl(h), FGFR2(h), FGFR3(h), Fltl(h) (VEGFrl), Flt4(h) (VEGFr3), KDR(h) (VEGFr2), Aurora-B PDGFRα(h), PDGFRα(h) and PDGFRβ(h).

12. A compound having the structure represented by 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]-heptan-7-ol according to claim 1, its stable crystalline form, its salt form, and / or its stable crystalline salt form, and / or its enantiomer, which inhibits common cancer cell lines of pancreas, prostate, sarcoma, thyroid, colon, ovary, blood, breast, brain, NSCLC, SCLC, liver, kidney, colon, cervix and stomach, but not limited to, a compound that inhibits cancer cell lines selected from the group consisting of PANC-1, NCI-H157, MDA-MB-231, HeLa, PC-3, BEL7404, MKN45, Ishikawa, Saos-2, SKOV3, SW579, NCI-H1436, Bel-7402, U87, and HCT116.

13. The compound according to claim 1, having a representative structure selected from S-5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]-heptan-7-ol and R-(2-(4-(4-fluoro-2-methyl-lH-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]-heptan-7-ol and their phosphate forms.

14. A pharmaceutical composition comprising an active ingredient which is a compound according to any one of claims 1, 6 or 13, and a pharmaceutically acceptable carrier and / or a pharmaceutically acceptable excipient.

15. A pharmaceutical composition comprising the compound of claim 1, comprising 10 mg or 30 mg of a stable crystalline form (Compound A-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]-heptan-7-ol (Compound A), and a pharmaceutically acceptable carrier selected from the group consisting of, but not limited to, lactose T80, microcrystalline cellulose (PH101), sodium carboxymethyl starch, silicon dioxide, magnesium stearate, and Opadry enteric coating powder, thereby forming tablets.

16. A method for treating a neoplastic disease, comprising administering to a subject in need thereof a pharmaceutical composition comprising a stable crystalline form (Compound A-Xln) of 5-(2-(4-(4-fluoro-2-methyl-1H-indol-5-yloxy)-6-methoxyquinolin-7-yloxy)ethyl)-5-azaspiro[2.4]-heptan-7-ol (Compound A) or a stable crystalline form (Compound A-Xln) and a pharmaceutically acceptable excipient.

17. The treatment method according to claim 16, further comprising administering an additional agent which is a chemotherapeutic compound and / or an immunotherapeutic agent.

18. The combination chemotherapy agent according to claim 17, selected from the group consisting of, but not limited to, platinum-based or taxane-based or topoisomerase 1 inhibitor or alkaloid agent or alkylating agent groups, cisplatin, carboplatin, paclitaxel, or cisplatin / paclitaxel, carboplatin / paclitaxel, or carboplatin / etoposide, or topotecan, irinotecan, or lomustine.

19. The combined immunotherapy agent is selected from the group of PD-1 or PD-L1 antibodies consisting of, but not limited to, nivolumab, pembrolizumab, ipilimumab, blinatumomab, elotuzumab, daratumumab, semipilimab, avelumab, durvalumab, atezolizumab, tripalimab, sintilimab, camrelizumab, tislelizumab, AK104, AK105, pemprilumab, KN035, CS1001, talimogene laherparepvec, the treatment method according to claim 17.

20. The neoplastic disease is a solid tumor selected from the group of NSCLC, SCLC, mesothelioma, renal cancer, colorectal cancer, gastric cancer, melanoma, head and neck cancer, thyroid cancer, pancreatic cancer, liver cancer, prostate cancer, bladder cancer, brain cancer, sarcoma, breast cancer, ovarian cancer, cervical cancer and endometrial cancer, but not limited thereto, and a hematological cancer selected from the group of ALL, CLL, AML, CML and multiple myeloma, but not limited thereto, the treatment method according to claim 16.

21. The neoplastic disease is small cell lung cancer, non-small cell lung cancer, mesothelioma, ovarian cancer, brain tumor, and sarcoma, the treatment method according to claim 16.

22. The treatment uses Compound A-Xln tablets for maintenance treatment of patients undergoing primary treatment or treatment of patients after secondary or tertiary treatment, the treatment method according to claim 16.

23. The treatment dosage is 90 mg or 80 mg or 70 mg or 60 mg or 50 mg or 40 mg or 30 mg or 20 mg once a day, the treatment method according to claims 16 and 17.

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