Crystalline salt forms of SHP2 inhibitors

Abstract

The present invention relates to a crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1. The present invention also relates to pharmaceutical compositions comprising this crystalline form, and methods of using this crystalline form and such compositions for the treatment of abnormal cell growth, such as cancer, in a mammal. [Figure 1] TIFF2025509818000021.tif85166

Description

[Technical field]

[0001] The present invention relates to crystalline salts of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine, pharmaceutical compositions comprising the crystalline salts, and methods of using the crystalline salts in the treatment of abnormal cell growth, such as cancer, in mammals, particularly humans. One crystalline salt is a crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt ("Form 1"), pharmaceutical compositions comprising succinate salt Form 1, and methods of using succinate salt Form 1 and such compositions in the treatment of abnormal cell growth, such as cancer, in mammals, particularly humans. [Background technology]

[0002] The compound (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine is a SHP2 inhibitor having the formula (I).

[0003] [ka] The preparation of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine is disclosed in International Patent Publication WO2020 / 201991 (International Application No. PCT / IB2020 / 053019 published on October 8, 2020, see also Example 1), the contents of which are incorporated herein by reference in their entirety.

[0004] The preparation of the hemihydrate crystalline form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base Form 1 is disclosed in U.S. Provisional Patent Application No. 63 / 169,340 (see also Example 2). Summary of the Invention [Problem to be solved by the invention]

[0005] Thus, there remains a need for improved forms of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine that have desirable properties such as high crystallinity, high purity, stability, and solubility. [Means for solving the problem]

[0006] The present invention provides, in part, crystalline salt forms of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine. The salt forms may be useful in the treatment of abnormal cell growth. Also provided are pharmaceutical compositions comprising the salt forms, alone or in combination with additional therapeutic agents. The present invention also provides, in part, methods of preparing such compounds, and methods of using the foregoing. This Summary is provided to introduce a selection of concepts in a simplified form that are further described below in the Detailed Description. This Summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used alone as an aid in determining the scope of the claimed subject matter. Each of the embodiments described below may be combined with any other embodiment described herein so long as it is not inconsistent with the embodiment with which it is combined.

[0007] One embodiment of the present invention provides, in part, a crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinate Form 1. The succinate Form 1 compound may be useful in the treatment of abnormal cell growth. Also provided are pharmaceutical compositions comprising succinate Form 1, alone or in combination with an additional therapeutic agent. The invention also provides, in part, methods of preparing such compounds, and methods of using the foregoing.

[0008] According to one embodiment of the present invention, there is provided a crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1. Succinic acid salt Form 1 has the following properties as measured by powder X-ray diffraction ("PXRD") (2θ), Raman spectrum (cm -1 ), and / or 13 It can be characterized by C solid state nuclear magnetic resonance ("NMR") (ppm).

[0009] According to another embodiment of the present invention, there is provided a pharmaceutical composition comprising a crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1.

[0010] According to another embodiment of the present invention, there is provided a crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 for use as a pharmaceutical.

[0011] In accordance with another embodiment of the present invention, there is provided a method of treating abnormal cell growth comprising administering an effective amount of the crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1.

[0012] According to another embodiment of the present invention, there is provided a crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 for use in a method of treating abnormal cell growth.

[0013] According to another embodiment of the present invention, there is provided the use of the crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 for the manufacture of a medicament.

[0014] Various embodiments of the present invention will be described below, starting with embodiment 1 (E1).

[0015] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed. [Brief description of the drawings]

[0016] [Figure 1] FIG. 1 shows the PXRD pattern of the crystalline anhydrous form of (S)-1′-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4′-piperidine]-1-amine monosuccinic acid salt Form 1. [Diagram 2]FIG. 13C ssNMR spectrum of the crystalline anhydrous form of (S)-1′-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4′-piperidine]-1-amine monosuccinic acid salt Form 1. [Diagram 3] FIG. 1 shows the Raman spectrum of the crystalline anhydrous form of (S)-1′-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4′-piperidine]-1-amine monosuccinic acid salt Form 1. [Figure 4] FIG. 1 shows the PXRD pattern (Pattern 1) of the crystalline form of (S)-1′-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4′-piperidine]-1-amine phosphate. [Diagram 5] FIG. 1 shows the PXRD pattern (Pattern 2) of the crystalline form of (S)-1′-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4′-piperidine]-1-amine phosphate. [Figure 6] FIG. 1 shows the PXRD pattern of the crystalline form of (S)-1′-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4′-piperidine]-1-amine fumarate. [Figure 7] FIG. 1 shows the PXRD pattern of the crystalline form of (S)-1′-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4′-piperidine]-1-amine tartrate. [Figure 8]FIG. 1 shows the PXRD pattern of the crystalline form of (S)-1′-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4′-piperidine]-1-amine hydrochloride. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] The present invention can be more readily understood by referring to the following detailed description of the embodiments of the present invention and the examples contained herein. It should be understood that the present invention is not limited to a specific method of preparation and synthesis, which may of course vary. It should also be understood that the terminology used herein is for the purpose of describing specific embodiments only, and is not intended to be limiting.

[0018] A crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinate Form 1 was found. Figure 1 shows the PXRD pattern of succinate Form 1. Figure 2 shows the PXRD pattern of succinate Form 1. 13 Figure 3 shows the CssNMR spectrum. Figure 4 shows the Raman spectrum of succinate salt Form 1. As described and claimed herein, the peak positions (°2θ), wavenumbers (cm -1), and resonance values ​​(ppm) should be essentially the same. As used herein, the term "essentially the same" means that the inherent variability of a particular method is taken into account. For example, with respect to X-ray diffraction peak positions, the term "essentially the same" means that the inherent variability in peak positions and intensities is taken into account. Those skilled in the art will recognize that peak positions (2θ) typically exhibit variability on the order of ±0.2°. Furthermore, those skilled in the art will recognize that relative peak intensities exhibit instrument-to-instrument variability, as well as variability due to crystallinity, preferred orientation, sample surface preparation, and other factors well known to those skilled in the art, and should be considered merely a qualitative measure. With respect to wavenumber values, those skilled in the art will recognize that wavenumber values ​​exhibit some variability, typically ±2 cm. -1 With respect to resonance values, those skilled in the art will recognize that resonance values ​​will exhibit some degree of variation, typically on the order of ±0.2 ppm.

[0019] E1 Crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt.

[0020] E2 (1) a powder X-ray diffraction pattern containing peaks at 2θ values ​​of 19.7, 24.3, 9.3, and 16.6 ± 0.2°2θ; (2) 1041 and 1217 cm -1 ±2cm -1 Wave number (cm -1 ) value, or (3) a Raman spectrum containing a resonance (ppm) value of 179.0 ppm ± 0.2 ppm. 13 The compound described in E1, characterized by C solid state NMR spectrum.

[0021] E3 (1) a powder X-ray diffraction pattern containing peaks at 2θ values ​​of 19.7, 24.3, 9.3, and 16.6 ± 0.2° 2θ; (2) 1041 and 1217 cm -1 ±2cm -1or (3) a Raman spectrum containing a resonance value of 179.0 ppm ± 0.2 ppm. 13 The compound according to E1 or E2, characterized by two of the C solid state NMR spectra.

[0022] E4 (1) a powder X-ray diffraction pattern containing peaks at 2θ values ​​of 19.7, 24.3, 9.3, and 16.6 ± 0.2° 2θ; (2) 1041 and 1217 cm -1 ±2cm -1 and (3) a Raman spectrum containing a resonance value of 179.0 ppm ± 0.2 ppm. 13 The compound of any one of embodiments E1 to E3, characterized by a C solid state NMR spectrum.

[0023] E5 The compound of any one of embodiments E1 to E4 having an X-ray powder diffraction pattern comprising peaks at 2θ values ​​of 19.7, 24.3, 9.3, and 16.6±0.2 degrees 2θ.

[0024] E6 The compound of embodiment E6 having an X-ray powder diffraction pattern further comprising a peak at a 2θ value of 9.0±0.2 degrees 2θ.

[0025] E7 The compound of any one of embodiments E5 to E6 having an X-ray powder diffraction pattern further comprising a peak at a 2θ value of 28.0±0.2 degrees 2θ.

[0026] E8. The compound of any one of embodiments E5 to E7 having an X-ray powder diffraction pattern further comprising a peak at a 2θ value of 18.8±0.2 degrees 2θ.

[0027] E9. The compound of any one of embodiments E5 to E8 having an X-ray powder diffraction pattern further comprising a peak at a 2θ value of 15.7±0.2 degrees 2θ.

[0028] E10 The compound of any one of embodiments E5 to E9 having an X-ray powder diffraction pattern further comprising a peak at a 2θ value of 12.1±0.2 degrees 2θ.

[0029] E11. The compound of any one of embodiments E5 to E10 having an X-ray powder diffraction pattern further comprising peaks at 2θ values ​​of 4.5, 8.4, 13.9, 16.5, 17.1, 17.6, 17.8, 18.0, 19.9, 21.1, 21.3, 21.7, 21.8, 22.8, 23.0, 23.5, 24.0, 24.4, 25.6, 26.0, 26.2, 26.9, 28.3, 28.7, 29.1, 29.9, 30.5, 31.7, 31.8, 32.8, 33.4, 33.8, 35.0, 35.9, 36.1, 36.4, 36.8, 37.1, 38.0, 38.7, and 39.3±0.2 degrees 2θ.

[0030] E12. A compound according to any one of embodiments E5 to E11 having a PXRD essentially the same as in FIG.

[0031] E13 1041 and 1217 cm -1 ±2cm -1 Wave number (cm -1 The compound of any one of embodiments E1 to E12 having a Raman spectrum comprising:

[0032] E14 1026 and 1555 cm -1 ±2cm -1 Wave number (cm -1 The compound of embodiment E13 having a Raman spectrum further comprising a ) value.

[0033] E15. A compound according to any one of embodiments E13 to E14 having a Raman spectrum essentially the same as in FIG.

[0034] E16 Includes resonance (ppm) value of 179.0 ppm ± 0.2 ppm 13 A compound according to any one of embodiments E1 to E15 having a C solid state NMR spectrum.

[0035] E17 further includes a resonance (ppm) value of 46.4 ppm ± 0.2 ppm 13The compound of any one of embodiments E1 to E16 having a C solid state NMR spectrum.

[0036] E18 further includes a resonance (ppm) value of 38.0 ppm ± 0.2 ppm 13 The compound of any one of embodiments E1 to E17 having a C solid state NMR spectrum.

[0037] E19 further includes a resonance (ppm) value of 141.7 ppm ± 0.2 ppm 13 The compound of any one of embodiments E1 to E18 having a C solid state NMR spectrum.

[0038] E20 also includes a resonance (ppm) value of 27.1 ppm ± 0.2 ppm 13 A compound according to any one of embodiments E16 to E19 having a C solid state NMR spectrum.

[0039] E21 Essentially the same as Figure 3 13 A compound according to any one of embodiments E16 to E20 having a C solid state NMR spectrum.

[0040] E22. A compound according to any one of embodiments E1 to E21 which is substantially pure and free from other forms.

[0041] E23 The compound according to any one of embodiments E1 to E22, wherein the succinate salt Form 1 is greater than 95% substantially pure.

[0042] E24. The compound according to any one of embodiments E1 to E23, wherein the succinate salt Form 1 is greater than 97% substantially pure.

[0043] E25. The compound according to any one of embodiments E1 to E24, wherein the succinate salt Form 1 is substantially greater than 99% pure.

[0044] Also contemplated by the present invention is a pharmaceutical composition comprising succinate Form 1. A typical formulation or composition is prepared by mixing the compound described herein and a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, for example, Ansel, Howard C. et al., Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R. et al., Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C., Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005, the disclosures of which are incorporated herein by reference.

[0045] E26. A pharmaceutical composition comprising a compound according to any one of embodiments E1 to E25 and at least one pharma- ceutically acceptable excipient.

[0046] The compositions of the present invention can be in various forms. These include, for example, liquid, semi-solid, and solid dosage forms, such as liquid solutions (e.g., injectable and infusible solutions), dispersions or suspensions, tablets, capsules, pills, powders, liposomes, and suppositories. The form depends on the intended mode of administration and therapeutic application.

[0047] Typical compositions are in the form of injectable or infusible solutions, such as compositions similar to those generally used for passive immunization of humans with antibodies. One mode of administration is parenteral (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). In another embodiment, the compound is administered by intravenous infusion or injection. In yet another embodiment, the compound is administered by intramuscular or subcutaneous injection.

[0048] Oral administration of solid dosage forms can be provided, for example, as separate units, such as hard or soft capsules, pills, cachets, lozenges, or tablets, each of which contains a predetermined amount of at least one compound of the present invention.In another embodiment, oral administration can be in the form of powder or granules.In another embodiment, oral dosage forms are sublingual, such as lozenges.In such solid dosage forms, the compound of the present invention is usually combined with one or more adjuvants.Such capsules or tablets can include controlled release formulations.In the case of capsules, tablets, and pills, dosage forms can also include buffering agents or be formulated with enteric coatings.

[0049] Other excipients and modes of administration known in the pharmaceutical art can also be used.The pharmaceutical composition of the present invention can be prepared by any of the well-known pharmaceutical techniques, such as effective formulation and administration procedures.The above considerations regarding effective formulation and administration procedures are well-known in the art and are described in standard textbooks.Drug formulation is described, for example, in Ansel (supra) and Gennaro (supra).

[0050] The present invention also contemplates a crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 for use as a pharmaceutical.

[0051] E27 A compound according to any one of embodiments E1 to E26 for use as a medicament.

[0052] The present invention also contemplates a method of treatment comprising administering a crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 according to any of the embodiments described herein. In particular, the method of treatment is to a subject in need thereof. In most embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0053] E28 A method of treating abnormal cell growth comprising the step of administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of embodiments E1 to E26.

[0054] E29 A method of treating abnormal cell growth in a mammal, comprising the step of administering to the mammal a therapeutically effective amount of a compound according to any one of embodiments E1 to E26.

[0055] E30 The method of embodiment E29, wherein the mammal is a human.

[0056] E31 The method of any one of embodiments E28 to E30, wherein the abnormal cell growth is cancer.

[0057] The present invention also contemplates a crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 for use in the treatment of abnormal cell growth.

[0058] E32 A compound according to any one of embodiments E1 to E26 for use in the treatment of abnormal cell growth.

[0059] E33 A compound according to any one of embodiments E1 to E26 for use in the treatment of abnormal cell growth in a mammal.

[0060] E34 A compound according to any one of embodiments E1 to E26 for use in the treatment of abnormal cell growth in a human.

[0061] E35 The compound according to any one of embodiments E32 to E34, wherein the abnormal cell growth is cancer.

[0062] The present invention also contemplates the use of the crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 for the manufacture of a medicament.

[0063] E36 The use of a compound according to any one of embodiments E1 to E26 for the manufacture of a medicament.

[0064] E37 The use of a compound according to any one of embodiments E1 to E26 for the manufacture of a medicament for treating abnormal cell growth.

[0065] E38 The use of a compound according to any one of embodiments E1 to E26 for the manufacture of a medicament for treating abnormal cell growth in a mammal.

[0066] E39 The use of a compound according to any one of embodiments E1 to E26 for the manufacture of a medicament for the treatment of abnormal cell growth in a human.

[0067] E40 The use of any one of embodiments E36 to E39, wherein the abnormal cell growth is cancer.

[0068] In frequent embodiments of the methods provided herein, the abnormal cell growth is cancer. "Cancer," as used herein, refers to a physiological condition in a mammal that is typically characterized by abnormal or unregulated cell growth. Cancer includes solid tumors, named for the type of cells that form them, cancers of the blood, bone marrow, or lymphatic system. Examples of solid tumors include sarcomas and carcinomas. Blood cancers include, but are not limited to, leukemia, lymphoma, and myeloma. Cancer also includes primary cancers that begin at a specific site in the body, metastatic cancers that have spread from where they began to other parts of the body, recurrences from the original primary cancer after remission, and second primary cancers, which are new primary cancers in people with a history of a different type of previous cancer than the later cancer.

[0069] The provided methods result in one or more of the following effects: (1) inhibition of cancer cell proliferation, (2) inhibition of cancer cell invasiveness, (3) induction of apoptosis in cancer cells, (4) inhibition of cancer cell metastasis, or (5) inhibition of angiogenesis.

[0070] The treatment provided by the crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 is defined by reference to any of the following: partial response (PR), complete response (CR), overall response (OR), progression-free survival (PFS), disease-free survival (DFS), and overall survival (OS). PFS, also known as "time to tumor progression," refers to the length of time that the cancer does not grow during and after treatment, and includes the period during which the patient experienced a CR or PR, as well as the period during which the patient experienced stable disease (SD). DFS refers to the length of time that the patient remains disease-free during and after treatment. OS refers to an increase in life expectancy compared to naive or untreated controls or patients. In some embodiments, the response to the combination of the present invention is either PR, CR, PFS, DFS, OR, or OS as assessed using Response Evaluation Criteria in Solid Tumors (RECIST) 1.1 response criteria.

[0071] E41 The method, compound or use of any one of embodiments E31, E35, or E40, wherein the cancer is selected from the group consisting of melanoma, juvenile myelomonocytic leukemia, neuroblastoma, Philadelphia chromosome positive chronic myelogenous leukemia, Philadelphia chromosome positive acute lymphoblastic leukemia, acute myelogenous leukemia, myeloproliferative neoplasms (such as polycythemia vera, essential thrombocythemia, and primary myelofibrosis), breast cancer, lung cancer, liver cancer, colorectal cancer, esophageal cancer, gastric cancer, squamous cell carcinoma of the head and neck, glioblastoma, anaplastic large cell lymphoma, thyroid cancer, and spitzoid neoplasm.

[0072] E42 The method, compound, or use of embodiment E41, wherein the cancer is melanoma.

[0073] E43 The method, compound, or use of embodiment E41, wherein the cancer is juvenile myelomonocytic leukemia.

[0074] E44 The method, compound or use of embodiment E41, wherein the cancer is neuroblastoma.

[0075] E45 The method, compound, or use of embodiment E41, wherein the cancer is Philadelphia chromosome positive chronic myeloid leukemia.

[0076] E46 The method, compound, or use of embodiment E41, wherein the cancer is Philadelphia chromosome positive acute lymphoblastic leukemia.

[0077] E47 The method, compound, or use of embodiment E41, wherein the cancer is acute myeloid leukemia.

[0078] E48 The method, compound, or use of embodiment E41, wherein the cancer is a myeloproliferative neoplasm (such as polycythemia vera, essential thrombocythemia, and primary myelofibrosis).

[0079] E49 The method, compound, or use of embodiment E48, wherein the cancer is polycythemia vera.

[0080] E50 The method, compound, or use of embodiment E48, wherein the cancer is essential thrombocythemia.

[0081] E51 The method, compound, or use of embodiment E48, wherein the cancer is primary myelofibrosis.

[0082] E52 The method, compound, or use of embodiment E41, wherein the cancer is breast cancer.

[0083] E53 The method, compound, or use of embodiment E41, wherein the cancer is lung cancer.

[0084] E54 The method, compound, or use of embodiment E41, wherein the cancer is liver cancer.

[0085] E55 The method, compound, or use of embodiment E41, wherein the cancer is colorectal cancer.

[0086] E56 The method, compound, or use of embodiment E41, wherein the cancer is esophageal cancer.

[0087] E57 The method, compound, or use of embodiment E41, wherein the cancer is gastric cancer.

[0088] E58 The method, compound, or use of embodiment E41, wherein the cancer is head and neck squamous cell carcinoma.

[0089] E59 The method, compound, or use of embodiment E41, wherein the cancer is glioblastoma.

[0090] E60 The method, compound, or use of embodiment E41, wherein the cancer is anaplastic large cell lymphoma.

[0091] E61 The method, compound, or use of embodiment E41, wherein the cancer is thyroid cancer.

[0092] E62 The method, compound, or use of embodiment E41, wherein the cancer is a Spitzoid neoplasm.

[0093] E63 The method, compound, or use of any one of embodiments E31, E35, or E40, wherein the cancer is selected from the group consisting of non-small cell lung cancer ("NSCLC"), colon cancer, esophageal cancer, rectal cancer, juvenile myelomonocytic leukemia ("JMML"), breast cancer, melanoma, and pancreatic cancer.

[0094] E64 The method, compound, or use of embodiment E63, wherein the cancer is NSCLC.

[0095] E65 The method, compound, or use of embodiment E63, wherein the cancer is colon cancer.

[0096] E66 The method, compound, or use of embodiment E63, wherein the cancer is esophageal cancer.

[0097] E67 The method, compound, or use of embodiment E63, wherein the cancer is rectal cancer.

[0098] E68 The method, compound, or use of embodiment E63, wherein the cancer is JMML.

[0099] E69 The method, compound, or use of embodiment E63, wherein the cancer is breast cancer.

[0100] E70 The method, compound, or use of embodiment E63, wherein the cancer is melanoma.

[0101] E71 The method, compound, or use of embodiment E63, wherein the cancer is pancreatic cancer.

[0102] E72 The method, compound, or use of any one of embodiments E31, E35, or E40, wherein the cancer is selected from the group consisting of ALK-positive NSCLC, ROS1-positive NSCLC, BRAF V600E mutated colorectal cancer, RAS mutated solid tumor, NF1 mutated solid tumor, and BRAF class 3 mutated solid tumor.

[0103] E73 The method, compound, or use of embodiment E72, wherein the cancer is ALK-positive NSCLC.

[0104] E74 The method, compound, or use of embodiment E73, wherein the cancer being treated is with prior lorlatinib treatment and prior platinum-based chemotherapy treatment.

[0105] E75 The method, compound, or use of embodiment E73, wherein the cancer being treated has prior lorlatinib treatment and no prior platinum-based chemotherapy treatment.

[0106] E76 The method, compound, or use of embodiment E73, wherein the cancer being treated does not involve prior lorlatinib treatment.

[0107] E77 The method, compound, or use of embodiment E72, wherein the cancer is ROS1-positive NSCLC.

[0108] E78 The method, compound, or use of embodiment E72, wherein the cancer is BRAF V600E mutated colorectal cancer.

[0109] E79 The method, compound, or use of embodiment E78, wherein the cancer being treated is resistant to BRAF inhibitor ("BRAFi") + epidermal growth factor receptor inhibitor ("EGFRi") treatment.

[0110] E80 The method, compound, or use of embodiment E78, wherein the cancer being treated is refractory to BRAFi+EGFRi treatment.

[0111] E81 The method, compound, or use of embodiment E78, wherein the cancer being treated does not involve prior BRAFi+EGFRi treatment.

[0112] E82 The method, compound, or use of embodiment E72, wherein the cancer is a RAS mutated solid tumor.

[0113] E83 The method, compound, or use of embodiment E81, wherein the cancer being treated has previously received standard therapy.

[0114] E84 The method, compound, or use of embodiment E72, wherein the cancer is an NF1 mutated solid tumor.

[0115] E85 The method, compound, or use of embodiment E84, wherein the cancer being treated has previously undergone standard therapy.

[0116] E86 The method, compound, or use of embodiment E72, wherein the cancer is a BRAF class 3 mutated solid tumor.

[0117] The method, compound, or use of embodiment E86, wherein the BRAF class 3 mutation is selected from one or more of the following amino acid substitutions in human BRAF: D287H; P367R; V459L; G466V; G466E; G466A; S467L; G469E; N581S; N581I; D594N; D594G; D594A; D594H; F595L; G596D; G596R; and A762E.

[0118] E88 The method, compound, or use of embodiment E86 or E87, wherein the cancer being treated has previously undergone standard therapy.

[0119] E89 The method, compound or use of any one of embodiments E31, E35, or E40, wherein the cancer is a KRAS mutant cancer.

[0120] E90 Cancer, KRAS G12A Mutation, KRAS G12C Mutation, KRAS G12D Mutation, KRAS G12F Mutation, KRAS G12l Mutation, KRAS G12L Mutation, KRAS G12R Mutation, KRAS G12S Mutation, KRAS G12V Mutations, and KRAS G12Y The method, compound or use of embodiment E89, wherein the mutation is selected from:

[0121] The present invention also contemplates that the crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 be used in combination with an additional therapeutic compound.

[0122] Administering two or more compounds "in combination" means that all of the compounds are administered close enough in time to affect the treatment of the subject. Two or more compounds can be administered simultaneously or sequentially, via the same or different routes of administration, on the same or different administration schedules, with or without specific time restrictions depending on the treatment plan. Furthermore, simultaneous administration can be performed by mixing the compounds before administration, or by administering the compounds at the same time, but in different dosage forms, to the same or different administration sites. Examples of "combination" include, but are not limited to, "concurrent administration", "co-administration", "simultaneous administration", "sequential administration", and "administered simultaneously".

[0123] The crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 and one or more other therapeutic agents can be administered as a fixed or non-fixed combination of active ingredients. The term "fixed combination" means that the crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 and one or more other therapeutic agents are both administered to a subject simultaneously in a single composition or dosage. The term "non-fixed combination" refers to the crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 and one or more therapeutic agents are formulated as separate compositions or dosages such that they can be administered to a subject in need thereof at the same time or at different times separated by variable time limits, such administration providing efficacious levels of the two or more compounds to the subject's body.

[0124] E91 The method, compound, or use of any one of embodiments E27 to E90, wherein the compound of any one of embodiments E1 to E26 is used in combination with an additional therapeutic compound.

[0125] E92 The method, compound, or use of embodiment E91, wherein the additional therapeutic compound is selected from the group consisting of lorlatinib, binimetinib, cetuximab, and encorafenib.

[0126] E93 The method, compound, or use of embodiment E91 or E92, wherein the additional therapeutic compound is lorlatinib.

[0127] E94 The method, compound, or use of embodiment E91 or E92, wherein the additional therapeutic compound is binimetinib.

[0128] E95 The method, compound, or use of embodiment E91 or E92, wherein the additional therapeutic compounds are cetuximab and encorafenib.

[0129] E96 The method, compound, or use of embodiment E91 or E92, wherein the additional therapeutic compound is cetuximab.

[0130] E97 The method, compound, or use of embodiment E91 or E92, wherein the additional therapeutic compound is encorafenib.

[0131] definition As used herein, the singular forms "a," "an," and "the" include plural references unless otherwise indicated. For example, "a" substituent includes one or more substituents.

[0132] The term "about" means having a value that falls within the acceptable standard error of the mean as would be considered by one of ordinary skill in the art.

[0133] The term "crystalline" as used herein means a regularly repeating arrangement of molecules or external face planes. Crystalline forms may differ with respect to thermodynamic stability, physical parameters, X-ray structure, and preparation method.

[0134] The invention described herein may suitably be practiced in the absence of any element not specifically disclosed herein. Thus, for example, in each instance herein, any of the terms "comprising," "consisting essentially of," and "consisting of" can be replaced with either of the other two terms.

[0135] The term "substantially pure" means that a particular crystalline form contains less than 10% by weight of any other physical form of the compound, preferably less than 5%, preferably less than 3%, preferably less than 1%.

[0136] "Pharmaceutical composition" as used herein means a crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 according to any of the embodiments described herein as an active ingredient, and at least one pharma-ceutically acceptable excipient.

[0137] "Pharmaceutically acceptable carrier," as used herein, means a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

[0138] As used herein, the term "excipient" is used to represent any component other than the crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1. The choice of excipient will depend largely on factors such as the mode of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form. Excipients include any physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonicity and absorption delaying agents, carriers, diluents, and the like. Examples of excipients include one or more of water, saline, phosphate buffer solutions, dextrose, glycerol, ethanol, and the like, and combinations thereof, and may include isotonicity agents, such as sugars, sodium chloride, or polyalcohols such as mannitol or sorbitol, in the composition. Examples of excipients also include various organic solvents (such as hydrates and solvates). Pharmaceutical compositions may optionally contain additional excipients such as flavorings, binders / binding agents, lubricants, disintegrants, sweeteners or flavorings, colorants or dyes. For example, for oral administration, tablets containing various excipients such as citric acid can be used with various disintegrants such as starch, alginic acid, and certain complex silicates, and with binders such as sucrose, gelatin, and acacia. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and various starch types, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate, and talc are often useful for tableting. Similar types of solid compositions can also be used in soft and hard-filled gelatin capsules. Thus, non-limiting examples of excipients also include lactose or milk sugar and high molecular weight polyethylene glycols.When aqueous suspensions or elixirs are desired for oral administration, the active compound therein can be combined with additional excipients such as water, ethanol, propylene glycol, glycerin, or combinations thereof, along with various sweetening or flavoring agents, coloring agents or dyes, and optionally, emulsifying or suspending agents.

[0139] Examples of excipients include pharma- ceutically acceptable substances, such as wetting agents, which enhance the shelf life or effectiveness of the compound, or minor amounts of auxiliary substances such as wetting or emulsifying agents, preservatives, or buffers.

[0140] As used herein, an "effective dosage" or "effective amount" of a drug, compound, or pharmaceutical composition is an amount sufficient to have any one or more beneficial or desired effects, including the biochemical, histological, and / or behavioral symptoms of a disease, its complications, and intermediate pathological phenotypes manifested during the development of the disease. In the case of therapeutic use, a "therapeutically effective amount" refers to an amount of the compound administered that relieves to some extent one or more symptoms of the disorder being treated. In the context of treating cancer, a therapeutically effective amount refers to an amount that has the following effects: (1) reducing tumor size; (2) inhibiting (i.e., slowing to some extent, preferably stopping) tumor metastasis; (3) inhibiting (i.e., slowing to some extent, preferably stopping) tumor growth or tumor invasiveness; (4) reducing to some extent (or preferably eliminating) one or more signs or symptoms associated with cancer; (5) reducing the dose of other drugs required to treat the disease; and / or (6) enhancing the effects of other drugs; and / or (7) slowing the progression of the disease in a patient.

[0141] "Mammal" as used herein means a warm-blooded animal having or at risk of developing a disease as described herein, including, but not limited to, guinea pigs, dogs, cats, rats, mice, hamsters, and primates, including humans.

[0142] "Subject" as used herein means a human or animal subject. In certain embodiments, the subject is a mammal. In certain preferred embodiments, the subject is a human.

[0143] "Treat" or "treating" as used herein means administering a crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 to a subject having a condition to be treated to achieve at least one positive therapeutic effect. For example, treating cancer means administering crystalline anhydrous form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 to a subject having or diagnosed with cancer to achieve at least one positive therapeutic effect, such as a reduction in the number of cancer cells, a reduction in tumor size, a slowing down of cancer cell invasion into peripheral organs, or a slowing down of tumor metastasis or tumor growth, reversing, alleviating, inhibiting progression, or preventing the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition. The term "treatment", as used herein, unless otherwise indicated, refers to the act of treating as defined immediately above as "treating". The term "treating" also includes adjuvant and neoadjuvant treatment of a subject. EXAMPLES

[0144] The examples and preparations provided below further describe and illustrate certain aspects and embodiments of the present invention. It should be understood that the scope of the present invention is not limited by the scope of the following examples. The compounds and intermediates described herein are named using the naming conventions provided by ChemDraw Professional, version 19.0.0.22 (Perkin Elmer Informatics, Inc., Waltham, Massachusetts).

[0145] Example 1 Amorphous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base (R)-N-((S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-yl)-2-methylpropane-2-sulfinamide (20 mg, 0.037 mmol) was diluted with dioxane (1 mL) followed by the addition of HCl (92 μL, 0.37 mmol). After stirring for 30 minutes, the reaction was diluted with dichloromethane ("DCM") and saturated aqueous sodium bicarbonate. The mixture was stirred for 10 minutes before the layers were separated and the DCM was dried over MgSO4, filtered and concentrated. This material was purified on silica gel eluting with 20% methanol / ethyl acetate to give amorphous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base (15 mg, 0.034 mmol, 93% yield). 1 H NMR (400 MHz, CDCl3) δ 8.2 (s, 1H), 7.76 (d, 1H, J = 5.2 Hz), 7.2-7.35 (m, 4H), 6.15 (d, 1H, J = 5.2 Hz), 4.92 (br, 2H), 4.78 (br, 1H), 4.01 (s, 1H), 3.37 (m, 2H), 3.14 (d, 1H, J = 15.6 Hz), 2.78 (d, 1H, J = 15.6), 1.3-1.91 (m, 7H); m / z (esi / APCI) M + 1 = 440.1.

[0146] Example 2 Crystalline (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base hemihydrate (Form 1) (S)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine dihydrochloride (3.89 Kg) and 3,6-dibromo-1,2,4-triazine (3.71 Kg) were reacted with triethylamine (6.45 Kg) in 1,4-dioxane (34.3 Kg) at 20-30°C to give (S)-1'-(6-bromo-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine in situ. (S)-1'-(6-bromo-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine was reacted with sodium 2-amino-3-chloropyridine-4-thiolate (3.08 Kg) and triethylamine (6.45 Kg) in 1,4-dioxane (2.05 Kg) at 20-30°C, and the mixture was heated to 70-75°C for about 12.5 hours to give (S)-1'-(6-((2-amino-3-chloropyridine-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine. The mixture was cooled to about 15-30°C. Celite® (1.95 Kg) was added to the reaction at 15-30° C., stirred for about 1 hour, and the slurry was filtered and rinsed with 1,4-dioxane. A solvent exchange was performed replacing 1,4-dioxane with tetrahydrofuran. The crude (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine was purified by chromatography on silica gel (pretreated with isopropyl alcohol) using a mixture of dichloromethane, methanol, ethanol, n-hexane, and triethylamine as eluents. After chromatography, the product (1.57 Kg) was combined with methanol (24.0 Kg) and water (1.6 Kg) at 15-30° C. The mixture was heated to 40°C and the pressure was reduced until approximately 22-25 L of mixture remained. Additional methanol (24.0 Kg) and water (1.6 Kg) were added at 15-30°C. The mixture was heated to 40°C and the pressure was reduced until approximately 22-25 L of mixture remained. Additional water (15.70 Kg) was added at 15-30°C.The mixture was stirred at 15-25° C. for 12 hours to induce crystallization. The product was collected by filtration and the filter cake was rinsed with a mixture of methanol (3.7 Kg) and water (1.6 Kg) to give purified crystalline (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base hemihydrate (Form 1) (1.39 Kg).

[0147] Powder X-ray diffraction analysis was performed using a Bruker AXS D8 Endeavor diffractometer equipped with a copper (Cu) radiation source. The divergence slit was set to 15 mm continuous irradiation. Diffracted radiation was detected with a PSD-Lynx Eye detector with the detector PSD aperture set to 2.99 degrees. X-ray tube voltage and amperage were set to 40 kV and 40 mA, respectively. Data were collected at Cu wavelengths (CuK) using a step size of 0.00998 degrees and a step time of 1.0 seconds on a θ-θ goniometer. α = 1.5418 λ) from 3.0 to 40.0 degrees 2θ. The anti-scatter screen was set at a fixed distance of 3.0 mm. Samples were rotated at 15 / min during collection. Samples were prepared by placing them on a silicon low background sample holder and rotated during collection. Data were collected using Bruker DIFFRAC Plus software and analyzed with EVA diffract plus software. No processing of the PXRD data files was performed prior to peak searching. Preliminary peak assignments were made using the peak search algorithm in the EVA software, with peaks selected at a threshold of 1. To ensure certainty, adjustments were made manually, the output of the automatic assignment was visually inspected, and peak positions were adjusted to the peak maximum. Peaks with a relative intensity of 3% or greater were generally selected. Peaks that were unresolved or consistent with noise were typically not selected. Typical errors associated with peak positions from PXRD are listed in the USP as a maximum of ±0.2 degrees 2θ (USP-941).

[0148] The PXRD peak list and relative intensity data for the compound of Example 2, free base hemihydrate Form 1 (2θ°) is shown in Table 1 below.

[0149] [Table 1]

[0150] The characteristic PXRD peak list and relative intensity data for the compound of Example 2, free base hemihydrate Form 1 (2θ°), is shown in Table 2 below.

[0151] [Table 2]

[0152] Solid-state NMR ("ssNMR") analysis was performed using a Bruker BioSpin Avance III 500 MHz ( 1 H frequency NMR spectrometer. The materials were packed into a 4 mm ZrO2 rotor. A magic angle spinning rate of 14.0 kHz was used. Spectra were collected at ambient temperature (temperature uncontrolled).

[0153] 13 CssNMR spectra were collected using a proton decoupled CPMAS experiment. A phase modulated proton decoupling magnetic field of 80-100 kHz was applied during acquisition of the spectra. The cross-polarization contact time was set to 2 ms and the recycle delay to 11.4 s. The number of scans was adjusted to obtain a suitable signal-to-noise ratio. 13 The C chemical shift scale is 13 A C CPMAS experiment was used, referenced to the external standard crystalline adamantane, setting its upfield resonance to 29.5 ppm.

[0154] Automated peak picking was performed using Bruker-BioSpin TopSpin version 3.6 software. A threshold of 5% relative intensity was generally used for preliminary peak selection. The output of the automated peak picking was visually inspected to ensure robustness and manually adjusted as necessary. Although specific solid-state NMR peak values ​​are reported herein, there is a range of these peak values ​​due to differences in instruments, samples, and sample preparation. This is common practice in the field of solid-state NMR, as variability in peak positions is inevitable. 13 Typical variations in the x-axis values ​​of the C chemical shifts are on the order of plus or minus 0.2 ppm for crystalline solids. Solid-state NMR peak heights reported herein are relative intensities. Solid-state NMR intensities may vary depending on the actual setting of the CPMAS experimental parameters and the thermal history of the sample.

[0155] Crystalline (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base hemihydrate (Form 1) had five characteristic peaks: 44.4, 47.7, 149.2, 36.6, and 26.9 ± 0.2 ppm. 13 C chemical shifts were identified. These characteristic peaks were selected because they are intense, have narrow peak shapes, are typical of solid forms, and are less likely to be hidden in the spectrum of a drug product.

[0156] Free Base Hemihydrate Form 1 of Example 2 13 The CssNMR spectra (ppm) are shown in Table 3 below.

[0157] [Table 3]

[0158] Raman spectra were collected using a Thermo Scientific iS50 FT-Raman accessory mounted on an FT-IR bench. The FT-Raman configuration utilizes a CaF2 beam splitter. The spectrometer is equipped with a 1064 nm diode laser and a room temperature InGaAs detector. Prior to data acquisition, instrument performance and calibration verification was performed using polystyrene. Samples were analyzed in glass NMR tubes either as tablets or in a suitable sample holder that was kept stationary during data collection. Spectra were collected using 0.1-0.5 W laser power and 512 simultaneous additive scans. The collection range was 3700-100 cm. -1 The API spectrum was measured using a 2 cm -1 Resolution was used and Happ-Genzel apodization was used for all spectra. Multiple spectra were recorded and the spectra reported are representative of two spots.

[0159] Prior to peak picking, the intensity scale was normalized to 1. Peaks were manually identified using Thermo Nicolet Omnic 9.7.46 software. Peak locations were picked at the peak maximum, therefore peaks were identified only if there was a slope on either side, and shoulders on the peak were not included. For neat free base hemihydrate form 1, an absolute threshold of 0.006 and a sensitivity of 75 were utilized during peak picking. Peak locations were rounded to whole numbers using standard techniques (0.5 rounded up, 0.4 rounded down). Peaks with normalized peak intensities between (1-0.75), (0.74-0.30), and (0.29-0) were classified as strong, medium, and weak, respectively.

[0160] The characteristic peaks of free base hemihydrate Form 1 were selected based on intensity as well as peak position. To confirm the uniqueness of free base hemihydrate Form 1, a comparison was made with free base hemihydrate Form 1 for placebo and active blends (6 and 15% DL).

[0161] FT-Raman Spectral Peak List and Normalized Intensity Data (cm) for the Compound of Example 2, Free Base Hemihydrate Form 1 -1 ) are shown in Table 4 below.

[0162] [Table 4-1]

[0163] [Table 4-2]

[0164] Characteristic FT-Raman peak list and normalized intensity data (cm) for the compound of Example 2, free base hemihydrate Form 1. -1 ) are shown in Table 5 below.

[0165] [Table 5]

[0166] Example 3

[0167] [ka] Crystalline anhydrous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinate Form 1 Crystalline (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base hemihydrate (Form 1) (Example 2; 8.99 Kg) was suspended in MeOH (500 L) at about 55-65°C. Succinic acid (2.52 Kg) dissolved in MeOH (17.8 Kg) was added to the suspension. The solution was stirred at about 55-65°C for about 2-10 hours, allowing a solid to appear in the solution. The solution was cooled to about 40-50°C. Methyl tert-butyl ether ("MTBE", 33.3 Kg) was added dropwise at about 40-50°C. After addition, the mixture was cooled to about 2-15°C and stirred at about 2-15°C for an additional 2 hours. The slurry was wet milled for about 25-90 minutes at about 2-15° C. The slurry was filtered, washed with MTBE (40 Kg) and dried to obtain crystalline anhydrous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 (9.4 Kg, 81% yield, 99.6% purity).

[0168] Powder X-ray diffraction analysis was performed using a Bruker AXS D8 Endeavor diffractometer equipped with a copper (Cu) radiation source. The divergence slit was set to 15 mm continuous irradiation. Diffracted radiation was detected with a PSD-Lynx Eye detector with the detector PSD aperture set to 2.99 degrees. X-ray tube voltage and amperage were set to 40 kV and 40 mA, respectively. Data were collected at Cu wavelengths (CuK) using a step size of 0.00998 degrees and a step time of 1.0 seconds on a θ-θ goniometer. α= 1.5418 Å) from 3.0 to 40.0 degrees 2θ. The anti-scatter screen was set at a fixed distance of 3.0 mm. Samples were rotated at 15 / min during collection. Samples were prepared by placing them on a silicon low background sample holder and rotated during collection. Data were collected using Bruker DIFFRAC Plus software and analyzed with EVA diffract plus software. No processing of the PXRD data files was performed prior to peak searching. Preliminary peak assignments were made using the peak search algorithm in the EVA software, with peaks selected at a threshold of 1. To ensure certainty, adjustments were made manually, the output of the automatic assignment was visually inspected, and peak positions were adjusted to the peak maximum. Peaks with a relative intensity of 3% or greater were generally selected. Peaks that were unresolved or consistent with noise were typically not selected. Typical errors associated with peak positions from PXRD are listed in the USP as a maximum of ±0.2 degrees 2θ (USP-941).

[0169] The PXRD pattern of Example 3, Form 1, anhydrous succinate salt is shown in Figure 1. The PXRD peak list and relative intensity data for the compound of Example 3, Form 1, anhydrous succinate salt (2θ°) is shown in Table 6 below.

[0170] [Table 6]

[0171] The characteristic PXRD peak list and relative intensity data for the compound of Example 3, Form 1, anhydrous succinate salt (2θ°) is provided in Table 7 below.

[0172] [Table 7]

[0173] Solid-state NMR (ssNMR) analysis was performed on a CPMAS probe placed on a Bruker-BioSpin Avance III 500 MHz (1H frequency) NMR spectrometer. Materials were packed into a 4 mm ZrO2 rotor. A magic angle spinning speed of 14.0 kHz was used. Spectra were collected at ambient temperature (temperature uncontrolled).

[0174] 13 CssNMR spectra were collected using proton decoupled cross-polarization magic angle spinning (CPMAS) experiments. A phase-modulated proton decoupling magnetic field of 80-100 kHz was applied during the acquisition of the spectra. The cross-polarization contact time was set to 2 ms and the recycle delay was set to 11.4 s (hemihydrate crystalline form of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base (form 1)) or 10.5 s (crystalline anhydrous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinate salt form 1). The number of scans was adjusted to give an adequate signal-to-noise ratio. 13 The C chemical shift scale is 13 A C CPMAS experiment was used, referenced to the external standard crystalline adamantane, setting its upfield resonance to 29.5 ppm.

[0175] Automated peak picking was performed using Bruker-BioSpin TopSpin version 3.6 software. A threshold of 5% relative intensity was generally used for preliminary peak selection. The output of the automated peak picking was visually inspected to ensure robustness and manually adjusted as necessary. Although specific solid-state NMR peak values ​​are reported herein, there is a range of these peak values ​​due to differences in instruments, samples, and sample preparation. This is common practice in the field of solid-state NMR, as variability in peak positions is inevitable. 13Typical variations in the x-axis values ​​of the C chemical shifts are on the order of plus or minus 0.2 ppm for crystalline solids. Solid-state NMR peak heights reported herein are relative intensities. Solid-state NMR intensities may vary depending on the actual setting of the CPMAS experimental parameters and the thermal history of the sample.

[0176] Example 3, Form 1 of the anhydrous succinate salt 13 The ssNMR spectrum is shown in Figure 2. Peaks marked with a hash mark are spinning sidebands. The ssNMR peak list and relative intensity data for the compound of Example 3, Form 1, anhydrous succinate salt, is shown in Table 8 below.

[0177] [Table 8]

[0178] Characteristic ssNMR peak list data for the compound of Example 3, Form 1, anhydrous succinate salt, is shown in Table 9 below.

[0179] [Table 9]

[0180] Raman spectra were collected using a Thermo Scientific iS50 FT-Raman accessory mounted on an FT-IR bench. The FT-Raman configuration utilizes a CaF2 beam splitter. The spectrometer is equipped with a 1064 nm diode laser and a room temperature InGaAs detector. Prior to data acquisition, instrument performance and calibration verification was performed using polystyrene. Samples were analyzed in glass NMR tubes either as tablets or in a suitable sample holder that was kept stationary during data collection. Spectra were collected using 0.1-0.5 W laser power and 512 simultaneous additive scans. The collection range was 3700-100 cm. -1 The API spectrum was measured using a 2 cm -1Resolution was used and Happ-Genzel apodization was used for all spectra. Multiple spectra were recorded and the spectra reported are representative of two spots.

[0181] Prior to peak picking, the intensity scale was normalized to 1. Peaks were manually identified using Thermo Nicolet Omnic 9.7.46 software. Peak positions were picked at the peak maximum, therefore peaks were identified only if there was a slope on either side, and shoulders on the peak were not included. For neat crystalline anhydrous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1, an absolute threshold of 0.06 and a sensitivity of 75 were utilized during peak picking. Peak positions were rounded to the nearest whole number using standard techniques (0.5 rounded up, 0.4 rounded down). Peaks with normalized peak intensities between (1–0.75), (0.74–0.30), and (0.29–0) were classified as strong, medium, and weak, respectively. Relative peak intensity values ​​are also presented in this report.

[0182] Characteristic peaks of crystalline anhydrous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt form 1 were selected based on intensity as well as peak position. To confirm the uniqueness of crystalline anhydrous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinate Form 1, a comparison was made with crystalline anhydrous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinate Form 1 in the case of placebo and active blends (6 and 15% DL).

[0183] A full Raman spectrum of the anhydrous succinate salt of Example 3, Form 1, is shown in Figure 3. A full Raman peak listing of the anhydrous succinate salt of Example 3, Form 1, is shown in Table 10 below.

[0184] [Table 10]

[0185] Characteristic Raman peak list data for the compound of Example 3, Form 1, anhydrous succinate salt is provided below in Table 11.

[0186] [Table 11]

[0187] Example 4 Crystalline (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine phosphate (Pattern 1) (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base (200 mg) was weighed into a 20 mL glass scintillation vial. 90% ethanol / 10% water (5 mL; % v / v) was added to the scintillation vial to form a slurry. Phosphoric acid (65.2 μL; 2.1 equiv.) was added neat to the slurry. The slurry was then temperature cycled between ambient and 40° C. (4 hour cycles) with rocking for 18 hours. A subsample was isolated by centrifugal filtration. XPRD analysis showed a phosphate pattern of 1. The remaining slurry was isolated using a Buchner funnel containing Whatmann grade 1 filter paper. The isolated solid was then dried under vacuum at 40° C. for about 24 hours.

[0188] Example 5 Crystalline (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine phosphate (Pattern 2) (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base (200 mg) was weighed into a 20 mL glass scintillation vial. 90% ethanol / 10% water (5 mL; % v / v) was added to the scintillation vial to form a slurry. Phosphoric acid (32.6 μL; 1.05 equiv.) was added neat to the slurry. The slurry was then temperature cycled between ambient and 40° C. (4 hour cycles) with rocking for 18 hours. The solvent was removed using a rotary evaporator. 90% methyl ethyl ketone / 10% water (% v / v) was added to the dry solid to form a slurry. The material was isolated using a Buchner funnel containing Whatmann grade 1 filter paper. The isolated solid was dried under vacuum at 40° C. for approximately 24 hours. This procedure produces a mixture of Pattern 1 and Pattern 2 phosphates.

[0189] Example 6 Crystalline (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine fumarate (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base (200 mg) was weighed into a 20 mL glass scintillation vial. Acetone (5 mL) was added to the scintillation vial to form a slurry. Fumaric acid (55.96 mg; 1.05 equiv.) was added neat to the slurry. The slurry was then temperature cycled between ambient and 40°C (4 hour cycles) with rocking for 18 hours. Centrifugal filtration was used for isolation.

[0190] Example 7 Crystalline (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine tartrate (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base (200 mg) was weighed into a 20 mL glass scintillation vial. 90% ethanol / 10% water (5 mL; % v / v) was added to the scintillation vial to form a slurry. L-Tartatic acid (72.36 mg; 1.05 equiv.) was added neat to the slurry. The slurry was then temperature cycled between ambient and 40°C (4 hour cycles) with rocking for 18 hours. Centrifugal filtration was used for isolation.

[0191] Example 8 Crystalline (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine hydrochloride (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base (40 mg) was weighed into an HPLC vial. 90% 2-propanol / 10% water (1 mL; % v / v) was added to the vial to form a slurry. Hydrochloric acid (16.0 μL; 2.1 equiv.) was added neat to the slurry. The slurry was then temperature cycled between ambient and 40° C. (4 hour cycles) with rocking for 96 hours. Centrifugal filtration was used for isolation.

[0192] Example 9 Comparative analysis between salt forms of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine solubility Solubility (Tables 12 and 13) was determined by adding 3 mg of compound (Examples 2 or 3) to 500 μL of each appropriate buffer (HCl at pH 2, acetate at pH 4, and phosphate at pH 6). After addition of the buffer, the samples were vortexed. The samples were rotated in an Envirogenie thermomixer at 40° C. for 8 hours, 15° C. for 5 hours, and 25° C. for 12 hours. After the thermomixing cycles, the slurry was isolated by centrifugal filtration and the isolated mother liquor was analyzed by HPLC.

[0193] HPLC method (Tables 12 and 13): An Agilent 1290 UPLC instrument was used with a Waters XSelect HSS T3 column (100 Å, 4.6×150 mm, 5 μm). The column temperature was set at 20° C., UV wavelength at 254 nm, injection volume at 10 μL, and flow rate at 1.0 mL / min. 10 mM ammonium acetate adjusted to pH 4.8 was used as mobile phase A and acetonitrile was used as mobile phase B. The gradient method was set to achieve 15% mobile phase B at 0 min, 95% mobile phase B at 17 min - hold to 22 min, and 15% mobile phase B at 22.1 min - hold to 28 min.

[0194] Solubility (Table 14) was determined by adding 20 mg of salt (Examples 3-8) to three HPLC vials, each containing a magnetic stir bar along with 1 mL of the appropriate buffer system (HCl at pH 1.2, sodium acetate at pH 4.5, or phosphate at pH 6.8). Samples were rocked at 37°C for 18 hours. After 24 hours, the slurry was isolated by centrifugal filtration and the isolated mother liquor was analyzed by HPLC.

[0195] HPLC method (Table 14): A Dionex Ultimate 3000 HPLC instrument was used with a Waters XSelect HSS T3 column (100 Å, 4.6×150 mm, 5 μm). The column temperature was set at 20° C., UV wavelength at 254 nm, injection volume at 10 μL, and flow rate at 1.0 mL / min. 10 mM ammonium acetate adjusted to pH 4.8 was used as mobile phase A and acetonitrile was used as mobile phase B. The gradient method was set to achieve 15% mobile phase B at 0 min, 95% mobile phase B at 17 min - hold to 22 min, and 15% mobile phase B - hold to 28 min at 22.1 min.

[0196] Crystalline Polarized light microscopy ("PLM") (Table 12) was performed using a Leitz Orthoplan optical microscope. Samples were dispersed in immersion oil (Type A) on a flat glass substrate with a glass cover plate. Samples were viewed under cross polarized light with a 530 nm waveplate.

[0197] Powder X-ray diffraction ("PXRD") (Table 12) analysis was performed using a Bruker AXS D8 Endeavor diffractometer equipped with a Cu radiation source. The divergence slit was set to 10 mm continuous irradiation. Diffracted radiation was detected with a PSD-Lynx Eye detector with the detector PSD aperture set to 2.99 degrees. The X-ray tube voltage and amperage were set to 40 kV and 40 mA, respectively. Data was collected at Cu wavelength from 3.0 to 40.0 degrees 2θ using a step size of 0.02 degrees and a step time of 0.3 seconds in a theta-theta goniometer. The anti-scatter screen was set at a fixed distance of 1.5 mm. Samples were rotated at 15 / min during collection. Samples were prepared by placing them in a silicon low background sample holder and rotated during collection. Data was collected using Bruker DIFFRAC Plus software and analyzed with EVA diffract plus software. A sample holder with a small indentation was used.

[0198] PLM (Table 13) was performed by adding less than 1 mg of API material to a microscope slide, using a drop of high viscosity immersion oil, and covering with a cover slip. The slide was then viewed under cross-polarized light with a 530 nm waveplate in an Olympus BX53 polarizing microscope. Sample images were collected with a built-in Olympus camera, and photos were taken using Olympus Stream Software.

[0199] PXRD (Table 13): Instrumentation parameters: A Rigaku MiniFlex 6G diffractometer equipped with a Cu radiation source was used. Diffracted radiation was detected by a D / teX Ultra2 detector. X-ray tube voltage and amperage were set at 40 kV and 15 mA, respectively. Data were collected on a Miniflex goniometer from 3.0 to 45.0° 2θ at Cu wavelength using a step size of 0.01° and a step rate of 3.00° / min. The entrance slit box was set at 1.25° and the length limiting slit was set at 10 mm. The sample was rotated at 10 RPM during collection. Data was analyzed using Rigaku software SmartLab Studio II. Sample preparation: Samples were prepared by placing them in a silicon low background sample holder (2 mm×0.5 mm well). The sample was ensured to be level and filled prior to measurement. Samples were analyzed using the above instrument parameters.

[0200] PLM (Table 14): The presence of crystallinity was measured using an Olmypus BX50 microscope equipped with cross-polarized lenses and a Motic camera. Images were captured using Motic Images Plus 2.0. All images were recorded using a 20x objective.

[0201] PXRD (Table 14) analysis was performed in a PANalytical X'pert pro equipped with a PIXcel detector (128 channels) by scanning samples between 3 and 35°2θ. The material was gently ground to break up aggregates and mounted in a multiwell plate with a Kapton or Mylar polymer film to support the sample. The multiwell plate was then placed in the diffractometer and analyzed using Cu K radiation (α1 λ = 1.54060 Å; α2 = 1.54443 Å; β = 1.39225 Å; α1:α2 ratio = 0.5) operated in transmission mode using a 40 kV / 50 mA generator setting. Data were visualized and images were generated using the HighScore Plus 4.7 desktop application (PANalytical, 2017).

[0202] stability Sample Preparation (Table 12): Control of crystalline (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base hemihydrate (Form 1): Compound (15.46 mg) was weighed into a 50 mL amber volumetric flask. Methanol (approximately 35 mL) was added to the flask, sonicated for 4 minutes with intermittent swirling, equilibrated to room temperature, made up to volume with methanol, and mixed well by repeated inversion of the flask. Crystalline (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base hemihydrate (Form 1) at 70°C / 5% relative humidity ("RH") / 1 week: Compound (14.83 mg) was weighed into a 50 mL amber volumetric flask. Methanol (approximately 35 mL) was added to the flask, sonicated for 4 minutes with intermittent swirling, equilibrated to room temperature, made up to volume with methanol, and mixed well by repeated inversion of the flask. Crystalline (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine free base hemihydrate (Form 1) at 70°C / 75% RH / 1 week: Compound (14.99 mg) was weighed into a 50 mL amber volumetric flask. Methanol (approximately 35 mL) was added to the flask, sonicated for 4 minutes with intermittent swirling, equilibrated to room temperature, made up to volume with methanol, and mixed well by repeated inversion of the flask.

[0203] Chemical Stability (HPLC) (Table 12): A Waters Acquity H-Class instrument was used with a Waters XSelect HSS T3 column (100 Å, 4.6×150 mm, 5 μm). The column temperature was set at 20° C., UV wavelength at 254 nm, injection volume at 10 μL, and flow rate at 1.0 mL / min. 10 mM ammonium acetate adjusted to pH 4.8 was used as mobile phase A and acetonitrile was used as mobile phase B. The gradient method was set to achieve 15% mobile phase B at 0 min, 95% mobile phase B at 17 min-hold until 22 min, 15% mobile phase was achieved.

[0204] Physical stability (PXRD) (Table 12): Powder X-ray diffraction analysis was performed using a Bruker AXS D8 Endeavor diffractometer equipped with a Cu radiation source. The divergence slit was set to 10 mm continuous irradiation. The detector PSD aperture was set to 2.99 degrees and the diffracted radiation was detected with a PSD-Lynx Eye detector. The X-ray tube voltage and amperage were set to 40 kV and 40 mA, respectively. Data was collected at Cu wavelength from 3.0 to 40.0 degrees 2θ using a step size of 0.02 degrees and a step time of 0.3 seconds in a theta-theta goniometer. The anti-scatter screen was set at a fixed distance of 1.5 mm. During collection, the sample was rotated at 15 / min. The sample was prepared by placing it in a silicon low background sample holder and rotated during collection. Data was collected using Bruker DIFFRAC Plus software and analyzed with EVA diffract plus software. A sample holder with a small indentation was used.

[0205] Light Stability (Table 12): Sample Preparation: Control- Approximately 1 g of sample was transferred to a weighing bottle and covered with a glass cover. The sample was wrapped in aluminum foil to protect it from light and placed in a light stability chamber. Light Stable Samples: Approximately 1 g of sample was transferred to a weighing bottle and then placed in a light stability chamber. Storage Conditions: Light stable samples were stored in compliance with the following ICH Light Stability Option 2: Cool white fluorescent lamps designed to produce an output similar to that specified in ISO 10977 (1993), near ultraviolet fluorescent lamps having a spectral distribution of 320 nm to 400 nm with maximum energy emission between 350 nm and 370 nm, and a significant portion of the UV should be in both the 320-360 nm and 360-400 nm bands. Light stable samples and dark control samples were stored in 5000 ± 500 Lx (near ultraviolet energy 0.84 W / m 2 ±10%) for 12 days (samples were exposed to light providing an overall illumination of ≥1.2 million lux hours and an integrated near-ultraviolet energy of ≥200 watt-hours per square meter).

[0206] Photostability (HPLC) (Table 12): An Agilent 1100 HPLC was used with a Waters XSelect HSS T3 column (100 Å, 4.6×150 mm, 5 μm). The column temperature was set at 20° C., UV wavelength at 254 nm, injection volume at 10 μL, and flow rate at 1.0 mL / min. 10 mM ammonium acetate adjusted to pH 4.8 was used as mobile phase A and acetonitrile was used as mobile phase B. The gradient method was set to 15% mobile phase B at 0 min, 95% mobile phase B at 17 min-hold until 22 min, achieving 15% mobile phase.

[0207] Sample preparation (Table 13): 70° C. / 75% RH Sample: Approximately 20 mg of crystalline anhydrous (S)-1′-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4′-piperidine]-1-amine monosuccinic acid salt Form 1 was placed in an open amber vial in a 70° C. / 75% RH chamber. 70° C. / 5% RH Sample: Approximately 20 mg of crystalline anhydrous (S)-1′-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4′-piperidine]-1-amine monosuccinic acid salt Form 1 was placed in an amber vial with the lid taped on in a non-humidity controlled 70° C. chamber. Control Sample: Approximately 20 mg of crystalline anhydrous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 was placed in a capped amber vial in the refrigerator. Each succinate Form 1 sample was prepared at approximately 0.3 mg / mL by transferring 1 mg of material to an amber vial, adding 3 mL of MeOH and vortexing to dissolve.

[0208] Chemical Stability (HPLC) (Table 13): A Waters Acquity UPLC I-Class instrument was used with a Waters XSelect HSS T3 column (100 Å, 4.6×150 mm, 5 μm). The column temperature was set at 20° C., UV wavelength at 254 nm, injection volume at 10 μL, and flow rate at 1.0 mL / min. Ammonium acetate (10 mM) adjusted to pH 4.8 was used as mobile phase A, and acetonitrile was used as mobile phase B. The gradient method was set to achieve 15% mobile phase B at 0 min, 95% mobile phase B at 17 min; hold until 22 min, 15% mobile phase was reached.

[0209] Physical stability (PXRD) (Table 13): A Rigaku MiniFlex 6G diffractometer equipped with a Cu radiation source was used. Diffracted radiation was detected by a D / teX Ultra2 detector. X-ray tube voltage and amperage were set at 40 kV and 15 mA, respectively. Data were collected on a Miniflex goniometer from 3.0 to 45.0° 2θ at Cu wavelength using a step width of 0.02° and a step rate of 2.00° / min. The entrance slit box was set at 1.25° and the longitudinal limiting slit was set at 10 mm. The sample was rotated at 10 RPM during collection. Data were analyzed using Rigaku software SmartLab Studio II.

[0210] The samples were prepared by placing them in a silicon low background sample holder (0.2 indentation plate). The samples were ensured to be level and packed before measurement. The samples were not ground (by mortar and pestle, or any other means) before being placed in the sample holder.

[0211] Photostability (Table 13): Crystalline anhydrous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 - Photolysis Control (foil wrapped): 1.716 mg was weighed into a 5 mL volumetric flask and approximately 3 mL of methanol was added to the flask, sonicated for 3 minutes with intermittent swirling, equilibrated to room temperature, made up to volume with methanol and mixed well by repeated inversion of the flask.

[0212] 2X ICH Photolysis: 1.569 mg was weighed into a 5 mL volumetric flask, approximately 3 mL of methanol was added to the flask, sonicated for 3 minutes with intermittent swirling, equilibrated to room temperature, made up to volume with methanol, and mixed well by repeated inversion of the flask.

[0213] Chemical Stability (HPLC) (Table 13): A Waters Acquity H-Class instrument was used with a Waters XSelect HSS T3 column (100 Å, 4.6×150 mm, 5 μm). The column temperature was set at 20° C., UV wavelength at 254 nm, injection volume at 10 μL, and flow rate at 1.0 mL / min. Ammonium acetate (10 mM) adjusted to pH 4.8 was used as mobile phase A, and acetonitrile was used as mobile phase B. The gradient method was set to achieve 15% mobile phase B at 0 min, 95% mobile phase B at 17 min; hold to 22 min, 15% mobile phase.

[0214] Physical (PXRD) (Table 13): Powder X-ray diffraction analysis was performed using a Bruker AXS D8 Endeavor diffractometer equipped with a Cu radiation source. The divergence slit was set to 10 mm continuous irradiation. The detector PSD aperture was set to 2.99 degrees and the diffracted radiation was detected by a PSD-Lynx Eye detector. The X-ray tube voltage and amperage were set to 40 kV and 40 mA, respectively. Data was collected at Cu wavelength from 3.0 to 40.0 degrees 2θ using a step size of 0.02 degrees and a step time of 0.3 seconds on a theta-theta goniometer. The anti-scatter screen was set at a fixed distance of 1.5 mm. During collection, the sample was rotated at 15 / min. The sample was prepared by placing it on a silicon low background sample holder and rotated during collection. Data was collected using Bruker DIFFRAC Plus software and analyzed with EVA diffract plus software.

[0215] Thermogravimetric Analysis ("TGA") (Table 12): Thermogravimetric analysis was performed using a Discovery TGA (TA instruments) thermogravimetric analyzer. Approximately 10 mg of sample was weighed into an aluminum pan and heated from ambient temperature to 300°C at a heating rate of 10°C / min under a nitrogen purge (10 mL / min in both the sample chamber and the balance).

[0216] Differential Scanning Calorimetry ("DSC") (Table 12): DSC measurements were performed using a Discovery DSC (TA instruments) equipped with a refrigerated cooling attachment. All experiments were performed in standard / Tzero aluminum pans. Cell constants were determined using indium and temperature calibration was performed using indium and tin as standards. All measurements were performed under a continuous dry nitrogen purge (50 mL / min). Approximately 1-5 mg of solid sample was weighed into a Tzero aluminum pan, non-hermetically sealed, and heated from 0°C to 250°C at a heating rate of 10°C / min. Experimental data was analyzed using commercially available software (TA Universal Analysis 2000 / Trios software, TA Instruments).

[0217] Dynamic Vapor Sorption ("DVS") (Table 12): The microbalance was calibrated using a 100 mg standard weight. The relative humidity sensor was calibrated using saturated saline at 5.0, 11.3, 32.8, 52.8, 75.3, and 84.3% RH (25°C) and polyvinylpyrrolidone at 80% RH (25°C). 8-10 mg of powdered sample was placed in a platinum sample pan. There is no drying step in this method. The RH was first maintained at 30% RH, then reduced to 10% RH, then reduced to 3% RH, using the following criteria: when the sample weight change was less than 0.001 wt% for 5 minutes or after the maximum equilibration time of 60 minutes was reached, the RH was then gradually increased to 90% in 10% increments, followed by a final reduction in the RH to 10% RH in 10% increments. Again, equilibrium was assumed to be reached when the sample weight change was less than 0.001 wt % for 5 min or the maximum equilibration time was 120 min.

[0218] DVS (Table 13): Water sorption and desorption were analyzed with DVS-Resolution. The microbalance is calibrated monthly with a 100 mg standard weight. Approximately 5 mg of API was added to a translucent sample pan and placed in chamber A of the DVS-Resolution. The relative humidity was held at 0% for 1 hour, increased to 90% in 10% increments, and then decreased back to 0% in 10% increments. A step was considered complete when the change in dm / dt was observed to be less than 0.001% for a minimum of 30 minutes or when the maximum step time reached 120 minutes.

[0219] TGA / DSC (Table 13): A simultaneous DSC / TGA instrument was used for thermal analysis. Approximately 3.2 mg of API was placed in an aluminum Tzero DSC pan and loaded into the instrument with an autosampler. Samples were equilibrated to 35° C. in the instrument prior to analysis and heated to 400° C. at a ramp rate of 10° C. / min. Dry nitrogen gas from an in-house supply was used.

[0220] TGA / DSC (Table 14): Approximately 5-10 mg of material was added to a pre-tared aluminum pan without a lid and loaded into a TA Instruments Discovery SDT 650 Auto-Simultaneous DSC and held at room temperature. The sample was then heated at a rate of 10 °C / min from 20 °C (ambient temperature) to 400 °C, during which the change in sample weight was recorded along with the heat flow response (DSC). Nitrogen was used as the sample purge gas at a flow rate of 200 cm3 / min.

[0221] DSC (Table 14): Approximately 1-5 mg of material was weighed into an aluminum DSC pan and non-hermetically sealed with an aluminum lid. The sample pan was then loaded into a TA Instruments Discovery DSC 2500 Differential Scanning Calorimeter equipped with an RC90 cooler. The samples and standards were heated to 310°C at a scan rate of 10°C / min and the resulting heat flow response was monitored. The samples were re-cooled to 20°C and then re-heated to 205°C, all at 10°C / min. 50 cm of nitrogen was used as the purge gas. 3 A flow rate of 1 / min was used.

[0222] DVS (Table 14): Approximately 10-20 mg of sample was placed on a mesh vapor sorption balance pan and loaded into a DVS Intrinsic dynamic vapor sorption balance from Surface Measurement Systems. At 25°C, the sample was subjected to a profile increasing from 40 to 90% relative humidity (RH) in 10% increments, with the sample held at each step until a stable weight was achieved (dm / dt of 0.004%, minimum step length of 30 minutes, maximum step length of 500 minutes). After completion of the sorption cycle, the sample was dried to 0% RH using the same procedure and then returned to 40% RH for a second sorption cycle. Two cycles were performed. The weight change during the sorption / desorption cycle was plotted to allow the hygroscopicity of the sample to be determined. XRPD analysis was then performed on the retained solid. Approximately 10-20 mg of sample was placed on a mesh vapor sorption balance pan and loaded into a DVS Advantage dynamic vapor sorption balance from Surface Measurement Systems. At 25°C, the samples were subjected to a relative humidity (RH) profile increasing from 40 to 90% in 10% increments, with the sample held at each step until a stable weight was achieved (dm / dt of 0.004%, minimum step length of 30 min, maximum step length of 500 min). After completion of the sorption cycle, the sample was dried to RH 0% using the same procedure and then returned to RH 40% for a second sorption cycle. Two cycles were performed. The weight change during the sorption / desorption cycle was plotted to allow the hygroscopicity of the sample to be determined. XRPD analysis was then performed on the retained solids.

[0223] NMR (Table 14): NMR experiments were performed on a Bruker AVIIIHD spectrometer equipped with a DCH cryoprobe operating at 500.12 MHz for protons. Experiments were performed in deuterated dimethylsulfoxide ((CD3)2SO) and methanol (CD3OD), and each sample was prepared to a concentration of approximately 10 mM.

[0224] Table 12 shows the properties of the crystalline free base hemihydrate Form 1 of Example 2. Table 13 shows the properties of the crystalline monosuccinate salt Form 1 of Example 3.

[0225] [Table 12]

[0226] [Table 13]

[0227] Table 14 shows comparative data for salts of (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine.

[0228] [Table 14]

[0229] Example 10 Pharmaceutical formulation of crystalline anhydrous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt form 1 (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 is formulated to make 10 mg, 25 mg (the 10 mg and 25 mg tablets use a common blend of 50 mg / g), and 100 mg immediate release tablets. The formulations shown in Tables 15 and 16 are composed of microcrystalline cellulose and anhydrous dibasic calcium phosphate (diluents), crospovidone (disintegrant), and sodium stearyl fumarate (lubricant). (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 tablet cores are manufactured using a dry granulation manufacturing process.

[0230] The tablet compositions for 10 mg and 25 mg tablets prepared using a typical blend of 50 mg / g of active are shown in Table 15 and for 100 mg tablets in Table 16.

[0231] [Table 15]

[0232] [Table 16]

[0233] (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinic acid salt Form 1 immediate release 10, 25, and 100 mg tablet cores are manufactured using a platform designed and developed for batch dry granulation of solid oral immediate release tablets.

[0234] First, the diluent (one of the two splits of microcrystalline cellulose) and disintegrant are blended, followed by the addition and blending of the active ingredient. The remaining microcrystalline cellulose (the other of the two splits) and the active blend (previous step) are co-milled to deagglomerate any agglomerates. This is followed by blending to improve homogeneity after milling. A homogeneous blend is prepared by lubrication with sodium stearyl fumarate. This blend is dry granulated (using roller compaction or slugging) and the resulting slugs are milled (using an in-line mill or co-mill). The milled granules are blended to obtain homogeneously distributed granule particles. The granules are lubricated to obtain homogeneously lubricated granules. Finally, the homogeneous blend is conveyed to a tablet press for compression of tablet cores.

[0235] All publications and patent applications cited herein are hereby incorporated by reference in their entirety.

[0236] All publications and patent applications cited herein are incorporated by reference in their entirety. It will be apparent to one of ordinary skill in the art that certain changes and modifications can be made without departing from the spirit or scope of the appended claims.

Claims

1. Crystals of anhydrous (S)-1'-(6-((2-amino-3-chloropyridin-4-yl)thio)-1,2,4-triazin-3-yl)-1,3-dihydrospiro[indene-2,4'-piperidine]-1-amine monosuccinate.

2. (1) a powder X-ray diffraction pattern including peaks at 2θ values ​​of 19.7, 24.3, 9.3, and 16.6±0.2°2θ; (2) peaks at 1041 and 1217 cm －１ ±2cm －１ or (3) a Raman spectrum containing a resonance value of 179.0 ppm ± 0.2 ppm. １３ 2. The crystal of claim 1, characterized by one of the following C solid-state NMR spectra:

3. (1) a powder X-ray diffraction pattern including peaks at 2θ values ​​of 19.7, 24.3, 9.3, and 16.6±0.2°2θ; (2) peaks at 1041 and 1217 cm －１ ±2cm －１ or (3) a Raman spectrum containing a resonance value of 179.0 ppm ± 0.2 ppm. １３ 2. The crystal of claim 1, characterized by two of the following C solid-state NMR spectra:

4. (1) a powder X-ray diffraction pattern containing peaks at 2θ values ​​of 19.7, 24.3, 9.3, and 16.6±0.2°2θ; (2) peaks at 1041 and 1217 cm －１ ±2cm －１ and (3) a Raman spectrum containing a resonance value of 179.0 ppm ± 0.2 ppm. １３ The crystal of claim 1 characterized by a C solid-state NMR spectrum.

5. 2. The crystal of claim 1, having a powder X-ray diffraction pattern including peaks at 2θ values ​​of 19.7, 24.3, 9.3, and 16.6±0.2°2θ.

6. 2. The crystal of claim 1, having a powder X-ray diffraction pattern further including a peak at a 2θ value of 9.0±0.2°2θ.

7. 2. The crystal of claim 1, having a powder X-ray diffraction pattern further including a peak at a 2θ value of 28.0±0.2°2θ.

8. 2. The crystal of claim 1, having a powder X-ray diffraction pattern further including a peak at a 2θ value of 18.8±0.2°2θ.

9. 2. The crystal of claim 1, having a powder X-ray diffraction pattern further including a peak at a 2θ value of 15.7±0.2°2θ.

10. 2. The crystal of claim 1, having a powder X-ray diffraction pattern further including a peak at a 2θ value of 12.1±0.2°2θ.

11. 1041 and 1217 cm －１ ±2cm －１ Wave number (cm －１ 2. The crystal of claim 1, having a Raman spectrum comprising a .alpha.-.alpha.) value.

12. 1026 and 1555 cm －１ ±2cm －１ Wave number (cm －１ 12. The crystal of claim 11, having a Raman spectrum comprising a .alpha.-.alpha.) value.

13. Contains a resonance (ppm) value of 179.0 ppm ± 0.2 ppm １３ The crystal of claim 1, having a C solid-state NMR spectrum.

14. Contains a resonance (ppm) value of 46.4 ppm ± 0.2 ppm １３ The crystal of claim 1, having a C solid-state NMR spectrum.

15. Contains a resonance (ppm) value of 38.0 ppm ± 0.2 ppm １３ The crystal of claim 1, having a C solid-state NMR spectrum.

16. Contains a resonance (ppm) value of 141.7 ppm ± 0.2 ppm １３ The crystal of claim 1, having a C solid-state NMR spectrum.

17. Contains a resonance (ppm) value of 27.1 ppm ± 0.2 ppm １３ The crystal of claim 13, having a C solid-state NMR spectrum.

18. 10. The crystal of claim 1, which is substantially pure and free of alternative forms.

19. A pharmaceutical composition comprising a crystal according to any one of claims 1 to 18 and at least one pharmaceutically acceptable excipient.

20. The pharmaceutical composition of claim 19 for use in treating abnormal cell proliferation in a mammal.

21. 21. The pharmaceutical composition of claim 20, wherein the mammal is a human.

22. 21. The pharmaceutical composition of claim 20, wherein the abnormal cell growth is cancer.

23. 23. The pharmaceutical composition of claim 22, wherein the cancer is selected from the group consisting of melanoma, juvenile myelomonocytic leukemia, neuroblastoma, Philadelphia chromosome positive chronic myeloid leukemia, Philadelphia chromosome positive acute lymphoblastic leukemia, acute myeloid leukemia, myeloproliferative neoplasms (such as polycythemia vera, essential thrombocythemia, and primary myelofibrosis), breast cancer, lung cancer, liver cancer, colorectal cancer, esophageal cancer, gastric cancer, head and neck squamous cell carcinoma, glioblastoma, anaplastic large cell lymphoma, thyroid cancer, and spitzoid neoplasm.

24. 23. The pharmaceutical composition of claim 22, wherein the cancer is selected from the group consisting of non-small cell lung cancer, colon cancer, esophageal cancer, rectal cancer, juvenile myelomonocytic leukemia, breast cancer, melanoma, and pancreatic cancer.

25. 23. The pharmaceutical composition of claim 22, wherein the cancer is selected from the group consisting of ALK-positive NSCLC, ROS1-positive NSCLC, BRAF V600E-mutated colorectal cancer, RAS-mutated solid tumors, NF1-mutated solid tumors, and BRAF class 3-mutated solid tumors.

26. 26. The pharmaceutical composition of claim 25, wherein the cancer is ALK-positive NSCLC.

27. 27. The pharmaceutical composition of claim 26, wherein the cancer being treated is with prior lorlatinib treatment and prior platinum-based chemotherapy.

28. 27. The pharmaceutical composition of claim 26, wherein the cancer being treated has had prior treatment with lorlatinib and no prior treatment with platinum-based chemotherapy.

29. 27. The pharmaceutical composition of claim 26, wherein the cancer being treated is without prior treatment with lorlatinib.

30. 26. The pharmaceutical composition of claim 25, wherein the cancer is BRAF V600E mutant colorectal cancer.

31. 31. The pharmaceutical composition of claim 30, wherein the cancer being treated is resistant to BRAF inhibitor plus epidermal growth factor receptor inhibitor treatment.

32. 31. The pharmaceutical composition of claim 30, wherein the cancer being treated is resistant to BRAFi+EGFRi therapy.

33. 31. The pharmaceutical composition of claim 30, wherein the cancer being treated is without prior BRAFi+EGFRi treatment.

34. 26. The pharmaceutical composition of claim 25, wherein the cancer is a RAS-mutated solid tumor.

35. 35. The pharmaceutical composition of claim 34, wherein the cancer being treated has previously received standard treatment.

36. 26. The pharmaceutical composition of claim 25, wherein the cancer is an NF1-mutated solid tumor.

37. 37. The pharmaceutical composition of claim 36, wherein the cancer being treated has previously received standard treatment.

38. 26. The pharmaceutical composition of claim 25, wherein the cancer is a BRAF class 3 mutant solid tumor.

39. The pharmaceutical composition of claim 38, wherein the BRAF class 3 mutation is selected from one or more of the following amino acid substitutions in human BRAF: D287H; P367R; V459L; G466V; G466E; G466A; S467L; G469E; N581S; N581I; D594N; D594G; D594A; D594H; F595L; G596D; G596R; and A762E.

40. 39. The pharmaceutical composition of claim 38, wherein the cancer being treated has previously received standard treatment.

41. 23. The pharmaceutical composition of claim 22, wherein the cancer is a KRAS mutant cancer.

42. 42. The pharmaceutical composition of claim 41, wherein the KRAS mutation is selected from a KRASG12A mutation, a KRASG12C mutation, a KRASG12D mutation, a KRASG12F mutation, a KRASG12I mutation, a KRASG12L mutation, a KRASG12R mutation, a KRASG12S mutation, a KRASG12V mutation, and a KRASG12Y mutation.

43. The pharmaceutical composition of claim 20, used in combination with an additional therapeutic compound.

44. 44. The pharmaceutical composition of claim 43, wherein the additional therapeutic compound is selected from the group consisting of lorlatinib, binimetinib, cetuximab, and encorafenib.

45. 45. The pharmaceutical composition of claim 44, wherein the additional therapeutic compound is lorlatinib.

46. 45. The pharmaceutical composition of claim 44, wherein the additional therapeutic compound is binimetinib.

47. 45. The pharmaceutical composition of claim 44, wherein the additional therapeutic compounds are cetuximab and encorafenib.

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