TWI765848B - Crystalline forms of quinolone analogs and their salts - Google Patents

Abstract

The present invention includes crystalline forms of 2-(4-Methyl-[1,4]diazepan-1-yl)-5-oxo-5H-7-thia-1,11b-diaza-benzo[c]fluorene-6-carboxylic acid (5-methyl-pyrazin-2-ylmethyl)-amide and crystalline forms of salts and/or solvates of 2-(4-Methyl-[1,4]diazepan-1-yl)-5-oxo-5H-7-thia-1,11b-diaza-benzo[c]fluorene-6-carbox ylic acid (5-methyl-pyrazin-2-ylmethyl)-amide. Furthermore, the present invention provides compositions comprising the crystalline forms and therapeutic use of the crystalline forms and the compositions thereof.

Description

Crystalline forms of quinolone analogs and their salts

The present invention relates to crystalline forms of tetracyclic quinolone compounds or salts and/or solvates of tetracyclic quinolone compounds, pharmaceutical compositions comprising these crystalline forms, and methods of using these crystalline forms.

Various tetracyclic quinolone compounds have been proposed to function by interacting with quadruplex-forming regions of nucleic acids and regulating ribosomal RNA transcription. See, eg, US Pat. Nos. 7,928,100 and 8,853,234. Specifically, tetracyclic quinolone compounds can stabilize G-quadruplexes (G4s) of DNA in tumor cells, thereby inducing synthetic lethality in cancer cells. Since treatment of cells with G4 stabilizers can lead to the formation of DNA double-strand breaks (DSBs), treatment with G4 stabilizing ligands/agents (eg, tetracyclic quinolones) to induce DSB formation is more pronounced for cells that include non- Both the homologous end joining (NHEJ) and homologous recombination repair (HRR) repair pathways are either genetically deficient or chemically inhibited. In addition, tetracyclic quinolone compounds selectively inhibit rRNA synthesis of polymerase I in the nucleolus, but not mRNA synthesis of RNA polymerase II (Pol II), and do not inhibit DNA replication or protein synthesis. Targeting RNA polymerase I (Pol I) to activate p53 through the nucleolar stress pathway has been proposed to result in selective p53 activation in tumor cells. By causing cancer cells to self-destruct, the p53 protein The usual function is to suppress tumors. Activation of p53 protein to kill cancer cells is a well-validated anticancer strategy, and many approaches have been employed to exploit this pathway. The selective activation of p53 in tumor cells would be an interesting approach in the treatment, control, and remission of tumor cells without affecting normal healthy cells. The aforementioned tetracyclic quinolones are disclosed in US Pat. Nos. 7,928,100 and 8,853,234, the disclosures of which are hereby incorporated by reference in their entirety for all intended purposes.

Those skilled in the art of medicine will understand that crystallization of an active pharmaceutical ingredient provides the best control over important biochemical properties such as stability, solubility, bioavailability, particle size, bulk density, fluidity, polymorph content, and other properties. method. Accordingly, there is a need for crystalline forms of tetracyclic quinolones and methods of making such forms. These crystalline forms should be suitable for medicinal use.

或其在醫藥上可接受的鹽、酯及/或溶劑合物。在一具體實施例中，化合物I之結晶形式是化合物I之游離鹼。在另一具體實施例中，化合物I之結晶形式是化合物 I的鹽及/或溶劑合物。在一具體實施例中，化合物I之結晶形式是化合物I之酸式鹽。 In a specific embodiment, the present invention provides a crystalline form of a tetracyclic quinolone compound or a pharmaceutically acceptable salt, ester and/or solvate thereof. In a specific embodiment, the crystalline form of the tetracyclic quinolone compound is Compound I:

or a pharmaceutically acceptable salt, ester and/or solvate thereof. In a specific embodiment, the crystalline form of Compound 1 is the free base of Compound 1. In another specific embodiment, the crystalline form of Compound 1 is a salt and/or solvate of Compound 1. In a specific embodiment, the crystalline form of Compound 1 is an acid salt of Compound 1.

In some embodiments, the crystalline form of Compound I exhibits an X-ray powder diffraction (XRPD) pattern comprising peaks at about 7.730±0.3, 22.050±0.3, and 24.550±0.3 degrees 2Θ. In yet another embodiment, the crystalline form of Compound I further exhibits XRPD peaks at about 9.410 ± 0.3 and 27.700 ± 0.3 degrees 2Θ. In another embodiment, the crystalline form of Compound I further exhibits XRPD peaks at about 17.950 ± 0.3 and 25.400 ± 0.3 degrees 2Θ. In one embodiment, the crystalline form of Compound I further exhibits at least one XRPD peak at about 11.230±0.3, 11.630±0.3, 16.900±0.3, 18.580±0.3, 23.300±0.3, and 26.700±0.3 degrees 2Θ. In a specific embodiment, the crystalline form of Compound 1 exhibits an XRPD pattern comprising three or more peaks selected from the group consisting of: peaks at about 7.730±0.3, 9.410±0.3, 11.230±2Θ 0.3, 11.630±0.3, 16.900±0.3, 17.950±0.3, 18.580±0.3, 22.050±0.3, 23.300±0.3, 24.550±0.3, 25.400±0.3, 26.700±0.3 and 27.700±0.3 degrees. In another embodiment, the crystalline form of Compound 1 exhibits an XRPD pattern substantially similar to Figure 1 . In a specific embodiment, the crystalline form of Compound I exhibits a differential scanning calorimetry (DSC) pattern with peak characteristic values at about 215.41±2.0°C.

In a specific embodiment, the crystalline form of Compound I is Polymorph A. In some embodiments, the crystalline form of Compound 1 and/or the crystalline form of Polymorph A of Compound 1 has a purity of about 95%, about 97%, about 99%, about 99.5% or higher.

In another embodiment, the crystalline form of Compound I exhibits an XRPD pattern comprising a peak at about 5.720 ± 0.3 degrees 2Θ. In another embodiment, the crystalline form of Compound 1 exhibits an XRPD pattern substantially similar to FIG. 7 . In a specific embodiment, the crystalline form of Compound I exhibits a DSC pattern with a peak characteristic value at about 246.47±2.0°C.

In a specific embodiment, the crystalline form of Compound I is polymorph C. In some embodiments, the crystalline form of Compound 1 and/or the crystalline form of Polymorph C of Compound 1 has a purity of about 95%, about 97%, about 99%, about 99.5% or higher.

In some embodiments, the crystalline form of Compound I exhibits an XRPD pattern comprising a peak at about 5.680 ± 0.2 degrees 2Θ. In a specific embodiment, the crystalline form of Compound I further exhibits XRPD peaks at about 12.200±0.2, 12.600±0.3, 25.360±0.3, and 27.560±0.3 degrees 2Θ. In a specific embodiment, the crystalline form of Compound 1 exhibits an XRPD pattern comprising two or more peaks selected from the group consisting of: peaks at about 5.680±0.2, 12.200±0.2, 12.600±2Θ 0.3, 25.360±0.3 and 27.560±0.3. In another embodiment, the crystalline form of Compound 1 shows an XRPD pattern substantially similar to Figure 11 . In a specific embodiment, the crystalline form of Compound I exhibits a DSC pattern with peak characteristic values at about 231.99±2.0°C.

In a specific embodiment, the crystalline form of Compound I is polymorph E. In some embodiments, the crystalline form of Compound 1 and/or the crystalline form of Polymorph E of Compound 1 has a purity of about 95%, about 97%, about 99%, about 99.5% or higher.

In one embodiment, the crystalline form of Compound I exhibits an XRPD pattern comprising peaks at about 5.000±0.3 and 6.060±0.4 degrees 2Θ. In another embodiment, the crystalline form of Compound 1 shows an XRPD pattern substantially similar to Figure 16 . In another specific embodiment, the crystalline form of Compound I exhibits a DSC pattern with a peak characteristic value at about 222.11±2.0°C.

In a specific embodiment, the crystalline form of Compound I is polymorph G. In some embodiments, the crystalline form of Compound 1 and/or the crystalline form of Polymorph G of Compound 1 has a purity of about 95%, about 97%, about 99%, about 99.5% or higher.

In a specific embodiment, the crystalline form of Compound 1 exhibits a purity of Compound 1 greater than about 90%, 95%, 97%, 98%, 99%, or 95%.

In a specific embodiment, the crystalline form of Compound 1 is an acid salt of Compound 1 selected from the group consisting of: hydrochloride, maleate, fumarate, citrate, malic acid Salt, acetate, sulfate, phosphate, L-(+)-tartrate, D-glucuronate, benzoate, succinate, ethanesulfonate, mesylate, p-toluene Sulfonate, propionate, benzenesulfonate and 1-hydroxy-2-naphthoate. In another embodiment, the crystalline form of the salt of Compound I is selected from the group consisting of hydrochloride, maleate, fumarate, citrate and L-malate .

In a specific embodiment, the crystalline form of Compound 1 is a solvate of Compound 1. In some embodiments, the crystalline form of Compound 1 is a NMP (N-methylpyrrolidone) solvate of Compound 1.

In a specific embodiment, the crystalline form of Compound 1 is the crystalline form of the hydrochloride salt of Compound 1. In some embodiments, the crystalline form of the hydrochloride salt of Compound I exhibits an X-ray powder diffraction (XRPD) pattern comprising peaks at about 4.660±0.3 and 24.540±0.3 degrees 2Θ. In a specific embodiment, the crystalline form of the hydrochloride salt of Compound I further exhibits one or more XRPD peaks at about 19.260±0.4, 20.160±0.4, 24.920±0.3, and 26.360±0.5 degrees 2Θ. In another embodiment, the crystalline form of the hydrochloride salt of Compound I further exhibits one or more XRPD peaks at about 13.980±0.4, 14.540±0.3, 25.380±0.3, and 28.940±0.3 degrees 2Θ. In another embodiment, the crystalline form of the hydrochloride salt of Compound I exhibits an XRPD pattern substantially similar to Figure 21 . In another specific embodiment, the crystalline form of the hydrochloride salt of Compound I exhibits a DSC pattern with a peak characteristic value at about 266.27±2.0°C.

In a specific embodiment, the crystalline form of Compound 1 is the crystalline form of the maleate salt of Compound 1. In some embodiments, the crystalline form of the maleate salt of Compound I exhibits an X-ray powder diffraction (XRPD) pattern comprising peaks at about 7.400±0.3, 18.440±0.5 and 26.500±0.4 degrees 2Θ. In another embodiment, the crystalline form of the maleate salt of Compound I further exhibits one or more XRPD peaks at about 22.320±0.4, 23.920±0.3, 24.300±0.4, and 25.240±0.7 degrees 2Θ. In another embodiment, the crystalline form of the maleate salt of Compound I further exhibits one or more XRPD peaks at about 5.040±0.3, 15.080±0.3, 15.880±0.4, 20.860±0.4, and 28.540±0.3 degrees 2Θ . In another embodiment, the crystalline form of the maleate salt of Compound 1 exhibits an XRPD pattern substantially similar to FIG. 26 . In another specific embodiment, the crystalline form of the maleate salt of Compound I exhibits a DSC pattern with a peak characteristic value at about 217.32±2.0°C.

In a specific embodiment, the crystalline form of Compound 1 is the crystalline form of the fumarate salt of Compound 1. In some embodiments, the crystalline form of the fumarate salt of Compound I exhibits an X-ray powder diffraction (XRPD) pattern comprising peaks at about 6.360±0.3 and 24.800±0.3 degrees 2Θ. In another embodiment, the crystalline form of the fumarate salt of Compound I further exhibits one or more XRPD peaks at about 19.660±0.3, 20.420±0.3, and 26.860±0.3 degrees 2Θ. In another embodiment, the crystalline form of the fumarate salt of Compound I further exhibits one or more XRPD peaks at about 12.680±0.3, 17.020±0.2, 25.180±0.2, and 28.280±0.3 degrees 2Θ. In another embodiment, the crystalline form of the fumarate salt of Compound 1 exhibits an XRPD pattern substantially similar to FIG. 31 . In another specific embodiment, the crystalline form of the fumarate salt of Compound I exhibits a DSC pattern with a peak characteristic value at about 222.40±2.0°C.

In a specific embodiment, the crystalline form of Compound 1 is the crystalline form of Compound 1 citrate salt. In some of the embodiments, the crystalline form of the citrate salt of Compound 1 shows an X-ray The powder diffraction (XRPD) pattern contained peaks at about 4.900±0.3, 25.380±0.3 and 27.500±0.4 degrees 2Θ. In another embodiment, the crystalline form of the citrate salt of Compound I further exhibits one or more XRPD peaks at about 15.360±0.3, 18.100±0.3, 19.300±0.3, and 26.140±0.4 degrees 2Θ. In another embodiment, the crystalline form of the citrate salt of Compound I further exhibits one or more XRPD peaks at about 17.400±0.3, 18.680±0.4, 24.040±0.4, and 26.740±0.3 degrees 2Θ. In another embodiment, the crystalline form of the citrate salt of Compound 1 exhibits an XRPD pattern substantially similar to FIG. 36 . In another specific embodiment, the crystalline form of the citrate salt of Compound I exhibits a DSC pattern with peak characteristic values at about 196.86±2.0°C.

In a specific embodiment, the crystalline form of Compound 1 is the crystalline form of the L-malate salt of Compound 1. In some embodiments, the crystalline form of the L-malate salt of Compound I exhibits an X-ray powder diffraction (XRPD) pattern comprising peaks at about 6.580±0.2, 6.780±0.3, and 25.560±0.4 degrees 2Θ. In another embodiment, the crystalline form of the L-malate salt of Compound I further exhibits one or more XRPD peaks at about 19.560±0.4, 23.660±0.4, 26.060±0.7, and 26.960±0.7 degrees 2Θ. In another specific embodiment, the crystalline form of the L-malate salt of Compound I further exhibits one or more XRPD peaks at about 8.800±0.3, 11.800±0.3, 18.600±0.3, 24.460±0.5, and 25.080±0.3 2Θ Spend. In another embodiment, the crystalline form of the L-malate salt of Compound 1 exhibits an XRPD pattern substantially similar to Figure 41 . In another specific embodiment, the crystalline form of the L-malate salt of Compound I exhibits a DSC pattern with a peak characteristic value at about 209.67±2.0°C.

In a specific embodiment, there is provided a composition comprising a crystalline form of Compound I as described herein, or a pharmaceutically acceptable salt, ester and/or solvate thereof. In another specific embodiment, a composition comprising a crystalline form of the free base of Compound I is provided. In a specific embodiment, a composition is provided comprising a crystalline form of a salt or solvate of Compound I. In a specific embodiment In , there is provided a composition comprising a crystalline form of an acid salt of Compound I. In a specific embodiment, there is provided a composition comprising one or more crystalline forms of any of Compound I as described herein. In some embodiments, any of the compositions described herein comprise at least one pharmaceutically acceptable carrier.

In a specific embodiment, there is provided a method of stabilizing G-quadruplexes (G4s) in an individual, the method comprising administering to the individual a therapeutically effective amount of a crystalline form of Compound I as described herein, or Pharmaceutically acceptable salts, esters and/or solvates.

In a specific embodiment, there is provided a method of modulating p53 activity in an individual, the method comprising administering to the individual a therapeutically effective amount of a crystalline form of Compound I as described herein, or a pharmaceutically acceptable salt thereof , esters and/or solvates.

In a specific embodiment, there is provided a method of treating or ameliorating a cell proliferative disorder in an individual, the method comprising administering to an individual in need thereof a therapeutically effective amount of a crystalline form of Compound I as described herein, or pharmaceutically Acceptable salts, esters and/or solvates. In a specific embodiment, the method is provided to treat or alleviate cancer.

In a specific embodiment, there is provided a method for treating or alleviating cancer, the method comprising administering to an individual in need thereof a therapeutically effective amount of a crystalline form of Compound I as described herein, or a pharmaceutically acceptable form thereof Salts, esters and/or solvates, wherein the cancer is selected from the group consisting of: blood cancer, colorectal cancer, breast cancer, lung cancer, liver cancer, ovarian cancer, cervical cancer, Ewing's sarcoma, pancreas cancer, lymph node cancer, colon cancer, prostate cancer, brain cancer, head and neck cancer, skin cancer, kidney cancer and heart cancer. In a specific embodiment, the cancer treated or in remission by the method is a blood cancer selected from the group consisting of leukemia, lymphoma, myeloma, and multiple myeloma. In a specific embodiment, the cancer treated or ameliorated by the method is homologous recombination (HR) DNA double-strand break (DSB) repair deficient cancer, or non-homologous end joining (NHEJ) DNA double-strand break repair Defective cancer. exist In another specific embodiment, the cancer treated or ameliorated by the method comprises cancer cells with defects in breast cancer susceptibility gene 1 (BRCA1), breast cancer susceptibility gene 2 (BRCA2) and/or homologs Other members of the sexual recombination pathway. In another specific embodiment, the cancer cells are deficient in BRCA1 and/or BRCA2. In another specific embodiment, the cancer cells are homozygous for the genotype with BRCA1 and/or BRCA2 mutation. In another specific embodiment, the cancer cells are heterozygous for the genotype with BRCA1 and/or BRCA2 mutation.

In a specific embodiment, the methods provided herein further comprise administering one or more additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents are anticancer agents or immunotherapeutic agents. In a specific embodiment, the one or more therapeutically active agents are immunotherapeutic agents. In some embodiments, the one or more immunotherapeutic agents include, but are not limited to, monoclonal antibodies, immune effector cells, adoptive cell transfer, immunotoxins, vaccines, or cytokines.

In a specific embodiment, there is provided a method for reducing or inhibiting cell proliferation, the method comprising contacting the cell with a therapeutically effective amount of Compound I or a pharmaceutically acceptable salt, ester and/or solvate thereof. In a specific embodiment, the cell is a cancer cell line or a tumor cell in a subject. In a specific embodiment, the cancer cell line in the above method is selected from the group consisting of: blood cancer, colorectal cancer, breast cancer, lung cancer, liver cancer, ovarian cancer, cervical cancer, Ewing's sarcoma, pancreas cancer, lymph node cancer, colon cancer, prostate cancer, brain cancer, head and neck cancer, skin cancer, kidney cancer and heart tumor. In a specific embodiment, the blood cancer cell line described in the above method is selected from the group consisting of leukemia, lymphoma, myeloma, and multiple myeloma. In a specific embodiment, the cancer cell line described in the above method is a cancer cell deficient in homologous recombination (HR) DNA double-strand break (DSB) repair or non-homologous end joining (NHEJ) DNA double-strand break repair. In a specific embodiment, the cancer cell line is a cancer cell with a defect in breast cancer susceptibility gene 1 (BRCA1), breast cancer susceptibility gene 2 (BRCA2), and/or other members of the homologous recombination pathway. In another specific embodiment, the cancer cells are deficient in BRCA1 and/or BRCA2. In another specific embodiment, the cancer cells are homozygous for the genotype with BRCA1 and/or BRCA2 mutation. In another specific embodiment, the cancer cells are heterozygous for the genotype with BRCA1 and/or BRCA2 mutation.

Fig. 1 is the X-ray powder diffraction (XRPD) spectrum of polymorph A of compound I (free base); Fig. 2 is the differential scanning calorimetric analysis (DSC) spectrum of polymorph A of compound I (free base). Fig. 3 is the thermogravimetric analysis (TGA) spectrum of the polymorphic form A of compound I (free base); Fig. 4 is the overlay of the DSC spectrum and TGA spectrum of the polymorphic form A of compound I (free base); Figure 5 is a hydrogen nuclear magnetic resonance ( ¹ H NMR) spectrum of polymorph A of compound I (free base); Figure 6A is a photomicrograph of polymorph A of compound I (free base) using cross-polarized light and Figure 6B is a micrograph of polymorph A of compound I (free base) using unpolarized light; Figure 7 is the XRPD pattern of polymorph C of compound I (free base); Figure 8 is compound I (free base) DSC spectrum of polymorph C of compound I (free base); Figure 9 is the ¹ H NMR spectrum of polymorph C of compound I (free base); Figure 10A is polymorph C of compound I (free base) using cross Polarized light photomicrograph and Figure 10B is a photomicrograph of polymorph C of compound I (free base) using unpolarized light; Figure 11 is an XRPD pattern of polymorph E of compound I (free base); Figure 12 is the DSC spectrum of polymorph E of compound I (free base); Figure 13 is the ¹ H NMR spectrum of polymorph E of compound I (free base); Figure 14A is of compound I (free base) Figure 14B is a photomicrograph of Polymorph E using cross polarized light and Figure 14B is a photomicrograph of Polymorph E of Compound I (free base) using unpolarized light; Figure 15 is a polymorph of Compound I (free base) Thermogravimetric analysis (TGA) pattern of Form E.

Figure 16 is the XRPD pattern of polymorph G of compound I (free base); Figure 17 is the DSC pattern of polymorph G of compound I (free base); Figure 18 is the polymorph of compound I (free base) ¹ H NMR spectra of Form G; Figure 19A is a photomicrograph of Polymorph G of Compound I (free base) using cross-polarized light and Figure 19B is a photomicrograph of Polymorph G of Compound I (free base) using no Micrographs of polarized light; Figure 20 depicts the solubility of several polymorphic forms of Compound I (free base) in methanol and THF; Figure 21 is the XRPD pattern of the hydrochloride salt of Compound I; Figure 22 is the hydrochloric acid of Compound I The DSC spectrum of the salt; Figure 23 is the ¹ H NMR spectrum of the hydrochloride of compound I; Figure 24 is the photomicrograph of the hydrochloride of compound I using polarized light; Figure 25 is the TGA spectrum of the hydrochloride of compound I; Figure 26 is the XRPD spectrum of the maleate of compound I; Figure 27 is the DSC spectrum of the maleate of compound I; Figure 28 is the ¹ H NMR spectrum of the maleate of compound I; Figure 29 is the horse of the compound I Figure 30 is the TGA pattern of the maleate of compound I; Figure 31 is the XRPD pattern of the fumarate of compound I; Figure 32 is the fumarate of compound I DSC spectrum; Figure 33 is the ¹ H NMR spectrum of the fumarate of compound I; Figure 34 is the photomicrograph of the fumarate of compound I using polarized light; Figure 35 is the TGA spectrum of the fumarate of compound I ; Figure 36 is the XRPD spectrum of the citrate of compound I; Figure 37 is the DSC spectrum of the citrate of compound I; Figure 38 is the ¹ H NMR spectrum of the citrate of compound I; Figure 39 is the citric acid of compound I Figure 40 is the TGA pattern of the fumarate salt of compound I; Figure 41 is the XRPD pattern of the L-malate salt of compound I; Figure 42 is the L-malate salt of compound I. DSC spectrum; Figure 43 is the ¹ H NMR spectrum of L-malate of Compound 1; Figure 44 is a photomicrograph of L-malate of Compound 1 using polarized light; and Figure 45 is L-malic acid of Compound 1 TGA spectrum of salt.

The present invention relates to crystalline forms of tetracyclic quinolone compounds that stabilize G-quadruplexes (G4s) and/or inhibit polymerase I (Pol I), and also to salts and/or solvates of tetracyclic quinolone compounds. crystalline form. These crystalline substances can be formulated into pharmaceutical compositions and used to treat diseases characterized by cell proliferation.

Definition :

It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, representative methods and materials are described herein.

In accordance with long-standing patent law practice, the terms "a," "an," and "the" are used in this application (including in the claims) to mean "one or more." Thus, for example, a reference to "a carrier" includes a mixture of one or more carriers, two or more carriers, or the like.

Unless otherwise stated, all numbers used in the specification and claims to represent quantities of ingredients, reaction conditions, etc., should be understood to mean adding "about" in all cases. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and claimed claims are approximations that may vary depending upon the desired properties sought to be obtained by the present invention. In general, the term "about" as used herein, when referring to a measurement, such as weight, time, dose, etc., is intended to encompass ±15% relative to the specified amount, in one example or ±10% variance, in another example ±5% variance, in another example ±1% variance, and in yet another example ±0.1% variance, as long as the above Variations are appropriate to implement the disclosed methods.

The term "compound of the present invention" or "compound" refers to 2-(4-methyl-[1,4]diazepan-1-yl)-5-oxo-5H-7-thia 1,11b Diaza-benzo[c]fluorene-6-carboxylic acid (5-methyl-pyridin-2-ylmethyl)-amine (Compound I) or its isomers, salts, esters, N - Crystalline forms of oxides or solvates. Alternatively, the above terms may refer to a salt or solvate of Compound I or both forms. The crystalline forms of Compound 1 described in this application include crystalline forms of any single isomer of Compound 1 as well as mixtures of any number of isomers of Compound 1.

The term "co-administered" refers to (a) a crystalline form of Compound 1, or a crystalline form of a pharmaceutically acceptable salt, ester, solvate and/or prodrug of Compound 1; and (b) one or more additional The therapeutic agents and/or radiation therapy are administered in combination, that is, administered together in a complementary manner.

The term "isomers" refers to compounds having the same chemical formula, but may have different stereochemical formulas, structural formulas, or particular atomic arrangements. Examples of isomers include stereoisomers, diastereoisomers, enantiomers, conformers, rotamers, geometric isomers, and atropisomers.

"N-oxide", also known as amine oxide or amine-N-oxide, means a compound derived from a compound of the present invention via oxidation of the amine group of the compound of the present invention. N-oxides typically contain functional groups _R3N ^{+ -O-} (sometimes denoted as _R3N =O or _R3N →O).

Polymorphism can be characterized by the ability of a compound to crystallize into different crystalline forms while maintaining the same chemical formula. Crystalline polymorphs of a given drug substance that are chemically identical Any other crystalline polymorph of the drug substance with the same atoms bonded to each other in the same way, because of its different crystalline form, can affect one or more physical properties, such as stability, solubility, melting point, bulk density, mobility and bioavailability.

The term "composition" refers to a physical form of one or more substances, such as solids, liquids, gases, or mixtures thereof. An example of a composition is a pharmaceutical composition, that is, a composition pertaining to, prepared for, or used in medicine.

The term "carboxylic acid" refers to an organic acid characterized by one or more carboxyl groups, such as acetic acid and oxalic acid. "Sulfonic acid" refers to organic acids having the general formula R-(S(O) ₂ -OH) _n , where R is an organic hydrocarbyl moiety and n is an integer greater than zero, such as 1, 2, and 3. The term "polyhydroxy acid" refers to a carboxylic acid containing two or more hydroxyl groups. Examples of polyhydroxy acids include, but are not limited to, lactobionic acid, gluconic acid, and galactose.

"Pharmaceutically acceptable" as used herein means without undue toxicity, irritation, allergic reactions, and other similar effects, suitable for use in contact with human and animal tissues, with a reasonable benefit/risk ratio, and in It is valid for the intended use within the scope of sound medical judgment.

"Salt" includes derivatives of active agents wherein the active agent has been modified by making acid or base addition salts thereof. Preferably, the salt is a pharmaceutically acceptable salt. Such salts include, but are not limited to, pharmaceutically acceptable acid addition salts, pharmaceutically acceptable base addition salts, pharmaceutically acceptable metal salts, and ammonium and alkylated ammonium salts. Acid addition salts include salts of inorganic acids as well as organic acids. Representative examples of suitable inorganic acids include hydrochloric, hydrobromic, hydroiodic, phosphoric, sulfuric, nitric, and the like. Representative examples of suitable organic acids include formic acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, propionic acid, benzoic acid, cinnamic acid, citric acid, fumaric acid, glycolic acid, lactic acid, maleic acid, malic acid, malonic acid acid, mandelic acid, oxalic acid, picric acid, pyruvic acid, salicylic acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, tartaric acid, ascorbic acid, pamoic acid, bismethylenesalicylic acid, ethanedisulfonic acid, glucose acid, citraconic acid, aspartic acid, hard Fatty acid, palmitic acid, ethylenediaminetetraacetic acid (EDTA), glycolic acid, p-aminobenzoic acid, glutamic acid, benzenesulfonic acid, p-toluenesulfonic acid, sulfate, nitrate, phosphate, perchlorate, Borate, acetate, benzoate, hydroxynaphthalene, glycerophosphoric acid, salt ketoglutarate, and the like. Basic addition salts include, but are not limited to, ethylenediamine, N-methyl-glucosamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chlorine Procaine, Diethanolamine, Procaine, N-Benzylphenethylamine, Diethylamine, Piperazine, Tris(hydroxymethyl)-aminomethane, Tetramethyl Hydroxide, Triethylamine, Dibenzyl Amine, diphenylhydroxymethylamine, dehydroabietylamine, N-ethylpiperidine, benzylamine, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, ethylamine, basic amino acids such as Amino acids and arginine dicyclohexylamine and the like. Examples of metal salts include salts of lithium, sodium, potassium, magnesium, and the like. Examples of ammonium and alkylated ammonium salts include salts of ammonium, methylammonium, dimethylammonium, trimethylammonium, ethylammonium, hydroxyethylammonium, diethylammonium, butylammonium, tetramethylammonium and the like. Examples of organic bases include lysine, arginine, guanidine, diethanolamine, choline, and the like. Standard methods for the preparation of pharmaceutically acceptable salts and formulations thereof are well known in the art and are disclosed in various references including, for example, "Remington: The Science and Practice of Pharmacy", A. Gennaro, ed., 20th edition, Lippincott, Williams & Wilkins, Philadelphia, PA.

As used herein, "solvate" refers to a complex that is solvated (combination of a co-solubilizing molecule with a molecule or ion of an active agent of the present invention), or a complex composed of an ion or molecule (an active agent of the present invention) in a solvent. agent) and one or more solubilizing molecules. In the present invention, preferred solvates are hydrates. Examples of hydrates include, but are not limited to, hemihydrates, monohydrates, dihydrates, trihydrates, hexahydrates, and the like. It will be understood by those of ordinary skill in the art that pharmaceutically acceptable salts of the present compounds may also exist in the form of solvates. The solvates are typically formed by hydration, either as part of the preparation of the present compound, or as a natural consequence of the anhydrous compound of the present invention. Moisture absorption. Solvates, including hydrates, may be composed of stoichiometric ratios, eg, two, three, or four salt molecules per solvate or per hydrate molecule. Another possibility, for example, the stoichiometry of two salt molecules involves three, penta, heptasolvent or hydrate molecules. Solvents used for crystallization, such as alcohols, especially methanol and ethanol; aldehydes; ketones, especially acetone; esters, such as ethyl acetate; can be embedded in the crystal lattice. Preferred are pharmaceutically acceptable solvates.

The term "substantially similar" as used herein means that analytical spectra, such as XRPD spectra, Raman spectra, etc., are largely similar to a reference spectrum, both in peak position and in intensity.

The terms "excipient," "carrier," and "vehicle" are used interchangeably in this application and refer to a substance with which a compound of the present invention is administered.

A "therapeutically effective amount" means an amount of a crystalline form that, when administered to a patient to treat a disease or other adverse physical condition, is sufficient to produce a beneficial effect on the disease or condition. A therapeutically effective amount will vary depending on the crystalline form, the disease or disorder and its severity, and the age, weight, etc. of the patient being treated. Determination of a therapeutically effective amount of a particular crystalline form is within the ordinary skill in the art without routine experimentation.

The terms "diseases characterized by cell proliferation" or "disorders characterized by cell proliferation" as used herein include, but are not limited to, cancer, benign and malignant tumors. Examples of cancers and tumors include, but are not limited to, proliferations in the large intestine, breast, lung, liver, pancreas, lymph nodes, colon, prostate, brain, head and neck, skin, kidney, blood, and heart (eg, leukemia, lymphoma, and malignant tumors). cancer or tumor.

The term "treating" corresponds to a particular disease or disorder, and includes preventing the disease or disorder and/or alleviating, ameliorating, alleviating or eliminating the symptoms and/or pathology of the disease or disorder. Generally, the term is used herein to refer to ameliorating, alleviating, alleviating, and removing symptoms of a disease or disorder. this article The described candidate molecules or compounds can be present in a formulation or drug in a therapeutically effective amount that results in a biological effect, such as apoptosis of specific cells (eg, cancer cells), reduction in proliferation of specific cells, or, for example, can lead to a disease or disorder improvement, alleviation, alleviation or removal of symptoms. The term can also refer to reducing or stopping the rate of cell proliferation (eg, slowing or stopping tumor growth) or reducing the number of proliferating cancer cells (eg, removing part or all of a tumor). These terms also apply to a reduction in microbial titer, a reduction in the rate of microbial reproduction, a reduction in the number of symptoms, or the effect of symptoms associated with a microbial infection in a microbially infected system (ie, a cell, tissue or individual) and/or a Removal of detectable amount of microorganisms. Examples of microorganisms include, but are not limited to, viruses, bacteria, and fungi.

As used herein, the terms "inhibit", "reduce" or "reduce" cell proliferation mean, when measured using methods known to those of ordinary skill in the art, slow, reduce or, for example, stop the amount of cell proliferation, For example, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100% relative to proliferating cells that did not receive the methods and compositions of the present application.

As described herein, "apoptosis" refers to the self-destruction or suicide process inherent in a cell. In response to a triggering stimulus, cells undergo a cascade of events including cell shrinkage, membrane blebbing, and chromatin condensation and fragmentation. The final stage of this process involves the formation of multiple particles with intact membrane structures (apoptotic bodies), which are subsequently phagocytosed by macrophages.

Crystal material:

In a specific embodiment, the present invention provides a crystalline form of Compound I (free base). In another specific embodiment, the present invention provides crystalline forms of salts and/or solvates of Compound I. In a specific embodiment, the salt is a hydrochloric acid addition salt. In a specific embodiment, the salt is a sulfuric acid addition salt. In a specific embodiment, the salt is a sulfonic acid addition salt. In a specific embodiment, the salt is a carboxylic acid addition salt. In specific embodiments, the salt is a polyhydroxy acid addition salt.

Examples of crystalline salts include, but are not limited to, hydrochloride, maleate, fumarate, citrate, malate, sulfate, acetate, phosphate, L-(+)-tartrate, D- Glucuronate, benzoate, succinate, ethanesulfonate, mesylate, p-toluenesulfonate, malonate, benzenesulfonate and 1-hydroxy-2-naphthalene formate. In a specific embodiment, the ratio of Compound I to acid in the crystalline salt is from about 1:0.5 to about 1:3.

Examples of crystalline solvates include, but are not limited to, NMP (N-methyl-2-pyrrolidone) solvates of Compound 1.

In a specific embodiment, the crystalline form is characterized by the spacing between lattice planes as determined by X-ray powder diffraction (XRDP). XRDP spectra are typically presented as a graph of peak intensity versus peak position (ie, the angle of diffraction angle 2Θ). Intensities are generally shown in parentheses with the following abbreviations: very strong=vst, strong=st; moderate=m; weak=w; and very weak=vw. A particular XRDP characteristic peak can be selected based on the location of the peak and its relative intensities in order to distinguish this crystal structure from other crystal structures. The peak intensity percentage relative to the strongest peak can be expressed as I/Io.

Those skilled in the art recognize that measurements of XRDP peak positions and/or intensities for a given crystalline form of the same compound will vary within a margin of error. The value of the angle 2θ allows for an appropriate margin of error. Typically, the margin of error is expressed as "±". For example, a 2θ angle of about "8.716±0.3" represents a range from about 8.716+0.3 (ie, about 9.016) to about 8.716-0.3 (ie, about 8.416). Depending on sample preparation techniques, applied instrument calibration techniques, human manipulation variations, etc., those skilled in the art recognize that the appropriate XRDP error range may be about ±0.7; ±0.6; ±0.5; ±0.4; ±0.3; ±0.2; ±0.1 ; ±0.05; or less.

Details of other methods and equipment for XRDP analysis are described in the "Examples" section.

In a specific embodiment, the crystalline form is characterized by Differential Scanning Thermal Analysis (DSC). A DSC pattern is typically a graphical representation of normalized heat flow in watts per gram ("W/g") plotted against the measured sample temperature in degrees Celsius. DSC spectra are commonly used to estimate extrapolated onset and outset temperatures, peak temperatures, and heat of fusion. The peak characteristic value of the DSC spectrum is usually used as the characteristic peak to distinguish this crystal structure from other crystal structures.

Those skilled in the art recognize that measurements of DSC spectra for a given crystalline form of the same compound will vary within a margin of error. The values of the single peak eigenvalues (expressed in degrees Celsius) allow for an appropriate margin of error. Typically, the margin of error is expressed as "±". For example, a single peak characteristic value of about "53.09±2.0" represents a range from about 53.09+2 (ie, about 55.09) to about 53.09-2 (ie, about 51.09). Depending on sample preparation techniques, applied instrument calibration techniques, human manipulation variations, etc., the error range for a single peak characteristic value that is known to those skilled in the art as appropriate may be about ±2.5; ±2.0; ±1.5; ±1.0; ±0.5; or smaller.

Details of other methods and apparatus for DSC pattern analysis are described in the "Examples" section.

In a specific embodiment, the crystalline form is characterized by Raman spectroscopy. Raman spectra are typically represented as a graph of the Raman intensity of a peak versus the Raman shift of the peak. The "peaks" in the Raman spectrum are also called "absorption bands". Intensities are usually shown in parentheses with the following abbreviations: strong=st; moderate=m; weak=w. The characteristic peaks of a given Raman spectrum can be selected based on the location of the peaks and their relative intensities, in order to distinguish this crystal structure from other crystal structures.

Those skilled in the art recognize that measurements of Raman peak shift and/or intensity for a given crystalline form of the same compound will vary within a margin of error. The value of the peak displacement, expressed in inverse wavenumber (cm ^-1 ), allows for an appropriate margin of error. Typically, the margin of error is expressed as "±". For example, a representation of a Raman shift of about "1310±10" ranges from about 1310+10 (ie, about 1320) to about 1310-10 (ie, about 1300). Depending on the sample preparation technique, applied instrument calibration techniques, human manipulation variation, etc., the error range of the appropriate single peak characteristic value for those skilled in the art may be ±12; ±10; ±8; ±5; ±3; ±1 ; or less.

Details of other methods and apparatus for Raman spectroscopy analysis are described in the "Examples" section.

In a specific embodiment, the crystalline form of Compound I (free base). In some embodiments, the crystalline form of Compound 1 (free base) exhibits different polymorphs. Examples of crystalline forms of Compound I (free base) include, but are not limited to, polymorphs A, C, E, and G, as defined in the following sections.

In some embodiments, one form of a polymorph is more stable than other forms. In a specific embodiment, Polymorph A, the crystalline form of Compound I (free base) exhibits high stability. In some specific embodiments, different polymorphic forms of Compound 1 (free base) are converted to another polymorphic form under certain conditions. In some embodiments, polymorphs C, E and/or G are converted to polymorph A under suitable conditions. In a specific embodiment, each of polymorphs C, E, and G independently converts to polymorph A in solution at room temperature or at elevated temperature. Polymorph A is the most thermodynamically stable form; however, other forms may be formed or kinetically favorable under certain conditions.

In one embodiment of the present invention, the crystalline form of Compound 1 may comprise a mixture of one or more forms of the polymorph of Compound 1. In some embodiments, the crystalline form of Compound 1 may comprise a single polymorphic form in substantially pure form. In a specific embodiment, the crystalline form of Compound 1 may comprise more than about 99.9%, about 99.8%, about 99.7%, about 99.6%, about 99.5%, about 99.4%, about 99.3%, about 99.2%, about 99.1% or about 99.0% of the single polymorph of Compound I. In another specific embodiment, the crystalline form of Compound 1 may comprise more than about 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91% or 90% of the single polymorph of Compound I. In a specific embodiment, the crystalline form of Compound 1 may comprise more than about 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, or 40% of the compound Single polymorph of I.

In an embodiment of the present invention, the crystalline form of Compound I may comprise at least about 99.9%, about 99.8%, about 99.7%, about 99.6%, about 99.5%, about 99.4%, about 99.3%, about 99.2%, About 99.1%, about 99.0%, about 98%, about 97%, about 96%, about 95%, about 94%, about 93%, about 92%, about 91%, about 90%, or about 85% of Compound 1 Polymorph A (free base).

In one embodiment of the present invention, the crystalline form of Compound I may be the polymorph A of Compound I (free base), which includes about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6% , 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5 %, 8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% polymorphs Substances C, E or G or mixtures thereof.

In an embodiment of the present invention, the crystalline form of Compound I may be the polymorph C of Compound I (free base), which includes about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6% , 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5 %, 8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% polymorphs item A.

In one embodiment of the present invention, the crystalline form of Compound I may be the polymorph E of Compound I (free base), which includes about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6% , 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5 %, 8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% polymorphs item A.

In an embodiment of the present invention, the crystalline form of Compound I may be the polymorph G of Compound I (free base), which includes about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6% , 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5 %, 8%, 8.5%, 9%, 9.5%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% polymorphs item A.

In a specific embodiment, the crystalline form of Compound I provided by the present invention is of high purity. In an embodiment of the invention, the crystalline form of Compound 1 may be at least 99.9%, about 99.8%, about 99.7%, about 99.6%, about 99.5%, about 99.4%, about 99.3%, about 99.2% pure , about 99.1%, about 99.0%, about 98%, about 97%, about 96%, about 95%, about 94%, about 93%, about 92%, about 91%, or about 90% pure compound I.

Other methods for characterizing the crystalline forms of the present invention are described in the "Examples" section.

Polymorph A

In a specific embodiment, Polymorph A of Compound 1 (free base) exhibits an XRDP comprising peaks at about 7.730, 22.050 and 24.550 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In another specific embodiment, the XRDP of crystalline polymorph A of Compound 1 (free base) further comprises peaks at about 9.410 and 27.700 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.3; 0.2; about ±0.1; about ±0.05; or less. In yet another embodiment, the crystalline form of Polymorph A of Compound 1 (free base) further comprises peaks at about 17.950 and 25.400 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.3; 0.2; about ±0.1; about ±0.05; or less. In another embodiment, the crystalline form of Polymorph A of Compound I (free base) further comprises at least one peak at about 11.230, 11.630, 16.900, 18.580, 23.300, and 26.700 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In yet another embodiment, the crystalline form of Polymorph A of Compound 1 (free base) exhibits an XRDP comprising the peaks shown in Table 1 below:

In a specific embodiment, the crystalline form of Polymorph A of Compound I (free base) exhibits an XRDP substantially similar to that of FIG. 1 .

In a specific embodiment, the crystalline form of Polymorph A of Compound I (free base) exhibits a DSC pattern comprising a sharp endothermic peak at about 215.41°C, with a margin of error of about ±2.5; about ±2.0; about ±2.0; 1.5; about ±1.0; about ±0.5; or less. In another embodiment, the crystalline form of Polymorph A of Compound I (free base) further exhibits a DSC pattern comprising one or more of the following peaks: an exothermic peak at about 216.92±2.0°C, an exothermic peak at about 230.56 Endothermic peak at ±2.0°C, exothermic peak at about 232.84±2.0°C, and endothermic peak at about 241.56±2.0°C. In a specific embodiment, the crystalline form of Polymorph A of Compound I (the free base) exhibits a DSC pattern substantially similar to that of FIG. 2 .

In one embodiment, the crystalline form of Polymorph A of Compound I (the free base) exhibits a ¹ H NMR spectrum substantially similar to that of FIG. 5 . In another embodiment, the crystalline form of Polymorph A of Compound I (free base) may have a fine needle-like crystal phase on a macroscopic scale that forms a loosely bound mass, which is substantially Similar to Figures 6A and 6B.

Polymorph C

In a specific embodiment, polymorph C of Compound 1 (free base) exhibits an XRDP comprising a peak at about 5.720 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In another specific embodiment, the crystalline form of crystalline polymorph C of Compound 1 (free base) exhibits an XRDP comprising the peaks shown in Table 2 below:

In a specific embodiment, the crystalline form of polymorph C of Compound I (free base) exhibits an XRDP substantially similar to that of FIG. 7 .

In a specific embodiment, the crystalline form of Polymorph C of Compound I (the free base) exhibits a DSC pattern comprising peak characteristic values at about 246.47°C with a margin of error of about ±2.5; about ±2.0; about ±1.5; about ±1.0; about ±0.5; or less. In a specific embodiment, the crystalline form of polymorph C of Compound I (free base) exhibits a DSC pattern substantially similar to that of FIG. 8 .

In one embodiment, the crystalline form of polymorph C of Compound I (the free base) exhibits a ¹ H NMR spectrum substantially similar to FIG. 9 . In another embodiment, the crystalline form of polymorph C of Compound I (free base) may consist of needle-like agglomerates on a macroscopic scale, which are substantially similar to Figures 10A and 10B.

Polymorph E

In a specific embodiment, Polymorph E of Compound 1 (free base) exhibits an XRDP comprising a peak at about 5.680 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In another embodiment, the XRDP of the crystalline polymorph E of Compound I (free base) further comprises at least one peak at 2θ 12.200, 12.600, 25.360 and 27.560 degrees, with an error range of about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In yet another specific embodiment, the crystalline form of Polymorph E of Compound I (free base) exhibits an XRDP comprising the peaks shown in Table 3 below:

In a specific embodiment, the crystalline form of polymorph E of Compound I (free base) exhibits an XRDP substantially similar to that of FIG. 11 .

In a specific embodiment, the crystalline form of Polymorph E of Compound I (the free base) exhibits a DSC pattern comprising peak characteristic values at about 231.99°C with a margin of error of about ±2.5; about ±2.0; about ±1.5; about ±1.0; about ±0.5; or less. In a specific embodiment, the crystalline form of polymorph E of Compound I (free base) exhibits a DSC pattern substantially similar to that of FIG. 12 .

In one embodiment, the crystalline form of polymorph E of Compound I (free base) exhibits a ¹ H NMR spectrum substantially similar to that of FIG. 13 . In another embodiment, the crystalline form of Polymorph E of Compound I (free base) is substantially similar to Figures 14A and 14B.

Polymorph G

In a specific embodiment, polymorph G of Compound I (free base) exhibits an XRDP comprising peaks at about 5.000 and 6.060 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.2 ; about ±0.1; about ±0.05; or less. In another specific embodiment, the crystalline form of Polymorph G of Compound I (free base) exhibits an XRDP comprising the peaks shown in Table 4 below:

In a specific embodiment, the crystalline form of polymorph G of Compound I (free base) exhibits an XRDP substantially similar to that of FIG. 16 .

In a specific embodiment, the crystalline form of polymorph G of Compound I (free base) exhibits a DSC pattern comprising peak characteristic values at about 222.11°C, with a margin of error of about ±2.5; about ±2.0; about ±1.5; about ±1.0; about ±0.5; or less. In a specific embodiment, the crystalline form of polymorph G of Compound I (free base) exhibits a DSC pattern substantially similar to that of FIG. 17 .

In one embodiment, the crystalline form of polymorph G of Compound I (free base) exhibits a ¹ H NMR spectrum substantially similar to FIG. 18 . In another embodiment, the crystalline form of Polymorph G of Compound I (free base) is substantially similar to Figures 19A and 19B.

Compound I hydrochloride

In a specific embodiment, the crystalline form of the hydrochloride salt of Compound 1 exhibits an XRDP comprising peaks at about 4.660 and 24.540 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.2; 0.1; about ±0.05; or less. In another embodiment, the XRDP of the crystalline form of the hydrochloride salt of Compound I further comprises one, two, three or four peaks at 2θ selected from the group consisting of: 19.260, 20.160, 24.920 and 26.360 degrees, with a margin of error of approx. ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In another specific embodiment, the crystalline form of the hydrochloride salt of Compound I further comprises one, two or three peaks at 2θ selected from the group consisting of: 13.980, 14.540, 25.380 and 28.940 degrees with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In yet another embodiment, the crystalline form of the hydrochloride salt of Compound 1 exhibits an XRDP comprising the peaks shown in Table 6 below:

In a specific embodiment, the crystalline form of the hydrochloride salt of Compound 1 exhibits an XRDP substantially similar to FIG. 21 .

In a specific embodiment, the crystalline form of the hydrochloride salt of Compound I exhibits a DSC pattern comprising peak characteristic values at about 266.27°C, with a margin of error of about ±2.5; about ±2.0; about ±1.5; about ±1.0; 0.5; or less. In a specific embodiment, the crystalline form of the hydrochloride salt of Compound 1 shows The resulting DSC spectrum is substantially similar to Figure 22. In a specific embodiment, the crystalline form of the hydrochloride salt of Compound I does not have a distinct melting point.

In one embodiment, the crystalline form of the hydrochloride salt of Compound 1 exhibits a ¹ H NMR spectrum substantially similar to that of FIG. 23 . In another embodiment, the crystalline form of the hydrochloride salt of Compound I consists of cohesive aggregates formed in small needles on a macroscopic scale, which is substantially similar to FIG. 24 . In a specific embodiment, the ratio of hydrochloric acid and Compound I in the hydrochloride salt of Compound I is about 1:1.

Maleate of Compound I

In a specific embodiment, the crystalline form of the maleate salt of Compound I exhibits an XRDP comprising peaks at about 7.400, 18.440, and 26.500 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.2 ; about ±0.1; about ±0.05; or less. In another embodiment, the XRDP of the crystalline maleate salt of Compound I further comprises one, two, three or four peaks at 2θ selected from the group consisting of: 22.320, 23.920, 24.300 and 25.240 degrees, with an error range of about ± about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In another embodiment, the crystalline maleate salt of Compound 1 further comprises one, two, three, or four peaks selected from the group consisting of 2Θ selected from about 5.040, 15.080, 15.880, 20.860, and 28.540 degrees, The margin of error is about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In yet another embodiment, the crystalline form of the maleate salt of Compound 1 exhibits an XRDP comprising the peaks shown in Table 7 below:

In a specific embodiment, the crystalline form of the maleate salt of Compound 1 exhibits an XRDP substantially similar to that of FIG. 26 .

In a specific embodiment, the crystalline form of the maleate salt of Compound I exhibits a DSC pattern comprising peak characteristic values at about 217.32°C, with a margin of error of about ±2.5; about ±2.0; about ±1.5; about ±1.0; about ±0.5; or less. In a specific embodiment, the crystalline form of the maleate salt of Compound 1 exhibits a DSC pattern substantially similar to that of FIG. 27 .

In one embodiment, the crystalline form of the maleate salt of Compound 1 exhibits a &lt ^;1 >H NMR spectrum substantially similar to Figure 28. In another embodiment, the crystalline form of the maleate salt of Compound I consists of small needle-like cohesive aggregates on a macroscopic scale, which is substantially similar to FIG. 29 . In a specific embodiment, the ratio of maleic acid and Compound I in the maleate salt of Compound I is about 1:1.

Fumarate of compound I

In a specific embodiment, the crystalline form of the fumarate salt of Compound I exhibits an XRDP comprising peaks at about 6.360 and 24.800 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In another specific embodiment, the XRDP of the crystalline fumarate salt of Compound I further comprises at least one, two or three peaks selected from about 19.660, 20.420 and 26.860 degrees, with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In another embodiment, the XRDP of the crystalline fumarate salt of Compound I further comprises one, two, three or four peaks at about 12.680, 17.020, 25.180 and 28.280 degrees 2θ with a margin of error of about ±0.5; about ±0.4 about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In yet another embodiment, the crystalline form of the fumarate salt of Compound 1 exhibits an XRDP comprising the peaks shown in Table 8 below:

In a specific embodiment, the crystalline form of the fumarate salt of Compound 1 exhibits an XRDP substantially similar to that of FIG. 31 .

In a specific embodiment, the crystalline form of the fumarate salt of Compound I exhibits a DSC pattern comprising peak characteristic values at about 222.40°C, with a margin of error of about ±2.5; about ±2.0; about ±1.5; about ±1.0; about ±0.5; or less. In a specific embodiment, the crystalline form of the fumarate salt of Compound 1 exhibits a DSC pattern substantially similar to that of FIG. 32 .

In one embodiment, the crystalline form of the fumarate salt of Compound 1 exhibits a ¹ H NMR spectrum substantially similar to FIG. 33 . In another embodiment, the crystalline form of the fumarate salt of Compound I consists of small needle-like cohesives on a macroscopic scale, which is substantially similar to FIG. 34 . In a specific embodiment, the ratio of fumaric acid and Compound I in the fumarate salt of Compound I is about 1:1.

Citrate of compound I

In a specific embodiment, the crystalline form of the citrate salt of Compound 1 exhibits an XRDP comprising peaks at about 4.900, 25.380 and 27.500 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In another embodiment, the XRDP of the crystalline citrate salt of Compound I further comprises at least one, two, three or four peaks at about 15.360, 18.100, 19.300 and 26.140 degrees 2Θ with a margin of error of about ±0.5; about ±0.4 about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In another specific embodiment, the crystalline citrate salt of Compound I further comprises at least one, two, three or four peaks at about 17.400, 18.680, 24.040 and 26.740 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In yet another embodiment, the crystalline form of the citrate salt of Compound 1 exhibits an XRDP comprising the peaks shown in Table 9 below:

In a specific embodiment, the crystalline form of the citrate salt of Compound 1 exhibits an XRDP substantially similar to FIG. 36 .

In a specific embodiment, the crystalline form of the citrate salt of Compound I exhibits a DSC pattern comprising peak characteristic values at about 196.86°C, with an error range of about ±2.5; about ±2.0; about ±1.5; about ±1.0; about ±1.0; 0.5; or less. In a specific embodiment, the crystalline form of the citrate salt of Compound 1 exhibits a DSC pattern substantially similar to that of FIG. 37 .

In one embodiment, the crystalline form of the citrate salt of Compound 1 exhibits a &lt ^;1 >H NMR spectrum substantially similar to Figure 38. In another embodiment, the crystalline form of the citrate salt of Compound I consists of cohesive aggregates formed in small needles on a macroscopic scale, which is substantially similar to FIG. 39 . In a specific embodiment, the ratio of citric acid and compound I in the citrate salt of compound I is about 1:1.

L-malate of compound I

In a specific embodiment, the crystalline form of the L-malate salt of Compound 1 exhibits an XRDP comprising peaks at about 6.580, 6.780 and 25.560 degrees 2Θ with a margin of error of about ±0.5; about ±0.4; about ±0.3; about ±0.3; 0.2; about ±0.1; about ±0.05; or less. In another specific embodiment, the crystalline L-apple of Compound I The XRDP of the fruit salt further comprises one, two, three, or four peaks at 2Θ selected from the group consisting of: about 19.560, 23.660, 26.060, and 26.960 degrees, with a margin of error of about ±0.7; about ±0.6; about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less. In another specific embodiment, the crystalline L-malate salt of Compound I further comprises one, two, three, four or five peaks at 2Θ selected from the group consisting of: about 8.800, 11.800, 18.600, 24.460 and 25.080 degrees, The margin of error is about ±0.5; about ±0.4; about ±0.3; about ±0.2; about ±0.1; about ±0.05; or less.

In yet another embodiment, the crystalline form of the L-malate salt of Compound 1 exhibits an XRDP comprising the peaks shown in Table 10 below:

In a specific embodiment, the crystalline form of the L-malate salt of Compound 1 exhibits an XRDP substantially similar to that of FIG. 41 .

In a specific embodiment, the crystalline form of the L-malate salt of Compound I exhibits a DSC pattern comprising peak characteristic values at about 209.67°C, with a margin of error of about ±2.5; about ±2.0; about ±1.5; about ±1.0; about ±0.5; or less. In a specific embodiment, the crystalline form of the L-malate salt of Compound 1 exhibits a DSC pattern substantially similar to that of FIG. 42 .

In one embodiment, the crystalline form of the L-malate salt of Compound 1 exhibits a ¹ H NMR spectrum substantially similar to FIG. 43 . In another embodiment, the crystalline form of the L-malate salt of Compound I consists of small needle-like agglomerates on a macroscopic scale, which is substantially similar to FIG. 44 . In a specific embodiment, the ratio of L-malic acid and Compound I in the L-malate salt of Compound I is about 1:1.

Pharmaceutical preparations

In another specific embodiment, the present invention provides a pharmaceutical composition comprising, as an active ingredient, a therapeutically effective amount of a crystalline form of Compound I as described herein, or a pharmaceutically acceptable salt, ester and/or solvate thereof , in combination with a pharmaceutically acceptable excipient or carrier. The excipients are added to formulations for various purposes.

Diluents may be added to the formulations of the present invention. Diluents can increase the bulk of a solid pharmaceutical composition and can make it easier for patients and caregivers to handle the pharmaceutical dosage form containing the composition. for solid state combinations Diluents for compounds include, for example, microcrystalline cellulose (such as AVICEL), microfine cellulose, lactose, starch, pregelatinized starch, calcium carbonate, calcium sulfate, sugar, glucose binders, dextrin, glucose, phosphoric acid dihydrate Calcium hydrogen, tricalcium phosphate, kaolin, magnesium carbonate, magnesium oxide, maltodextrin, mannitol, polymethacrylates (eg, EUDRAGIT®)), potassium chloride, powdered cellulose, sodium chloride, sorbitol and talc.

Solid pharmaceutical compositions compressed into dosage forms, such as lozenges, may contain excipients whose functions include, after compression, aiding in binding of the active ingredient with other excipients. Binders used in solid pharmaceutical compositions include gum arabic, alginic acid, carbomer (eg, carbopol), sodium carboxymethyl cellulose, dextrin, ethyl cellulose, gelatin, guar gum, Gum tragacanth, hydrogenated vegetable oil, hydroxyethyl cellulose, hydroxypropyl cellulose (eg, KLUCEL), hydroxypropyl methylcellulose (eg, METHOCEL), liquid glucose, magnesium aluminum silicate, maltodextrin, methyl methacrylate Base cellulose, polymethacrylate, povidone (eg, KOLLIDON, PLASDONE), pregelatinized starch, sodium alginate, and starch.

The rate of dissolution of the compressed solid pharmaceutical composition in the patient's stomach can be increased by adding a disintegrant to the composition. Disintegrants include alginic acid, calcium carboxymethylcellulose, sodium carboxymethylcellulose (eg, AC-DI-SOL and PRIMELLOSE), colloidal silica, croscarmellose sodium, crospovidone (eg, AC-DI-SOL and PRIMELLOSE) , KOLLIDON and POLYPLASDONE), guar gum, magnesium aluminum silicate, methylcellulose, microcrystalline cellulose, polacrilin potassium, powdered cellulose, pregelatinized starch, sodium alginate, sodium starch glycolate (such as , EXPLOTAB), potato starch and starch.

Glidants can be added to improve the flow of uncompressed solid compositions and to improve the precision of administration. Excipients that function as glidants include colloidal silica, magnesium trisilicate, powdered cellulose, starch, talc, and tricalcium phosphate.

When a dosage form such as a lozenge is made by compressing a powdered composition, the composition is subjected to the pressure of a punch and die. Some excipients and active ingredients have a tendency to stick to punch and die surfaces, which can result in pits and other surface irregularities in the finished product. Lubricants can be added to the composition to reduce sticking and make the finished product easier to release from the die. Lubricants include magnesium stearate, calcium stearate, glyceryl monostearate, glyceryl palmitate stearate, hydrogenated castor oil, hydrogenated vegetable oil, mineral oil, polyethylene glycol, sodium benzoate, sodium lauryl sulfate , Sodium stearoyl fumarate, stearic acid, talc and zinc stearate.

Flavoring and aroma enhancing agents make the dosage form more palatable to the patient. The compositions of the present invention may contain flavoring agents and flavor enhancers commonly used in pharmaceutical products, including maltol, vanillin, ethyl vanillin, menthol, citric acid, fumaric acid, ethyl maltol, and tartaric acid .

Solid and liquid compositions may also be colored with any pharmaceutically acceptable colorant to improve their appearance and/or to facilitate patient identification of the product and unit dose.

Liquid pharmaceutical compositions can be prepared using the crystalline forms of the present invention as well as any other solid excipients wherein the ingredients are dissolved or suspended in a liquid carrier such as water, vegetable oils, alcohols, polyethylene glycol, propylene glycol or glycerol.

Liquid pharmaceutical compositions may contain emulsifiers to uniformly disperse active ingredients or other excipients that are insoluble in the liquid carrier throughout the composition. Emulsifiers useful in the liquid compositions of the present invention include, for example, gelatin, egg yolk, casein, cholesterol, acacia, tragacanth, carrageen, pectin, methylcellulose, carbomer, cetearyl alcohol and cetyl alcohol.

Liquid pharmaceutical compositions may also contain viscosity enhancers to improve the mouthfeel of the product and/or to form a film in the gastrointestinal tract. Viscosity enhancers include gum arabic, bentonite alginate, carbomer, calcium or sodium carboxymethyl cellulose, cetearyl alcohol, methyl cellulose, ethyl cellulose, gelatin guar, hydroxyethyl cellulose , hydroxypropyl cellulose, hydroxypropyl methylcellulose, maltodextrin, polyvinyl alcohol, povidone, Propylene carbonate, propylene glycol alginate, sodium alginate, sodium starch glycolate, starch tragacanth and xanthan gum.

Sweeteners such as aspartame, lactose, sorbitol, saccharin, sodium saccharin, sucrose, aspartame, fructose, mannitol and invert sugar may be added to improve taste.

Safe intake levels of preservatives and chelating agents such as alcohol, sodium benzoate, butylated hydroxytoluene, butylated hydroxyanisole, and EDTA may be added to improve storage stability.

Liquid compositions may also contain buffers such as glycolic, lactic, citric or acetic acid, sodium gluconate, sodium lactate, sodium citrate or sodium acetate. The choice and amount of excipients can be easily determined by pharmaceutical scientists based on experience and consideration of standard procedures and reference practices in the field.

Solid compositions of the present invention include powders, granules, aggregates and compressed compositions. Dosage forms include those suitable for oral, buccal, enteral, parenteral (including subcutaneous, intramuscular, and intravenous), inhalation, and ocular administration. Although the most appropriate mode of administration in any given situation will depend on the nature and severity of the condition being treated, the preferred route of the present invention is oral. The dosage forms may conveniently be presented in a single dosage form and prepared by any of the methods known in the art of medicine.

Dosage forms include solid dosage forms such as lozenges, powders, capsules, suppositories, sachets, lozenges and lozenges, as well as liquid syrups, suspensions, aerosols and elixirs.

The dosage form of the present invention can be a capsule containing the composition in a hard or soft shell, and the composition is preferably a powder or granular solid of the composition of the present invention. The shell can be made of gelatin and optionally contains plasticizers such as glycerol and sorbitol and opacifying or coloring agents.

Compositions for tableting or capsule filling can be prepared by wet granulation. In wet granulation, some or all of the active ingredients and excipients are mixed in powder form and then further mixed in a liquid (usually water), which causes the powder to agglomerate into granules. Sieve and/or grind and dry the particles, It is then screened and/or ground to the desired particle size. The granules can be tableted, or other excipients can be added prior to tableting, such as glidants and/or lubricants.

Tablet compositions can be conventionally prepared by dry blending. For example, a mixed composition of active substance and excipients can be compressed into blocks or flakes and then comminuted into compressed granules. The compressed granules can then be compressed into lozenges.

As an alternative to dry granulation, the blended composition can be directly compressed into a compressed dosage form using direct compression techniques. Direct compression produces a more uniform tablet free of particles. Excipients particularly suitable for direct compression tableting include microcrystalline cellulose, spray dried lactose, dicalcium phosphate dihydrate and colloidal silica. Those experienced and skilled in the art with specific formulation challenges for direct compression tableting are known to use these and other excipients appropriately in direct compression tableting.

The capsule fills of the present invention may contain any of the aforementioned mixing and granulation (described with reference to tableting); however, they do not have a final tableting step.

Active ingredients and excipients can be formulated into compositions and dosage forms according to methods known in the art.

In one embodiment, the pharmaceutical dosage form can be provided as a kit of reagents comprising, as individual components, the crystalline form of Compound I, together with pharmaceutically acceptable excipients and carriers thereof. In some embodiments, the dosage form kit allows physicians and patients to formulate oral or injectable solutions by dissolving, suspending or mixing the crystalline form of Compound I with pharmaceutically acceptable excipients and carriers prior to use. In one embodiment, a pharmaceutical dosage form kit is provided of a crystalline form of Compound 1 having improved stability of Compound 1 as compared to preformulated liquid formulations of Compound 1.

The formulations of the present invention do not necessarily contain only one crystalline form of Compound I. The crystalline form of the present invention is used as a single ingredient or in admixture with other crystalline forms of Compound I in pharmaceutical formulations or formulations in the compound. In a specific embodiment, the pharmaceutical preparation or composition of the present invention contains 25-100% or 50-100% by weight of at least one crystalline form of Compound I according to the present invention in the preparation or composition.

therapeutic use

The present invention also provides therapies for cell proliferation-related diseases. In one aspect, the present invention provides a method for selectively activating a p53 protein comprising contacting a cell affected by a cell proliferation-related disease with the compound. In a specific embodiment, the method comprises contacting cancer and/or tumor cells with a crystalline form of Compound I as described herein, or a pharmaceutically acceptable salt, ester and/or solvate thereof. In another specific embodiment, the method of contacting cancer and/or tumor cells with a crystalline form of Compound I or a pharmaceutically acceptable salt, ester and/or solvate thereof of the present invention can induce apoptosis or alleviating or preventing disease progression.

In addition, methods for treating cancer, cancer cells, tumors or tumor cells are disclosed. Non-limiting examples of cancers that can be treated by the methods of the present invention include cancers or cancer cells of the following: colon, breast, lung, liver, pancreas, lymph nodes, colon, prostate, brain, head and neck, skin, ovary, cervix, Thyroid, bladder, kidney, and blood and heart (eg, leukemias, lymphomas, and malignancies). Non-limiting examples of tumors that can be treated by the methods of the invention include the following tumors and tumor cells: large intestine, breast, lung, liver, pancreas, lymph nodes, colon, prostate, brain, head and neck, skin, kidney, and blood and heart ( For example, leukemias, lymphomas, and malignancies).

The present invention also provides methods of treating, preventing, alleviating and/or reducing the progression of a disease or disorder characterized by cell proliferation in an individual. More specifically, the methods of the present invention involve administering to an individual an effective amount of a crystalline form of a quinolone compound as described herein to treat a disease or disorder characterized by cell proliferation. Administration of the crystalline form is effective in selectively activating p53 in cancer and/or tumor cells The amount of protein that can lead to cell death or apoptosis. The terms "individual" and "patient" are used interchangeably herein.

As described herein, drug administration may be accomplished or performed using any of a variety of methods known to those skilled in the art. Crystalline forms may be formulated in dosage forms containing generally non-toxic, physiologically acceptable carriers or vehicles, and administered, for example, subcutaneously, intravenously, parenterally, intraperitoneally, intradermally, intramuscularly, topically, Enteral (eg, oral), rectal, nasal, buccal, sublingual, vaginal, by inhalation spray, by drug pump, or via implanted reservoir.

In addition, the crystalline forms disclosed herein can be administered to a localized area in need of treatment. This can be accomplished by, for example, but not limited to, local infusion during surgery, topical administration, transdermal patches, by injection, by catheter, by suppository, or by implant (implant optional). porous, non-porous or gelatinous materials) including membranes, such as silicone rubber membranes or fibers.

The form in which the crystalline form is administered (eg, syrup, elixirs, capsules, lozenges, foams, emulsions, gels, etc.) will depend in part on the route of administration. For example, for mucosal (eg, oral mucosa, rectal, intestinal mucosa, bronchial mucosa) administration, nasal drops, aerosols, inhalants, nebulizers, eye drops, or suppositories can be used. Crystalline forms can also be used to coat bioimplantable materials to enhance neurite outgrowth, nerve survival, or cell interaction with the implant surface. The crystalline forms disclosed herein can be administered with other biologically active agents that control the symptoms or causes of one or more diseases or disorders characterized by cell proliferation, such as anticancer agents, analgesics, anti-inflammatory agents , anesthetics and other drugs.

In a specific embodiment, a crystalline form of Compound 1 or a crystalline form of a pharmaceutically acceptable salt, ester and/or solvate of Compound 1 as described herein can be administered in combination with one or more therapeutically active agents . In a specific embodiment, the one or more therapeutically active agents are anticancer agents. in part In specific embodiments, the one or more therapeutically active anticancer agents include, but are not limited to, paclitaxel, vinblastine, vincristine, etoposide, doxorubicin, Herceptin, lapatinib, gefitinib, Erlotinib, Tamoxifen, Fulvestrant, Anastrozole, Letrozole, Exemestane, Fadrozole, Cyclophosphamide, Taxotere, Melphalan, Benbutyrate Nitrogen mustard, nitrogen mustard, chlorambucil, phenylalanine, mustard, cyclophosphamide, ifosfamide, carmustine (BCNU), lomustine (CCNU), streptozotocin, white Paraben, Thiatepa, Cisplatin, Carboplatin, Actinomycin D, Doxorubicin (Adriamycin), Daunorubicin, Idarubicin, Mitoxantrone, Pukamycin, Silk Schizomycin, C bleomycin, combinations thereof, and analogs thereof. In another specific embodiment, the one or more therapeutically active anticancer agents include, but are not limited to, PARP (poly(DP-ribose) polymerase) inhibitors. Suitable PARP inhibitors include, but are not limited to, 4-(3-(1-(cyclopropanecarbonyl)piperidine-4-carbonyl)-4-fluorobenzyl)phthalazin-1(2H)-one (olaparib, AZD2281, Ku-0059436), 2-[(2R)-2-methylpyrrolidin-2-yl]-1H-benzimidazole-4-carboxamide (Veliparib, ABT-888), (8S, 9R)-5- Fluoro-8-(4-fluorophenyl)-9-(1-methyl-1H-1,2,4-triazol-5-yl)-8,9-dihydro-2H-pyrido[4, 3,2-De]phthalazinyl-3H-(7H)-one (talazoparib, BMN 673), 4-iodo-3-nitrobenzamide (iniparib, BSI-201), 8-fluoro-5-( 4-((Dimethylamino)methyl)phenyl)-3,4-dihydro-2H-aza[5,4,3-cd]indol-1(6H)-one phosphate (Rucaparib, AG-014699, PF-01367338), 2-[4-[(dimethylamino)methyl]phenyl]-5,6-dihydro-imidazo[4,5,1-JK][1,4 ] Benzodiazepine 7(4H)-one (AG14361), 3-aminobenzamide (INO-1001), 2-(2-fluoro-4-((S)-pyrrolidin-2-yl)benzene yl)-3H-benzo[d]imidazole-4-carboxamide (A-966492), N-(5,6-dihydro-6-oxo-2-phenanthridine)-2-acetamide hydrochloride (PJ34, PJ34 HCl), MK-4827, 3,4-dihydro-4-oxo-3,4-dihydro-4-oxo-N-[(1S)-1-phenethyl]-2 - Quinazoline propane (ME0328), 5-(2-oxo-2-phenethyl)-1(2H)-isoquinoline (UPF-1069), 4-[[4-fluoro-3-[( 4-Methoxy-1-piperidinyl)carbonyl]phenyl]methyl]-1(2H)-phthalazinone (AZD 2461) and analogs thereof. exist In another specific embodiment, the one or more therapeutically active agents are immunotherapeutic agents. In some embodiments, the one or more immunotherapeutic agents include, but are not limited to, monoclonal antibodies, immune effector cells, adoptive cell transfer, immunotoxins, vaccines, cytokines, and the like.

In another specific embodiment, a crystalline form of Compound 1 or a crystalline form of a pharmaceutically acceptable salt, ester and/or solvate of Compound 1 as described herein can be administered in combination with radiation therapy.

Furthermore, administering may comprise administering to the individual multiple doses over a suitable period of time. Such drug administration regimens can be determined according to conventional methods after reading this document.

The crystalline forms of the present invention are generally administered at a dose of about 0.01 mg/kg/dose to about 100 mg/kg/dose. Alternatively, the dosage may be from about 0.1 mg/kg/dose to about 10 mg/kg/dose; or about 1 mg/kg/dose to 10 mg/kg/dose. Time-controlled release formulations may be employed or, where convenient, the medicament may be administered in multiple doses. When using other methods (eg, intravenous administration), the crystalline form is administered to the affected tissue at a rate of from about 0.05 to 10 mg/kg/hour, or from about 0.1 to about 1 mg/kg/hour. This rate can be easily maintained when the crystalline form is administered intravenously as discussed herein. In general, formulations for topical administration are administered in doses ranging from about 0.5 mg/kg/dose to about 10 mg/kg/dose. Alternatively, topical formulations are administered at a dose of about 1 mg/kg/dose to about 7.5 mg/kg/dose, or even at a dose of about 1 mg/kg/dose to about 5 mg/kg/dose.

A range from about 0.1 to about 100 mg/kg is a suitable single dose. A suitable continuous administration range is from about 0.05 to about 10 mg/kg.

Drug doses can also be given in milligrams per square meter of body surface area rather than body weight, as this method correlates well with specific metabolic and excretory functions. In addition, body surface area can be used as a common denominator for drug doses in adults and children as well as between different animal species (Freireich et al., (1966) Cancer Chemother Rep. 50, 219-244). Briefly, the dose in mg/kg can be expressed as the equivalent mg/sq m dose in any given species by multiplying the dose by the appropriate km factor. In adults, 100 mg/kg is equivalent to 100 mg/kg x 37 kg/sq m = 3700 mg/m ² .

The dosage forms of the present invention may contain Compound I as described herein, or a pharmaceutically acceptable salt, ester and/or solvate thereof, in an amount from about 5 mg to about 500 mg. That is, the dosage form of the present invention may contain Compound I in an amount of about 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90mg, 95mg, 100mg, 110mg, 120mg, 125mg, 130mg, 140mg, 150mg, 160mg, 170mg, 175mg, 180mg, 190mg, 200mg, 210mg, 220mg, 225mg, 230mg, 240mg, 250mg, 260mg, 270mg, 275mg, 280mg, 290mg, 300mg, 310mg, 320mg, 325mg, 330mg, 340mg, 350mg, 360mg, 370mg, 375mg, 380mg, 390mg, 400mg, 410mg, 420mg, 425mg, 430mg, 440mg, 450mg, 460mg, 470mg, 475mg, 480mg, 490mg or 500mg. In a specific embodiment, the above-described doses are administered to a patient, either as a daily dose in a single dose, or in multiple daily divided doses, such as two, three, or four times daily.

The dosage forms of the present invention can be administered hourly, daily, weekly or monthly. The dosage forms of the present invention may be administered twice daily or once daily. The dosage forms of the present invention may be administered with or without food.

While the crystalline forms disclosed herein may take the form of mimetics or fragments thereof, it should be understood that the potency, and thus the dosage of the effective amount, may vary. However, one skilled in the art can readily analyze the efficacy of the type of crystalline form contemplated by the present invention.

In the case of progressive diseases or disorders characterized by cell proliferation, the crystalline forms of the present invention are administered on a consistent basis. In certain cases, it may be prescribed before symptoms develop The crystalline form of the present invention is initially administered as part of a strategy to delay or prevent disease. In other instances, the crystalline forms of the invention may be administered after the onset of disease symptoms, as part of a strategy to slow or reverse disease progression, and/or as part of a strategy to improve cellular function and alleviate symptoms.

It will be understood by those skilled in the art that dosage ranges will depend on the particular crystalline form and its potency. It should be understood that the dosage range is large enough to produce the desired effect, which relieves neurodegenerative or other diseases and their associated symptoms, and/or cell survival thereof, but is not so large as to cause unmanageable adverse side effects . It should be understood, however, that the specific dosage for any particular patient will depend on a variety of factors, including the activity of the particular crystalline form used; the age, weight, health, sex and diet of the subject being treated; time and route of administration; rate of excretion ; other previously administered drugs; and the severity of the particular disease being treated, as well understood by those skilled in the art. The dose may also be adjusted by the individual physician in case of any complications. No unacceptable toxicological effects are expected when the crystalline forms disclosed herein are used in accordance with the present invention.

Effective amounts of the crystalline forms disclosed herein include amounts sufficient to produce a measurable biological response. The actual dosage of the active ingredient of the therapeutic crystalline form of the present invention may vary in order to administer to a particular individual and/or application an amount of the active crystalline form effective to achieve the desired therapeutic response. Preferably, the minimum dose is administered and the dose is escalated in the absence of dose-limiting toxicity to the minimum effective dose. Determination and adjustment of therapeutically effective amounts, as well as assessment of when and how to make such adjustments, are well known to those of ordinary skill in the art.

Further in relation to the method of the present invention, the preferred subject is a vertebrate subject. Preferred vertebrates are warm-blooded; a preferred warm-blooded vertebrate is a mammal. The subject to be treated by the methods disclosed herein is preferably a human, but it is understood that the principles of the present invention are intended to be effective against all vertebrate species, which are included in the term "subject". In this regard, vertebrate shall be understood as any Vertebrate species that are interested in the treatment of neurodegenerative disorders. The term "individual" as used herein includes human and animal individuals. Thus, according to the present invention, veterinary therapeutic use is also provided.

Thus, the therapy provided by the present invention can be directed to mammals such as humans, as well as to mammals of endangered importance, such as the Siberian tiger; animals of economic importance, such as farm-raised human food animals. ; and animals of social importance to humans, such as animals kept as pets or in zoos or farms. Examples of such animals include, but are not limited to: carnivores, such as cats and dogs; pigs, including domestic pigs, hogs, and wild boars; ruminants and/or ungulates, such as livestock, bulls, sheep, giraffes, deer, goats, Bison and camels; and horses. Also provides therapy for birds, including those that are endangered and/or kept in zoos, and birds, especially domesticated birds, i.e. poultry such as turkeys, chickens, ducks, geese, guinea fowls, etc., Because they are also of economic importance to human beings. Accordingly, therapy is also provided for livestock including, but not limited to, domesticated pigs, ruminants, ungulates, horses (including racehorses), poultry, and the like.

The following examples may further illustrate the present invention, but should not be construed in any way to limit its scope.

Example

Analytical Methods - Various analytical methods, described below, are applicable to the crystalline forms of the present invention and their precursors, respectively, to characterize their physiochemical properties.

Differential Scanning Thermal Analysis (DSC):

DSC data were obtained on a DSC-system (DSC 822e-Mettler Toledo) or on a TA Instruments Q2000. In general, samples in the mass range of 1 to 5 mg were placed in dry aluminum crucibles and closed using an aluminum upper lid with a hole. In general, the starting temperature was 20°C, the heating rate was 10°C/min, and the final temperature was 300°C, using a nitrogen purge flow of 20 mL/min.

Thermogravimetric Analysis (TGA):

TGA data were collected on a TGA 851e device. In general, samples in the mass range of about 10 mg were placed in lean and dry alumina pans or aluminum pans. Typically, the sample pan was scanned between -30°C to about 350°C at 5°C/min or at 10°C/min using a nitrogen purge flow rate of 50mL/min.

TGA analysis was also performed using a TA Instruments Discovery IR thermogravimetric analyzer. The temperature was calibrated using nickel and Alumel (Alumel ^™ ). Place the sample in an aluminum pan. The sample is hermetically sealed, the lid pierced, and inserted into the thermogravimetric oven. The furnace is heated by nitrogen. The samples for analysis were heated from room temperature to 350°C at a heating rate of 10°C/min. Trios software v3.1.2.3591 was used to generate maps.

Hydrogen NMR ( ¹ H NMR):

¹ H NMR spectra were acquired on a Bruker AVANCE 400 MHz nuclear magnetic resonance apparatus in DMSO-d ₆ or CDCl ₃ using tetramethylsilane as an internal standard.

Hot Stage Optical Microscope (HSM):

Olympus BX41 with Di-Li 5MP camera and grab&measure software, used with Hostage Mettler Toledo FP90 with FP 82 heating stage. The sample is prepared as a brush and placed in the object holder. Observe using unpolarized light or polarized light using both polarized lights at 40, 100, 200 or 400 x magnification. Photos taken with software and exported as JPEGs, dimensions are approximate and not verified.

X-ray Powder Diffraction (XRPD):

XRPD patterns were acquired using a MiniFlex (Rigaku Corporation) using a silicon low background sample holder (24 mm diameter, 0.2 mm aperture). Collection time is typically 75 minutes. The samples were irradiated with a light source of Cu Kα radiation of 1.5406 Angstroms operating at 15 kV. The effective 2Θ range is about 2 to 40°C with a sampling width of 0.02°C. The samples were ground in a mortar and pestle. The solid was placed in a greased sample holder and flattened with a glass plate.

High performance liquid chromatography (HPLC) analysis:

Crystalline forms (ie, salts and free bases) of the present invention were analyzed by total area normalization (TAN).

HPLC conditions:

HPLC column: Phenomenex Luna8, 3μm C18, 4.6 x 50mm

Flow rate: 1.0mL/min

Injection volume: 5 μL

Detection: DAD detector, recording at 240nm

Mobile phase: AH ₂ O+0.05% CF ₃ COOH

B-CAN+0.05% CF ₃ COOH

Gradient pump program:

Example 1. 2-(4-Methyl-[1,4]diazepan-1-yl)-5-oxo-5H-7-thia 1,11bdiaza-benzo[c] Solubility of Fluorene-6-carboxylic acid (5-methyl-pyridin-2-ylmethyl)-amide (Compound I, free base):

The solubility of Compound 1 was determined by suspending Compound 1 in various solvents heated at elevated temperature and then cooled to room temperature. The mother liquor was sampled and the concentration of Compound I was determined by HPLC.

Example 2. Preparation of polymorph E of compound I (free base):

Compound I (free base) was dissolved in the solvent mixture to form a solution, which was then concentrated to a suspension. The suspension was then concentrated to dryness. An anti-solvent was added to the suspension, which was concentrated to dryness again. XRPD analysis confirmed the formation of polymorph E and showed that polymorph E was crystalline (Figure 11). The dry content was measured to be 99.97% w/w. ¹ H NMR (Figure 13) confirmed the structure of this material to be the free base of the compound.

Example 3. Preparation of Polymorph A of Compound 1 (free base):

Polymorph E of Compound I (free base), prepared as in Example 2, was suspended in the solvent mixture. The suspension was heated and then cooled to room temperature. The suspension was filtered and the resulting filter cake was dried to give polymorph A. XRPD analysis confirmed the formation of the polymorph and showed that polymorph A was crystalline (Figure 1). The dry content was measured to be 99.86% w/w. ¹ H NMR (Figure 5) confirmed the structure of this material to be the free base of the compound.

The DSC pattern (Figure 2) exhibited a sharp endothermic peak at about 215°C, followed by an exothermic peak at about 217°C, an endothermic peak at about 231°C, an exothermic peak at about 233°C, and an endothermic peak at about 242°C. Without being bound by any theory, these events are consistent with one melting and subsequent recrystallization, another melting and recrystallization, and a final melting event. The TGA profile (Figure 3) shows a weight loss of about 0.3% from ambient temperature to 300°C. Figure 4 shows an overlay of the DSC spectrum and the TGA spectrum.

Example 4. Preparation of Polymorph C of Compound 1 (free base):

Polymorph A of Compound I was suspended in an organic solvent at elevated temperature, the mother liquor was filtered, cooled to form a suspension, which was filtered again to obtain a white needle-like solid. ^<1 >H NMR (Figure 9) confirmed the structure of this material to be the free base of compound I.

XRPD analysis showed that polymorph C was crystalline, as shown in FIG. 7 .

Example 5. Preparation of Polymorph G of Compound I (free base):

Polymorph A of Compound I is suspended in another organic solvent at elevated temperature. The suspension was filtered and the mother liquor was evaporated to form a suspension which was filtered to give a pale yellow solid. ^<1 >H NMR (Figure 18) confirmed the structure of this material to be the free base of compound I.

XRPD analysis showed that polymorph G was crystalline, as shown in FIG. 16 .

Example 6. Solubility and Stability Testing of Polymorph Forms:

The solubility of polymorphs A, C, E and G of Compound I (free base) was determined by suspending 25 mg of each polymorph in an organic solvent and refrigerating the resulting mixture at room temperature or elevated temperature Stir for 1 hour. The sample was filtered and concentrated and determined by HPLC. The remaining suspension was evaporated and the solids were analyzed by XRPD to confirm that the polymorph form in the suspension was still the same as the polymorph form used in the experiments.

The solubility of each polymorph is shown in Figure 20. Polymorph A exhibited the lowest solubility and was confirmed to be the most stable.

Example 7. Preparation of the hydrochloride salt of compound I:

Compound I was dissolved in the solvent mixture at elevated temperature, followed by addition of hydrochloric acid. The suspension was stirred overnight, then filtered and washed to obtain the hydrochloride salt of Compound I as a white powder (62% yield). The resulting salt was characterized ^by1H NMR (FIG. 23), XRPD (FIG. 21), and TGA (FIG. 25). TGA showed that the hydrochloride salt of Compound I exhibited a loss of up to 0.9 wt% at 140°C.

Example 8. Preparation of the maleate salt of compound I:

Compound I was dissolved in the solvent mixture at elevated temperature and maleic acid was then added to the mixture. The resulting suspension was cooled and stirred, filtered and washed to give the maleate salt of compound I as a white powder (86% yield). The resulting salt was characterized as shown by ¹ H NMR (FIG. 28), XRPD (FIG. 26) and TGA (FIG. 30). TGA showed that the maleate salt of Compound I exhibited a loss of up to 2.9 wt% at 140°C. Without being bound by any theory, this weight loss may be due to the loss of solvent.

Example 9. Preparation of the Fumarate Salt of Compound I:

Compound I was dissolved in the solvent mixture at elevated temperature and fumaric acid was then added to the mixture. The resulting suspension was cooled and stirred, filtered and washed to give the fumarate salt of compound I as a white powder (104% yield). The resulting salt was characterized as shown ^by1H NMR (Figure 33), XRPD (Figure 31) and TGA (Figure 35). TGA showed that the fumarate salt of Compound I exhibited a loss of up to 6.6 wt% at 140°C.

Example 10. Preparation of the Citrate Salt of Compound 1:

Compound I was dissolved in the solvent mixture at elevated temperature and citric acid was then added to the mixture. The resulting suspension was cooled and stirred, filtered and washed to give the citrate salt of compound I as a white powder (93% yield). The resulting salt was characterized as shown ^by1H NMR (Figure 38), XRPD (Figure 36) and TGA (Figure 40). TGA showed that the citrate salt of Compound I exhibited a loss of up to 5.2 wt% at 140°C.

Example 11. Preparation of the L-malate salt of compound I:

Compound I was dissolved in the solvent mixture at elevated temperature, and then L-malic acid was added to the mixture. The resulting suspension was cooled and stirred, filtered and washed to give the citrate salt of compound I as a white powder (59% yield). The resulting salt was characterized as shown ^by1H NMR (Figure 43), XRPD (Figure 41) and TGA (Figure 45). TGA showed that the L-malate salt of Compound I exhibited a loss of up to 4.7 wt% at 140°C.

Example 12. Solubility and Stability Test of Acid Salt of Compound I:

The solubility of the hydrochloride, maleate, fumarate, citrate and L-malate salts of Compound I was determined by suspending approximately 50 mg of each salt in 0.25 mL of water (x2, added in portions). . The mixture was stirred at 42°C for 3 days and each solution was analyzed for concentration and purity by HPLC. After 3 days of testing, the solubility and purity of various salts are shown in Table 11 below.

The stability of the acid salts was also tested by storing the salts (solid form) at elevated temperatures of 45°C and 80°C. The results of the stability test are shown in Table 12 below.

NT=未測試

NT=Not Tested

The patents and publications listed herein are based on the ordinary skill in the art to which the invention pertains. All patents and publications are hereby incorporated by reference to the same extent as if each individual publication were specifically and individually indicated to be incorporated by reference. . In the event of any conflict between the cited reference and this specification, the present specification shall control. In describing specific embodiments of the present invention, specific terminology is employed for clarity. However, the invention is not intended to be limited to the specific terminology so selected. Nothing in this specification should be construed as limiting the scope of the invention. All examples presented are representative and not limiting. The specific embodiments described above may be modified or varied without departing from the invention in light of the above teachings and as understood by those skilled in the art. It is therefore to be understood that within the scope of the patent application and its equivalents, the invention may be practiced otherwise than as specifically described herein.

Claims (23)

a crystalline form of Compound I,

Wherein, the crystalline form is the polymorph A of compound I, and the X-ray powder diffraction pattern (XRDP) displayed by it comprises peaks at 2θ of about 7.730±0.3, 22.050±0.3 and 24.550±0.3 degrees, and each peak has The intensity is greater than 50% of the strongest peak. The crystalline form of claim 1, wherein the X-ray powder diffraction pattern further comprises peaks at (a) about 9.410 ± 0.3 and 27.700 ± 0.3 degrees 2Θ; and/or (b) about 17.950 ± 0.3 and 25.400 ± 0.3 degrees 2Θ . The crystalline form of claim 1 or 2, wherein the X-ray powder diffraction pattern further comprises one or more peaks selected from the group consisting of peaks at about 11.230±0.3, 11.630±0.3, 16.900±0.3, 18.580±0.3 at 2θ , 23.300±0.3 and 26.700±0.3 degrees. The crystalline form of claim 1 or 2, which exhibits an X-ray powder diffraction pattern comprising three or more peaks selected from the group consisting of: peaks at about 7.730±0.3, 9.410±0.3 at 2θ , 11.230±0.3, 11.630±0.3, 16.900±0.3, 17.950±0.3, 18.580±0.3, 22.050±0.3, 23.300±0.3, 24.550±0.3, 25.400±0.3, 26.700±0.3 and 27.700±0.3 degrees. The crystalline form of claim 3, which exhibits a differential scanning calorimetry (DSC) pattern having a peak characteristic value at about 215.41±2.0°C. The crystalline form of claim 1 having a purity of about 95% or greater. A crystalline form as claimed in claim 1 which exhibits an X-ray powder diffraction pattern substantially similar to that of FIG. 1 . a crystalline form of the hydrochloride salt of Compound I,

Among them, the X-ray powder diffraction pattern (XRDP) displayed by the crystalline form includes peaks at 2θ of about 4.660±0.3 and 24.540±0.3, and each peak has an intensity greater than 50% of the strongest peak. The crystalline form of claim 8, wherein the displayed DSC pattern has a peak characteristic value at about 266.27±2.0°C; and/or the displayed X-ray powder diffraction pattern is substantially similar to FIG. 21 . The crystalline form of claim 8, wherein the X-ray powder diffraction pattern further comprises: (a) one or more peaks at about 19.260±0.4, 20.160±0.4, 24.920±0.3 and 26.360±0.35 degrees 2θ; and/or (b) One or more peaks at about 13.980±0.4, 14.540±0.3, 25.380±0.3 and 28.940±0.3 degrees 2θ. The crystalline form of claim 8, having a purity of about 95% or greater. A pharmaceutical composition comprising the crystalline form of any one of claims 1-11 and at least one pharmaceutically acceptable carrier. A use of the crystalline form of any one of claims 1-11 for: (a) the preparation of a medicament that stabilizes G-quadruplexes (G4s); and/or (b) the preparation of a medicament that modulates p53 activity. A use of the crystalline form of any one of claims 1-11 for the manufacture of a medicament for the treatment or amelioration of cancer. The use of claim 14, wherein the cancer is selected from the group consisting of: blood cancer, colorectal cancer, breast cancer, lung cancer, liver cancer, ovarian cancer, cervical cancer, Ewing's sarcoma, pancreatic cancer, lymph node cancer , colon cancer, prostate cancer, brain cancer, head and neck cancer, skin cancer, kidney cancer and heart cancer. The use of claim 15, wherein the blood cancer is selected from the group consisting of leukemia, lymphoma, myeloma and multiple myeloma. The use of claim 15, wherein the cancer is a homologous recombination (HR) DNA double-strand break (DSB) repair-deficient cancer, or a non-homologous end joining (NHEJ) DNA double-strand break repair-deficient cancer. The use of claim 15, wherein the cancer is contained in cancer cells with defects in breast cancer susceptibility gene 1 (BRCA1), breast cancer susceptibility gene 2 (BRCA2) and/or other members of the homologous recombination pathway . The use of claim 18, wherein the cancer cells are deficient in BRCA1 and/or BRCA2. The use of claim 19, wherein the cancer cells are homozygous for the genotype with BRCA1 and/or BRCA2 mutation. The use according to claim 19, wherein the cancer cells are heterozygous for the genotype with BRCA1 and/or BRCA2 mutation. The use of any of claims 14-18, wherein the medicament is for co-administration with one or more additional therapeutic agents and/or radiation therapy. The use of claim 22, wherein the one or more additional therapeutic agents are anticancer or immunotherapeutic agents.

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