CN101048398A - Polymorphic forms of 6-[2-(methylcarbamoyl)phenylsulfanyl]-3-E-[2-(pyridin-2-yl)ethenyl]indazole - Google Patents

Abstract

The present invention relates to a novel polymorphic form of 6-[2-(methylcarbamoyl)phenylsulfanyl]-3-E-[2-(pyridin-2-yl)vinyl]indazole and a process for its preparation . Such polymorphic forms may be components of pharmaceutical compositions and may be used to treat hyperproliferative disorders or mammalian conditions mediated by protein kinase activity.

Description

Polymorphs of 6-[2-(methylcarbamoyl)phenylsulfanyl]-3-E-[2-(pyridin-2-yl)vinyl]indazole

This application claims priority to US Provisional Application No. 60/624,665, filed November 2, 2004, which is hereby incorporated by reference in its entirety.

Technical field

The present invention relates to novel polymorphic forms of 6-[2-(methylcarbamoyl)phenylsulfanyl]-3-E-[2-(pyridin-2-yl)vinyl]indazole and processes for their preparation . The invention also relates to pharmaceutical compositions containing at least one polymorphic form and the therapeutic or prophylactic use of said polymorphic forms and compositions.

Background technique

Compound 6-[2-(methylcarbamoyl)phenylsulfanyl]-3-E-[2-(pyridin-2-yl)vinyl]indazole (also known as "compound 1"),

and pharmaceutically acceptable salts thereof are described in U.S. Patent No. 6,534,524 issued March 18, 2003 and U.S. Patent No. 6,531,491 issued March 11, 2003, the disclosures of which are incorporated by reference for all purposes The contents of which are incorporated in this application in their entirety. This compound is a protein kinase receptor inhibitor and represents a synthetic, small molecule inhibitor of angiogenic receptor signaling.

Protein kinases are a family of enzymes that catalyze the hydroxyl phosphorylation of specific tyrosine, serine, or threonine residues in proteins. Often, this phosphorylation significantly perturbs protein function and thus protein kinases are critical in regulating a wide variety of cellular processes including metabolism, cell proliferation, cell differentiation, and cell survival. Among the many different cellular functions known to require protein kinase activity, some processes represent attractive targets for therapeutic intervention in certain disease conditions. Two examples are angiogenesis and cell cycle control, in which protein kinases play a key role.

Excess angiogenesis is a hallmark of several diseases such as retinopathy, psoriasis, rheumatoid arthritis, age-related macular degeneration (AMD), and cancer (including solid tumors) Folkman, Nature Med., 1, 27- 31 (1995). Protein kinases that have been shown to be involved in the angiogenic process include VEGF-R2 (vascular endothelial growth factor receptor 2, also known as KDR (kinase insert domain receptor) and FLK-1). Thus, direct inhibition of the kinase activity of VEGF-R2 can lead to reduced angiogenesis even in the presence of exogenous VEGF (see Strawn et al., Cancer Research, 56, 3540-3545 (1996)).

Effective inhibitors of protein kinases are therefore needed. Furthermore, as will be appreciated by those skilled in the art, kinase inhibitors are required to have physical properties amenable to reliable formulation. These properties include stability to heat, moisture and light.

Crystalline polymorphs are different crystal forms of the same compound. The term polymorph may or may not include other solid molecular forms, including hydrates (eg, bound water present in a crystalline structure) and solvates (eg, bound solvents other than water) of the same compound. Different crystalline polymorphs have different crystal structures due to the different packing of molecules in the crystal lattice. This results in different crystal symmetry and/or unit cell parameters, which directly affect their physical properties, such as the X-ray diffraction characteristics of the crystal or powder. Different polymorphs, for example, will generally diffract at different sets of angles and will give different values of intensity. Thus, X-ray powder diffraction can be used in a reproducible and reliable manner to identify different polymorphs or solid forms comprising more than one polymorph.

Crystalline polymorphs are of interest to the pharmaceutical industry and especially those involved in the development of suitable dosage forms. If the polymorphic form does not remain constant during clinical or stability studies, the precise dosage form used or studied is not comparable between batches. It is also desirable to have a method of producing a compound in a selected polymorph of high purity when the compound is used in clinical research or in a commercial product due to the presence of impurities that can produce undesired toxicological effects. Certain polymorphic forms may exhibit enhanced thermodynamic stability or may be easier to manufacture in large quantities with high purity, and thus be more suitable for inclusion in pharmaceutical formulations. Certain polymorphs may exhibit other advantageous physical properties, such as lack of hygroscopic tendency, improved solubility, and enhanced dissolution rates due to different lattice energies.

A discussion of the Background of the Invention is included in this application for the purpose of explaining the context of the invention. It should not be taken as an admission that any of the material referred to was published, known or part of the common general knowledge in any country before the priority date of any claim.

Contents of invention

The present invention relates to 6-[2-(methylcarbamoyl)phenylsulfanyl]-3-E-[2-(pyridin-2-yl)vinyl]indazole (also known as "compound 1") ) novel polymorphic form

Compound 1 is a potent inhibitor of VEGF-R2 and exhibits very favorable toxicological and pharmacological properties. The present invention also relates to processes for the preparation of different polymorphic forms of compound 1, their use in pharmaceutical compositions and their use in the treatment of disease conditions associated with unwanted angiogenesis and/or cell proliferation.

In one embodiment, the present invention provides a crystal form of Compound 1 or a pharmaceutically acceptable salt thereof.

In another embodiment, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a polymorph designated as Form I. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a substantially pure polymorph of Form I. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a powder X-ray diffraction (PXRD) pattern comprising peaks at diffraction angles (2θ) of about 8.1 and about 29.8 )pattern. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of 8.1 ± 0.1 and 29.8 ± 0.1. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 8.1, about 18.2, about 18.5, and about 29.8 . Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a crystal form comprising diffraction angles (2θ) of about 8.1±0.1, 18.2±0.1, 18.5±0.1 and 29.8±0.1 PXRD pattern of the peaks. Still more specifically, the present invention provides a crystal form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of about 8.1, about 9.1, about 10.6, about 15.4, about 16.3, about 17.4 , about 18.2, about 18.5, about 20.0, about 20.8, about 23.2, about 24.0, about 25.9, about 27.4, and about 29.8 PXRD patterns of peaks. Still more specifically, the present invention provides a crystal form of compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 8.1±0.1, 9.1±0.1, 10.6±0.1, 15.4±0.1, 16.3 PXRD patterns of peaks at ±0.1, 17.4±0.1, 18.2±0.1, 18.5±0.1, 20.0±0.1, 20.8±0.1, 23.2±0.1, 24.0±0.1, 25.9±0.1, 27.4±0.1 and 29.8±0.1. Still more specifically, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at substantially the same diffraction angles (2Θ) as shown in Figure 1A. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, characterized by a differential scanning calorimetry (DSC) thermogram substantially the same as that shown in Figure IB.

In another embodiment, is a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2θ) of 8.1±0.1 and 29.8±0.1 . Even more particularly, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has an and PXRD patterns of peaks at 29.8 ± 0.1. Still more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 8.1±0.1, 9.1±0.1, 10.6±0.1, Peaks at 15.4±0.1, 16.3±0.1, 17.4±0.1, 18.2±0.1, 18.5±0.1, 20.0±0.1, 20.8±0.1, 23.2±0.1, 24.0±0.1, 25.9±0.1, 27.4±0.1 and 29.8±0.1 The PXRD pattern.

In another embodiment, a method for the manufacture of polymorph I of Compound 1, comprising preparing a polymorph comprising 6-[2-(methylcarbamoyl)phenylsulfanyl]-3-E-[2-( A slurry of pyridin-2-yl)vinyl]indazole and an alcohol such as methanol, heating the slurry between about 40°C and about 60°C, adding water to the slurry, cooling the slurry and separating the solid portion with The other components of the slurry are separated.

In another embodiment, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a polymorph designated as Form II. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a substantially pure polymorph of Form II. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 8.5 and about 18.8. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of 8.5±0.1 and 18.8±0.1. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 8.5, about 10.9, about 14.8, and about 18.8 . Even more specifically, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form comprising: PXRD patterns of peaks. Still more specifically, the present invention provides a crystal form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of about 8.5, about 10.9, about 14.8, about 16.2, about 18.8, about 21.5 , PXRD patterns of peaks at about 24.8, about 25.9, about 30.3, and about 32.2. Still more specifically, the present invention provides a crystal form of compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 8.5±0.1, 10.9±0.1, 14.8±0.1, 16.2±0.1, 18.8 PXRD patterns of peaks at ±0.1, 21.5±0.1, 24.8±0.1, 25.9±0.1, 30.3±0.1 and 32.2±0.1. Still more specifically, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at substantially the same diffraction angles (2Θ) as shown in Figure 2A. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, characterized by a differential scanning calorimetry (DSC) thermogram substantially the same as that shown in Figure 2B.

In another embodiment, is a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2θ) of 8.5±0.1 and 18.8±0.1 . Even more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form comprising diffraction angles (2θ) of 8.5±0.1, 10.9±0.1, 14.8±0.1 and PXRD pattern of peaks at 18.8±0.1. Still more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 8.5±0.1, 10.9±0.1, 14.8±0.1, PXRD patterns of peaks at 16.2±0.1, 18.8±0.1, 21.5±0.1, 24.8±0.1, 25.9±0.1, 30.3±0.1 and 32.2±0.1.

[ 2-(Pyridin-2-yl)vinyl]indazole was exposed to moisture. In another aspect, the humidity is at least 80% relative humidity.

In another embodiment, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a polymorph designated as Form III. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a substantially pure polymorph of Form III. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 13.0 and about 24.1. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of 13.0±0.1 and 24.1±0.1. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 13.0, about 13.3, about 21.7, and about 24.1 . Even more specifically, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form comprising: PXRD patterns of peaks. Still more specifically, the present invention provides a crystal form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of about 10.5, about 13.0, about 13.3, about 15.8, about 16.4, about 17.5 , about 19.5, about 20.1, about 21.4, about 21.7, about 24.1, about 25.0 and about 26.9 PXRD patterns of peaks. Still more specifically, the present invention provides a crystal form of compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 10.5±0.1, 13.0±0.1, 13.3±0.1, 15.8±0.1, 16.4 PXRD patterns of peaks at ±0.1, 17.5±0.1, 19.5±0.1, 20.1±0.1, 21.4±0.1, 21.7±0.1, 24.1±0.1, 25.0±0.1 and 26.9±0.1. Still more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at substantially the same diffraction angles (2Θ) as shown in Figure 3A. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, characterized by a differential scanning calorimetry (DSC) thermogram substantially the same as that shown in Figure 3B.

In another embodiment, is a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2θ) of 13.0±0.1 and 24.1±0.1 . Even more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 13.0±0.1, 13.3±0.1, 21.7±0.1 and PXRD pattern of peaks at 24.1±0.1. Still more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form at a diffraction angle (2θ) of 10.5±0.1, 13.0±0.1, 13.3±0.1, PXRD patterns of peaks at 15.8±0.1, 16.4±0.1, 17.5±0.1, 19.5±0.1, 20.1±0.1, 21.4±0.1, 21.7±0.1, 24.1±0.1, 25.0±0.1 and 26.9±0.1.

In another embodiment, is the method for the manufacture of polymorph III of compound 1, it comprises the preparation comprising 6-[2-(methylcarbamoyl)phenylsulfanyl]-3-E-[2 - a slurry of a pharmaceutically acceptable salt of (pyridin-2-yl)vinyl]indazole, a base and an aprotic solvent, heating and stirring the slurry to a temperature between about 45°C and about 80°C and The solid portion is separated from the other components of the slurry. In another aspect, the aprotic solvent is ethyl acetate. In another aspect, the base is NaHCO ₃ .

In another embodiment, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a polymorph designated as Form IV. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a substantially pure polymorph of Form IV. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 8.9 and about 15.7. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of 8.9±0.1 and 15.7±0.1. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 8.9, about 14.6, about 15.7, and about 19.2 . Even more specifically, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form comprising: PXRD patterns of peaks. Still more specifically, the present invention provides a crystal form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of about 8.9, about 12.0, about 14.6, about 15.2, about 15.7, about 17.8 , about 19.2, about 20.5, about 21.6, about 23.2, about 24.2, about 24.8, about 26.2, and about 27.5 PXRD patterns of peaks. Still more specifically, the present invention provides a crystal form of compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 8.9±0.1, 12.0±0.1, 14.6±0.1, 15.2±0.1, 15.7 PXRD patterns of peaks at ±0.1, 17.8±0.1, 19.2±0.1, 20.5±0.1, 21.6±0.1, 23.2±0.1, 24.2±0.1, 24.8±0.1, 26.2±0.1 and 27.5±0.1. Still more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at substantially the same diffraction angles (2Θ) as shown in Figure 4A. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, characterized by a differential scanning calorimetry (DSC) thermogram substantially the same as that shown in Figure 4B.

In another embodiment, a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof having a PXRD pattern comprising peaks at diffraction angles (2θ) of 8.9±0.1 and 15.7±0.1 . Even more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form comprising diffraction angles (2θ) of 8.9±0.1, 14.6±0.1, 15.7±0.1 and PXRD pattern of peaks at 19.2±0.1. Still more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 8.9±0.1, 12.0±0.1, 14.6±0.1, PXRD patterns of peaks at 15.2±0.1, 15.7±0.1, 17.8±0.1, 19.2±0.1, 20.5±0.1, 21.6±0.1, 23.2±0.1, 24.2±0.1, 24.8±0.1, 26.2±0.1 and 27.5±0.1.

In another embodiment, different poly(s) from 6-[2-(methylcarbamoyl)phenylsulfanyl]-3-E-[2-(pyridin-2-yl)vinyl]indazole Crystal Forms A method of making polymorph Form IV of Compound 1 comprising heating a different polymorph, wherein the different polymorph is hydrated or solvated. In another aspect, heating is performed under vacuum. In another aspect, the heating is between about 110°C and about 135°C. And in yet another aspect, the solvates of the different polymorphs are selected from the group consisting of methanol solvates, ethanol solvates, and ethyl acetate solvates. In yet another aspect, the different polymorph is Compound 1 polymorph III.

Another aspect of this embodiment is a method for converting polymorph VI of compound 1 to polymorph IV of compound 1 comprising heating a slurry of polymorph VI of compound 1 in an aromatic solvent and The solid portion is separated from the other components of the slurry. In another aspect, the heating step is performed at a temperature of at least 110°C.

Another aspect of this embodiment is a method for making polymorph Form IV of Compound 1 comprising heating a slurry comprising a hydrated form of Compound 1 and an aromatic solvent between about 110 and about 140° C., and The solid portion is separated from the other components of the slurry. In another aspect, the aromatic solvent is toluene or xylene. In yet another aspect, the hydrated form of Compound 1 is Compound 1 polymorph III.

Another aspect of this embodiment is a method for making polymorph Form IV of Compound 1 comprising recrystallizing Compound 1 to form a recrystallized product, heating between about 110°C and about 150°C comprising the recrystallized product, and Slurry of aromatic solvent and separation of solid portion from other components of the slurry. In yet another aspect, Compound 1 is recrystallized from a solution comprising dichloromethane and methanol. In yet another aspect, the aromatic solvent is toluene or xylene.

Another aspect of this embodiment is a process for the manufacture of polymorphic Form IV of Compound 1 comprising recrystallizing 6-[2-(methylcarbamoyl)phenylsulfane from a solution of a water-soluble polymer yl]-3-E-[2-(pyridin-2-yl)vinyl]indazole, adding water to the solution to precipitate a solid and separating the precipitated solid from the water-soluble polymer and water. In yet another aspect, the water soluble polymer is (poly)ethylene glycol. In yet another aspect, the (poly)ethylene glycol is PEG-400.

In another embodiment, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a polymorph designated Form VI. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a substantially pure polymorph of Form VI. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 9.6 and about 18.1. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of 9.6±0.1 and 18.1±0.1. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 9.6, about 11.6, about 18.1, and about 25.2 . Even more specifically, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form comprising: PXRD patterns of peaks. Still more specifically, the present invention provides a crystal form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of about 9.6, about 11.6, about 17.5, about 18.1, about 19.9 and about 25.2 PXRD pattern of the peak at . Still more specifically, the present invention provides a crystal form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 9.6±0.1, 11.6±0.1, 17.5±0.1, 18.1±0.1, 19.9 PXRD patterns of peaks at ±0.1 and 25.2±0.1. Still more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at substantially the same diffraction angles (2Θ) as shown in Figure 5A. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, characterized by a differential scanning calorimetry (DSC) thermogram substantially the same as that shown in Figure 5B.

In another embodiment is a pharmaceutical composition comprising a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of 9.6±0.1 and 18.1±0.1. Even more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form comprising diffraction angles (2θ) of 9.6±0.1, 11.6±0.1, 18.1±0.1 and PXRD pattern of peaks at 25.2±0.1. Still more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 9.6±0.1, 11.6±0.1, 17.5±0.1, PXRD patterns of peaks at 18.1±0.1, 19.9±0.1 and 25.2±0.1.

In another embodiment is a method for the manufacture of polymorph VI of Compound 1 comprising the preparation of a compound comprising 6-[2-(methylcarbamoyl)phenylsulfanyl]-3-E-[2- A slurry of a pharmaceutically acceptable salt of (pyridin-2-yl)vinyl]indazole, a base, and a protic solvent, heating and stirring the slurry between about 45°C and about 80°C and combining the solid portion with the The other components of the slurry are separated. In yet another aspect, the protic solvent is an alcohol. In yet another aspect, the protic solvent is ethanol. In another aspect, the base is NaHCO ₃ .

In another embodiment, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a polymorph designated as Form VII. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a substantially pure polymorph of Form VII. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 9.4 and about 17.0. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of 9.4±0.1 and 17.0±0.1. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 9.4, about 17.0, about 23.6, and about 25.1 . Even more specifically, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form comprising: PXRD patterns of peaks. Still more specifically, the present invention provides a crystal form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of about 9.4, about 10.2, about 16.2, about 17.0, about 18.9, about 19.7 , about 21.5, about 22.7, about 23.6, about 25.1, about 26.2, about 27.4 and about 29.3 PXRD patterns of peaks. Still more specifically, the present invention provides a crystal form of compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 9.4±0.1, 10.2±0.1, 16.2±0.1, 17.0±0.1, 18.9 PXRD patterns of peaks at ±0.1, 19.7±0.1, 21.5±0.1, 22.7±0.1, 23.6±0.1, 25.1±0.1, 26.2±0.1, 27.4±0.1 and 29.3±0.1. Still more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at substantially the same diffraction angles (2Θ) as shown in Figure 6A. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, characterized by a differential scanning calorimetry (DSC) thermogram substantially the same as that shown in Figure 6B.

In another embodiment is a pharmaceutical composition comprising a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of 9.4±0.1 and 17.0±0.1. Even more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form comprising diffraction angles (2θ) of 9.4±0.1, 17.0±0.1, 23.6±0.1 and PXRD pattern of peaks at 25.1±0.1. Still more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 9.4±0.1, 10.2±0.1, 16.2±0.1, PXRD patterns of peaks at 17.0±0.1, 18.9±0.1, 19.7±0.1, 21.5±0.1, 22.7±0.1, 23.6±0.1, 25.1±0.1, 26.2±0.1, 27.4±0.1 and 29.3±0.1.

In another embodiment is a method for the manufacture of polymorph VII of Compound 1, which comprises preparing a polymorph comprising 6-[2-(methylcarbamoyl)phenylsulfanyl]-3-E-[2- A slurry of (pyridin-2-yl)vinyl]indazole or a solvate thereof and a protic solvent; heating and stirring the slurry between about 45°C and about 80°C; and mixing the solid portion with the slurry other components are separated. In yet another aspect, the protic solvent is isopropanol.

In another embodiment, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a polymorph designated Form VIII. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, wherein the crystalline form is a substantially pure polymorph of Form VIII. In another embodiment, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 24.6 and about 26.3. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 24.6±0.1 and 26.3±0.1. Even more particularly, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of about 24.6, about 25.9, about 26.3, and about 32.0 . Even more specifically, the present invention provides a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form comprising 24.6±0.1, 25.9±0.1, 26.3±0.1 and 32.0±0.1 at diffraction angles (2θ). PXRD patterns of peaks. Still more specifically, the present invention provides a crystal form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of about 10.7, about 15.5, about 15.9, about 20.6, about 22.7, about 24.6 , PXRD patterns of peaks at about 25.9, about 26.3, and about 32.0. Still more specifically, the present invention provides a crystal form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a diffraction angle (2θ) of 10.7±0.1, 15.5±0.1, 15.9±0.1, 20.6±0.1, 22.7 PXRD patterns of peaks at ±0.1, 24.6±0.1, 25.9±0.1, 26.3±0.1 and 32.0±0.1. Still more specifically, the present invention provides a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at substantially the same diffraction angles (2Θ) as shown in FIG. 7 .

In another embodiment is a pharmaceutical composition comprising a crystalline form of Compound 1, or a pharmaceutically acceptable salt thereof, having a PXRD pattern comprising peaks at diffraction angles (2Θ) of 24.6±0.1 and 26.3±0.1. Even more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form comprising diffraction angles (2θ) of 24.6±0.1, 25.9±0.1, 26.3±0.1 and PXRD pattern of peaks at 32.0±0.1. Still more specifically, the present invention provides a pharmaceutical composition comprising a crystalline form of Compound 1 or a pharmaceutically acceptable salt thereof, which has a crystal form at a diffraction angle (2θ) of 10.7±0.1, 15.5±0.1, 15.9±0.1, PXRD patterns of peaks at 20.6±0.1, 22.7±0.1, 24.6±0.1, 25.9±0.1, 26.3±0.1 and 32.0±0.1.

In another embodiment is a method for the manufacture of polymorph VIII of Compound 1 comprising 6-[2-(methylcarbamoyl)phenylsulfanyl]-3-E-[2-( Pyridin-2-yl)vinyl]indazole is dissolved in a minimal amount of refluxing aprotic solvent to form a solution; the solution is cooled, thereby forming a crystalline form; and the crystalline product is isolated. In yet another aspect, the aprotic solvent is dioxane.

In another embodiment of the present invention is a solid form of Compound 1 or a pharmaceutically acceptable salt thereof, wherein the solid form comprises at least two of the following crystal forms: Polymorph I, II, III, IV, VI , VII and VIII.

In yet another aspect of the invention is a pharmaceutical composition comprising polymorph I of Compound 1 . In another aspect is a method of treating a disease condition in a mammal mediated by protein kinase activity comprising administering a therapeutically effective amount of Compound 1 polymorph I. In yet another aspect is a method of treating a hyperproliferative disorder, such as tumor growth, cell proliferation, or angiogenesis in a mammal comprising administering a therapeutically effective amount of Compound 1 polymorph I. In another aspect is a method of treating a disease condition mediated by VEGF activity in a mammal comprising administering to a mammal in need of such treatment a therapeutically effective amount of Compound 1 polymorph I.

In yet another aspect of the invention is a pharmaceutical composition comprising polymorph II of Compound 1 . In another aspect is a method of treating a disease condition in a mammal mediated by protein kinase activity comprising administering a therapeutically effective amount of Compound 1 polymorph II. In yet another aspect is a method of treating a hyperproliferative disorder, such as tumor growth, cell proliferation, or angiogenesis in a mammal comprising administering a therapeutically effective amount of Compound 1 polymorph II. In another aspect is a method of treating a disease condition in a mammal mediated by VEGF activity comprising administering to a mammal in need of such treatment a therapeutically effective amount of Compound 1 polymorph II.

In yet another aspect of the invention is a pharmaceutical composition comprising polymorphic Form III of Compound 1 . In another aspect is a method of treating a disease condition in a mammal mediated by protein kinase activity comprising administering a therapeutically effective amount of Compound 1 polymorph III. In yet another aspect is a method of treating a hyperproliferative disorder, such as tumor growth, cell proliferation, or angiogenesis in a mammal comprising administering a therapeutically effective amount of Compound 1 polymorph III. In another aspect is a method of treating a disease condition in a mammal mediated by VEGF activity comprising administering to a mammal in need of such treatment a therapeutically effective amount of Compound 1 polymorph III.

In yet another aspect of the invention is a pharmaceutical composition comprising polymorphic Form IV of Compound 1 . In another aspect is a method of treating a disease condition in a mammal mediated by protein kinase activity comprising administering a therapeutically effective amount of Compound 1 polymorph IV. In yet another aspect is a method of treating a hyperproliferative disorder, such as tumor growth, cell proliferation, or angiogenesis in a mammal comprising administering a therapeutically effective amount of Compound 1 polymorph IV. In another aspect is a method of treating a disease condition in a mammal mediated by VEGF activity comprising administering to a mammal in need of such treatment a therapeutically effective amount of Compound 1 polymorph IV.

In yet another aspect of the invention are pharmaceutical compositions comprising polymorphic Form VI of Compound 1 . In another aspect is a method of treating a disease condition in a mammal mediated by protein kinase activity comprising administering a therapeutically effective amount of Compound 1 polymorph VI. In yet another aspect is a method of treating a hyperproliferative disorder, such as tumor growth, cell proliferation or angiogenesis in a mammal comprising administering a therapeutically effective amount of Compound 1 polymorph VI. In another aspect is a method of treating a disease condition in a mammal mediated by VEGF activity comprising administering to a mammal in need of such treatment a therapeutically effective amount of Compound 1 polymorph VI.

In yet another aspect of the invention are pharmaceutical compositions comprising polymorphic Form VII of Compound 1 . In another aspect is a method of treating a disease condition in a mammal mediated by protein kinase activity comprising administering a therapeutically effective amount of Compound 1 polymorph VII. In yet another aspect is a method of treating a hyperproliferative disorder, such as tumor growth, cell proliferation or angiogenesis in a mammal comprising administering a therapeutically effective amount of Compound 1 polymorph VII. In another aspect is a method of treating a disease condition in a mammal mediated by VEGF activity comprising administering to a mammal in need of such treatment a therapeutically effective amount of Compound 1 polymorph VII.

In yet another aspect of the invention is a pharmaceutical composition comprising the polymorphic Form VIII of Compound 1 . In another aspect is a method of treating a disease condition in a mammal mediated by protein kinase activity comprising administering a therapeutically effective amount of Compound 1 polymorph VIII. In yet another aspect is a method of treating a hyperproliferative disorder, such as tumor growth, cell proliferation, or angiogenesis in a mammal comprising administering a therapeutically effective amount of Compound 1 polymorph VIII. In another aspect is a method of treating a disease condition in a mammal mediated by VEGF activity comprising administering to a mammal in need of such treatment a therapeutically effective amount of Compound 1 polymorph VIII.

The invention further relates to the modulation or inhibition of, for example, protein kinase activity (e.g. VEGF, VEGF, FGF, CDK complexes, TEK, CHK1, LCK, among others) in mammalian tissues by administering at least one polymorphic form of Compound 1 , FAK and phosphorylase kinase receptors) method.

The present invention also relates to a method of combination therapy for the treatment of hyperproliferative disorders or conditions mediated by VEGF activity comprising administering to a mammal in need of such treatment a therapeutically effective amount of any of the polymorphic forms discussed above A pharmaceutical composition, or multiple pharmaceutical compositions, combined with a therapeutically effective amount of one or more substances selected from antineoplastic agents, antiangiogenic agents, signal transduction inhibitors and antiproliferative agents.

The term "active agent" or "active ingredient" refers to a polymorphic form of Compound 1 or a solid form comprising two or more polymorphic forms of Compound 1 .

The term "ambient temperature" refers to temperature conditions typically encountered in a laboratory environment. This includes an approximate temperature range of about 20 to about 30°C.

The term "aqueous base" refers to any organic or inorganic base. Aqueous bases include, by way of example only, metal bicarbonates such as sodium bicarbonate, potassium bicarbonate, cesium bicarbonate, and the like.

The term "aromatic solvent" refers to an organic solvent having an aromatic moiety, including, by way of example only, benzene, toluene, xylene isomers or mixtures thereof, and the like.

The term "chemical stability" refers to a class of stability in which a particular compound retains its chemical integrity and includes, but is not limited to, thermal stability, light stability, and humidity stability.

The term "detectable amount" refers to an amount or amount per unit volume that can be detected using conventional techniques such as X-ray powder diffraction, differential scanning calorimetry, HPLC, FT-IR, Raman spectroscopy, and the like.

The term "exposed to moisture" refers to the method of exposing a substance to water vapor in a humidifier, humidity cabinet, or any device that can control relative humidity. The term can also describe the method by which a substance is exposed to ambient moisture during storage.

The term "inert solvent" refers to any solvent or liquid component of a slurry that does not chemically react with other components of the solution or slurry. Inert solvents include, by way of example only, aprotic solvents such as aromatic solvents, ethyl acetate, acetone, methyl tert-butyl ether, dioxane, THF, and the like. Protic solvents include, by way of example only, methanol, ethanol, propanol isomers, butanol isomers, and the like.

The term "mediated by VEGF activity" refers to a biological or molecular process that is regulated, modulated or inhibited by VEGF protein kinase activity. For certain applications, those that inhibit the activity of protein kinases associated with CDK complexes and inhibit angiogenesis and/or inflammation are preferred, among others. The present invention includes methods of modulating or inhibiting, for example, protein kinase activity in mammalian tissues by administering a polymorphic form of Compound 1. The activity of antiproliferative agents is readily measured by known methods (eg, by using whole cell cultures in an MTT assay). The activity of a polymorphic form of Compound 1 as a mediator of protein kinase activity (eg, kinase activity) can be measured by any method available to one of skill in the art, including in vivo and/or in vitro assays.

The term "minimum amount" refers to the minimum amount of solvent required to completely dissolve a substance at a given temperature.

The term "pharmaceutically acceptable salt" refers to salts that retain the biological effectiveness of the free acids and bases of the specified compound and are not biologically or otherwise undesirable. The compounds of the present invention may possess sufficiently acidic, sufficiently basic, or both functional groups and thus react with any of a number of inorganic or organic bases and inorganic and organic acids to form pharmaceutically acceptable salts. Exemplary pharmaceutically acceptable salts include those salts prepared by reacting a compound of the invention with an inorganic or organic acid or inorganic base, such as salts including sulfates, pyrosulfates, bisulfates, sulfites , bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, caprate, caprylic acid Salt, acrylate, p-toluenesulfonate (tosylate), formate, isobutyrate, hexanoate, heptanoate, propiolate, oxalate, malonate, succinate Salt, Suberate, Sebacate, Fumarate, Maleate, Butyne-1,4-dioate, Hexyne-1,6-dioate, Benzene salt, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, methoxybenzoate, phthalate, sulfonate, dinitrobenzoate Tosylate, Phenylacetate, Phenylpropionate, Phenylbutyrate, Citrate, Lactate, Gamma-Hydroxybutyrate, Glycolate, Tartrate, Methanesulfonate, Propane sulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate and mandelate.

When the compound of the present invention is a base, the desired pharmaceutically acceptable salt can be prepared by any suitable method available in the art, for example, by treating the free base with an inorganic acid such as hydrochloric acid, hydrobromic acid, Sulfuric acid, nitric acid, phosphoric acid and similar or organic acids such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranic acid Glycosidic acids such as glucuronic acid or galacturonic acid, alpha-hydroxy acids such as citric acid or tartaric acid, amino acids such as aspartic acid or glutamic acid, aromatic acids such as benzoic acid or cinnamic acid, sulfonic acids such as p-toluene Sulfonic acid or ethanesulfonic acid and similar.

When the compound of the invention is an acid, the desired pharmaceutically acceptable salt can be prepared by any suitable method, for example, by treating the free acid with an inorganic base or an organic base such as an amine (primary, secondary or tertiary), Alkali metal hydroxide or alkaline earth metal hydroxide or the like. Illustrative examples of suitable salts include organic salts derived from amino acids such as glycine and arginine, ammonia, primary, secondary and tertiary amines and cyclic amines such as piperidine, morpholine and piperazine, and those derived from sodium, calcium, , potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium inorganic salts.

The term "polymorph" refers to different crystal forms of the same compound and includes, but is not limited to, other solid molecular forms, including hydrates (such as bound water present in the crystalline structure) and solvates of the same compound (such as non-aqueous binding solvents).

The term "peak intensity" refers to the relative signal intensity within a given X-ray diffraction pattern. Factors that can affect the relative peak intensities are sample thickness and preferred orientation (ie, the crystalline particles are non-randomly distributed).

The term "peak position" as used in this application refers to the position of the X-ray reflection as measured and observed in an X-ray powder diffraction experiment. The peak position is directly related to the size of the unit cell. Peaks identified by their respective peak positions have been extracted from the diffraction patterns of various polymorphic forms I, II, III, IV, VI, VII and VIII of Compound 1.

The term "PEG" refers to poly(ethylene glycol). PEGs are commercially available and come in varying ranges of polymer chain lengths and thus viscosities. PEG 400 is soluble in alcohols, acetone, benzene, chloroform, acetic acid, CCl ₄ and water.

The term "pharmaceutically acceptable carrier, diluent or vehicle" refers to a material (or materials) that can be included with a particular agent to form a pharmaceutical composition and which can be solid or liquid. Exemplary solid carriers are lactose, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid, and the like. Exemplary liquid carriers are syrup, peanut oil, olive oil, water and the like. Similarly, the carrier or diluent may include time delay or time release materials known in the art, such as waxes, ethyl cellulose, hydroxypropyl methyl cellulose, methyl methacrylate and Glyceryl monostearate or glyceryl distearate together with analogues.

The term "pharmaceutical composition" refers to one or more compounds or polymorphs described in this application or their physiologically/pharmaceutically acceptable salts or solvates together with other chemical components such as physiologically/pharmaceutical Mixtures of acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate the administration of a compound to an organism.

The term "recrystallization" refers to the process of completely dissolving a solid in a first solvent (with heating if necessary), and then inducing precipitation, typically by cooling the solution or by adding a second solvent in which the solid is poorly soluble.

The term "relative humidity" means the ratio, expressed as a percentage, of the amount of water vapor in air at a given temperature to the maximum amount of water vapor that can be maintained at that temperature and pressure.

The term "relative intensity" refers to the intensity value resulting from the X-ray diffraction pattern of a sample. The full ordinate scale of the diffraction pattern is assigned a value of 100. Peaks with intensities falling between about 50% and about 100% of the intensity on this scale were defined as very intense (vs); peaks with intensities falling between about 50% and about 25% were defined as strong (s). Additional weaker peaks are present in the general diffraction pattern and are also characteristic of a given polymorph.

The term "slurry" refers to a solid substance suspended in a liquid medium, typically water or an organic solvent.

The term "separated from" refers to a step in a synthesis to separate the desired species from other undesired species, including, but not limited to, any of the following steps: filtration, washing with additional solvent or water, in Dry with heat and or vacuum.

The term "substantially pure" for a particular polymorphic form of Compound 1 means that the polymorphic form comprises less than 10%, preferably less than 5%, preferably less than 3%, preferably less than 1% by weight of impurities (including compound 1 other polymorphs). The purity can be determined, for example, by X-ray powder diffraction.

"Effective amount" is intended to mean a demonstrated agent that significantly inhibits proliferation and/or prevents dedifferentiation of eukaryotic cells, e.g. mammalian, insect, plant or fungal cells, and is effective for e.g. specific therapeutic treatment amount.

The term "therapeutically effective amount" refers to the amount of a compound or polymorph to be administered that will alleviate to some extent one or more symptoms of the disease being treated. For the treatment of cancer, a therapeutically effective amount refers to an amount that has at least one of the following effects:

(1) Reduce tumor size;

(2) Inhibit (that is, slow down to a certain extent, preferably stop) tumor metastasis;

(3) to some extent inhibit (i.e. to some extent slow down, preferably stop) tumor growth, and

(4) Alleviate (or preferably eliminate) one or more symptoms related to cancer to a certain extent.

The term "2Θ value" or "2Θ" refers to the peak position based on the experimental setup of an X-ray diffraction experiment and is a common abscissa unit in the diffraction pattern. The experimental setup entails recording the reflected beam at an angle 2Θ (2Θ) if the reflection is diffracted when the incident beam forms an angle Θ (Θ) with a particular lattice plane.

The term "treatment" refers to a method of alleviating or eliminating the hyperproliferative disorder and/or its accompanying symptoms. With regard to cancer in particular, these terms simply mean that the life expectancy of an individual with cancer will be increased or that one or more symptoms of the disease will be reduced.

The term "under vacuum" refers to the typical pressure obtainable by a laboratory oil or oil-free diaphragm vacuum pump.

The term "X-ray powder diffraction pattern" refers to an experimentally observed diffraction pattern or a parameter resulting therefrom. The X-ray powder diffraction pattern is characterized by peak position (abscissa) and peak intensity (ordinate).

The term "xylenes" refers to any xylene isomers or mixtures thereof.

A brief description of the drawings

FIG. 1A is an X-ray powder diffraction pattern of polymorph I of Compound 1. FIG.

FIG. 1B is a differential scanning calorimetry (DSC) curve of polymorph I of Compound 1. FIG. A typical curve shows an endotherm onset at 183-190°C at a scan rate of 10°C/min.

FIG. 2A is an X-ray powder diffraction pattern of polymorph II of Compound 1. FIG.

FIG. 2B is a differential scanning calorimetry (DSC) curve of polymorph II of Compound 1. FIG. A typical curve shows an endotherm onset at 102, 152 and 202°C at a scan rate of 10°C/min, followed by an exotherm at 206°C and another exotherm at 210°C.

FIG. 3A is an X-ray powder diffraction pattern of polymorph III of Compound 1. FIG.

FIG. 3B is a differential scanning calorimetry (DSC) curve of polymorph III of Compound 1. FIG. A typical curve shows an endotherm onset at 125-129°C followed by another endotherm at 210°C at a scan rate of 10°C/min.

Figure 3C is a thermogravimetric analysis (TGA) profile of polymorph III. Desolvation was shown by 10% sample weight loss at 125-129°C at a scan rate of 10°C/min.

FIG. 4A is an X-ray powder diffraction pattern of Compound 1 polymorph IV.

FIG. 4B is a differential scanning calorimetry (DSC) curve of polymorphic Form IV of Compound 1. FIG. A typical curve shows an endotherm onset at 216°C at a scan rate of 10°C/min.

FIG. 5A is an X-ray powder diffraction pattern of polymorph VI of Compound 1. FIG.

5B is a differential scanning calorimetry (DSC) curve of Compound 1 polymorph VI. Typical curves show endotherms onset at about 197°C and about 209°C at a scan rate of 10°C/min.

FIG. 6A is an X-ray powder diffraction pattern of polymorph VII of Compound 1. FIG.

6B is a differential scanning calorimetry (DSC) curve of Compound 1 polymorph VII. Typical curves are sample dependent. A typical sample isolated from refluxing THF has an endotherm at 105°C followed by an exotherm at 115°C and then endotherms at 137 and 175°C at a scan rate of 10°C/min.

FIG. 7 is an X-ray powder diffraction pattern of Compound 1 polymorph VIII.

Figure 8 is a schematic diagram showing the structures of several human metabolites of Compound 1.

Detailed ways

It has surprisingly been found that the substance compound 1 can exist in more than one polymorphic crystal form. These forms are useful in formulated products for the treatment of hyperproliferative indications including cancer. Each form may have advantages over other forms in terms of bioavailability, stability, or manufacturability. It has been found that the crystalline polymorph of Compound 1 may be more suitable for large-scale preparation and handling than other polymorphs. Methods for making these polymorphs in high purity are described in this application. Another object of the present invention is to provide a process for preparing each polymorphic form of Compound 1 substantially free of other polymorphic forms of Compound 1 . Furthermore, it is an object of the present invention to provide pharmaceutical formulations comprising the different polymorphic forms of Compound 1 as discussed above and methods of treating hyperproliferative conditions by administering such pharmaceutical formulations.

I. Polymorphic Forms of Compound 1

Each crystalline form of Compound 1 can be characterized by one or more of the following features: X-ray powder diffraction pattern (i.e., X-ray diffraction peaks at different diffraction angles (2θ)), by differential scanning calorimetry Onset of melting point (and onset of dehydration in hydrated form) as illustrated by endotherm of (DSC) thermogram, Raman spectral pattern, water solubility, under high-intensity light conditions according to the International Conference on Harmonization (ICH) Light stability and physical and chemical storage stability. For example, samples of polymorphic forms I, II, III, IV, VI, VII, and VIII of Compound 1 are each characterized by the position and relative intensity of peaks in their X-ray powder diffraction patterns. X-ray powder diffraction parameters are different for each polymorphic form I, II, III, IV, VI, VII and VIII of Compound 1. These polymorphic forms of Compound 1 can therefore be distinguished using X-ray powder diffraction.

The X-ray powder diffraction patterns of each polymorph or amorphous form of Compound 1 are determined by operating at 40kV and 50mA on a Shimadzu XRD-6000X-ray diffractometer equipped with a Cu Kα X-ray radiation source (1.5406 Ȧ) of. Samples were placed in sample holders and then filled and leveled with glass slides. During analysis, samples were rotated at 60 rpm and analyzed at angles from 4 to 40 degrees (θ-2Θ) at 5 degrees per minute in 0.04 degree steps or 2 degrees per minute in 0.02 degree steps. If limited material is available, samples are placed on silicon plates (0 background) and analyzed without rotation. The X-ray diffraction peaks characterized by peak positions and intensity assignments were extracted from the X-ray powder diffraction patterns of each polymorphic form of Compound 1. Those skilled in the art will appreciate that peak positions (2Θ) will show some device-to-device variability, typically as much as 0.1 degrees. Thus, when reporting peak positions (2Θ), those skilled in the art will recognize that such numbers are intended to account for variability between the devices. Furthermore, when describing a crystalline form of the present invention as having substantially the same powder X-ray diffraction pattern as shown in a given figure, the term "substantially the same" is also intended to encompass the inter-equipment differences in the diffraction peak positions. variability. Furthermore, those skilled in the art will appreciate that relative peak intensities will show inter-equipment variability as well as variability due to crystallinity, preferred orientation, sample surface prepared, and other factors known to those skilled in the art and should be It is considered a qualitative measure only.

Different polymorphic forms of compound 1 were also distinguished using differential scanning calorimetry (DSC). DSC measures the difference in thermal energy uptake between a sample solution and an appropriate reference solvent with increasing temperature. DSC thermograms are characterized by endotherms (indicating energy uptake) and also exotherms (indicating energy release), typically when a sample is heated. DSC thermograms were acquired using a Mettler Toledo DSC821 instrument at a scan rate of 10°C/min over the temperature range of 30-250°C. Samples were weighed into 40 μl sealed aluminum crucibles scored with a single hole. The inferred onset of the calculation of the melting temperature and, when applicable, the onset of the dehydration temperature is also calculated. Depending on the heating rate at which the DSC analysis is performed (i.e., the scan rate), the manner in which the DSC onset temperature is defined and determined, the calibration standards used, the instrumental calibration and the relative humidity and chemical purity of the sample, the endotherm exhibited by the compounds of the invention can vary. Varies between above and below the endotherm (approximately 0.01-5°C for crystalline polymorph melting and approximately 0.01-20°C for polymorph dehydration). Endotherms observed for any given instance may also vary from instrument to instrument; however, they will generally be within the limits defined herein, provided the instrument is similarly calibrated.

Different polymorphic forms of compound 1 were also distinguished using thermogravimetric analysis (TGA). TGA was performed on a Mettler Toledo TGA 500 instrument. TGA is a test procedure that records the change in weight of a sample as the sample is heated in air or in a controlled atmosphere such as nitrogen. The thermogravimetric curve (thermogram) provides information on the solvent and water content and the thermal stability of the material.

Different polymorphic forms of compound 1 can also be distinguished by different stability and different solubility.

In one embodiment, the polymorphs of the invention are substantially pure, meaning that each polymorph of Compound 1 comprises less than 10%, such as less than 5% or such as less than 3% or even further such as less than 1% by weight impurities, including other polymorphic forms of compound 1.

The solid forms of the invention may also contain more than one polymorph. Those skilled in the art will recognize that a crystalline form for a given compound can exist in substantially pure form as a single polymorph and also as a crystalline form comprising two or more different polymorphs. When the solid form contains two or more polymorphs, the X-ray diffraction pattern will have peaks characteristic of each individual polymorph of the invention. For example, a solid form comprising two polymorphs will have a powder X-ray diffraction pattern that is a convolute of two X-ray diffraction patterns corresponding to the substantially pure polymorph. In one embodiment, for example, a solid form of the invention comprising a first and a second polymorph contains at least 10% of the first polymorph. In another embodiment, the solid form contains at least 20% of the first polymorph. Yet another embodiment contains at least 30%, at least 40%, or at least 50% of the first polymorph. Those skilled in the art will recognize that many such combinations of several individual polymorphs in varying amounts are possible.

A. Polymorph I

Polymorph I of Compound 1 can be produced by direct crystallization of Compound 1 from methanol and water with stirring at elevated temperature. Polymorph I of Compound 1 is chemically stable at 80°C and stable at 40°C at 75% relative humidity for at least 13 days. Polymorph I of Compound 1 has an aqueous solubility of 179 μg/mL at pH 2 and 9 μg/mL at pH 6.5.

Form I is characterized by an X-ray powder diffraction pattern with peaks at the following approximate diffraction angles (2θ): 8.1, 9.1, 10.6, 15.4, 16.3, 17.4, 18.2, 18.5, 20.0, 20.8, 23.2, 24.0, 25.9, 27.4 and 29.8. Figure IA provides the X-ray powder diffraction pattern of Form I. The DSC thermogram for Form I shown in Figure IB shows an endotherm onset at 183-190°C at a scan rate of 10°C/min.

B. Polymorph II

Polymorph II of Compound 1 is a hydrate. Polymorph II of Compound 1 can be produced by exposing Polymorph I of Compound 1 to 93% relative humidity at room temperature for 6 days.

Form II is characterized by an X-ray powder diffraction pattern with peaks at the following approximate diffraction angles (2Θ): 8.5, 10.9, 14.8, 16.2, 18.8, 21.5, 24.8, 25.9, 30.3, and 32.2. Figure 2A provides the X-ray powder diffraction pattern of Form II. The DSC thermogram for Form II shown in Figure 2B shows an endotherm onset at 102, 152 and 202°C at a scan rate of 10°C/min, followed by an exotherm at 206°C and another onset at 210°C. Exothermic.

C. Polymorph III

Polymorph III of Compound 1 can be produced by neutralizing the p-toluenesulfonate derivative of Compound 1 in ethyl acetate with NaHCO ₃ solution. Polymorph III of Compound 1 is generally ethyl acetate solvate.

Form III is characterized by an X-ray powder diffraction pattern with peaks at the following approximate diffraction angles (2Θ): 10.5, 13.0, 13.3, 15.8, 16.4, 17.5, 19.5, 20.1, 21.4, 21.7, 24.1, 25.0, and 26.9. Figure 3A provides the X-ray powder diffraction pattern of Form III. The DSC thermogram for Form III shown in Figure 3B shows the onset of an endotherm at 125-129°C followed by another endotherm at 210°C at a scan rate of 10°C/min. Form III of Compound 1 was further characterized by thermogravimetric analysis (TGA). Figure 3C is a thermogravimetric analysis (TGA) curve of a sample of polymorph III. A typical TGA thermogram of a sample of Compound 1 polymorph III shows desolvation. The loss of ethyl acetate is indicated by a 10% sample weight loss at 125-129°C at a scan rate of about 10°C/min.

D. Polymorph IV

Polymorphic Form IV of Compound 1 can be prepared in several different ways: (i) direct vacuum desolvation of Polymorphic Form III of Compound 1 at 110-135°C; Solid state conversion of polymorph III by slurrying polymorph III in toluene or xylene; (iii) via recrystallization of compound 1 from a dichloromethane/methanol solution followed by slurrying the precipitate in toluene at 140 °C (iv) by solid state conversion of polymorph VI by refluxing polymorph VI as a toluene slurry at 140°C; and (v) by precipitation of compound 1 in PEG-400 solution with water. Polymorph Form IV has an aqueous solubility of about 550 μg/mL at about pH 1, about 157 μg/mL at about pH 2, about 6 μg/mL at about pH 4, and about 2 μg/mL at about pH 6.5 And about 2 μg/mL at about pH 8.

Polymorphic Form IV is physically and chemically stable at 80°C and 40°C at 75% relative humidity for at least 30 days. Polymorph Form IV is considered to be the most thermodynamically stable form of Compound 1 .

Form IV is further characterized by an X-ray powder diffraction pattern with peaks at the following approximate diffraction angles (2Θ): 8.9, 12.0, 14.6, 15.2, 15.7, 17.8, 19.2, 20.5, 21.6, 23.2, 24.2, 24.8, 26.2 and 27.5. Figure 4A provides the X-ray powder diffraction pattern of Form IV. The DSC thermogram for Form IV shown in Figure 4B shows an endotherm onset at 216°C at a scan rate of 10°C/min.

E. Polymorph VI

Polymorph VI of Compound 1 can be prepared by direct crystallization of Compound 1 from NaHCO ₃ solution with ethanol. Form VI is characterized by an X-ray powder diffraction pattern with peaks at the following approximate diffraction angles (2Θ): 9.6, 11.6, 17.5, 18.1, 19.9, and 25.2. Figure 5A provides an X-ray powder diffraction pattern of Form VI. The DSC thermogram for Form VI shown in Figure 5B shows an endotherm onset at 197°C at a scan rate of 10°C/min.

F. Polymorphic Form VII

Polymorph VII of compound 1 can be prepared by refluxing a suspension of polymorph VI of compound 1 in isopropanol, tetrahydrofuran or methyl tert-butyl ether.

Form VII is characterized by an X-ray powder diffraction pattern with peaks at the following approximate diffraction angles (2Θ): 9.4, 10.2, 16.2, 17.0, 18.9, 19.7, 21.5, 22.7, 23.6, 25.1, 26.2, 27.4, and 29.3. Figure 6A provides an X-ray powder diffraction pattern of Form VII. The DSC thermogram for Form VII shown in Figure 6B shows an endotherm onset at 105°C, followed by an exotherm at 115°C and subsequent endotherms at 137 and 175°C at a scan rate of 10°C/min.

G. Polymorphic Form VIII

Polymorph VIII of Compound 1 can be produced by refluxing a suspension of Polymorph VI of Compound 1 in dioxane.

Form VIII is characterized by an X-ray powder diffraction pattern with peaks at the following approximate diffraction angles (2Θ): 10.7, 15.5, 15.9, 20.6, 22.7, 24.6, 25.9, 26.3, and 32.0. Figure 7 provides the X-ray powder diffraction pattern of Form VIII.

II. The pharmaceutical composition of the present invention

The active agent of the invention (i.e., a polymorph of Compound 1 described herein or a solid form comprising two or more such polymorphs) can be formulated as a medicament suitable for medical use in mammals combination. Any suitable route of administration may be employed to provide an effective dose of any polymorphic Forms I, II, III, IV, VI, VII and VIII of Compound 1 or a pharmaceutically acceptable salt thereof to a patient. For example, oral or parenteral formulations and the like may be employed. Dosage forms include capsules, tablets, dispersions, suspensions and the like, such as enteric-coated capsules and/or tablets, capsules and/or tablets containing enteric-coated pellets of Compound 1 or a pharmaceutically acceptable salt thereof . In all dosage forms, polymorphic Form IV of Compound 1 or a pharmaceutically acceptable salt thereof may be admixed with other suitable ingredients. Such compositions may conveniently be presented in unit dosage form and prepared by any methods known in the art of pharmacy. The pharmaceutical compositions of the present invention comprise a therapeutically effective amount of an active agent together with one or more inert, pharmaceutically acceptable carriers and optionally any other therapeutic ingredients, stabilizers or the like. The carrier must be pharmaceutically acceptable in the sense of being compatible with the other ingredients of the formulation and not unduly deleterious to the recipient thereof. Such compositions may further include diluents, buffers, binders, disintegrants, thickeners, lubricants, preservatives (including antioxidants), flavoring agents, taste-masking agents, inorganic salts (such as sodium chloride ), antimicrobial agents (such as benzalkonium chloride), sweeteners, antistatic agents, surfactants (such as polysorbates available from BASF such as "Tween 20" and "Tween 80" and poloxamers such as F68 and F88), sorbitan esters, lipids (such as phospholipids, such as lecithin and other phosphatidylcholines, phosphatidylethanolamines, fatty acids and fatty esters, steroids (such as cholesterol)) and chelating agents (such as EDTA, zinc and other such appropriate cations). "Remington: The Science & Practice of Pharmacy , 19th ed., Williams & Williams, (1995) and Physician's Desk Reference , 52nd ed., Medical Economics, Montvale, NJ (1998) and Handbook of Pharmaceutical Excipients , 3rd ed. , AHKibbe ed., Pharmaceutical Press, 2000" list other pharmaceutical excipients and/or additives suitable for the composition according to the invention. The active agents of the invention may be formulated in compositions comprising those suitable for oral, rectal, topical, nasal, ophthalmic or parenteral (including intraperitoneal, intravenous, subcutaneous or intramuscular injection) administration.

The amount of active agent in a formulation will vary depending on a variety of factors including dosage form, the condition being treated, target patient population and other considerations and will generally be readily judged by one skilled in the art. A therapeutically effective amount will be that amount required to modulate, regulate or inhibit a protein kinase. In practice, this amount will vary widely depending on the particular active agent, the severity of the condition being treated, the patient population, the stability of the formulation, and the like. The composition will generally contain from about 0.001% to about 99% by weight active agent, preferably from about 0.01% to about 5% by weight active agent and more preferably from about 0.01% to 2% by weight active agent, and will also depend on the composition The relative amount of excipients/additives contained in.

The pharmaceutical composition of the present invention is administered in a conventional dosage form prepared by mixing a therapeutically effective amount of the active agent as the active ingredient with one or more suitable pharmaceutical carriers according to conventional procedures. These procedures may include mixing, granulating and compressing or dissolving the ingredients required for the desired formulation.

The pharmaceutical carrier used can be solid or liquid. Exemplary solid carriers include lactose, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid and the like. Exemplary liquid carriers include syrup, peanut oil, olive oil, water and the like. Similarly, the carrier may include time delay or time release materials known in the art, such as, alone or with waxes, ethylcellulose, hydroxypropylmethylcellulose, methylmethacrylate, and the like. Glyceryl monostearate or glyceryl distearate.

Various drug forms are available. Thus, if a solid carrier is used, the preparation can be tableted, placed in a hard gelatin capsule in powder or pill form or in the form of a troche or lozenge. The amount of solid carrier will vary but will generally vary from about 25 mg to about 1 g. If a liquid carrier is used, the preparation can be in the form of a syrup, emulsion, soft gelatin capsule, sterile injectable solution or suspension in ampoules or vials, or suspension in a non-aqueous liquid.

To obtain stable water-soluble dosage forms, a pharmaceutically acceptable salt of the active agent can be dissolved in an aqueous solution of an organic or inorganic acid, such as a 0.3M solution of succinic or citric acid. If a water-soluble salt form is not available, the active agent can be dissolved in an appropriate co-solvent or combination of co-solvents. Examples of suitable co-solvents include, but are not limited to, alcohols, propylene glycol, polyethylene glycol 300, polysorbate 80, glycerol, and the like in concentrations varying from 0-60% total volume. The composition may also be in the form of a solution of a salt form of the active agent in a suitable aqueous vehicle such as water or isotonic saline or dextrose solution.

It will be appreciated that the actual dosage of the active agent employed in the compositions of the invention will vary depending on the particular crystalline form employed, the particular composition formulated, the mode and particular site of administration, the host and disease being treated. Optimal dosages for a given set of conditions can be determined by those skilled in the art, from experimental data for the agents, using routine dosimetry experiments. For oral administration, an exemplary daily dosage generally used is about 0.001 to about 1000 mg/kg body weight, more preferably about 0.001 to about 50 mg/kg body weight, with the course of treatment repeated at appropriate intervals. Administration of prodrugs is generally dosed at a weight level that is chemically equivalent to that of the fully active form. In the practice of this invention, the most appropriate route of administration and therapeutic dosage level will depend upon the nature and severity of the disease being treated. Dosage and dosage frequency may also vary according to the age, weight and response of the individual patient. In general, suitable oral dosage forms may encompass doses in the range of 5 mg to 250 mg total daily dose, administered as a single dose or in divided doses. A preferred dosage range is 10 mg to 80 mg.

The compositions of the invention can be manufactured in a manner generally known for the preparation of pharmaceutical compositions. For example using conventional techniques such as mixing, dissolving, granulating, dragee-making, milling, emulsifying, encapsulating, entrapping or lyophilizing. Pharmaceutical compositions may be formulated in conventional manner using one or more physiologically acceptable carriers selected from excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically.

For oral administration, such compounds can be formulated readily by combining the active agent with pharmaceutically acceptable carriers known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, lozenges, capsules, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use can be obtained using a solid excipient in which the active agent is mixed, optionally grinding the resulting mixture, and processing the mixture of granules, after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients include: fillers such as sugars, including lactose, sucrose, mannitol or sorbitol; and cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, gum, methylcellulose hydroxypropylmethylcellulose, sodium carboxymethylcellulose, or polyvinylpyrrolidone (PVP). If desired, disintegrants such as cross-linked polyvinylpyrrolidone, agar or alginic acid or a salt thereof such as sodium alginate may be added.

Provide suitable coating for the lozenge core. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, polyvinylpyrrolidone, carbopol gel, polyethylene glycol and/or titanium dioxide, lacquers and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings to distinguish or to characterize different combinations of active agents.

Pharmaceutical preparations which can be used orally include snap-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The snap-fit capsules can contain the active ingredients in admixture with filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In soft capsules, the active agents may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. Furthermore, stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration. For buccal administration, the compositions may take the form of tablets or lozenges formulated in conventional manner.

For intranasal or administration by inhalation, the compounds used according to the present invention can be self-activated using suitable propellants such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases. Delivery is conveniently in the form of an aerosol spray presented in pressurized packs or nebulizers. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve for delivering the dose. Capsules and cartridges of gelatin for use in an inhaler or insufflator and the like may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.

The active agent can be formulated for parenteral administration, eg, by injection by bolus injection or continuous infusion. Formulations for injection may be presented in unit dosage form, eg, in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.

Pharmaceutical formulations for parenteral administration include suspensions of the active agents and appropriate oily injection suspensions may be prepared. Suitable lipophilic solvents or vehicles include fatty oils, such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the active agents to allow for the preparation of highly concentrated solutions.

For ocular administration, the active agent is delivered in a pharmaceutically acceptable ophthalmic vehicle such that the compound remains in contact with the surface of the eye for a period of time sufficient to allow the compound to penetrate the cornea and intraocular regions, including (e.g. ) anterior chamber, posterior chamber, vitreous, aqueous humor, vitreous humor, cornea, iris/ciliary body, lens, choroid/retina and sclera. Pharmaceutically acceptable ophthalmic vehicles can be, for example, ointments, vegetable oils, or encapsulating materials. The active agents of the invention may also be injected directly into the vitreous and aqueous humor or subtenon.

Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, eg sterile pyrogen-free water, before use. The compounds may also be formulated in rectal or vaginal compositions such as suppositories or enemas, eg, containing conventional suppository bases such as cocoa butter or other glycerides.

In addition to the formulations described above, polymorphic forms may also be formulated as depot preparations. Such long-acting formulations may be administered by implantation (eg, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, polymorphic forms may be formulated with suitable polymeric or hydrophobic materials (e.g., as emulsions in acceptable oils) or ion exchange resins or in the form of sparingly soluble derivatives (e.g., as sparingly soluble salts) .

Additionally, the active agent can be delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained release materials have been identified and are known to those skilled in the art. Sustained-release capsules can, depending on their chemical nature, release compounds for several weeks to over 100 days. Depending on the chemical nature and biological stability of the therapeutic agent, additional strategies for protein stabilization may be used.

The pharmaceutical composition may also contain suitable solid or gel phase carriers or excipients. Examples of such carriers or excipients include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives, gelatin and polymers such as polyethylene glycol.

III. Methods of using the polymorphs of the invention

The polymorphic forms of Compound 1 of the present invention are suitable for modulating the activity of protein kinases. More particularly, such polymorphs are useful as anti-angiogenic agents and for modulating and/or inhibiting the activity of protein kinases, e.g. (among others) in complex with VEGF, FGF, CDK, TEK, CHK1, LCK , FAK, and phosphorylated kinase-related activities are therefore provided for the treatment of cancer or other diseases associated with cell proliferation mediated by protein kinases in mammals, including humans.

A therapeutically effective amount of an agent of the invention can be administered, usually in the form of a pharmaceutical composition, to treat diseases mediated by modulating or modulating protein kinases. "Effective amount" is intended to mean an amount of an agent sufficient to effect treatment of a disease mediated by the activity of one or more protein kinases (eg, tyrosine kinases) when administered to a mammal in need of such treatment. Accordingly, a therapeutically effective amount of a compound of the invention is an amount sufficient to modulate, modulate or inhibit the activity of one or more protein kinases to reduce or alleviate a disease condition mediated by said activity. The effective amount of a given compound will vary depending on factors such as the disease condition and its severity and the identity and condition (eg body weight) of the mammal in need of treatment, but can still be routinely determined by one skilled in the art. "Treatment" is intended to mean at least amelioration of a disease condition in a mammal, such as a human, that is affected at least in part by the activity of one or more protein kinases, such as tyrosine kinases, and includes preventing the occurrence of a disease condition in a mammal , especially when the mammal is found to be predisposed to the disease condition but has not yet been diagnosed as having the disease condition; modulating and/or suppressing the disease condition; and/or alleviating the disease condition. Exemplary disease conditions include diabetic retinopathy, neovascular glaucoma, rheumatoid arthritis, psoriasis, age-related macular degeneration (AMD), and cancer (solid tumors).

The activity of polymorphic forms of Compound 1 as modulators of protein kinase activity can be measured by any method available to those skilled in the art, including in vivo and/or in vitro assays. Examples of suitable assays for activity measurement include those described in Parast C. et al., Biochemistry, 37, 16788-16801 (1998); Jeffrey et al., Nature, 376, 313-320 (1995); WIPO International Publication No. WO 97/34876 and those in WIPO International Publication No. WO 96/14843.

The present invention also relates to methods of combination therapy for the treatment of hyperproliferative disorders or conditions mediated by VEGF activity comprising administering to a mammal in need of such treatment a therapeutically effective amount of a pharmaceutical composition comprising any of the polymorphic forms or polymorphic forms discussed above. A pharmaceutical composition, combined with a therapeutically effective amount of one or more substances selected from antineoplastic agents, antiangiogenic agents, signal transduction inhibitors and antiproliferative agents. Such materials are included in PCT Publication Nos. WO 00/38715, WO 00/38716, WO 00/38717, WO 00/38718, WO 00/38719, WO 00/38730, WO 00/38665, WO 00/37107 and WO 00/38786, the disclosures of which are incorporated herein by reference in their entirety.

Examples of antineoplastic agents include mitotic inhibitors, such as vinca alkaloid derivatives, such as vinblastine, vinorelbine, vindesine, and vincristine; colchicine, allochochine, halocolchicine ( halichodrine), N-benzoyltrimethyl-methyl ether colchicic acid, dolastatin10, maytansine, rhizocin, taxanes such as paclitaxel, docetaxel (Taxotere ), 2′-N-[3-(dimethylamino)propyl]glucose-paclitaxel ester (glutaramate) (paclitaxel derivative), thiocolchicine, tritylcysteine, teniposide , methotrexate, azathioprine, fluorouracil, cytarabine, 2'2'-difluorodeoxycytidine (gemcitabine)), doxorubicin and mitomycin. Alkylating agents such as cisplatin, carboplatin, oxiplatin, isoproplatin, ethyl ester of N-acetyl-DL-sarcosyl-L-leucine (Asaley or Asalex), 1,4- Cyclohexadiene-1,4-dicarbamic acid, 2,5-bis(1-aziridine)-3,6-dioxo-diethyl ester (deacroquinone), 1,4-bis (Methylsulfonyloxy)butane (bisulfan or leucosulfan) chlorurecin, clomesone, cyanomorpholino doxorubicin, cyclodisone, didehydrodulcitol , Fluorodopan (fluorodopan), hepsulfam, mitomycin C, oxymethylsulfanthone mitomycin C, mitozolomide, 1-(2-chloroethyl)-4-(3-chloropropyl )-piperazine dihydrochloride, piperazine diketone, propiperazine bromide, porphyrin, spirohydantoin mustard, epoxytriazinone, tetraplatinum, thiotepa, triimazine , uracil mustard, bis(3-methanesulfonyloxypropyl)amine hydrochloride, mitomycin, nitrosourea agents such as cyclohexyl-chloroethylnitrosourea, methylcyclohexyl -Chloroethylnitrosourea, 1-(2-chloroethyl)-3-(2,6-dioxo-3-piperidinyl)-1-nitroso-urea, bis(2-chloro Ethyl)nitrosourea, procarbazine, dacarbazine, nitrogen mustard related compounds such as nitrogen mustard, cyclophosphamide, ifosfamide, melphalan, chlorambucil, estramustine phosphate , streptozoin and temozolomide. DNA anti-metabolites such as 5-fluorouracil, cytarabine, hydroxyurea, 2-[(3-hydroxy-2-indenyl)methylene]-hydrazine carbothioamide, deoxyfluridine, 5-Hydroxy-2-formylpyridinethiosemicarbazide, α-2′-deoxy-6-thioguanosine, aphidicolin glycine, 5-azadeoxycytidine, β-thioguanine deoxy Nucleosides, cyclic cytosine, guanidine, inosine glyceredialdehyde, macbecin II, pyrazolimidazole, cladribine, pentostatin, thioguanine, mercaptopurine, bleomycin , 2-chlorodeoxyadenosine, thymidylate synthase inhibitors, such as raltitrexed and pemetrexed disodium, clofarabine, floxuridine and fludarabine. DNA/RNA anti-metabolites such as L-alanosine, 5-azacytidine, acivicin, aminopterin and their derivatives such as N-[2-chloro-5-[[(2,4 -Diamino-5-methyl-6-quinazolinyl)methyl]amino]benzoyl]-L-aspartic acid, N-[4-[[(2,4-diamino-5- Ethyl-6-quinazolinyl)methyl]amino]benzoyl]-L-aspartic acid, N-[2-chloro-4-[[(2,4-diaminopteridinyl)methyl Base]amino]benzoyl]-L-aspartic acid, soluble Baker's folic acid antagonist, dichloroallyl 2-hydroxy-1,4-naphthoquinone, buquinal, tegafur, Dihydro-5-azacytidine, methotrexate, N-(phosphonacetyl)-L-aspartic acid tetrasodium salt, pyrazolofuran, trimethopterin, plicamycin, actin Bacterin D, cryptophycin and analogues, such as cryptophycin-52 or one of the preferred antimetabolites such as disclosed in European Patent Application No. 239362, such as N- (5-[ N- ( 3,4-Dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N -methylamino]-2-thienoyl)-L-glutamic acid; growth factor inhibitor cell cycle inhibitors; intercalating antibiotics such as doxorubicin and bleomycin; proteins such as interferons; and antihormonal agents such as anti-estrogens such as Novaldex (tamoxifen) or For example anti-androgens such as Casodex ^™ (4'-cyano-3-(4-fluorophenylsulfonyl)-2-hydroxy-2-methyl-3'-(trifluoromethyl)propionanilide) . Such combination therapy may be achieved by simultaneous, sequential or separate administration of the individual components of the treatment.

Anti-angiogenic agents include MMP-2 (matrix metalloproteinase 2) inhibitors, MMP-9 (matrix metalloproteinase 9) inhibitors and COX-II (cyclooxygenase II) inhibitors. Examples of suitable COX-II inhibitors include CELEBREX ^™ (alecoxib), valdecoxib and rofecoxib. Examples of suitable matrix metalloproteinase inhibitors are described in WO 96/33172 (published October 24, 1996), WO 96/27583 (published March 7, 1996), European Patent Application No. 97304971.1 (published July 1997 February 8), European Patent Application No. 99308617.2 (applied on October 29, 1999), WO 98/07697 (published on February 26, 1998), WO 98/03516 (published on January 29, 1998), WO 98/34918 (published on August 13, 1998), WO 98/34915 (published on August 13, 1998), WO 98/33768 (published on August 6, 1998), WO 98/30566 (published on July 1998) published on July 16), European Patent Publication 606,046 (published on July 13, 1994), European Patent Publication 931,788 (published on July 28, 1999), WO 90/05719 (published on May 31, 1990), WO 99 /52910 (published on October 21, 1999), WO 99/52889 (published on October 21, 1999), WO 99/29667 (published on June 17, 1999), PCT International Application No. PCT/IB98/01113 (filed 21 July 1998), European Patent Application No. 99302232.1 (filed 25 March 1999), British Patent Application No. 9912961.1 (filed 3 June 1999), US Patent Provisional Application No. 60/148,464 (applied August 12, 1999), US Patent 5,863,949 (issued January 26, 1999), US Patent 5,861,510 (issued January 19, 1999) and European Patent Publication 780,386 (issued June 25, 1997 ), all documents are incorporated by reference in their entirety into this application. Preferred MMP-2 and MMP-9 inhibitors are those that have little or no activity against MMP-1. More preferred are those relative to other matrix metalloproteases (i.e. MMP-1, MMP-3, MMP-4, MMP-5, MMP-6, MMP-7, MMP-8, MMP-10, MMP-11, MMP- 12 and MMP-13) substances that selectively inhibit MMP-2 and/or MMP-9.

Examples of MMP inhibitors include AG-3340, RO 32-3555, RS 13-0830, and the following compounds: 3-[[4-(4-fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxyaminomethyl Acyl-cyclopentyl)-amino]-propionic acid; 3-exo-3-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane Alkane-3-carboxylic acid hydroxyamide; (2R,3R)1-[4-(2-Chloro-4-fluoro-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine -2-carboxylic acid hydroxyamide; 4-[4-(4-fluoro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-4-carboxylic acid hydroxyamide; 3-[[4-(4 -fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-cyclobutyl)-amino]-propionic acid; 4-[4-(4-chloro-phenoxy)-benzenesulfonyl Amino]-tetrahydro-pyran-4-carboxylic acid hydroxyamide; 3-[4-(4-chloro-phenoxy)-benzenesulfonylamino]-tetrahydro-pyran-3-carboxylic acid hydroxyamide; (2R, 3R) 1-[4-(4-fluoro-2-methyl-benzyloxy)-benzenesulfonyl]-3-hydroxy-3-methyl-piperidine-2-carboxylic acid hydroxyamide; 3-[[4-(4-Fluoro-phenoxy)-benzenesulfonyl]-(1-hydroxycarbamoyl-1-methyl-ethyl)-amino]-propionic acid; 3-[[4- (4-fluoro-phenoxy)-benzenesulfonyl]-(4-hydroxycarbamoyl-tetrahydro-pyran-4-yl)-amino]-propionic acid; 3-exo-3-[4-( 4-Chloro-phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylic acid hydroxyamide; 3-endo-3-[4-(4-fluoro- Phenoxy)-benzenesulfonylamino]-8-oxa-bicyclo[3.2.1]octane-3-carboxylic acid hydroxyamide; 3-[4-(4-fluoro-phenoxy)-benzenesulfonyl Amino]-tetrahydro-furan-3-carboxylic acid hydroxyamide; and pharmaceutically acceptable salts, solvates and hydrates thereof.

Examples of signal transduction inhibitors include agents that inhibit EGFR (epidermal growth factor receptor) responses, such as EGFR antibodies, EGF antibodies, and molecules that are EGFR inhibitors; VEGF (vascular endothelial growth factor) inhibitors; and erbB2 receptors Inhibitors, such as organic molecules or antibodies that bind to the erbB2 receptor, such as HERCEPTIN ^™ (Genentech, Inc. of South San Francisco, California, USA).

EGFR inhibitors are described, for example, in WO 95/19970 (published July 27, 1995), WO 98/14451 (published April 9, 1998), WO 98/02434 (published January 22, 1998) and U.S. In Patent 5,747,498 (issued May 5, 1998). EGFR-inhibitors include (but are not limited to) monoclonal antibody C225 and anti-EGFR 22Mab (ImClone Systems Incorporated of New York, New York, USA), compound ZD-1839 (AstraZeneca), BIBX-1382 (Boehringer Ingelheim), MDX- 447 (Medarex Inc. of Annandale, New Jersey, USA) and OLX-103 (Merck & Co. of Whitehouse Station, New Jersey, USA), VRCTC-310 (Ventech Research) and EGF fusion toxin (Seragen Inc. of Hopkinton, Massachusetts).

VEGF inhibitors, such as SU-5416 and SU-6668 (Sugen Inc. of South San Francisco, California, USA) may also be combined or co-administered with the composition. VEGF inhibitors are described, for example, in WO 99/24440 (published 20 May 1999), PCT International Application PCT/IB99/00797 (filed 3 May 1999), WO 95/21613 (published 17 August 1995) Publication), WO 99/61422 (published on December 2, 1999), U.S. Patent 5,834,504 (issued on November 10, 1998), WO 98/50356 (published on November 12, 1998), U.S. Patent 5,883,113 (1999 issued on March 16, 1999), US Patent 5,886,020 (issued on March 23, 1999), US Patent 5,792,783 (issued on August 11, 1998), WO 99/10349 (published on March 4, 1999) , WO 97/32856 (published on September 12, 1997), WO 97/22596 (published on June 26, 1997), WO98/54093 (published on December 3, 1998), WO 98/02438 (published on 1998 Published on April 22), WO 99/16755 (published on April 8, 1999) and WO 98/02437 (published on January 22, 1998), the entire texts of all documents are incorporated into this application by reference. Other examples of some specific VEGF inhibitors are IM862 (Cytran Inc. of Kirkland, Washington, USA); anti-VEGF monoclonal antibody bevacizumab (Genentech, Inc. of South San Francisco, California) and nuclear Angiozyme, a synthetic ribozyme from Ribozyme (Boulder, Colorado) and Chiron (Emeryville, California).

ErbB2 receptor inhibitors such as GW-282974 (Glaxo Wellcome plc) and monoclonal antibodies AR-209 (Aronex Pharmaceuticals Inc. of The Woodlands, Texas, USA) and 2B-1 (Chiron) can be administered in combination with the composition. Such erbB2 inhibitors include WO 98/02434 (published on January 22, 1998), WO 99/35146 (published on July 15, 1999), WO 99/35132 (published on July 15, 1999), WO 98 /02437 (published on January 22, 1998), WO 97/13760 (published on April 17, 1997), WO95/19970 (published on July 27, 1995), U.S. Patent 5,587,458 (issued on December 24, 1996 to) and those described in US Patent 5,877,305 (issued March 2, 1999), each of which is incorporated by application in its entirety into this application. ErbB2 receptor inhibitors suitable for use in the present invention are also described in U.S. Provisional Application No. 60/117,341, filed January 27, 1999, and U.S. Provisional Application No. 60/117,346, filed January 27, 1999, Both are incorporated into this application in their entirety by application.

Other antiproliferative agents that may be used include inhibitors of the enzyme farnesyl protein transferase and inhibitors of the receptor tyrosine kinase PDGFr, including compounds disclosed and claimed in the following U.S. patent application: 09/221,946 (December 1998 28); 09/454,058 (filed 2 December 1999); 09/501,163 (filed 9 February 2000); 09/539,930 (filed 31 March 2000); 09/202,796 (filed 1997 09/384,339 (filed August 26, 1999) and 09/383,755 (filed August 26, 1999) and compounds disclosed and claimed in the following U.S. provisional patent applications: 60/168207 (filed 30 November 1999); 60/170,119 (filed 10 December 1999); 60/177,718 (filed 21 January 2000); 60/168,217 (filed 30 November 1999) and 60 /200,834 (applied on May 1, 2000). The entirety of each of the aforementioned patent applications and provisional patent applications is incorporated into this application by application.

The compositions of the present invention may also be used with other agents useful in the treatment of abnormal cell growth or cancer, such other agents include, but are not limited to, agents that enhance anti-tumor immune responses, such as CTLA4 (cytotoxic lymphocyte antigen 4 ) antibodies and other agents that can block CTLA4; and antiproliferative agents, such as other farnesyl protein transferase inhibitors. Specific CTLA4 antibodies useful in the present invention include those described in US Provisional Application 60/113,647 (filed December 23, 1998), which is hereby incorporated by reference in its entirety.

Example

The following examples will further illustrate the preparation of unique polymorphs of the present invention (i.e. polymorphs I, II, III, IV, VI, VII and VIII of Compound 1) but are not intended to be limited as defined in this application or hereinafter scope of the claimed invention. All temperatures are listed in degrees Celsius and all parts and percentages are by weight unless otherwise indicated. HPLC data were obtained using a Hewlett Packard HP-1100 HPLC.

Example 1: Preparation and Characterization of Polymorph I of Compound 1

6-[2-(Methylcarbamoyl)phenylsulfanyl]- 3-E-[2-(Pyridin-2-yl)vinyl]indazole (4.6 g) was slurried in 50 mL of methanol at 50 °C for 15 minutes, followed by the addition of 50 mL of water. The slurry was stirred well and allowed to cool to room temperature. The solid was collected by filtration, washed with 50 mL of water and then 30 mL of ethyl acetate. The product was then dried under high vacuum. HPLC purity greater than 99%.

FIG. 1A is an X-ray powder diffraction pattern of polymorph I of Compound 1. FIG. Polymorph I of compound 1 was further characterized by differential scanning calorimetry. FIG. 1B is a differential scanning calorimetry (DSC) curve of a sample of polymorph I of Compound 1. FIG. At a scan rate of about 10°C/min, a sample of polymorph I of Compound 1 exhibited an endotherm onset at 183-190°C.

Example 2: Preparation and Characterization of Polymorphic Form II of Compound 1

Compound 1 polymorph II (as a hydrate) was generated by placing Compound 1 polymorph I (37 mg) in a 93% relative humidity cabinet for 6 days at room temperature. (HPLC purity >98.5%). FIG. 2A is an X-ray powder diffraction pattern of Compound 1 polymorph II. Polymorph II of compound 1 was further characterized by differential scanning calorimetry. FIG. 2B is a differential scanning calorimetry (DSC) curve of a polymorph II sample of Compound 1. FIG. At a scan rate of about 10°C/min, Form II exhibited an endotherm starting at 102, 152, and 202°C, followed by an exotherm at 206°C and another exotherm at 210°C.

Example 3: Preparation and Characterization of Polymorph III of Compound 1

Polymorph III of Compound 1 was prepared by neutralizing the p-toluenesulfonate derivative of Compound 1 in ethyl acetate, followed by vacuum drying at 65°C. The p-toluenesulfonate salt of compound 1 (421 g) was suspended in 1800 mL of 0.84M NaHCO ₃ and 1800 mL of ethyl acetate and stirred at 65 °C for 2 hours. The solid was collected by filtration, washed with 1800 mL of water and with 800 mL of ethyl acetate and dried overnight in laboratory vacuum at 50°C. Yield: 92% (HPLC purity greater than 99%). Polymorph III is the ethyl acetate solvate.

FIG. 3A is an X-ray powder diffraction pattern of Compound 1 polymorph III. Polymorph III of compound 1 was further characterized by differential scanning calorimetry. FIG. 3B is a differential scanning calorimetry (DSC) curve of a sample of polymorph III of Compound 1. FIG. At a scan rate of about 10°C/min, a sample of polymorph III of Compound 1 exhibited an endotherm onset at 125-129°C followed by another endotherm at 210°C.

Polymorph III was further characterized by thermogravimetric analysis (TGA). Figure 3C is a thermogravimetric analysis (TGA) curve of a sample of polymorph III. A typical TGA thermogram of a sample of polymorph III shows desolvation. The loss of ethyl acetate is indicated by a 10% sample weight loss at 125-129°C at a scan rate of about 10°C/min.

Example 4a: Preparation and Characterization of Polymorphic Form IV of Compound 1

Compound 1 polymorph IV was prepared from Compound 1 polymorph III. A sample (1.015 kg) of polymorph III of Compound 1 was dissolved in 3 L of methanol and 5 L of acetic acid at 60°C. The solution was then filtered and concentrated by moderate vacuum. 6 L of xylenes were added at 60°C and then removed by full vacuum. 4 L xylenes were added and then removed under full vacuum, followed by 4 L xylene additions to work up. The xylenes were then removed in full vacuum to yield polymorph Form IV of Compound 1 in 92% yield. HPLC analysis showed greater than 98.5% purity.

FIG. 4A is an X-ray powder diffraction pattern of Compound 1 polymorph IV. Polymorphic Form IV of Compound 1 was further characterized by differential scanning calorimetry. Figure 4B is a differential scanning calorimetry (DSC) curve of a polymorphic Form IV sample. At a scan rate of about 10°C/min, a sample of polymorphic Form IV of Compound 1 exhibited an endotherm onset at 216°C.

Example 4b: Preparation and Characterization of Polymorphic Form IV of Compound 1

After compound 1 was synthesized using palladium catalyst, the following procedure was performed to remove residual palladium and crystallize compound 1 as polymorph IV.

A 12 L 3-neck flask equipped with a mechanical stirrer was charged with 160.20 g of Compound 1 along with 1.6 L of DMA and 1.6 L of THF. After stirring for 20 minutes, the mixture became homogeneous. To the clear solution was charged 800.99 g of 10% cysteine-silica and the resulting mixture was allowed to stir overnight at room temperature. The mixture was filtered through a "medium" sintered fritted glass funnel and the filter cake was washed with a solution of 500 mL DMA and 500 mL THF. The filter cake was further washed with 2.0 L THF and the filtrate was collected in another flask. Volatile portions of the latter filtrate were removed in vacuo and the residue was combined with the main filtrate. The combined filtrates were refilled into the 12L flask, followed by the addition of 800 g of 10% cysteine-silica. The flask was equipped with a mechanical stirrer and stirred at room temperature over the weekend. The mixture was filtered through a "medium" sintered glass fritted funnel and the silica was washed with a solvent mixture of 500 mL DMA and 500 mL THF, followed by 3.0 L THF. Volatile portions of the filtrate were removed in vacuo and the residual solution was transferred to a 22L 3-neck flask and treated with 12L of water (added over 20 minutes), at which stage a thick precipitate formed. After stirring overnight, the mixture was filtered and the filter cake was washed with 2.0 L of water and suction dried.

The filter cake was packed into a 5L 3-neck flask, followed by 1.6L THF and 160mL DMF. The flask was equipped with a mechanical stirrer, reflux condenser and the mixture was heated at reflux for 8 hours. After cooling overnight, the mixture was filtered through sharkskin filter paper and suction dried. The filter cake was charged to a 5L 3-neck flask and 1.6L MeOH was added. The flask was equipped with a mechanical stirrer, water condenser and the contents were heated at reflux for 6 hours. After cooling overnight, the mixture was filtered through sharkskin filter paper and suction dried. The filter cake was dissolved in 1.6 L HOAc with the aid of gentle heating in a water bath on a rotary evaporator. The solution was filtered through #3 filter paper and the total volume of the filtrate was reduced to about 500 mL volume on a rotary evaporator at 60 °C/60 mmHg. At this stage, a large amount of the mixture remained a yellow solution and a small amount of precipitate formed. The flask was charged with 500 mL of xylene (precipitation formed) and the total volume was reduced to about 500 mL on a rotary evaporator at 60 °C/60 mmHg. The method was repeated two additional times. After cooling, the mixture was filtered, the filter cake was washed with 500 mL xylene and suction dried. The filter cake was transferred to a glass dish and further dried overnight at 80°C/27 inches of vacuum. The filter cake was off-white in color and weighed 108.38 g, as determined subsequently in the crystalline form of Form IV.

Example 5: Preparation and Characterization of Polymorphic Form VI of Compound 1

Polymorph III (2 g) of Compound 1 was suspended in 15 mL of ethanol. 4 g of p-toluenesulfonic acid monohydrate were added and the mixture was heated to 82°C for 14 hours. After cooling to room temperature, 25 mL of saturated NaHCO ₃ solution was added and the suspension was stirred for 2 hours. The solid was collected by filtration, washed with 50 mL of water and dried overnight in a laboratory vacuum at 45°C (HPLC purity >99%).

FIG. 5A is an X-ray powder diffraction pattern of polymorph VI of Compound 1. FIG. Polymorph VI of compound 1 was further characterized by differential scanning calorimetry. FIG. 5B is a differential scanning calorimetry (DSC) curve of a polymorph VI sample of Compound 1. FIG. At a scan rate of about 10°C/min, Form VI exhibited an onset endotherm at about 197°C followed by another endotherm at about 209°C.

Example 6: Preparation and Characterization of Polymorphic Form VII of Compound 1

Polymorph VI of Compound 1 (102 mg) was suspended in 20 mL of isopropanol, refluxed for 30 minutes and cooled to room temperature. The solid was collected by filtration, washed with isopropanol and dried in vacuo. Polymorph VII of Compound 1 is an isopropanol solvate.

FIG. 6A is an X-ray powder diffraction pattern of Compound 1 polymorph VII. Polymorphic Form VII of Compound 1 was further characterized by differential scanning calorimetry. 6B is a differential scanning calorimetry (DSC) curve of a polymorphic Form VII sample of Compound 1. Typical curves are sample dependent. At a scan rate of about 10°C/min, the sample isolated from refluxing THF showed an endotherm at 105°C, followed by an exotherm at 115°C and then endotherms at 137 and 175°C.

Example 7: Preparation and Characterization of Polymorphic Form VIII of Compound 1

Polymorph VII of Compound 1 was dissolved in a minimal amount of dioxane at reflux at about 100°C and then allowed to cool to room temperature overnight. Larger yellow crystals were collected by filtration, washed with dioxane and dried in vacuo. Polymorph VIII of Compound 1 is a dioxane solvate. Figure 7 shows the X-ray powder diffraction pattern of Compound 1 polymorph VIII.

Example 8: Application of Polymorph IV in Tablet Formulation

Povidone (4% w/w) was dissolved in water (5 times, w/w) to form a solution for granulation. The polymorph IV (37%, weight/weight) of compound 1 as prepared in Example 4 was mixed with lactose (25%, weight/weight), cornstarch (16%, weight/weight) and a part of cross-linked carboxyl Sodium methylcellulose (2%, w/w) was combined in a high clarity granulator. The mixture was dry blended and then granulated with povidone solution. The granules are first wetted for 2 minutes and dried at 60°C to a loss on drying value of 5% or less. The material was dry ground with a screen size 045R. The milled material was blended with the remaining croscarmellose sodium (3% w/w) and microcrystalline cellulose (12% w/w). The blended mixture was further blended with magnesium stearate (1%, w/w). The mixture was compressed on a tablet compression machine to yield tablets containing 160 mg of Compound 1 each.

Embodiment 9: the generation of the salt of compound 1

Compound 1 was subjected to a salt screen to improve its water solubility. Compound 1 was added to 7 different 100 mM acid solutions and stirred for 14 days to generate 7 salt forms of Compound 1 in situ. The seven acids used are as follows: methanesulfonic acid, sulfuric acid, hydrochloric acid, phosphoric acid, hydrobromic acid, maleic acid and benzenesulfonic acid. For each of these different acids, 20 mg of compound 1 was stirred with 1.6 mL of a 100 mM solution of the acid of interest in a sealed vial in the dark. To ensure that maximum solubility levels are achieved, samples are checked periodically to ensure that excess solids are present. After 8 days, 400 μL of the mixture was removed and centrifuged at 14,000 rpm for 5 minutes. Then 100 μL of the supernatant was removed, diluted with 900 μL of a 1:1 mixture of acetonitrile/methanol and then analyzed by HPLC. A second set of data was collected 14 days after the start of the experiment to observe any long-term changes in solubility. Samples at 14 days were prepared following the same procedure as described for the 8-day study. HPLC analysis was performed using a Primesphere column C ₁₈ , 5 μm, 150×4.6 mm, at a flow rate of 1.5 mL/min and an injection volume of 10 μL. The table below summarizes the solubility of 7 different salt forms of Compound 1 formed over 2 weeks. Typically solubility values show only small changes from 8 to 14 days. Salt Solubility at 8 days (μg/mL) Solubility at 14 days (μg/mL) Methanesulfonic acid 1970 1835 sulfuric acid 601 603 hydrochloric acid 576 549 phosphoric acid 295 292 hydrobromic acid 277 220 Maleic acid 69 68 Benzenesulfonic acid 10 11

The salt forms of Compound 1 showing the highest solubility (methanesulfonic acid, sulfuric acid and hydrochloric acid) were further characterized. Approximately 30 mg of each salt was placed in a vial in a cabinet at room temperature and 93% relative humidity. After 6 days, the weight percent water absorbed, X-ray powder diffraction pattern and differential scanning calorimetry data were obtained. Two polymorphic forms of the hydrochloride salt (Forms I and II), three polymorphic forms of the mesylate salt (Forms I, II and III) and three polymorphic forms of the sulfate salt (Forms I, II and III) were observed. I, II and III). The stability of these polymorphs to high intensity light was further analyzed. Approximately 0.4 mg of each salt was weighed into an HPLC vial. This step was repeated a total of 5 times for each salt to generate 4 samples and one standard. Samples were placed in a high intensity light chamber and illuminated for 0, 1, 2 and 6 hours. 1 mL of acetonitrile and 1 mL of methanol were added to dissolve each standard and sample prior to HPLC analysis. All samples were significantly degraded (14%-97%) after exposure to high intensity light, except Form II of the sulphate.

Example 10: Human Metabolites of Compound 1

Compound 1 undergoes extensive metabolism in humans into multiple metabolites, as shown in Figure 8. The chemical structures of the three oxidized metabolites, M12 (sulfoxide of compound 1), M15 (sulfone of compound 1), and M9 were confirmed based on comparison of the chromatographic retention times and mass spectra of the in vivo metabolites with their bona fide reference standards. (Mixed sulfoxide/and N-oxidation products of compound 1). The chemical structure of the glucuronide (M7) of compound 1 was confirmed by isolating metabolites followed by NMR measurements. Metabolite M5 demonstrated a [M+H] ⁺ ion at m/z 342. Interpretation of the MS ² and MS ³ product ion mass spectra of M5 indicated that M5 is the despyridylcarboxylic acid of compound 1. The proposed structures (or elemental compositions) of M5 and its major fragment ions (m/z 342, 311, 265, and 237) are all in good agreement with elemental compositions determined by accurate mass (all protons measured with an accuracy < 1.2 ppm). The definitive structures of the metabolites M8a, M12a and M14 are currently unknown.

Claims (10)

1. formula 16-[2-(methyl carbamyl) the phenyl sulfane base of expression]-3-E-[2-(pyridine-2-yl) vinyl] crystal formation of indazole or its pharmaceutically acceptable salt:

2. the crystal formation of claim 1, wherein this crystal formation is selected from the group of being made up of polymorphic I, type II, type III, type IV, type VI, type VII and type VIII.

3. the crystal formation of claim 1, wherein this crystal formation is the polymorphic form of type IV.

4. the crystal formation of claim 1, wherein this crystal formation has to comprise and is positioned at powder X-ray-optical diffraction pattern that diffraction angle (2 θ) is the peak at 8.9 ± 0.1 and 15.7 ± 0.1 places.

5. the crystal formation of claim 1, wherein this crystal formation has to comprise and is positioned at powder X-ray-optical diffraction pattern that diffraction angle (2 θ) is the peak at 8.9 ± 0.1,14.6 ± 0.1,15.7 ± 0.1 and 19.2 ± 0.1 places.

6. the crystal formation of claim 1, wherein this crystal formation has and comprises the powder X-ray-optical diffraction pattern that is positioned at diffraction angle (2 θ) and the peak at basic identical place shown in Fig. 4 A.

(7.6-[2-methyl carbamyl) phenyl sulfane base]-3-E-[2-(pyridine-2-yl) vinyl] solid form of indazole or its pharmaceutically acceptable salt, wherein this solid form comprises in the following crystal formation at least two kinds: polymorphic I, II, III, IV, VI, VII and VIII.

8. pharmaceutical composition, it comprises in the claim 1 to 7 each crystal formation.

9. a treatment is by the method for the mammalian diseases situation of the active mediation of protein kinase, and this method comprises the pharmaceutical composition of the claim 8 of the Mammals treatment significant quantity that needs this treatment.

10. the method for claim 9, wherein said mammalian diseases situation is relevant with tumor growth, cell proliferation or vasculogenesis.

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