Crystalline forms of 5-chloro-N4-[2-(dimethylphosphoryl) phenyl]-N2-{2-methoxy-4-[4-(4-methylpiperazin-1-yl) piperidin-1-yl] phenyl} pyrimidine-2, 4-diamine

Abstract

The present application relates to a crystalline form of 5-chloro-N4-[2-(dimethylphosphoryl) phenyl]-N2-{2-methoxy-4-[4-(4-methylpiperazin-1-yl) piperidin-1-yl] phenyl} pyrimidine-2, 4-diamine. Disclosed herein are crystalline forms of brutinib, pharmaceutical compositions comprising crystalline forms of brutinib, and methods of making and using the same.

Description

[0001] This application is a divisional application of an application filed on October 21, 2015, with application number 202010394526.8, and invention name “Crystalline form of 5-chloro-N4-[2-(dimethylphosphoryl)phenyl]-N2-{2-methoxy-4-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]phenyl}pyrimidine-2,4-diamine”.

[0002] The application date is October 21, 2015, the application number is 202010394526.8, and the invention name is "5-chloro-N4-[2-(dimethylphosphoryl)phenyl]-N2-{2-methoxy-4-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]phenyl}pyrimidine-2,4-diamine crystalline form" is the application date is October 21, 2015, the application number is 201580069762.X, and the invention name is Divisional application of application for “5-chloro-N4-[2-(dimethylphosphoryl)phenyl]-N2-{2-methoxy-4-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]phenyl}pyrimidine-2,4-diamine, crystalline form” (formerly named “5-chloro-N4-[2-(dimethylphosphoryl)phenyl]-N2-{2-methoxy-4-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]pyrimidine-2,4-diamine, crystalline form”)

[0003] The present application relates to novel crystalline forms of 5-chloro-N4-[2-(dimethylphosphoryl)phenyl]-N2-{2-methoxy-4-[4-(4-methylpiperazin-1-yl)piperidin-1-yl]phenyl}pyrimidine-2,4-diamine (also known as "AP26113" and "brigatinib"), compositions containing such crystalline forms, and methods of making and using the same.

[0004] Brigatinib has the chemical formula C 29 H 39 ClN7O2P, which corresponds to a formula weight of 584.09 g / mol. Its chemical structure is shown below:

[0005]

[0006] Brigatinib is a multi-targeted tyrosine kinase inhibitor that can be used to treat non-small cell lung cancer (NSCLC) and other diseases. It is a potent inhibitor of ALK (anaplastic lymphoma kinase) and is in clinical development for the treatment of adult patients with ALK-driven NSCLC. Crizotinib It is an FDA-approved drug for the first-line treatment of ALK-positive NSCLC. "Despite initial response to crizotinib, most patients relapse within 12 months due to the emergence of resistance." Shaw et al., New Eng. J. Med. 370: 1189-97 2014. Therefore, more and more cancer patients need new and effective ALK-positive cancer therapies.

[0007] Brigatinib may also potentially be used to treat other diseases or conditions involving ALK or other protein kinases inhibited by brigatinib. Such kinases and their associated disorders or conditions are disclosed in WO 2009 / 143389, both of which are hereby incorporated by reference for all purposes.

[0008] Knowledge of potential polymorphs of an active pharmaceutical ingredient (API), such as brigatinib, can be used to develop drugs, as can knowledge of the characteristics of those polymorphs. Not knowing the specific polymorphs present or desired in an API can lead to inconsistent manufacturing of the API, thus resulting in drugs that can potentially differ between batches of API. In addition, knowledge of the polymorphs of an API informs and allows long-term system stability assays of the API. Once a specific polymorph is selected for drug development, a method for reproducibly preparing the polymorph may be useful. It can also be used as a method for preparing an API (such as brigatinib) at a specified level or higher of chemical purity and / or polymorphic purity.

[0009] The chemical structure of brigatinib was first disclosed in WO 2009 / 143389, which is owned by the applicant (ARIAD Pharmaceuticals, Inc.) and is hereby incorporated by reference in its entirety for all purposes. Example 122 of WO 2009 / 143389 discloses the synthesis of brigatinib and indicates that the product is obtained as an off-white solid but the product does not provide additional characteristics, such as chemical purity or solid form. Example 122 does not indicate the extent to which its product is crystalline, if any.

[0010] Provided herein are certain crystalline and other polymorphs of brigatinib, some of which are suitable for pharmaceutical formulation development.

[0011] In some embodiments, the present disclosure relates to crystalline brigatinib. In some embodiments, the present disclosure relates to substantially pure crystalline brigatinib.

[0012] In one embodiment, the present disclosure relates to polymorphs of brigatinib. Polymorphs of brigatinib are designated herein as Form A, Form B, Form C, Form D, Form E, Form F, Form G, Form H, Form J, and Form K.

[0013] In another embodiment, the present disclosure relates to substantially pure crystalline forms of brigatinib. Substantially pure crystalline forms of brigatinib are designated herein as Form A, Form B, Form C, Form D, Form E, Form F, Form G, Form H, Form J, and Form K.

[0014] In another embodiment, the present disclosure relates to a pharmaceutical composition consisting essentially of a crystalline form of brigatinib disclosed herein and at least one additional component selected from a pharmaceutically acceptable carrier, a pharmaceutically acceptable solvent, and a pharmaceutically acceptable excipient. In another embodiment, the present disclosure relates to a pharmaceutical composition consisting of at least one polymorph of brigatinib disclosed herein and at least one additional component selected from a pharmaceutically acceptable carrier, a pharmaceutically acceptable solvent, and a pharmaceutically acceptable excipient.

[0015] In another embodiment, the disclosure relates to a method of treating a disorder and / or condition responsive to protein kinase inhibition in a subject by administering to the subject a polymorphic form of brigatinib disclosed herein. In certain embodiments, at least one disorder and / or condition is non-small cell lung cancer (NSCLC).

[0016] In another embodiment, the disclosure relates to a method of treating a disorder and / or condition responsive to protein kinase inhibition in a subject by administering to the subject a substantially pure crystalline form of brigatinib disclosed herein. In certain embodiments, when the protein kinase is ALK or a mutant form thereof, at least one disorder and / or condition is NSCLC.

[0017] This application provides the following:

[0018] 1) Crystalline brigatinib.

[0019] 2) Substantially pure crystalline brigatinib.

[0020] 3) Crystalline Form A of Brigatinib.

[0021] 4) The crystalline Form A as described in 3), wherein the crystalline Form A of brigatinib is substantially pure.

[0022] 5) The crystalline form A as described in 3) or 4) has substantially Figure 2 The x-ray powder diffraction spectrum is shown.

[0023] 6) The crystalline form A as described in 3) or 4), which has an x-ray powder diffraction spectrum comprising at least two peaks represented by degrees 2θ selected from 6.1, 8.6, 9.6, 10.8, 11.3, 13.5, 14.3, 15.9, 17.2, 18.9, 19.4, 20.1, 21.8, 22.6, 23.1, 23.9 and 27.7.

[0024] 7) The crystalline form A as described in 3) or 4), which has an x-ray powder diffraction spectrum comprising at least two peaks expressed in degrees 2θ selected from 9.6, 17.2, 19.4, 20.1, 23.1 and 27.7.

[0025] 8) The crystalline form A as described in any one of 3) to 7), wherein the crystalline form A is anhydrous.

[0026] 9) The crystalline form A according to any one of 3) to 7), which has an onset melting temperature of 214.5°C.

[0027] 10) The crystalline form A according to any one of 3) to 7), which has an FT-IR spectrum containing any at least one of the following bands:

[0028]

[0029]

[0030] 11) A method for preparing Form A of brigatinib, comprising raising the temperature of crystallization to 60°C and slowly adding a NaOH solution.

[0031] 12) Crystalline Form B of Brigatinib.

[0032] 13) The crystalline Form B as described in 14), wherein the crystalline Form B of brigatinib is substantially pure.

[0033] 14) The crystalline form B as described in 12) or 13), which has substantially Fig.14 The x-ray powder diffraction spectrum is shown.

[0034] 15) The crystalline form B as described in 12) or 13), which has an x-ray powder diffraction spectrum comprising at least four peaks represented by degrees 2θ selected from 5.7, 9.2, 11.5, 12.8, 14.5, 15.5, 16.9, 17.7, 19.2, 20.4, 21.8, 23.2 and 29.5.

[0035] 16) The crystalline form B as described in 12) or 13), which has an x-ray powder diffraction spectrum comprising at least two peaks expressed in degrees 2θ selected from 11.5, 14.5, 16.9, 19.2 and 23.2.

[0036] 17) Crystalline Form C of Brigatinib.

[0037] 18) The crystalline Form C as described in 17), wherein the crystalline Form C of brigatinib is substantially pure.

[0038] 19) The crystalline form C as described in 17) or 18) has substantially Fig.18The x-ray powder diffraction spectrum is shown.

[0039] 20) The crystalline form C as described in 17) or 18), which has an x-ray powder diffraction spectrum comprising at least four peaks represented by degrees 2θ selected from 2.1, 2.5, 5.4, 9.9, 10.9, 12.9, 14.9, 15.9, 16.6, 17.3, 17.9, 19.2, 20.6, 23.9, 26.8 and 27.4.

[0040] 21) The crystalline form C as described in 17) or 18), which has an x-ray powder diffraction spectrum comprising at least two peaks expressed in degrees 2θ selected from 5.4, 14.9, 15.9, 17.3, 19.2 and 23.9.

[0041] 22) Crystalline Form D of Brigatinib.

[0042] 23) The crystalline Form D as described in 22), wherein the crystalline Form D of brigatinib is substantially pure.

[0043] 24) The crystalline form D as described in 22) or 23), which has substantially Fig.19 The x-ray powder diffraction spectrum is shown.

[0044] 25) The crystalline form D as described in 22) or 23), which has an x-ray powder diffraction spectrum comprising at least four peaks expressed in degrees 2θ selected from 4.7, 9.2, 9.7, 11.1, 14.5, 17.4, 18.9, 22.4 and 23.7.

[0045] 26) The crystalline form D as described in 22) or 23), which has an x-ray powder diffraction spectrum comprising at least two peaks expressed in degrees 2θ selected from 9.7, 11.1, 17.4, 18.9 and 23.7.

[0046] 27) Crystalline Form E of Brigatinib.

[0047] 28) The crystalline Form E as described in 27), wherein the crystalline Form E of brigatinib is substantially pure.

[0048] 29) The crystalline form E as described in 27) or 28), which has substantially Fig.21 The x-ray powder diffraction spectrum is shown.

[0049] 30) The crystalline form E as described in 27) or 28), which has an x-ray powder diffraction spectrum comprising at least four peaks represented by degrees 2θ selected from 9.1, 10.2, 11.2, 12.0, 13.7, 14.4, 15.8, 16.5, 17.4, 18.3, 19.2, 21.6, 22.3, 23.1, 23.9, 26.0, 26.4, 25.8 and 29.3.

[0050] 31) The crystalline form E as described in 27) or 28), which has an x-ray powder diffraction spectrum comprising at least two peaks expressed in degrees 2θ selected from 9.1, 10.2, 15.8, 19.2 and 23.9.

[0051] 32) Crystalline Form F of Brigatinib.

[0052] 33) The crystalline Form F as described in 32), wherein the crystalline Form F of brigatinib is substantially pure.

[0053] 34) The crystalline form F as described in 32) or 33), which has substantially Fig.23 The x-ray powder diffraction spectrum is shown.

[0054] 35) The crystalline form F as described in 32) or 33), which has an x-ray powder diffraction spectrum comprising at least four peaks represented by degrees 2θ selected from 8.5, 9.8, 11.1, 16.3, 17.0, 17.6, 18.7, 19.4, 20.3, 22.0, 23.2, 23.9 and 27.1.

[0055] 36) The crystalline form F as described in 32) or 33), which has an x-ray powder diffraction spectrum comprising at least two peaks expressed in degrees 2θ selected from 9.8, 17.0, 19.4, 20.3 and 27.1.

[0056] 37) Crystalline Form G of Brigatinib.

[0057] 38) The crystalline Form G as described in 37), wherein the crystalline Form G of brigatinib is substantially pure.

[0058] 39) The crystalline form G as described in 37) or 38) has substantially Fig.24 The x-ray powder diffraction spectrum is shown.

[0059] 40) The crystalline form G as described in 37) or 38), which has an x-ray powder diffraction spectrum comprising at least four peaks represented by degrees 2θ selected from 7.2, 8.3, 9.7, 10.4, 12.9, 15.8, 18.1, 18.7, 20.7, 21.5, 22.8, 23.5, 24.5 and 26.8.

[0060] 41) The crystalline form G as described in 37) or 38), which has an x-ray powder diffraction spectrum comprising at least two peaks represented by degrees 2θ selected from 8.3, 9.7, 12.9, 15.8, 18.1, 20.7, 22.8 and 26.8.

[0061] 42) Crystalline Form H of Brigatinib.

[0062] 43) The crystalline Form H as described in 42), wherein the crystalline Form H of brigatinib is substantially pure.

[0063] 44) The crystalline form H as described in 42) or 43), which has substantially Fig.25 The x-ray powder diffraction spectrum is shown.

[0064] 45) The crystalline form H as described in 42) or 43), which has an x-ray powder diffraction spectrum comprising at least four peaks represented by degrees 2θ selected from 4.2, 5.2, 8.4, 10.9, 12.7, 15.0, 15.7, 16.5, 17.2, 18.4, 19.5 and 21.3.

[0065] 46) The crystalline form H as described in 42) or 43), which has an x-ray powder diffraction spectrum comprising at least two peaks expressed in degrees 2θ selected from 4.2, 5.2, 8.4, 10.9, 12.7 and 21.3.

[0066] 47) A pharmaceutical composition comprising at least one crystalline form of brigatinib and at least one component selected from a pharmaceutically acceptable carrier, a pharmaceutically acceptable solvent and a pharmaceutically acceptable excipient.

[0067] 48) The pharmaceutical composition of 47), wherein the at least one crystalline form of brigatinib is substantially pure.

[0068] 49) A pharmaceutical composition as described in 47) or 48), wherein the at least one crystalline form of brigatinib is selected from Form A, Form B, Form C, Form D, Form E, Form F, Form G and Form H.

[0069] 50) The pharmaceutical composition of 47) or 48), wherein the crystalline brigatinib is in Form A.

[0070] 51) A method for treating non-small cell lung cancer in a subject in need thereof, comprising administering to the subject at least one crystalline form of brigatinib.

[0071] 52) The method of 51), wherein the at least one crystalline form of brigatinib is substantially pure.

[0072] 53) The method of 51) or 52), wherein the at least one crystalline form of brigatinib is selected from Form A, Form B, Form C, Form D, Form E, Form F, Form G and Form H.

[0073] 54) The method of 51) or 52), wherein at least one crystalline form of brigatinib is Form A. BRIEF DESCRIPTION OF THE DRAWINGS

[0075] The following drawings form part of this specification and are included to further illustrate certain embodiments of the present disclosure. The present disclosure can be understood by reference to one or more of these drawings in conjunction with the detailed description of the embodiments disclosed herein.

[0076] Figure 1 This is the synthesis scheme of brigatinib.

[0077] Figure 2 is an X-ray powder diffraction (XRPD) pattern obtained from a sample of brigatinib Form A. Relative intensity (in counts) is shown on the vertical axis and angles (in degrees 2θ (° 2θ)) are shown on the horizontal axis.

[0078] Figure 3 is an adsorption / desorption plot of a dynamic vapor sorption (DVS) experiment for a sample of brigatinib Form A. Mass change (%) is shown on the vertical axis and target RH (%) is shown on the horizontal axis.

[0079] Figure 4 is a differential scanning calorimetry (DSC) scan obtained from a sample of brigatinib Form A. Heat flow (mW) is shown on the vertical axis and temperature (°C) is shown on the horizontal axis.

[0080] Figure 5A is a thermogravimetric analysis / single differential thermal analysis thermogram (TGA / SDTA) of a sample of brigatinib Form A.

[0081] Figure 5B is a thermogravimetric mass spectrometry (TGMS) thermogram of a sample of brigatinib Form A.

[0082] Figure 6 The obtained sample of brigatinib dissolved in CD3OD 1 H-NMR spectrum. Normalized intensity is shown on the vertical axis and chemical shift (ppm) is shown on the horizontal axis.

[0083] Figure 7 The sample of brigatinib dissolved in CDCl3 was obtained 13 C-NMR spectrum. Normalized intensity is shown on the vertical axis and chemical shift (ppm) is shown on the horizontal axis.

[0084] Figure 8 is a mass spectrometric fragmentation pattern of a sample of brigatinib Form A. Relative abundance is shown on the vertical axis and atomic weight (m / z) is shown on the horizontal axis.

[0085] Figure 9A-9E Depicted is a fragmentation pattern of ions of a sample of brigatinib Form A using collision activation measured using an electrospray time-of-flight mass spectrometer. Relative abundance is shown on the vertical axis and atomic weight (m / z) is shown on the horizontal axis.

[0086] Fig.10 is the crystal structure of brigatinib Form A as determined by single crystal X-ray diffraction.

[0087] Fig.11 is a differential scanning calorimetry (DSC) scan obtained from a sample of brigatinib Form B. Heat flow (mW) is shown on the vertical axis and temperature (°C) is shown on the horizontal axis.

[0088] Fig.12 is a cyclic differential scanning calorimetry (DSC) scan of a sample of brigatinib Form B; heating to 190° C. at 10° C. / min and cooling to 25° C. at the same rate. Heat flow (mW) is shown on the vertical axis and temperature (° C.) is shown on the horizontal axis.

[0089] Fig.13A is a cyclic differential scanning calorimetry (DSC) scan of a sample of brigatinib Form B; heating to 190° C. at 10° C. / min, cooling to 25° C. at the same rate, and then a second heating to 300° C. at the same rate. Heat flow (mW) is plotted on the vertical axis and temperature (° C.) is plotted on the horizontal axis.

[0090] Fig. 13B is a cyclic differential scanning calorimetry (DSC) scan of a sample of brigatinib Form B; heated to 190°C at 10°C / min, cooled to 25°C at the same rate, and then heated a second time to 300°C at the same rate. Heat flow (mW) is plotted on the vertical axis and time (min) is plotted on the horizontal axis. ;

[0091] Fig. 13C is a thermogravimetric analysis / single difference thermal analysis thermogram (TGA / SDTA) of a sample of brigatinib Form B.

[0092] Fig.13D is a thermogravimetric mass spectrometry (TGMS) thermogram of a sample of brigatinib Form B.

[0093] Fig.14is an X-ray powder diffraction (XRPD) pattern obtained from a sample of brigatinib Form B. Relative intensity (in counts) is shown on the vertical axis and angles (in degrees 2θ (° 2θ)) are shown on the horizontal axis.

[0094] Fig.15 is a differential scanning calorimetry (DSC) scan obtained from a sample of brigatinib Form C. Heat flow (mW) is shown on the vertical axis and temperature (°C) is shown on the horizontal axis.

[0095] Fig.16A is a thermogravimetric analysis / single differential thermal analysis (TGA / SDTA) thermogram of a sample of brigatinib Form C. A 4.25% water mass loss was observed up to approximately 75 °C, corresponding to 1.44 water molecules.

[0096] Fig. 16B is a thermogravimetric mass spectrometry (TGMS) thermogram of a sample of brigatinib Form C. A 4.25% water mass loss was observed up to approximately 75 °C, corresponding to 1.44 water molecules.

[0097] Fig.17A is a thermogravimetric analysis / single differential thermal analysis (TGA / SDTA) thermogram of a sample of brigatinib Form C. A 6.14% water mass loss was observed up to approximately 75 °C, corresponding to 2.12 water molecules.

[0098] Fig. 17B is a thermogravimetric mass spectrometry (TGMS) thermogram of a sample of brigatinib Form C. A 6.14% water mass loss was observed up to approximately 75 °C, corresponding to 2.12 water molecules.

[0099] Fig.18 is an X-ray powder diffraction (XRPD) pattern obtained from a sample of brigatinib Form C. Relative intensity (in counts) is shown on the vertical axis and angles (in degrees 2θ (°2θ)) are shown on the horizontal axis.

[0100] Fig.19 is an X-ray powder diffraction (XRPD) pattern obtained from a sample of brigatinib Form D. Relative intensity (in counts) is shown on the vertical axis and angles (in degrees 2θ (°2θ)) are shown on the horizontal axis.

[0101] Fig.19A is a thermogravimetric mass spectrometry (TGMS) thermogram of a sample of brigatinib Form D.

[0102] Fig. 20 is a thermogravimetric mass spectrometry (TGMS) thermogram of a sample of brigatinib Form E.

[0103] Fig.21is an X-ray powder diffraction (XRPD) pattern obtained from a sample of brigatinib Form E. Relative intensity (in counts) is shown on the vertical axis and angles (in degrees 2θ (°2θ)) are shown on the horizontal axis.

[0104] Fig. 22 is a thermogravimetric analysis / single differential thermal analysis (TGA / SDTA) of a sample of brigatinib Form F.

[0105] Fig.23 is an X-ray powder diffraction (XRPD) pattern obtained from a sample of brigatinib Form F. Relative intensity (in counts) is shown on the vertical axis and angles (in degrees 2θ (° 2θ)) are shown on the horizontal axis.

[0106] Fig.24 is an X-ray powder diffraction (XRPD) pattern obtained from a sample of brigatinib Form G. Relative intensity (in counts) is shown on the vertical axis and angles (in degrees 2θ (° 2θ)) are shown on the horizontal axis.

[0107] Fig.25 is an X-ray powder diffraction (XRPD) pattern obtained from a sample of brigatinib Form H. Relative intensity (in counts) is shown on the vertical axis and angles (in degrees 2θ (° 2θ)) are shown on the horizontal axis.

[0108] Fig.26 is an X-ray powder diffraction (XRPD) pattern obtained from a mixture of samples of brigatinib Form A and Form J. Relative intensity (in counts) is shown on the vertical axis and angles (in degrees 2θ (°2θ)) are shown on the horizontal axis.

[0109] Fig.27A is an overlay of X-ray powder diffraction (XRPD) spectra obtained from a sample of a mixture of brigatinib Form A and Form K, a sample of a mixture of brigatinib Form A and Form L, and a sample of brigatinib Form A. Relative intensity (in counts) is shown on the vertical axis and angles (in degrees 2θ (°2θ)) are shown on the horizontal axis.

[0110] Fig.27B yes Fig.27A of expansion.

[0111] Fig.28 Overlaid X-ray powder diffraction (XRPD) spectra of brigatinib Form A that have been subjected to milling experiments for various lengths of time. Relative intensity (in counts) is shown on the vertical axis and angles (in degrees 2θ (°2θ)) are shown on the horizontal axis.

[0112] Fig.29Depicted are solubility data for brigatinib Form A and Form B at various pH values ​​at 25°C and 37°C.

[0113] Fig. 30A yes Fig. 30B An expanded graph showing concentration versus time for brigatinib Forms A and B obtained from intrinsic dissolution rate (IDR) experiments, wherein concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0114] Fig. 30B ] is a concentration versus time curve of brigatinib Forms A and B obtained from intrinsic dissolution rate (IDR) experiments, wherein concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0115] Fig.31 5 is a plot of concentration versus time for brigatinib Forms A and B obtained from IDR experiments in pH 1.0 HCl buffer at 25° C. and 37° C. Concentration (mg / mL) is shown on the vertical axis and time (min) is shown on the horizontal axis.

[0116] Fig.32A yes Fig.32B Expanded graph showing concentration versus time curves of brigatinib Form A and Form B obtained from IDR experiments in pH 6.5 buffer at 25° C. and 37° C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0117] Fig.32B : is a concentration versus time curve of brigatinib Form A and Form B obtained from IDR experiments in pH 6.5 buffer at 25° C. and 37° C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0118] Fig.33A yes Fig.33B Expanded graph showing concentration versus time for brigatinib Form A and Form B obtained from IDR experiments in SGF at 25°C and 37°C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0119] Fig.33B : is a concentration versus time curve of brigatinib Form A and Form B obtained from IDR experiments in SGF at 25° C. and 37° C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0120] Fig.34≠ 4.5 and 6.5 are concentration versus time curves of brigatinib Form A obtained from IDR experiments in water and aqueous buffer at pH 1.0, 4.5 and 6.5 at 25°C.

[0121] Fig.35 is a plot of concentration of brigatinib Form A versus time obtained from IDR experiments in water and aqueous buffer at pH 1.0, 4.5, and 6.5 at 37°C.

[0122] Fig.36 ≠ 4.5 and 6.5 are concentration versus time curves of brigatinib Form B obtained from IDR experiments in water and aqueous buffer at pH 1.0, 4.5 and 6.5 at 25°C.

[0123] Fig.37 is a plot of brigatinib Form B concentration versus time obtained from IDR experiments in water and aqueous buffer at pH 1.0, 4.5, and 6.5 at 37°C.

[0124] Fig.38A yes Fig.38B Expanded graph showing concentration versus time of brigatinib Form A obtained from IDR experiments in water at 25°C and 37°C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0125] Fig.38B is a concentration versus time curve of brigatinib Form A obtained from IDR experiments in water at 25° C. and 37° C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0126] Fig.39A yes Fig.39B Expanded graph showing concentration versus time of brigatinib Form A obtained from IDR experiments in pH 6.5 buffer at 25°C and 37°C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0127] Fig.39B is a concentration versus time curve of brigatinib Form A obtained from IDR experiments in pH 6.5 buffer at 25° C. and 37° C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0128] Fig.40A yes Fig.40B Expanded graph showing concentration versus time of brigatinib Form B obtained from IDR experiments in pH 6.5 buffer at 25°C and 37°C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0129] Fig.40B is a concentration versus time curve of brigatinib Form B obtained from IDR experiments in pH 6.5 buffer at 25°C and 37°C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0130] Fig.41A yes Fig.41B Expanded graph showing concentration versus time for brigatinib Form A and Form B obtained from an IDR experiment in pH 6.5 buffer at 25°C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0131] Fig.41B 5 is a plot of concentration versus time for brigatinib Form A and Form B obtained from an IDR experiment in pH 6.5 buffer at 25° C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0132] Fig.42A yes Fig.42B Expanded graph showing concentration versus time for brigatinib Form A and Form B obtained from an IDR experiment in pH 6.5 buffer at 37° C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0133] Fig.42B TABLE 5 Concentration versus time curves of brigatinib Form A and Form B obtained from IDR experiments in pH 6.5 buffer at 37° C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0134] Fig.43A TABLE 5 Concentration versus time curves of brigatinib Form A and Form B obtained from dissolution rate (IDR) experiments in water at 37° C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0135] Fig.43B yes Fig.43A Expanded graph showing concentration versus time for brigatinib Form A and Form B obtained from dissolution rate (IDR) experiments in water at 37° C. Concentration (mg / mL) is plotted on the vertical axis and time (min) is plotted on the horizontal axis.

[0136] Fig.44 is the DVS curve of Form B, where the total mass increase at 95% RH corresponds to 2.26 water molecules.

[0137] Fig.45is the DVS curve of Form B, where the total mass increase at 85% RH corresponds to 5.6 water molecules.

[0138] Fig.46 is the DVS curve of Form B, where the total mass increase at 95% RH corresponds to 5.15 water molecules.

[0139] Fig.47 is the DVS curve of Form B, where the total mass increase at 95% RH corresponds to 7.2 water molecules.

[0140] Fig.48 is an overlay of the XRPD spectra of Forms A, B, C and D.

[0141] Fig.49 Overlay of the XRPD spectra of Forms A, B, C, D, E, F, G, H and A mixed with J.

[0142] Fig.50 is an interconversion scheme based on experiments for Forms A, B, C, and D. The dashed boxes show that increasing humidity at 30°C results in hydration of Form B to Form C, and ultimately to Form D. The changes are reversible after the temperature is lowered. The solid boxes show that increasing temperature at ambient humidity results in dehydration of Forms C and D to Form B (at about 40°C) and dehydration to Form A via a solid-solid transition at about 150°C. These transformations are irreversible: Form A remains stable after the temperature is lowered.

[0143] Disclosed herein are various crystalline forms of brigatinib. As used herein, the terms "crystalline form," "polymorphic form," and "polymorph" are used interchangeably and refer to a solid form of brigatinib that is different from an amorphous form of brigatinib and from other solid forms of brigatinib, as evidenced by certain properties, such as, for example, kinetic and / or thermodynamic stability, certain physical parameters, X-ray crystal structure, DSC, and / or manufacturing process. Polymorphs of a compound may have different chemical and / or physical properties, including, for example, stability, solubility, dissolution rate, optical properties, melting point, chemical reactivity, mechanical properties, vapor pressure, and / or density. These properties may affect, for example, the ability to process and / or manufacture drug substances and drug products, stability, dissolution, and / or bioavailability. Thus, polymorphism may affect at least one property of a drug, including, but not limited to, quality, safety, and / or efficacy.

[0144] Although polymorphism is generally the ability of a compound to crystallize in more than one crystalline form (having the same chemical structure), the term "pseudopolymorphism" is generally applied to both solvent and hydrate crystalline forms. However, for the purposes of this disclosure, both true polymorphs as well as pseudopolymorphs (i.e., hydrate and solvent forms) are included within the scope of the terms "crystalline form" and "polymorph". In addition, "amorphous form" refers to a non-crystalline solid state.

[0145] There may be variations in the angles of the peaks (XRPD maxima) in the XRPD diffractogram. One of ordinary skill in the art will recognize that variations in the 2-θ peak positions may be observed, such as, for example, a variation of ±0.2° 2θ or a variation of ±0.3° 2θ. In addition, one of ordinary skill in the art will recognize that the relative intensity (expressed in counts) of the peaks may vary between samples, for example due to a preferred orientation. See, e.g., U.S. Pharmacopeia <941> X-ray diffraction. Thus, the crystalline forms disclosed herein have X-ray powder diffraction patterns substantially as shown in certain figures, such as Forms AH, respectively, having X-ray powder diffraction patterns substantially as shown in certain figures. Figure 2 , Fig.14 , Fig.18 , Fig.19 and Fig.21 , Figure 23-Figure 25 Of course, one of ordinary skill in the art will recognize that any additional components in the XRPD sample may contribute peaks to the XRPD spectrum observed for the sample, and the peaks of the sample may mask or overlap (partially or completely) the peaks attributable to the crystalline form of brigatinib in the XRPD sample.

[0146] As used herein, the terms "isolated" and "substantially pure" mean that more than 50%, such as more than 60%, such as more than 70%, such as more than 80%, such as more than 85%, such as more than 90%, such as more than 95%, such as more than 99%, such as more than 99.5%, such as more than 99.8%, or such as more than 99.9% of the brigatinib present in a sample is in a single crystalline form (as can be determined according to methods described in the art). For example, some embodiments of the invention are substantially pure crystalline brigatinib Form A. In some embodiments, the substantially pure crystalline form of brigatinib contains less than 5%, such as less than 1%, such as less than 0.5%, such as less than 0.2%, or such as less than 0.1% of any other solid form of brigatinib (as can be determined according to methods described in the art, such as XPRD analysis, for example).

[0147] As used herein, "pure" when used in reference to the chemical purity of a compound such as brigatinib means that greater than 90%, such as greater than 95%, such as greater than 99%, such as greater than 99.5%, such as greater than 99.8%, or such as greater than 99.9% of the total amount of all chemical species present in a selected material (e.g., in a sample of the API) are brigatinib molecules (as can be determined according to methods described in the art).

[0148] The following solvent abbreviations may be used herein:

[0149] ·DCM Dichloromethane

[0150] DMA N,N-dimethylacetamide

[0151] ·DMF N,N-dimethylformamide

[0152] DMSO Dimethyl sulfoxide

[0153] ·EtOAc Ethyl acetate

[0154] EtOH Ethanol

[0155] IPA Isopropyl alcohol

[0156] ·LiHDMS Lithium bis(trimethylsilyl)amide ·MeCN Acetonitrile

[0157] MeOH Methanol

[0158] ·NMP N-Methylpyrrolidine

[0159] ·TFE 2,2,2-Trifluoroethanol

[0160] ·THF Tetrahydrofuran

[0161] ·2-MethylTHF 2-Methyltetrahydrofuran

[0162] Other abbreviations that may be used herein (in alphabetical order) include:

[0163] ·Am Amorphous

[0164] API Active Pharmaceutical Ingredient

[0165] AS Anti-Solvent

[0166] DSC Differential Scanning Calorimetry

[0167] DVS Dynamic Vapor Sorption

[0168] HPLC High Performance Liquid Chromatography

[0169] IDR Intrinsic Dissolution Rate

[0170] MS

[0171] NSCLC Non-small cell lung cancer

[0172] psi pounds per square inch

[0173] QSA Quantitative Solubility Assessment

[0174] RH Relative humidity

[0175] ·S solvent

[0176] SDTA Single Differential Thermal Analysis

[0177] ·SGF simulated gastric fluid

[0178] SM raw materials

[0179] TGA Thermogravimetric Analysis

[0180] TGMS coupled to mass spectrometry for thermogravimetric analysis

[0181] VH-XRPD Variable Humidity X-ray Powder Diffraction

[0182] VT-XRPD variable temperature X-ray powder diffraction

[0183] ·Xantphos 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene

[0184] ·XRPD X-ray powder diffraction

[0185] "Subjects" to which administration is contemplated include, but are not limited to, humans (i.e., males or females of any age group, such as pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or elderly adults)), another primate (e.g., cynomolgus monkeys, macaques), a mammal, including, but not limited to, cows, pigs, horses, sheep, goats, cats, and / or dogs; and / or birds, including, but not limited to, chickens, ducks, geese, quail, and / or turkeys.

[0186] The XRPD spectra disclosed herein were obtained using a Crystallics T2 high throughput XRPD setup. The plates were mounted on a Bruker GADDS diffractometer equipped with a Hi-Star area detector. The XRPD platform was calibrated using Silver Behenate for long d-spacings and Corundum for short d-spacings.

[0187] Data collection was performed at room temperature using monochromatic CuKα radiation in the 2θ region of 1.5° to 41.5°. Diffraction spectra of each hole in two 2θ ranges (1.5°≤2θ≤21.5° for the first frame and 19.5°≤2θ≤41.5° for the second frame) were collected with an exposure time of 90 seconds for each frame. No background subtraction or curve smoothing was applied to the XRPD spectra in the figures.

[0188] The support material used during XRPD analysis is transparent to X-rays.

[0189] The high-resolution X-ray powder diffraction patterns disclosed herein were collected in Brag-Brentano geometry on a D8 Advance system equipped with a LynxEye solid-state detector. The radiation used to collect the data was CuKα1 monochromatized by a germanium crystal. Without further processing, spectra were collected in the range of 4-41.5° 2θ with a step size of 0.016° 2θ. All spectra were acquired at room temperature of about 295K. The material was placed in a 0.3 mm diameter boron glass capillary. For the variable humidity and variable temperature experiments disclosed herein, an ANSYCO HT chamber was used. The material was placed on a fixed sample holder fixed in the chamber. Humidity was applied locally and varied from 10% to 80% (dew point). The temperature change rate was 10°C / min.

[0190] The step sizes used during the experiments were 0.016, 0.017 or 0.064° 2θ / sec.

[0191] The melting properties disclosed herein were obtained from DSC thermograms recorded with a heat flux DSC822e instrument (Mettler-Toledo GmbH, Switzerland). The DSC822e was calibrated for temperature and thermal enthalpy using a small piece of indium (mp = 156.6 °C; Hf=28.45Jg -1 The samples were sealed in standard 40 μL aluminum pans, drilled with needle holes, and run in a DSC at 10 °C min -1 The heating rate was from 25 °C to 300 °C. During the measurement, a flow rate of 50 mL min -1 The DSC equipment was purged with anhydrous N2 gas.

[0192] The mass loss due to solvent or water loss of various crystal solvents disclosed herein was determined by TGA / SDTA. During heating in a TGA / SDTA851e instrument (Mettler-Toledo GmbH, Switzerland), the sample weight was detected to generate a weight versus temperature curve. The TGA / SDTA851e was calibrated for temperature with indium and aluminum. The sample was weighed into a 100 μL aluminum crucible and sealed. The seal was drilled with a needle hole and the crucible was heated in the TGA at 10 °C min -1 The heating rate was from 25 to 300° C. Anhydrous N 2 gas was used for purging.

[0193] The gases evolved from the TGA samples were analyzed by a quadrupole mass spectrometer Omnistar GSD 301T2 (Pfeiffer Vacuum GmbH, Germany), which analyzed the mass in the range of 0-200 amu.

[0194] Electronic images disclosed herein were automatically collected of all wells of each plate using a Philips PCVC 840K CCD camera controlled by Avantium Photoslider software.

[0195] The HPLC analyses disclosed herein were performed using an Agilent 1200SL HPLC system equipped with UV and MS detectors according to the conditions presented below:

[0196]

[0197]

[0198] The integrity of the compounds disclosed herein is expressed as the "Peak-Area %" for each peak (not peaks due to injections), which is calculated as follows by dividing the peak area of ​​each peak in the chromatogram ("Peak-Area") by the total peak-area ("Total-Area") and multiplying by 100%:

[0199]

[0200] The peak-area percentage of the target compound can be used as an indication of the purity of the components in the sample.

[0201] The mass spectra disclosed herein were performed using a Finnigan ion-trap Mass Spectrometer Model LTQ XL. The samples were infused into an atmospheric pressure electrospray ionization (ESI) probe via a syringe pump. Ion fragmentation was achieved using collision activation, and mass spectral data were collected in full scan (MS1) and multistage MS modes (MS2 and MS3). The structures of the product ions were derived using established fragmentation rules and by using Mass Frontier software (High Chem Ltd., Slovak Republic, Version 5.1.0.3).

[0202] I. Polymorphs of Brigatinib

[0203] By the analysis disclosed herein, ten polymorphs of brigatinib were identified. The ten novel polymorphs are referred to herein as Form A, Form B, Form C, Form D, Form E, Form F, Form G, Form H, Form J, and Form K. Generally, compared with amorphous brigatinib, the crystalline form of brigatinib has physical properties (such as high stability, etc.) that are conducive to the commercial preparation of solid dosage forms. The difference between crystalline brigatinib and amorphous brigatinib can be easily seen with the same type of physicochemical data (such as DSC, XRPD, thermal analysis), which is used to distinguish the various crystalline forms of brigatinib disclosed herein.

[0204] Form A:

[0205] Form A is the major crystalline form identified in the experiments disclosed herein. Form A can be obtained from Figure 1 The final synthetic step in the synthesis of brigatinib shown in is obtained, for example, by raising the temperature of crystallization to 60° C. and adding NaOH solution at a slow rate. Form A is anhydrous and not hygroscopic. Form A is not converted to other forms via solvent-mediated or solid-solid transformations as disclosed herein or exposure to elevated temperature, elevated humidity, mechanical pressure, or grinding.

[0206] The chemical and crystal structure of Form A has been unambiguously determined by nuclear magnetic resonance spectroscopy (NMR), mass spectroscopy (MS) and X-ray powder diffraction (XRPD), single crystal X-ray crystallography in conjunction with data determined from elemental analysis (EA) and Fourier transform infrared (FT-IR) spectroscopy.

[0207] In some embodiments, the present disclosure relates to crystalline Form A of brigatinib. In some embodiments, the present disclosure relates to crystalline Form A of brigatinib, wherein the crystalline Form A of brigatinib is substantially pure. In some embodiments, the crystalline Form A is anhydrous.

[0208] Samples of Form A were analyzed by X-ray powder diffraction (XRPD). In some embodiments, the present disclosure relates to a composition having substantially Figure 2 The x-ray powder diffraction pattern of Form A is shown in FIG.

[0209] In some embodiments, the XRPD pattern of Form A has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen, at least fifteen, at least sixteen, or at least seventeen peaks expressed in degrees 2θ selected from 6.1, 8.6, 9.6, 10.8, 11.3, 13.5, 14.3, 15.9, 17.2, 18.9, 19.4, 20.1, 21.8, 22.6, 23.1, 23.9, and 27.7. As previously noted, in some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-theta peaks may be observed.

[0210] In some embodiments, the XRPD pattern of Form A has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen, at least fifteen, at least sixteen, or at least seventeen peaks expressed in degrees 2θ selected from 6.1, 8.58, 9.58, 10.78, 11.34, 13.46, 14.34, 15.9, 17.22, 18.86, 19.38, 20.1, 21.82, 22.58, 23.14, 23.86, and 27.66. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0211] In some embodiments, the present disclosure is directed to Form A having an x-ray powder diffraction pattern comprising at least one, at least two, at least three, at least four, at least five, or at least six peaks expressed in degrees 2θ selected from 9.6, 17.2, 19.4, 20.1, 23.1, and 27.7. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0212] In some embodiments, the present disclosure is directed to Form A having an x-ray powder diffraction pattern comprising at least one, at least two, at least three, at least four, at least five, or at least six peaks expressed in degrees 2θ selected from 9.58, 17.22, 19.38, 20.1, 23.14, and 27.66. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0213] In the differential vapor sorption (DVS) experiment using Form A, the sample was first dried at 0% RH for 6 hours. Then, at a constant temperature of 25°C, the relative humidity was cycled from 5% to 95% RH (adsorption) and then to 5% RH (desorption) with a holding time of 60 minutes per step. Figure 3 As shown, these results demonstrate that Form A is not hygroscopic.

[0214] refer to Figure 4 , the melting point of Form A was determined by differential scanning calorimetry (DSC). A sample of Form A was analyzed in a 40 μL aluminum pan with a needle hole in the range of 25°C to 300°C at a heating rate of 10°C / min. An endothermic peak at 214.5°C was observed. Therefore, in some embodiments, the present disclosure relates to a crystalline Form A having an onset melting temperature of 214.5°C. In some embodiments, the onset melting temperature of Form A is 214°C. In some embodiments, the onset melting temperature of Form A is 215°C.

[0215] 5 , thermogravimetric analysis / single differential thermal analysis (TGA / SDTA) and thermogravimetric mass spectrometry (TGMS) were performed on Form A. The sample contained in the drilled pinhole crucible was heated in a TGA instrument at 10° C. min -1 The heating rate was from 25°C to 300°C, where anhydrous N2 gas was used for purging. The gases evolved from TGA were analyzed using a quadrupole mass spectrometer. TGA / TGMS experiments showed that 0.23% mass loss (water) was observed in the temperature range of 30°C-100°C.

[0216] The elemental analysis of hydrogen, carbon, nitrogen, chlorine, phosphorus and oxygen was performed on the Form A sample. The results are shown in Table 1 and the molecular formula of brigatinib was determined to be C 29 H 40 ClN7O2P. The determined elemental composition is consistent with the molecular formula of brigatinib.

[0217] Table 1: Elemental analysis results

[0218]

[0219] Solution phase NMR studies were performed on Form A to obtain 1 H. 13 C and 31 The complete assignment of the P resonances and thus the chemical formula of brigatinib were determined. Samples of Form A dissolved in CD3OD solvent were subjected to 1 H NMR analysis was performed on a sample of Form A dissolved in CDCl3 solvent. 13 CNMR analysis. Figure 6 Provided 1D of Form A1 H-NMR spectrum. Figure 7 1D showing form A 13 C-NMR spectrum.

[0220] Table 2 summarizes the 1 H and 13 Relative chemical shift data of Form A obtained from C-NMR experiments. The number of signals and the relative intensity (integral) of the signals determine the number of protons and carbons in the structure of Form A of brigatinib. 31 The P-NMR chemical shift is 43.6 ppm. These are reported according to the atomic numbering scheme shown immediately below 1 H and 13 C-NMR chemical shift data:

[0221]

[0222] Table 2: Form A of brigatinib 1 H and 13 C chemical shift data (in ppm)

[0223]

[0224] refer to Figure 8 Mass spectrometry experiments of Form A were performed using an Agilent electrospray time-of-flight mass spectrometer (Model 6210) operating in positive ion mode using flow injection sample introduction. A sample of Form A was dissolved in methanol / water and analyzed, and the observed mass was m / z 584.263 (M+H + ), and the calculated accurate mass is 584.2664 (M+H + ). The observed molecular weight is consistent with the elemental composition calculated from the molecular formula of brigatinib.

[0225] Using the Finnigan ion trap mass spectrometer described above, ion fragmentation was achieved using collision activation and the Figure 9A-9E Mass spectrometry data were collected in full scan (MS1) and multistage MS modes (MS2 and MS3) as shown. The structures of the product ions were derived using established fragmentation rules and by using Mass Frontier software (High Chem Ltd., Slovak Republic, version 5.1.0.3) as shown in Table 3. The proposed structures of the key product ions were consistent with the brigatinib structure as shown in Table 4.

[0226] Table 3: Mass Spectrum Product Ions of Brigatinib

[0227]

[0228] Table 4: Mass spectral data of product ions of brigatinib

[0229]

[0230]

[0231]

[0232]

[0233]

[0234]

[0235]

[0236]

[0237] The crystal structure of Form A of brigatinib was solved by single crystal X-ray diffraction. Crystals of Form A of brigatinib were obtained from MeOH-toluene. The structure of Form A brigatinib is shown in Fig.10 , and the crystallization parameters are summarized in Table 5. The structure consists of a hydrogen-bonded dimer. Based on the structural solution, Form A was determined to be unsolvated. Some disorder in the crystals is associated with the terminal N-methylpiperidine moiety of brigatinib.

[0238] Table 5: Crystallization Data and Structural Elaboration of Brigatinib Form A

[0239]

[0240]

[0241]

[0242] The % transmittance FT-IR spectrum of brigatinib Form A is shown in Table 6, and a summary of selected IR band assignments is provided in Table 6. Data for Form A samples were collected in potassium bromide salt plates.

[0243] Table 6: Selected IR band assignments for brigatinib

[0244]

[0245] In some embodiments, the present disclosure relates to crystalline Form A having an FT-IR spectrum containing any at least one of the following bands:

[0246]

[0247]

[0248] Form B:

[0249] Form B is hygroscopic. Form B can be obtained indirectly, for example, by dehydration of hydrated forms C and D. A mixture of forms A, B and C can be formed by vapor diffusion onto a solid using water as solvent. None of the direct crystallization experiments disclosed herein yielded Form B.

[0250] Form B can be converted to hydrated forms C and D, depending on, for example, humidity levels (e.g., above 60% RH at 30°C). This conversion was determined to be reversible. As demonstrated by XRPD, Form B irreversibly converts to Form A via a solid-solid transition at about 150°C at ambient humidity. Form B also converts to Form A at elevated temperatures (e.g., at least 37°C) after slurrying in an aqueous medium. The solubility of Form B can be determined via a slurry of Form B converted to Form D and / or C (at 25°C) or Form A (at 37°C).

[0251] exist Fig.11 In the DSC thermogram shown in , a minimum endotherm is observed up to about 50°C, which corresponds to the loss of some small amount of water from Form C present in the sample. Thereafter, Form B converts to Form A via a solid-solid transition (exothermic at 171.8°C), which then melts (exothermic at 214.3°C). This series of events was confirmed by VT-XRPD experiments on Form B.

[0252] Two cyclic DSC experiments using Form B were performed. Fig.12 As shown in , in the first experiment, the temperature was increased to 190°C at 10°C / min and then decreased to 25°C at 10°C / min. Fig.12 The endotherm at about 70°C can be attributed to the presence of a small amount of Form C and its water loss. The exotherm at 161°C can be attributed to the solid-solid conversion of Form B to Form A. XRPD analysis of the solid at the end of the cyclic DSC experiment confirmed that the solid had converted to Form A.

[0253] The second cycle DSC experiment was performed using the following thermal profile: heating to 190°C at 10°C / min, cooling to 25°C at 10°C / min; second heating to 300°C at 10°C / min. The obtained thermograms are shown in FIG13 . The top thermogram is plotted against time, and the bottom thermogram is plotted against temperature. The behavior of the first cycle DSC experiment was described above for the first heating and cooling segments. After the second heating, only at T 峰 Melting of Form A was observed at 50°C = 214.0°C.

[0254] In some embodiments, the present disclosure relates to crystalline Form B of brigatinib. In some embodiments, the present disclosure relates to crystalline Form B of brigatinib, wherein the crystalline Form B of brigatinib is substantially pure.

[0255] Samples of Form B were analyzed by X-ray powder diffraction (XRPD). In some embodiments, the present disclosure relates to a Fig.14 The x-ray powder diffraction pattern of Form B is shown.

[0256] In some embodiments, the XRPD pattern of Form B has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, or at least thirteen peaks expressed in degrees 2θ selected from 5.7, 9.2, 11.5, 12.8, 14.5, 15.5, 16.9, 17.7, 19.2, 20.4, 21.8, 23.2, and 29.5. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0257] In some embodiments, the XRPD pattern of Form B has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, or at least thirteen peaks expressed in degrees 2θ selected from 5.74, 9.22, 11.46, 12.82, 14.5, 15.46, 16.94, 17.66, 19.22, 20.38, 21.78, 23.18, and 29.54. In some embodiments, a variation of ±0.30° 2θ in the position of one or more 2-θ peaks may be observed.

[0258] In some embodiments, the present disclosure is directed to Form A having an x-ray powder diffraction pattern comprising at least two peaks expressed in degrees 2θ selected from 11.5, 14.5, 16.9, 19.2, and 23.2. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0259] In some embodiments, the present disclosure is directed to Form A having an x-ray powder diffraction pattern comprising at least two peaks expressed in degrees 2θ selected from 11.46, 14.5, 16.94, 19.22, and 23.18. In certain embodiments, in some embodiments, a variation of ±0.30° 2θ in the position of one or more 2-θ peaks may be observed.

[0260] Form C:

[0261] For example, Form C can be obtained from partial dehydration of heptahydrate Form D or by hydration of Form B. Form C is a hydrate that dehydrates to Form B after exposure to a relative humidity level of less than 25% RH at 30°C. Form C converts to Form D after exposure to 90% RH at 30°C. These conversions are reversible and have hysteresis. As measured by XRPD, upon temperature increase at ambient humidity, Form C dehydrates to Form B, which irreversibly converts to Form A via a solid-solid transition. No direct crystallization experiments as described herein have yielded Form C.

[0262] Fig.15 The DSC thermogram in shows an endotherm corresponding to water loss (as determined by TGMS), through which the solid form converts to Form B. Form B converts to Form A via a solid-solid transition (exotherm at 159.6° C.), which in turn melts (endotherm at 214.3° C.). This series of events was confirmed by VT-XRPD experiments on Form C.

[0263] The TGMS thermograms of two different samples from Form C are shown in Fig.16A / 16B and Fig.17A / 17B, each containing a TGA / SDTA curve at the top and a TGMS curve at the bottom. These thermograms show a water mass loss of 4.25% and 6.14%, respectively. The corresponding numbers of water molecules are 1.44 and 2.12, indicating a hydration degree of 2.

[0264] Form C can be obtained as a mixture of Forms A, B, and C by vapor diffusion onto a solid using water as a solvent. A mixture of Forms A and C can be obtained by cooling crystallization using hot filtration using a solvent system (any one of acetone / water (50 / 50), water / methanol (50 / 50), and water / 1,4-dioxane (50 / 50)). Another route for the formation of Form C is by evaporation from an acetone / water (50 / 50) solvent.

[0265] In some embodiments, the present disclosure relates to crystalline Form C of brigatinib. In some embodiments, the present disclosure relates to crystalline Form C of brigatinib, wherein the crystalline Form C of brigatinib is substantially pure.

[0266] Samples of Form C were analyzed by X-ray powder diffraction (XRPD). In some embodiments, the present disclosure relates to a composition having substantially Fig.18 The x-ray powder diffraction pattern of Form C is shown.

[0267] In some embodiments, the XRPD pattern of Form C has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen, at least fifteen, at least sixteen peaks expressed in degrees 2θ selected from 2.1, 2.5, 5.4, 9.9, 10.9, 12.9, 14.9, 15.9, 16.6, 17.3, 17.9, 19.2, 20.6, 23.9, 26.8, and 27.4. As previously noted, in some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-theta peaks may be observed.

[0268] In some embodiments, the XRPD pattern of Form C has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen, at least fifteen, at least sixteen peaks expressed in degrees 2θ selected from 2.1, 2.54, 5.42, 9.9, 10.9, 12.86, 14.86, 15.94, 16.62, 17.26, 17.9, 19.18, 20.58, 23.94, 26.82, and 27.42. In some embodiments, a variation of ±0.30° 2θ in the position of one or more 2-θ peaks may be observed.

[0269] In some embodiments, the XRPD pattern of Form C has at least one, at least two, at least three, at least four, at least five, at least six peaks expressed in degrees 2θ selected from 5.4, 14.9, 15.9, 17.3, 19.2, and 23.9. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0270] Form D:

[0271] Form D is a heptahydrate that can be obtained directly from crystallization with methanol as solvent and water as antisolvent. Form D can also be obtained from Form B via Form C after slurrying in an aqueous medium and exposure to high relative humidity (90% or higher at 30°C). Form D dehydrates (partially) to Form C at about 80% RH at 30°C. Upon temperature increase at ambient humidity, Form D dehydrates to Form C as measured by XRPD.

[0272] In some embodiments, the present disclosure relates to crystalline Form D of brigatinib. In some embodiments, the present disclosure relates to crystalline Form D of brigatinib, wherein the crystalline Form D of brigatinib is substantially pure.

[0273] Samples of Form D were analyzed by X-ray powder diffraction (XRPD). In some embodiments, the present disclosure relates to a composition having substantially Fig.19 The x-ray powder diffraction pattern of Form D is shown.

[0274] In some embodiments, the XRPD pattern of Form D has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine peaks expressed in degrees 2θ selected from 4.7, 9.2, 9.7, 11.1, 14.5, 17.4, 18.9, 22.4, and 23.7. As previously noted, in some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0275] In some embodiments, the XRPD pattern of Form D has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine peaks expressed in degrees 2θ selected from 4.66, 9.22, 9.74, 11.06, 14.54, 17.38, 18.94, 22.42, and 23.66. As previously noted, in some embodiments, a variation of ±0.30° 2θ in the position of one or more 2-θ peaks may be observed.

[0276] In some embodiments, the XRPD pattern of Form D has at least one, at least two, at least three, at least four, at least five peaks expressed in degrees 2θ selected from 9.7, 11.1, 17.4, 18.9, and 23.7. As previously noted, in some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0277] In some embodiments, the XRPD pattern of Form D has at least one, at least two, at least three, at least four, at least five peaks expressed in degrees 2θ selected from 9.74, 11.06, 17.38, 18.94, and 23.66. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0278] Conversion of Form AD:

[0279] Once Form A is obtained, no common methods disclosed herein have been found to convert this form into another form. However, Forms B, C, and D are all interchangeable, depending on temperature and relative humidity conditions.

[0280] At 30°C, increasing humidity causes Form B to hydrate to Form C and ultimately to Form D. The change is reversible upon reduction of humidity and occurs with hysteresis: Form B converts to Form C at approximately 65% ​​RH while Form C dehydrates to Form B at 25% RH. Similarly, Form C converts to Form D at approximately 90% RH while Form D partially dehydrates to Form C at 80% RH.

[0281] At ambient humidity, increasing temperature causes Forms C and D to dehydrate to anhydrous Form B (at about 40°C) and to Form A via a solid-solid transition at about 150°C. These transformations are irreversible: Form A remains stable after the temperature is reduced.

[0282] Thermal stability and stability under humidity were evaluated after storage for up to 5 weeks at 50°C, 75°C (for Form A) and at 40°C / 75% relative humidity (for both Forms A and B). Samples were analyzed by XRPD and HPLC over this time period as follows: after 1 day, 3 days, 1 week, 2 weeks, 3 weeks, 4 weeks and 5 weeks. Form A was physically and chemically stable under all conditions tested. However, Form B converted to the hydrated Form C after 1 day in the climate chamber and subsequently to Form A (partially) (data up to 3 weeks).

[0283] Form E:

[0284] Form E can be obtained from freeze drying of chloroform and is a chloroform solvate. Form E can also be obtained as a mixture with Form A by slurrying with chloroform. After several weeks at ambient temperature, Form E can be converted to Form A as determined by XRPD. TGA / SDTA analysis showed a mass loss of 23.4% in the temperature range of 40-120°C, corresponding to 1.5 chloroform molecules / Brigatinib molecules. According to the SDTA signal and the indicated melting point, the solid present after desolvation is Form A.

[0285] In some embodiments, the present disclosure relates to crystalline Form E of brigatinib. In some embodiments, the present disclosure relates to crystalline Form E of brigatinib, wherein the crystalline Form E of brigatinib is substantially pure.

[0286] Samples of Form E were analyzed by X-ray powder diffraction (XRPD). In some embodiments, the present disclosure relates to a composition having substantially Fig.21 The x-ray powder diffraction pattern of Form E is shown.

[0287] In some embodiments, the XRPD pattern of Form E has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen, at least fifteen, at least sixteen, at least seventeen, at least eighteen, at least nineteen peaks expressed in degrees 2θ selected from 9.1, 10.2, 11.2, 12.0, 13.7, 14.4, 15.8, 16.5, 17.4, 18.3, 19.2, 21.6, 22.3, 23.1, 23.9, 26.0, 26.4, 25.8, and 29.3. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-theta peaks may be observed.

[0288] In some embodiments, the XRPD pattern of Form E has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen, at least fifteen, at least sixteen, at least seventeen, at least eighteen, at least nineteen peaks expressed in degrees 2θ selected from 9.06, 10.22, 11.18, 11.98, 13.66, 14.42, 15.82, 16.54, 17.42, 18.34, 19.22, 21.62, 22.3, 23.14, 23.9, 26.02, 26.42, 25.78, and 29.34. In some embodiments, a variation of ±0.30° 2θ in the position of one or more 2-theta peaks may be observed.

[0289] In some embodiments, the XRPD pattern of Form E has at least one, at least two, at least three, at least four, at least five peaks expressed in degrees 2θ selected from 9.1, 10.2, 15.8, 19.2, and 23.9. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0290] In some embodiments, the XRPD pattern of Form E has at least one, at least two, at least three, at least four, at least five peaks expressed in degrees 2θ selected from 9.06, 10.22, 15.82, 19.22, and 23.9. In some embodiments, a variation of ±0.30° 2θ in the position of one or more 2-θ peaks may be observed.

[0291] Form F:

[0292] Form F was obtained from a freeze-drying experiment using TFE / water and is a TFE solvate. Form F desolvates to give Form A after heating or storage at ambient conditions for 8 weeks as measured by XRPD. TGA / SDTA analysis ( Fig. 22 ) shows a mass loss of 17.5% in the temperature range of 40-160°C, corresponding to 1.24 trifluoroethanol molecules per brigatinib molecule. Based on the SDTA signal and the assigned melting point, the solid present after desolvation is Form A.

[0293] In some embodiments, the present disclosure relates to crystalline Form F of brigatinib. In some embodiments, the present disclosure relates to crystalline Form F of brigatinib, wherein the crystalline Form F of brigatinib is substantially pure.

[0294] Samples of Form F were analyzed by X-ray powder diffraction (XRPD). In some embodiments, the present disclosure relates to a composition having substantially Fig.23 The x-ray powder diffraction pattern of Form F is shown.

[0295] In some embodiments, the XRPD pattern of Form F has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen peaks expressed in degrees 2θ selected from 8.5, 9.8, 11.1, 16.3, 17.0, 17.6, 18.7, 19.4, 20.3, 22.0, 23.2, 23.9, and 27.1. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0296] In some embodiments, the XRPD pattern of Form F has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen peaks expressed in degrees 2θ selected from 8.46, 9.78, 11.14, 16.34, 17.02, 17.58, 18.74, 19.38, 20.34, 22.02, 23.22, 23.86, and 27.1. In some embodiments, a variation of ±0.30° 2θ in the position of one or more 2-θ peaks may be observed.

[0297] In some embodiments, the XRPD pattern of Form F has at least one, at least two, at least three, at least four, at least five peaks expressed in degrees 2θ selected from 9.8, 17.0, 19.4, 20.3, and 27.1. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0298] In some embodiments, the XRPD pattern of Form F has at least one, at least two, at least three, at least four, at least five peaks expressed in degrees 2θ selected from 9.78, 17.02, 19.38, 20.34, and 27.1. In some embodiments, a variation of ±0.30° 2θ in the position of one or more 2-θ peaks may be observed.

[0299] Form G:

[0300] Form G was obtained from shock crystallization experiments with chloroform as solvent and acetonitrile as antisolvent. Form G mixed with Form A was also obtained from two other experiments using chloroform (antisolvent addition and thermal cycling). After 5 weeks of storage of the measured plates at ambient conditions, re-measurement by XRPD of Form G showed that Form G had converted to Form A. Form G may be in an unstable form, and may, for example, be a chloroform solvate that desolvated and converted to Form A after storage at ambient conditions.

[0301] In some embodiments, the present disclosure relates to crystalline Form G of brigatinib. In some embodiments, the present disclosure relates to crystalline Form G of brigatinib, wherein the crystalline Form G of brigatinib is substantially pure.

[0302] A sample of Form G was analyzed by X-ray powder diffraction (XRPD). In some embodiments, the present disclosure relates to a composition having substantially Fig.24 The x-ray powder diffraction pattern of Form G is shown.

[0303] In some embodiments, the XRPD pattern of Form G has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen peaks expressed in degrees 2θ selected from 7.2, 8.3, 9.7, 10.4, 12.9, 15.8, 18.1, 18.7, 20.7, 21.5, 22.8, 23.5, 24.5, and 26.8. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0304] In some embodiments, the XRPD pattern of Form G has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve, at least thirteen, at least fourteen peaks expressed in degrees 2θ selected from 7.22, 8.34, 9.7, 10.38, 12.86, 15.78, 18.1, 18.7, 20.74, 21.46, 22.82, 23.54, 24.5, and 26.82. In some embodiments, a variation of ±0.30° 2θ in the position of one or more 2-θ peaks may be observed.

[0305] In some embodiments, the XRPD pattern of Form G has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight peaks expressed in degrees 2θ selected from 8.3, 9.7, 12.9, 15.8, 18.1, 20.7, 22.8, and 26.8. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0306] In some embodiments, the XRPD pattern of Form G has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight peaks expressed in degrees 2θ selected from 8.34, 9.7, 12.86, 15.78, 18.1, 20.74, 22.82, and 26.82. In some embodiments, a variation of ±0.30° 2θ in the position of one or more 2-θ peaks may be observed.

[0307] Form H:

[0308] Form H can be obtained in pure form or as a mixture with Form A from a variety of solvents such as, for example, ethanol / water, 1,4-dioxane / water, methanol, methanol / chloroform, and methanol / acetonitrile by the cool-evaporation method. Form H may be a solvate with small alcohols such as methanol, ethanol, and 1,4-dioxane. After storage at ambient conditions for 1-3 weeks, Form H has partially converted to Form A as determined by XRPD.

[0309] In some embodiments, the present disclosure relates to crystalline Form H of brigatinib. In some embodiments, the present disclosure relates to crystalline Form H of brigatinib, wherein the crystalline Form H of brigatinib is substantially pure.

[0310] Samples of Form H were analyzed by X-ray powder diffraction (XRPD). In some embodiments, the present disclosure relates to a composition having substantially Fig.25 The x-ray powder diffraction pattern of Form H is shown.

[0311] In some embodiments, the XRPD pattern of Form H has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve peaks expressed in degrees 2θ selected from 4.2, 5.2, 8.4, 10.9, 12.7, 15.0, 15.7, 16.5, 17.2, 18.4, 19.5, and 21.3. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0312] In some embodiments, the XRPD pattern of Form H has at least one, at least two, at least three, at least four, at least five, at least six, at least seven, at least eight, at least nine, at least ten, at least eleven, at least twelve peaks expressed in degrees 2θ selected from 4.22, 5.22, 8.38, 10.86, 12.66, 14.98, 15.74, 16.5, 17.18, 18.42, 19.5, and 21.3. In some embodiments, a variation of ±0.30° 2θ in the position of one or more 2-θ peaks may be observed.

[0313] In some embodiments, the XRPD pattern of Form H has at least one, at least two, at least three, at least four, at least five, at least six peaks expressed in degrees 2θ selected from 4.2, 5.2, 8.4, 10.9, 12.7, and 21.3. In some embodiments, a variation of ±0.3° 2θ in the position of one or more 2-θ peaks may be observed.

[0314] In some embodiments, the XRPD pattern of Form H has at least one, at least two, at least three, at least four, at least five, at least six peaks expressed in degrees 2θ selected from 4.22, 5.22, 8.38, 10.86, 12.66, and 21.30. In some embodiments, a variation of ±0.30° 2θ in the position of one or more 2-θ peaks may be observed.

[0315] Form J:

[0316] Form J was obtained on a μL scale from 2-methoxyethanol in a cool evaporation experiment as a mixture with Form A. Remeasurement by XRPD of the mixture of Forms A+J after 3 weeks of storage of the measurement plates at ambient conditions showed that the material was still a mixture of Forms A+J; however, the Form A component was clearly larger.

[0317] The mixture of Form A and Form J was analyzed by X-ray powder diffraction (XRPD) and the spectrum is shown in Fig.26In. The XRPD spectrum has at least one or all of the following peaks expressed in degrees 2θ (2θ), shown for Form A+J as: 5.3, 7.6, 11.2, 17.6, 18.5, 19.8, and 21.3. In certain embodiments, the mixture of Form A+J is characterized by an XRPD spectrum including one or more of the following peaks expressed in degrees 2θ (2θ): 7.6, 17.6, and 21.3. In certain embodiments, the XRPD spectrum of the mixture of Form A+J may have two peaks or three peaks of the peaks listed above.

[0318] Forms K and L:

[0319] Forms K and L were obtained as a mixture with Form A, and their XRPD spectra showed only minor differences from the XRPD spectrum of Form A. Form K was obtained on a μL scale as a mixture with Form A from a THF / NMP mixture in a cold evaporation experiment. After 3 weeks of storage of the measurement plates under ambient conditions, re-measurement by XRPD of Form A+K showed that the material was still a mixture of Forms A+K.

[0320] Form L was also obtained from slurry experiments using n-heptane, hexane or methylcyclohexane as a mixture with Form A. After 3 weeks storage of the measurement plates at ambient conditions, re-measurement by XRPD of the mixture A+L showed that the solid was still a mixture of A+L.

[0321] Fig.27A and Fig.27B XRPD spectra of the observed mixtures A+K and A+L are shown. The labels indicate the 2θ positions where additional intensity peaks occur. For Form K, as described above, additional peaks for Form A include expressed in degrees 2θ (2θ): 5.5, 7.7, and 12.3. For Form L, as described above, additional peaks for Form A include expressed in degrees 2θ (2θ): 18.2. In certain embodiments, the XRPD spectra of Form K or Form L may display two or three of the peaks listed above.

[0322] Amorphous form of brigatinib

[0323] Milling experiments were performed to obtain amorphous brigatinib. After milling samples of Form A for 30 and 60 minutes, XRPD studies showed that Fig.28 The increase of amorphous content shown. Purity was assessed by HPLC and it was confirmed that chemical degradation did not occur during the grinding process. In the mechanical stress test performed by grinding, the samples of Form A were ground for 2, 3, 4 and 5 hours. The recovered solid was analyzed by XRPD and HPLC. By 5 hours, the sample was almost completely amorphous.

[0324] II. Experiments to Identify Brigatinib Polymorphs

[0325] Initial efforts to identify polymorphs of brigatinib were divided into two phases. Phase 1 included raw material characterization, feasibility testing, solubility studies, compaction studies, and intrinsic dissolution rates to provide data for Phase 2 solvent selection. Phase 2 included polymorph screening experiments in milliliter (mL) and microliter (μL) scales. These efforts resulted in the identification of 10 polymorphs: Form A, Form B, Form C, Form D, Form E, Form F, Form G, Form H, Form J, and Form K.

[0326] Phase 1: Raw material characteristics

[0327] Raw material brigatinib is provided as an off-white solid, and its chemical purity is 99.9% as assessed by HPLC. Mass spectrometry data confirmed that the molecular weight of brigatinib is 584 g / mol. TGA and TGMS analysis showed a mass loss of 0.23% (corresponding to approximately 0.08 water molecules / Form A molecule) in the temperature interval of 30°C-100°C. DSC analysis showed a T associated with the melting of compound brigatinib. 峰 =214.5 ° C endothermic event. The water content of Form A was determined by coulometric titration Karl Fischer method. The average water content from two determinations was found to be 0.32%. Representative residual heavy metals in Brigatinib Form A were determined by ICP-MS. The elements detected included cadmium (0.02ppm), copper (0.14ppm), molybdenum (0.10ppm), palladium (0.087ppm) and silver (0.03ppm). The following metals were not detected: antimony, arsenic, bismuth, lead, mercury and tin.

[0328] NaOH titration experiments were performed to study the effects of NaOH addition rate and crystallization temperature on the isolated crystalline forms. The stock solution of brigatinib Form A was prepared by weighing 450 mg of Form A and slurrying in 9 mL of water for 10 min. 4.5 mL of 1M HCl was added to dissolve brigatinib (final API concentration 33.3 mg / mL). For each experiment, 3 mL of stock solution was added to an 8 mL vial containing a stirring bar, a pH probe, and a tube connected to a titrator (Titrino). The vial was placed in crystallization and the temperature was reached before starting NaOH titration. 3 mL of 0.1 M NaOH solution was titrated at a predetermined rate. During the experiment, a bottom stirring of 500 rpm was applied. Although brown solids appeared during titration; the color turned pink after stirring (10 min). Subsequently, all solids were separated from the solution by centrifugation, washed twice with 5 mL of water, and then dried.

[0329] Four sets of NaOH addition rate (mL / min) and temperature °C conditions were evaluated: 0.02 mL / min at 25 °C, 20 mL / min at 60 °C, 0.05 mL / min at 25 °C, and 20 mL / min at 60 °C. When the process occurred at 60 °C and slow NaOH addition was applied, direct formation of Form A from aqueous media was possible. Rapid NaOH addition produced a mixture of Form A and heptahydrate Form D, while at 25 °C the crystallized form was the heptahydrate, regardless of the NaOH addition rate.

[0330] Phase 1: Solubility studies

[0331] The brigatinib raw material was subjected to a quantitative solubility test using a set of 24 solvents (DMSO, heptane and water were performed in triplicate). In a vial, about 40 mg of raw material, 400 μL of solvent and a stirring bar were added. After stirring for 24 hours at 20 ° C for 24 hours, the liquid was retrieved, filtered and analyzed for API content by HPLC. The residual solid was characterized by XRPD and found to be Form A. The results are summarized in Table 7.

[0332] Table 7: Solubility of Brigatinib

[0333]

[0334]

[0335] 1 The samples were dissolved after 24 h equilibration time and no solids were harvested.

[0336] 2 In the range below the detection limit, the concentration was less than 0.22 mg / mL

[0337] 3 Data obtained from freeze-drying experiments

[0338] The solubility of Form A was also evaluated in simulated gastric fluid and observed to be 52 mg / ml. In aqueous buffer at 37°C, the solubility of Form A was observed to be 70 mg / mL (in pH 1.0), 26 mg / mL (in pH 4.5), and 6 mg / mL (in pH 6.5).

[0339] In the second solubility study, the solubility of forms A and B was determined in triplicate at 25°C and 37°C in water, pH 1.0 buffer (0.1N HCl), pH 4.5 acetate buffer, pH 6.5 phosphate buffer, and simulated gastric fluid SGF (at 37°C). For each medium, a standard 1.8mL screw cap vial was filled with about 40mg of raw material, 400μl solvent, and a magnetic stirring bar (in the case of chloroform and dichloromethane, 200μl solvent was used). The vial was then closed and balanced at the corresponding temperature under stirring for 24 hours. The liquid portion was retrieved with a syringe and filtered (0.5 micron filter); the separated mother liquor was diluted to two dilutions selected according to the calibration curve. The amount of API in the diluted solution was determined via HPLC analysis (DAD). A calibration curve was obtained from two independently prepared stock solutions of compound brigatinib in 50% water / 50% acetonitrile / 0.1% TFA. Subsequently, the isolated solid was measured to be wet by XRPD to confirm the solid form, and its solubility was measured.

[0340] In Table 8, the solid forms of the isolated slurries are listed. Form A remained stable in all media, while Form B converted to hydrated forms D and / or C in experiments at 25°C and to Form A in experiments at 37°C. At the latter temperature and in water, Form B converted to hydrates C and D, and did not convert to A in the remaining media. The solubility of Form B could not be measured because it converted to other solid forms. The average solubility values ​​shown in the same table refer to the solid form to which the initially placed Form B was converted. Therefore, it was not possible to measure the solubility of Form B but the solubility of Form C (and C+D) could be measured. The solubility values ​​were calculated by Fig.29 The solubility in acidic medium is better than that in alkaline medium.

[0341] Table 8. Forms obtained at the conclusion of the solubility studies

[0342]

[0343] In a third solubility study, Form A was measured in different buffer solutions as shown in Table 9.

[0344] Table 9: Solubility measurements of Form A in buffer

[0345]

[0346] Phase 1: Feasibility Study

[0347] Feasibility testing was performed to try to obtain amorphous material that could be used in some crystallization techniques in the Phase 2 portion of this study. Two techniques were employed, namely grinding and freeze drying. The results are presented below.

[0348] Grinding Two grinding experiments were performed on samples of Form A at a frequency of 30 Hz at two different durations (30 and 60 min). Their amorphous content increased over time, but their purity was stable at about 100%. Mechanical stress via grinding experiments were also performed, with grinding times of 2, 3, 4 and 5 hours. Similarly, the amorphous content increased without a decrease in chemical purity.

[0349] Freeze drying. Six freeze-drying experiments were performed with samples of Form A as described in Table 10. Samples 1, 2, and 4 remained mostly crystalline, but samples 3 and 5 were amorphous and contained approximately 15-16% residual solvent. Sample 6 was amorphous and contained approximately 7% residual solvent. Forms E and F were generated using this method. However, due to variable form and solvation, freeze drying was not additionally employed to obtain amorphous brigatinib.

[0350] Table 10: Freeze-drying feasibility study of brigatinib Form A

[0351]

[0352] Phase 1: Compression

[0353] Brigatinib Form A was subjected to compression testing to determine whether pressure-induced phase inversion or loss of crystallinity occurred. The press used was an Atlas Manual 25 Ton Hydraulic Press (from SPECAC). The pressure was adjusted to 3 and 6 tons / cm in each case. 2 The experiment was carried out for one minute. The pressed solid was measured by XRPD and no phase transition or peak shift was revealed in the XRPD spectrum. The HPLC purity of the two samples subjected to compression testing was determined to be comparable to that of the starting material.

[0354] Phase 1: Intrinsic dissolution rate

[0355] To measure the intrinsic dissolution rate (IDR), the raw materials were compressed into tablets using a mini-IDR compression system (pION / Heath Scientific). To prepare tablets, approximately 11 mg of the material was compressed into a uniform flat surface in a cylindrical cavity of a passivated stainless steel die with an exposed area of ​​0.072 cm 2 The pressure applied was about 50 bar for 3-5 min. The sample mold was inserted into a cyclindrical Teflon rotating disk carrier containing an embedded magnetic stir bar at its base. The mold / stirrer assembly was placed in a flat bottom glass vial ready for dissolution analysis.

[0356] The dissolution rate was measured in 20 mL of solvent (medium), and the path length of the UV meter was 2 mm. The stirring speed applied during the measurement was 100 rpm. The measurements were carried out at 20°C and 37°C.

[0357] To determine the dissolution rate from a powdered sample, weigh approximately 5 mg of brigatinib (Form A or B) into a 5 mL dissolution vial and insert a dissolution probe into the vial. Subsequently, 4 mL of water was added at the start of the measurement. The concentration was recorded for 20 hours.

[0358] In the first series, the IDR of Forms A and B was determined in monoplicate. Measurements were performed in water, pH 1.0 (0.1 N HCl) buffer, pH 6.8 phosphate buffer, and simulated gastric fluid SGF at 25°C and 37°C. Fig.37 In the , IDR is plotted for comparison between forms and the same medium and between various media and the same form. The IDR of each of Forms A and B in various media increases with increasing acidity of the medium (see Figure 34-Figure 37 ).

[0359] Intrinsic dissolution rate measurements of Form A in pH 1.0 and SGF showed that within 5 min, a concentration of approximately 0.25 mg / ml could be reached. This indicates that in the stomach, approximately 50 mg of Form A (the number is illustrative only) could be dissolved along with a 200 ml glass of water.

[0360] In addition, IDR experiments showed that Form A remained stable when slurried in water, SGF, and pH 1.0 aqueous buffer. Based on those results, no conversion was expected to occur in the stomach.

[0361] In several cases, the results are counterintuitive. These results are related to the following: (1) the dissolution rate of the compound at 25°C is higher (in the case of Form A in water and pH 6.5 buffer - for the first 3-4 min - and in the case of Form B in pH 6.5 buffer - over the entire range) compared to the dissolution rate at 37°C, while the IDR at 37°C is expected to be the highest; and (2) the IDR of Form A is higher compared to the IDR of Form B at pH 6.5 buffer, while the opposite is expected based on the relative stability of Forms A and B. To further investigate these results, two more series of experiments were performed: (a) the IDR of Forms A and B was measured (in duplicate) in water at 25°C and in pH 6.5 buffer at 25°C and 37°C; and (b) the IDR of Form A was measured in triplicate in water and pH 6.5 buffer at 25°C. The results of these additional experiments are plotted in Figures 38-42.

[0362] With regard to the first observation (IDR of Form A at 25°C compared to IDR at 37°C), the following comments can be made:

[0363] FIG30 : The IDR of Form A in water at 25° C. appears to be higher in the first 3 minutes. One possibility for this result is the detachment of tablet crystals, which increases the concentration. Thereafter, the concentration of both Forms A and B is higher at 37° C., as expected. However, re-measurements of the IDR of Form A in water at both temperatures show considerable variability ( Fig.38A / 38B). One possibility for this result is the low concentration, which makes the measurement result more sensitive to the measurement conditions.

[0364] Fig.31 : Similarly, the IDR in pH 1.0 buffer at 25°C appears to be higher for both Forms A and B, one possibility being detachment of tablet crystals, as indicated by the large increase in concentration (at about 1 min) in both cases. Concentrations above 0.25 mg / mL are not plotted because the detector saturates at about these values.

[0365] Fig.32A / 32B: In the first 4 min, the IDR of Form A at 25°C in pH 6.5 buffer appeared to be higher than that at 37°C, one possibility being the detachment of tablet crystals; after 4 min, the IDR at 37°C became even higher. However, re-measurement of the IDR of Form A in pH 6.5 buffer showed that the rate at 37°C was higher compared to the rate at 25°C ( Fig.39A / 39B). The IDR of Form B appears to be higher at 25°C, however the concentration of Form B appears to be stable at both temperatures. Where the IDR measurements of Form B in pH 6.5 buffer were repeated, in a second series of experiments the results showed that the IDR at 37°C was higher than that at 25°C ( Fig.40A / 40B). However, there may be variability in the measurements, which may also be due to the low concentrations, making the measured values ​​more sensitive to the measurement conditions.

[0366] The following observation was investigated in a second series of IDR measurements: Form A appears to dissolve faster than Form B ( Fig.32A / 32B). In Figure 41, all IDR measurements of Forms A and B at 25°C are plotted: The second series of experiments shows that after 3 min, the concentration of Form B is highest, which is expected. Before 3 min, a large increase in concentration was observed in several cases, indicating that the particles were detached from the tablet. Fig.42A In / 42B, all IDR measurements of Forms A and B at 37°C are plotted: The second series of experiments showed that the concentration of Form B was highest after approximately 1 min.

[0367] It should be noted that in the case of the IDRs of Forms A and B in water and pH 6.5 buffer, at both 25 and 37°C, the concentration values ​​are very low, making the values ​​recorded very sensitive to the measurement conditions. Measurements at these concentrations tend to vary to a greater extent than measurements at higher concentrations. These IDR values ​​should be considered indicative rather than absolute.

[0368] Fig.33A / 33B: For the first 5 min, the IDR of Form A at 25°C in SGF appears to be higher than that at 37°C, probably due to the detachment of tablet crystals at the beginning of the measurement, which increases the concentration. Thereafter, the IDR of Form A appears to be similar at both 25 and 37°C. As expected, the IDR of Form B is higher at 37°C than at 25°C. In pH 1.0 buffer and SGF, the IDRs of both Forms A and B and at both temperatures are comparable (see Fig.31 and Fig.33A / 33B). At a concentration of about 0.3 mg / mL, the detector was close to saturation.

[0369] Fig.34 : Curves of increasing concentrations of Form A versus time from IDR experiments in water and aqueous buffer at pH 1.0, 4.5 and 6.5 at 25°C.

[0370] Fig.35 : Curves of increasing concentrations of Form A versus time from IDR experiments in water and aqueous buffer at pH 1.0, 4.5 and 6.5 at 37°C.

[0371] Fig.36 : Curves of increasing concentrations of Form B versus time from IDR experiments in water and aqueous buffer at pH 1.0, 4.5 and 6.5 at 25°C.

[0372] Fig.37 : Curves of increasing concentrations of Form B versus time from IDR experiments in water and aqueous buffer at pH 1.0, 4.5 and 6.5 at 37°C.

[0373] To measure the dissolution rate from the powders, Forms A and B were tested only in water at 37°C because the solubility of Forms A and B was low enough to allow detection.

[0374] exist Fig.43AIn / 43B, the concentration of Form A and B is plotted against time. In both cases, within about 10 min, the concentration reaches a "maximum" and dissolution slows down thereafter. Between 10 min and 20 h, the concentration of Form A almost doubles (from 0.07 to 0.14 mg / mL). For Form B, between 10 min and 260 min, a decrease in concentration is observed; thereafter, the concentration increases again to reach a slightly higher value than the value at 10 min at the end of the experiment. The increase in concentration may be associated with the conversion of Form B into a resolvable Form D. Due to detector saturation at about 0.3 mg / mL, the maximum concentration of Form B is not convincingly identified.

[0375] Phase 2: Polymorph Identification

[0376] Brigatinib polymorph screening experiments were performed at the milliliter (mL) scale using nearly 300 different conditions, and also at the microliter scale using nearly 200 different conditions. Six different crystallization procedures were applied: (1) cooling-evaporation; (2) evaporative crystallization; (3) steam exposure; (4) cooling crystallization with hot filtration; (5) shock crystallization with antisolvent addition; (6) slurrying; (7) vapor diffusion into solution; (8) vapor diffusion onto solid; (9) grinding; (10) thermal cycling; (11) VT-XRPD; (12) VH-XRPD; (13) DVS; and (14) dehydration. After completion of the screening experiments, the material was collected and analyzed by XRPD and electron imaging.

[0377] Cooling-evaporation crystallization experiment

[0378] The μL-scale cooling-evaporation experiments shown in Tables 11-14 were performed in a 96-well plate using 24 different solvents and solvent mixtures, 2 concentrations, and 2 temperature profiles. In each well, 4 mg of Form A was weighed. Then, the screening solvent was added to achieve a concentration of approximately 40 mg / mL or 80 mg / mL. The plate, with each well individually sealed, was placed in a CrystalBreeder TM The plates were then placed under vacuum and evaporated at 200 mbar and / or 5 mbar for several days and then analyzed by XRPD and electron imaging. The final forms obtained are given in Tables 12-14.

[0379] Table 11: Cooling-evaporation crystallization parameters

[0380]

[0381] Table 12: Cooling-evaporation crystallization experimental results: T1 and T2 curves

[0382]

[0383]

[0384] Table 13: Cooling-evaporation crystallization experimental results: T curve 3

[0385]

[0386]

[0387]

[0388] Table 14: Cooling-evaporation crystallization experimental results: T curve 4

[0389]

[0390]

[0391] Evaporation crystallization experiment

[0392] Brigatinib Form A and 30 different solutions were used. In a vial, 20 mg of material was weighed and 1000 μL of a given solvent was added. After stirring at room temperature for up to 3 hours, the solvent was evaporated at room temperature (200 mbar for 120 h, then 5 mbar for 48 hours). As shown in Table 15, the dry solid obtained was analyzed by XRPD and electronic imaging.

[0393] Table 15: Evaporation crystallization experiment

[0394]

[0395]

[0396] Steam exposure experiment As shown in Table 16, the stability of Form A after exposure to solvent vapor was studied in twenty solvents. About 20 mg of brigatinib Form A was weighed into a 1.8 mL vial. The vial opening was prevented and placed in a sealed 40 mL vial containing 2 mL of solvent. The material was exposed to solvent vapor for two weeks at room temperature. At the end of the experimental time, wet and dry solids were harvested and analyzed by XRPD and electronic imaging.

[0397] Table 16: Steam exposure test

[0398]

[0399] Cooling crystallization using hot filtration experiments

[0400] The cooling crystallization method using hot filtration included 34 solvents and solvent mixtures. A supersaturated solution was prepared by stirring a slurry of brigatinib in 1300 μL of a given solvent or mixture at 60 °C for one hour. The liquid was then separated from the solid by filtration. The solution was placed on a crystallization plate for 16 TM The following cooling curve was experienced in the instrument. The sample was heated to 60 ° C and maintained for 60 min, then cooled at a rate of 1 ° C / h until 5 ° C was reached. The sample was then kept at this temperature for 48 h. In each experiment, at the end of the thermal cycle curve, no precipitation was observed. The solvent was evaporated for 104 hours at 200 mbar and for 70 hours at 5 mbar. In several cases, the evaporation at 5 mbar continued for about 400 hours, and in some other cases, no yield was obtained after the solvent was evaporated. All obtained solids were analyzed by XRPD and electronic imaging. Table 17 provides the applied crystallization conditions and the corresponding solid forms obtained.

[0401] Table 17: Cooling crystallization using hot filtration experiments

[0402]

[0403]

[0404] Shock crystallization using antisolvent addition

[0405] In the shock-crystallization experiments, 34 different crystallization conditions were applied using 6 different solvents and 24 different antisolvents (see Table 17). Antisolvent addition experiments were performed in the forward direction. For each solvent, a stock solution was prepared, and in each case the concentration of brigatinib reached saturation after 24 hours of equilibration at ambient temperature followed by filtration.

[0406] For each experiment, antisolvent was added to each solvent vial, and the ratio of solvent to antisolvent was 1:0.25. In the case of no precipitation, the ratio was increased to 1:1, and if no precipitation occurred again, the ratio was increased to 1:4, the waiting time between additions (until the third addition) was 60 minutes and the waiting time between the third addition and the fourth addition was 35 minutes. When no crystallization occurred or there was not enough solid to precipitate from separation, the sample was kept at 5 ° C for 17 hours. The precipitated solid was separated from the liquid by centrifugation and decantation. When decantation could not be applied, Pasteur's pipette was used to carefully remove the liquid. The solid was dried at 200 mbar for 17 hours and analyzed by XRPD and electronic imaging. In the case of no precipitation, the solvent was evaporated at 200 mbar for 17 hours, and then the vacuum was reduced to 5 mbar. All obtained solids were analyzed by XRPD and electronic imaging. The measuring plate containing the final solid was stored at ambient temperature for 5 weeks. The solid form was evaluated again by XRPD. The arrows in Table 18 indicate whether the form changed during storage.

[0407] Table 18. Shock crystallization experiments using antisolvent addition

[0408]

[0409]

[0410] *ML = from mother liquor; **No yield;

[0411] ***Apply two additions; ****Single crystals selected from liquid

[0412] Pulping experiment

[0413] A total of 68 slurry experiments were performed with brigatinib at room temperature (20°C) and 40°C using 34 solvents. In all cases, a solvent volume of 250 μL was used. The slurries were stirred for two weeks. After the slurry time was completed, the vials were centrifuged and the solids and mother liquor were separated. The wet and dry solids were analyzed by XRPD and electron imaging. The measurement plates were then stored at ambient conditions for 3-4 weeks and another XRPD of the solids was obtained, with any form changes shown by arrows. Tables 19a and 19b summarize the experimental conditions and the solid forms obtained

[0414] Table 19a: Slurrying experiment at 20°C

[0415]

[0416]

[0417] *In this experiment, the solid dissolved after 14 days

[0418] Table 19b: Slurrying experiment at 40°C

[0419]

[0420]

[0421] *In this experiment, the solid dissolved after 14 days

[0422] In the second group of slurry experiments, the same amount of Form A and Form B were weighed into 1.8mL bottles, and a stirring bar was loaded. After adding solvent, the slurries were placed at 25°C and 50°C under stirring. At 2, 4 and 14 days, the material from the slurries was sampled (sampling from the same bottle according to solvent and temperature). These materials were wet by XRPD and electronic imaging analysis. As shown in Table 20, Form B was converted to Form A at 37°C in all organic solvents and in water. Sampling after 2 and 4 days at 25°C in water showed that the solid was a mixture of Form A and Heptahydrate Form D. The observation indicated that Form B was converted to Form D and Form A remained stable in an aqueous environment. During the 14th day sampling period, only Form A was present, indicating that it had a higher stability in water compared to Form D.

[0423] Table 20: Slurrying experiment

[0424]

[0425]

[0426] Vapor Diffusion in Solution Experiment

[0427] For the experiment of vapor diffusion into solution, a saturated solution of brigatinib was exposed to antisolvent vapor for two weeks at room temperature. An aliquot of the saturated solution was transferred to a vial, its opening was placed and placed in a closed container containing an antisolvent (see Table 20). Two weeks later, the solid formation of the sample was evaluated. When a solid is present, the liquid is separated from the solid, and the solid is then dried under complete vacuum. In the case where no precipitation is observed, the dissolution is placed at 5 ° C overnight to promote precipitation. If no solid is present, the liquid is evaporated at 200 mbar for 75 hours, or if there is still no solid present, the liquid is evaporated at 10 mbar for up to 10 days. All obtained solids are analyzed by XRPD and electronic imaging. Table 21 provides experimental conditions and the corresponding solid forms obtained.

[0428] Table 21: Vapor Diffusion into Solution Experiment

[0429]

[0430]

[0431]

[0432] *No yield after evaporation

[0433] Vapor diffusion onto solids experiment

[0434] For 34 vapor diffusion experiments on solids, amorphous brigatinib was prepared by grinding the raw material for 4 hours. The vial containing amorphous brigatinib was opened and placed in a closed 40mL vial containing 2mL solvent (see Table 21). Amorphous brigatinib was exposed to solvent vapor for two weeks at room temperature. At the end of the experimental time, wet and dry solids were harvested and analyzed by XRPD and electronic imaging. See Table 22 for the applied crystallization conditions and the corresponding solid forms obtained.

[0435] Table 22. Vapor Diffusion onto Solids

[0436]

[0437]

[0438] Solvent-assisted grinding experiments

[0439] In the solvent-assisted grinding experiment, a small amount of solvent was added to the solid brigatinib, which was mechanically ground in a stainless steel vial containing 2 stainless steel grinding balls. In this way, 17 different solvents were studied. Typically, 30 mg of raw material was weighed into a grinding vial and 10 μL of solvent was added to the vial. The grinding experiment was performed at 30 Hz for 60 min. Subsequently, samples were collected and analyzed by XRPD and electron imaging. See Table 23 for the applied crystallization conditions and the corresponding solid forms obtained.

[0440] Table 23: Solvent-assisted grinding experiments

[0441]

[0442]

[0443] Thermal Cycling Experiment

[0444] A total of 33 slurries and 1 solution (chloroform) of the starting material in solvent were prepared at room temperature. The mixture was placed in a Crystal 16 TM The following temperature curve is experienced:

[0445] 1. Heat at a rate of 5°C / h with stirring (500 rpm) until reaching 40°C

[0446] 2. Cool at a rate of 5°C / h to 5°C while stirring (200 rpm)

[0447] 3. Aging at 5°C for 30 minutes

[0448] 4. Repeat 8 cycles

[0449] After the cycling procedure was completed, the solid was separated from the mother liquor by centrifugation, dried at 200 mbar for 48 hours (2-ethoxyethanol for 283 hours) and analyzed by XRPD and electron imaging. For the applied crystallization conditions and the corresponding solid forms obtained, see Table 24. The solid forms (or mixtures) behind the arrow (→) were obtained after re-measurement by XRPD after storage of the measured plates under ambient conditions for 5 weeks.

[0450] Table 24: Thermal Cycling Experiment

[0451]

[0452]

[0453]

[0454] Variable temperature XRPD experiments

[0455] Data for Formats A, B, C, and D were collected almost immediately after reaching the target temperature (within approximately 10 min).

[0456] For Form A, the temperatures used in the experiments were 25, 40, 60, 100, 120, 140, 150, 160, 170, 180, 190 and 200° C. Data collection by temperature lasted for 20 min, and the stabilization time in between was 10 min. The variable temperature XRPD data collected for Form A did not reveal any phase transformation. Only peak shifts contributing to thermal expansion were observed.

[0457] For Form B, the temperatures used in the experiments were 25, 40, 60, 100, 120, 140, 150, 155, 160, 165, 170, 180 and 190° C. Data collection by temperature lasted for 45 min, with stabilization times of 10 min in between. At 150° C., partial conversion to Form A was observed, and conversion at 155° C. was complete. Thereafter, Form A remained stable for the remainder of the temperature profile.

[0458] For Form C, the temperature used in the experiment was 25, 40, 60, 70, 80, 100, 120, 140, 150, 155, 160, 165, 170, 175, 180, 190 and 200 ° C. The data collection according to the temperature lasted for 40 min, and the stabilization time in between was 10 min. Form C is unstable at temperatures above 25 ° C. By the time of the first measurement, the material had been partially converted to dehydrated Form B. Thereafter, the solid form conversion was similar to those observed in the VT-XRPD experiment of Form B, with the difference that the conversion of Form B to Form A had already started at 120 ° C. Although the conversion was completed at the same temperature (155 ° C). Again, no phase transition was observed after cooling. Again, no phase transition was observed after cooling.

[0459] For Form D, the temperatures used in the experiments were 25, 35, 45, 55, 65, 75, 85, 100, 120, 140, 150, 155, 160, 165, 170, 175, 180, 190 and 200°C. Data collection lasted 10 min, with stabilization times of 1 min and 40 min for temperatures 25-85°C and 10 min for temperatures 100-25°C. Variable temperature XRPD confirmed that the heptahydrated Form D was unstable at temperatures above 25°C. Indeed, after the first measurement, Form D had been converted (partially) to the hydrated Form C (at 35°C) and to the dehydrated Form B at 45°C. Thereafter, solid form conversions were similar to those observed in the VT-XRPD experiments of Form C: at 120°C Form B was converted (partially) to Form A. The conversion was complete at 150°C. No phase transition was observed after cooling.

[0460] Variable humidity-XRPD experiments

[0461] The relative humidity was increased from the starting amount, raised to the maximum, and then dried back to the minimum value. The data collection time was 41 minutes per step, which started after the relative humidity equilibrated.

[0462] For Form A, the XRPD spectra of Form A collected do not show any phase transition, neither at 30°C nor at 60°C. In a specific peak, starting at about 60% RH, only some small but significant peak displacements (0.03° 2θ order of magnitude) are observed. The peak displacement is reversible at about 30% RH. The sample of Form A is then exposed to 80% RH for 15h. Peak displacement occurs after 90min, and the extent of displacement remains constant at 80% RH for 15h during the entire exposure period. After returning to 10% RH, the peak shifts to its original position. In order to study the amount of adsorbed water, a new sample of Form A is exposed to 80% for 2h and the TGMS of the sample is measured. The TGMS thermogram shows a mass loss of 0.35% corresponding to 0.1 water molecules.

[0463] For Form B, relative humidity was measured at 30°C. The RH% values ​​measured were 10, 30, 50, 60, 65, 70, 75, and 80%. After adsorption, Form B converted to hydrated Form C, starting at about 65% RH. At 80% RH, conversion to Form C was complete. After desorption, Form C dehydrated to Form B, starting at about 30% RH. At 10% RH, conversion to Form B was complete.

[0464] For Form C, a hydrate, relative humidity was measured at 30°C. The experiment was started at maximum RH and dehydrated, then rehydrated back to maximum. The RH% values ​​measured were 10, 15, 20, 25, 30, 35, 40, 60, and 80%. After desorption, Form C dehydrated to Form B, starting at about 25% RH. At 10% RH, the conversion to Form B was complete. After adsorption, Form B converted to Form C, starting at about 60% RH. At 80% RH, the conversion to Form C was complete. The results are consistent with the corresponding experiments of Form B.

[0465] For Form D, a hydrate, relative humidity was measured at 30°C. The experiment was started at maximum RH and dehydrated, then rehydrated back to maximum. The RH% values ​​measured were 10, 15, 20, 25, 30, 35, 40, 60 and 80%. Despite attempts to have freshly prepared Form D, even the first measurement at 80% RH showed that the solid had been partially converted to Form C. Thereafter, the solid was converted to hydrated Form C and eventually to anhydrous Form B, as observed in the VH-XRPD measurements of Forms B and C. After desorption, the heptahydrated Form D was converted to hydrated Form C. Form C was dehydrated to Form B, starting at about 20% RH. At 10% RH, the conversion of Form B was complete. After adsorption, Form B was converted to Form C, starting at about 40% RH. At 80% RH, the conversion of Form C was complete. The solid did not hydrate to Form D because 80% relative humidity was insufficient; for conversion to Form D, exposure to 95% relative humidity can be employed.

[0466] Dynamic vapor adsorption experiment

[0467] In the three DVS experiments, the relative humidity was varied as follows:

[0468] Experiment 1: 5% → 95% → 65% RH

[0469] Experiment 2: 5% → 95% → 5% RH

[0470] Experiment 3: 5% → 95% RH

[0471] Experiment 4: 0% for 6 h → 5% for 1 h → 15% for 1 h → 25-85% gradient over 2 h → 95 RH for 5 h

[0472] For Experiment 1, during adsorption, Form B adsorbed a water mass corresponding to 2.26 water molecules between 45 and 95% RH, as Fig.44 After desorption to 65% RH, the mass of water obtained remained almost constant. XRPD measurement of the solid showed that it was hydrated form C. The additional mass of water obtained may be due to adsorption on the surface of the material.

[0473] For Experiment 2, two steps of water mass increase were observed during adsorption, such as Fig.45As shown. In the first step, between 45-85% RH, a 6.45% mass change corresponding to 2.1 water molecules was observed. The data are consistent with the formation of hydrated form C at this stage. In the second step, between 85% and 95% RH, a total mass change of 16.7% was reached. During desorption, an additional increase of 17.4% in mass was observed at 85% RH. The continuous increase in mass during desorption shows that equilibrium is not reached within one hour at 95% RH, and water adsorption continues at least until 85% RH during humidity reduction. The maximum change in weight corresponds to 5.6 water molecules. The data are consistent with the formation of heptahydrate form D (partially) at maximum RH. During the two-step desorption, the mass change is roughly stable when reaching about 75% RH, and thereafter it is reduced to about 5.2%. The latter weight change corresponds to about 1.7 water molecules. At this stage, the data are consistent with the formation of hydrated form C. Thereafter and until about 25% RH, the increased mass is reduced to 4.2%, corresponding to 1.4 water molecules. This data is consistent with the formation of a mixture of hydrated Form C and anhydrous Form B. Thereafter, between 25% and 15% RH, an incremental loss of water was observed in one step. XRPD of the material at the end of the adsorption-desorption cycle showed it to be a mixture of Forms B and C.

[0474] For Experiment 3, the DVS instructions were as follows Fig.46 Two-step water adsorption is shown. The mass change during the first step (between 45-85% RH) is 5.59% corresponding to 1.8 water molecules. The total weight change at 95% RH is 15.88% corresponding to 5.15 water molecules. XRPD measurements of the solid after cycling showed that it was Form B+C.

[0475] For Experiment 4, the deviation between the measured values ​​of water mass gain or loss and the expected corresponding water molecules can be attributed to the fact that the measurements were made before the event equilibrium was reached. Therefore, in this experiment, the relative humidity curves were modified to study the effect of longer equilibrium times at each step. Fig.47 As shown, the maximum mass change is 22.2% corresponding to 7.2 water molecules. The XRPD spectrum of the material after cycling is Form C+D.

[0476] Hydration studies of Forms A and B

[0477] Slurries of Forms A and B (respectively) were performed in water, pH 1.0 HCl buffer (0.1N HCl) and SGF (for Form A) at room temperature. After 45 min, 1.5 h, 15 h, 48 h and 10 days (not in SGF), solids were harvested and wet solids were measured by XRPD. Form A remained stable even after slurrying in water and HCl buffer for 10 days or in SGF for 1.5 h. After 45 min, Form B converted to the heptahydrate Form D, which remained stable for at least 10 days. In a separate experiment, when Form B was exposed to 90% RH for one day, the material converted to a mixture of Forms C and D.

[0478] Dehydration of Forms C and D

[0479] In Table 25, a list of drying conditions for Form C is presented along with the final solid form. Form C appears to be stable after 1.5 hours at 30°C at ambient pressure, while at 40°C it converts to Form B within one hour.

[0480] In Table 26, a list of drying processes for Form D is presented. Form D at 5 mbar pressure and 60°C results in the formation of Form B after 24 h. In some cases, even after 5 days of drying, a small amount of Form C is visible on the XRPD spectra. This observation can be attributed to different particle morphologies (fine particles vs. agglomerates / aggregates). At 60°C and at 50 mbar pressure, Form D converts to a mixture of Form B+C after 86 h and to Form B after 110 h. Typically, Forms B and C occur, depending on time and pressure.

[0481] Table 25. Dehydration of Form C.

[0482]

[0483] Table 26. Dehydration of Form D

[0484]

[0485]

[0486] In Table 27, the appearance of solid forms of Brigatinib are given, along with the crystallization methods and associated solvents from which they were crystallized. The table provides the results of over 600 experiments for which wet and / or dry solid forms were measured by XRPD (as separate experiments, wet and dry counts). In eight cases, form assignment was not made due to low yields. The solid forms following the arrows were obtained after storing the measurement plates under ambient conditions for several weeks (2-5 weeks) and then re-measuring by XRPD.

[0487] Table 27: Summary of brigatinib solid forms

[0488]

[0489]

[0490] III. Pharmaceutical Compositions

[0491] In some embodiments, the present disclosure provides a pharmaceutical composition comprising at least one crystalline form of brigatinib and at least one component selected from a pharmaceutically acceptable carrier, a pharmaceutically acceptable solvent, and a pharmaceutically acceptable excipient. In some embodiments, at least one crystalline form of brigatinib is present in a therapeutically effective amount. In some embodiments, at least one crystalline form of brigatinib is substantially pure. In some embodiments, at least one crystalline form of brigatinib is selected from Form A, Form B, Form C, Form D, Form E, Form F, Form G, and Form H. In some embodiments, crystalline brigatinib is Form A.

[0492] In some embodiments, the unit dosage form of the pharmaceutical composition comprises a single crystalline form of brigatinib as an API. In some embodiments, the present disclosure provides a pharmaceutical composition consisting of a crystalline form of brigatinib. In some embodiments, the present disclosure provides a pharmaceutical composition consisting of a crystalline form of brigatinib and at least one component selected from a pharmaceutically acceptable carrier, a pharmaceutically acceptable solvent, and a pharmaceutically acceptable excipient. In some embodiments, the present disclosure provides a pharmaceutical composition consisting essentially of a crystalline form of brigatinib and optionally at least one component selected from a pharmaceutically acceptable carrier, a pharmaceutically acceptable solvent, and a pharmaceutically acceptable excipient.

[0493] In some embodiments, the present disclosure provides pharmaceutical compositions produced by combining at least one crystalline form of brigatinib and at least one component selected from a pharmaceutically acceptable carrier, a pharmaceutically acceptable solvent, and a pharmaceutically acceptable excipient.

[0494] In some embodiments, the unit dosage form of the pharmaceutical composition comprises more than one crystalline form of brigatinib. In some embodiments, more than 50%, more than 70%, more than 80%, more than 90%, more than 95%, or more than 99% of brigatinib in the composition is in a single crystalline form. In some embodiments, the single crystalline form of brigatinib is selected from Form A, Form B, Form C, Form D, Form E, Form F, Form G, and Form H. In some embodiments, the single crystalline form of brigatinib is Form A.

[0495] In some embodiments, one or all of the crystalline forms are substantially pure. For example, in some embodiments, the pharmaceutical composition comprises substantially pure Form A of brigatinib and at least one component selected from a pharmaceutically acceptable carrier, a pharmaceutically acceptable solvent, and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition comprises Form A and Form B of brigatinib and at least one component selected from a pharmaceutically acceptable carrier, a pharmaceutically acceptable solvent, and a pharmaceutically acceptable excipient. Other embodiments are variations of this theme, which will be apparent to those of ordinary skill in the art after reading this disclosure. For example, in some embodiments, the pharmaceutical composition may comprise Form A and at least one additional brigatinib crystalline form selected from Forms B, C, D, E, F, G, H, J, and K and at least one component selected from a pharmaceutically acceptable carrier, a pharmaceutically acceptable solvent, and a pharmaceutically acceptable excipient.

[0496] At least one component can be easily selected by a person of ordinary skill in the art and can be determined by the mode of administration. Illustrative and non-limiting examples of suitable modes of administration include oral, nasal, parenteral, topical, transdermal and rectal. The pharmaceutical compositions disclosed herein can take any pharmaceutical form that a person of ordinary skill in the art would consider suitable. Non-limiting examples of suitable pharmaceutical forms include solid, semisolid, liquid and lyophilized formulations, such as tablets, powders, capsules, suppositories, suspensions, liposomes and aerosols.

[0497] In some embodiments, the pharmaceutical composition also optionally comprises at least one additional therapeutic agent. In some embodiments, the compounds disclosed herein may be administered to subjects undergoing one or more other therapeutic interventions (such as crizotinib or other kinase inhibitors, interferons, bone marrow transplants, farnesyl transferase inhibitors, bisphosphonates, thalidomide, cancer vaccines, hormone therapy, antibodies, radiation, etc.). For example, in some embodiments, the compounds disclosed herein may be used as a component of a combination therapy with at least one additional therapeutic agent (such as, for example, an anticancer agent), wherein at least one additional therapeutic agent is formulated together with or separately from the compounds disclosed herein.

[0498] As used herein, the term "compound as disclosed herein" refers to at least one crystalline form of brigatinib (i.e., Forms A, B, C, D, E, F, G, H, J, and K) and amorphous brigatinib selected from those disclosed herein. A compound as disclosed herein may be present in a pharmaceutical composition as the sole active agent or may be combined with at least one additional active agent (which may be another form or amorphous brigatinib) or another non-brigatinib compound.

[0499] In some embodiments, the pharmaceutical compositions disclosed herein can be specially formulated for administration in solid or liquid form, including, as non-limiting examples, those suitable for the following: oral administration, such as infusions (aqueous or non-aqueous solutions or suspensions), tablets (such as those targeted for buccal, sublingual, and systemic adsorption), capsules, boluses, powders, granules, pastes (for application to the tongue and intraduodenal routes); parenteral administration, including intravenous, intraarterial, subcutaneous, intramuscular, intravascular, intraperitoneal, or infusion, as, for example, sterile solutions, sterile suspensions, or sustained-release preparations; topical application, such as creams, ointments, controlled-release patches, or sprays applied to the skin; vaginally or rectally, for example, with vaginal suppositories, creams, stents, or foams; sublingually; ocularly; pulmonary; local delivery by catheter or stent; intrathecally or nasally.

[0500] Non-limiting examples of suitable carriers that can be employed in the pharmaceutical compositions disclosed herein include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.), vegetable oils (such as olive oil), injectable organic esters (such as ethyl oleate), and mixtures thereof. For example, appropriate fluidity can be maintained by the use of coating materials (such as lecithin), by maintaining the desired particle size (in the case of dispersions), and by the use of surfactants.

[0501] In some embodiments, the compositions disclosed herein further comprise at least one adjuvant selected from preservatives, wetting agents, emulsifiers, dispersants, lubricants, antioxidants, antibacterial agents, antifungal agents (such as parabens, chlorobutanol, phenol sorbic acid, etc.), isotonic agents (such as sugars, sodium chloride, etc.) and agents capable of delaying adsorption (such as aluminum monostearate, gelatin, etc.).

[0502] Methods for preparing compositions disclosed herein may, for example, include associating at least one compound as disclosed herein with other components such as, for example, chemotherapeutic agents and / or carriers. In some embodiments, the compositions are prepared by uniformly and intimately associating a compound as disclosed herein with at least one carrier selected from a liquid carrier, and then shaping the product if desired.

[0503] The preparation of such pharmaceutical compositions is well known in the art. See, e.g., Anderson, Philip O.; Knoben, James E.; Troutman, William G, ed., Handbook of Clinical Drug Data, 10th ed., McGraw-Hill, 2002; Pratt and Taylor, ed., Principles of Drug Action, 3rd ed., Churchill Livingston, New York, 1990; Katzung, ed., Basic and Clinical Pharmacology, 9th ed., McGraw Hill, 2003; Goodman and Gilman, ed., The Pharmacological Basis of Therapeutics, 10th ed., McGraw Hill, 2001; Remington's Pharmaceutical Sciences, 20th ed., Lippincott Williams & Wilkins., 2000; Martindale, The Extra Pharmacopoeia, 32nd ed. (The Pharmaceutical Press, London, 1999); all of which are incorporated by reference in their entirety. Except insofar as any conventional media is incompatible with the compounds provided herein, such as by producing any undesirable biological effect or otherwise interacting in a deleterious manner with any other ingredient of the pharmaceutically acceptable composition, the use of excipients is contemplated as being within the scope of the present invention.

[0504] In some embodiments, the concentration of brigatinib in the disclosed pharmaceutical compositions is less than 100%, about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, about 19%, about 18%, about 17%, about 16%, about 15%, about 14%, about 13%, about 12%, about 11%, about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, about 2%, about 1%, about 0.5%, about 0.4%, about 0.3%, about 0.2%, about 0.1%, about 0.09%, about 0.08%, about 0.09%, about 0.10%, about 0.010%, about 0.011%, about 0.012%, about 0.013%, about 0.014%, about 0.015%, about 0.016%, about 0.017%, about 0.018%, about 0.019%, about 0.019%, about 0.011%, about 0.013%, about 0.014%, about 0.015%, about 0.016%, about 0.017%, about 0.018%, about 0.019%, about 0.019%, about 0.019%, about 0.011%, about As used herein, "about" means a value modified by ±10%.

[0505] In some embodiments, the concentration of brigatinib in the disclosed pharmaceutical compositions is greater than about 90%, about 80%, about 70%, about 60%, about 50%, about 40%, about 30%, about 20%, about 19.75%, about 19.50%, about 19.25%, about 19%, about 18.75%, about 18.50%, about 18.25%, about 18%, about 17.75%, about 17.50%, about 17.25%, about 17%, about 16.75%, about 16.50%, about 16.25%, about 16%, about 15.75%, about 15.50%, about 15.25%, about 15 %, about 14.75%, about 14.50%, about 14.25%, about 14%, about 13.75%, about 13.50%, about 13.25%, about 13%, about 12.75%, about 12.50%, about 12.25%, about 12%, about 11.75%, about 11.50%, about 11.25%, about 11%, about 10.75%, about 10.50%, about 10.25%, about 10%, about 9.75%, about 9.50%, about 9.25%, about 9%, about 8.75%, about 8.50%, about 8.25%, about 8%, about 7.75%, about 7.50% , about 7.25%, about 7%, about 6.75%, about 6.50%, about 6.25%, about 6%, about 5.75%, about 5.50%, about 5.25%, about 5%, about 4.75%, about 4.50%, about 4.25%, about 4%, about 3.75%, about 3.50%, about 3.25%, about 3%, about 2.75%, about 2.50%, about 2.25%, about 2%, about 1.75%, about 1.50%, about 1.25%, about 1%, about 0.5%, about 0.4%, about 0.3%, about 0.2%, about 0.1%, about 0.09%, about 0.08%, about 0. As used herein, "about" means a value modified by ±10%.

[0506] In some embodiments, the concentration of brigatinib in the disclosed pharmaceutical compositions is in the range of about 0.0001% to about 50%, about 0.001% to about 40%, about 0.01% to about 30%, about 0.02% to about 29%, about 0.03% to about 28%, about 0.04% to about 27%, about 0.05% to about 26%, about 0.06% to about 25%, about 0.07% to about 24%, about 0.08% to about 23%, about 0.09% to about 22%, about 0.1% to about 21%, about 0.2% to about 20%, about 0.3% to about 19%, about 0.4% to about 18%, about 0.5% to about 17%, about 0.6% to about 16%, about 0.7% to about 15%, about 0.8% to about 14%, about 0.9% to about 12%, about 1% to about 10% w / w, w / v or v / v, v / v. As used herein, "about" means a value modified by ±10%.

[0507] In some embodiments, the concentration of brigatinib in the disclosed pharmaceutical compositions is in the range of about 0.001% to about 10%, about 0.01% to about 5%, about 0.02% to about 4.5%, about 0.03% to about 4%, about 0.04% to about 3.5%, about 0.05% to about 3%, about 0.06% to about 2.5%, about 0.07% to about 2%, about 0.08% to about 1.5%, about 0.09% to about 1%, about 0.1% to about 0.9% w / w, w / v or v / v. As used herein, "about" means a value modified by ±10%.

[0508] In some embodiments, the amount of brigatinib in the disclosed pharmaceutical compositions is equal to or less than about 10 g, about 9.5 g, about 9.0 g, about 8.5 g, about 8.0 g, about 7.5 g, about 7.0 g, about 6.5 g, about 6.0 g, about 5.5 g, about 5.0 g, about 4.5 g, about 4.0 g, about 3.5 g, about 3.0 g, about 2.5 g, about 2.0 g, about 1.5 g, about 1.0 g, about 0.95 g, about 0.9 g, about 0.85 g, about 0.8 g, about 0.75 g, about 0.7 g, about 0.65 g, about 0.6 g, about 0.55 g, about 0.5 g, about 0.45 g, about 0.4 g, about 0.35 g, about 0.3 g, about 0.2 About 0.5g, about 0.2g, about 0.15g, about 0.1g, about 0.09g, about 0.08g, about 0.07g, about 0.06g, about 0.05g, about 0.04g, about 0.03g, about 0.02g, about 0.01g, about 0.009g, about 0.008g, about 0.007g, about 0.006g, about 0.005g, about 0.004g, about 0.003g, about 0.002g, about 0.001g, about 0.0009g, about 0.0008g, about 0.0007g, about 0.0006g, about 0.0005g, about 0.0004g, about 0.0003g, about 0.0002g or about 0.0001g.In some embodiments, the amount of one or more compounds as disclosed herein may exceed about 0.0001 g, about 0.0002 g, about 0.0003 g, about 0.0004 g, about 0.0005 g, about 0.0006 g, about 0.0007 g, about 0.0008 g, about 0.0009 g, about 0.001 g, about 0.0015 g, about 0.002 g, about 0.0025 g, about 0.003 g, about 0.0035 g. 0.004g, about 0.0045g, about 0.005g, about 0.0055g, about 0.006g, about 0.0065g, about 0.007g, about 0.0075g, about 0.008g, about 0.0085g, about 0.009g, about 0.0095g, about 0.01g, about 0.015g, about 0.02g, about 0.025g, about 0.03g, about 0.035g, about 0.04g, about 0.045g, about 0.05g, about 0.055g, about 0.06g, about 0.065g, about 0.07g, about 0.075g, about 0.08g, about 0.085g, about 0 As used herein, "about" means a value modified by ±10%.

[0509] In some embodiments, the amount of brigatinib in the disclosed pharmaceutical composition is in the range of about 0.0001 to about 10 g, about 0.0005 g to about 9 g, about 0.001 g to about 0.5 g, about 0.001 g to about 2 g, about 0.001 g to about 8 g, about 0.005 g to about 2 g, about 0.005 g to about 7 g, about 0.01 g to about 6 g, about 0.05 g to about 5 g, about 0.1 g to about 4 g, about 0.5 g to about 4 g, or about 1 g to about 3 g. As used herein, "about" means a value modified by ±10%.

[0510] In some embodiments, the present disclosure provides a pharmaceutical composition for oral administration, comprising at least one compound as disclosed herein and at least one pharmaceutically acceptable excipient suitable for oral administration. In some embodiments, the present disclosure provides a pharmaceutical composition for oral administration, comprising: (i) a therapeutically effective amount of at least one compound as disclosed herein; optionally (ii) an effective amount of at least one second agent; and (iii) at least one pharmaceutically acceptable excipient suitable for oral administration. In some embodiments, the pharmaceutical composition further comprises (iv) an effective amount of at least one third agent.

[0511] In some embodiments, the pharmaceutical composition may be a liquid pharmaceutical composition suitable for oral consumption. Pharmaceutical compositions suitable for oral administration may be presented, for example, in discrete dosage forms, such as capsules, cachets or tablets, or liquids or aerosol sprays (each containing a predetermined amount of active ingredients in powder or granules), solutions or suspensions in aqueous or non-aqueous liquids, oil-in-water emulsions or water-in-oil liquid emulsions. Such dosage forms may be prepared by any pharmaceutical method, but all methods include the step of associating the active ingredient with a carrier constituting one or more ingredients. Typically, pharmaceutical compositions are prepared by uniformly and closely combining the active compound with a liquid carrier or a finely divided solid carrier or both, and then, if necessary, forming the product into the desired presentation form. For example, tablets may be prepared by compression or molding, optionally with one or more auxiliary ingredients. Compressed tablets may be prepared by compressing an active ingredient in a free-flowing form (such as a powder or granules), optionally mixed with an excipient (such as, but not limited to, a binder, a lubricant, an inert diluent and / or a surfactant or a dispersant) in a suitable machine. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

[0512] Tablets may be uncoated or coated by known techniques to delay disintegration and adsorption in the gastrointestinal tract and thereby provide sustained action over a longer period of time. For example, time-delayed materials such as glyceryl monostearate or glyceryl distearate may be used. Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient may be mixed with an inert solid diluent (e.g., calcium carbonate, calcium phosphate, or kaolin), or as soft gelatin capsules in which the active ingredient may be mixed with water or an oil medium (e.g., peanut oil, liquid paraffin, or olive oil).

[0513] In some embodiments, the present disclosure also encompasses anhydrous pharmaceutical compositions and dosage forms comprising at least one active ingredient. Water can promote degradation of some compounds. For example, water (such as about 5%) in the pharmaceutical field can be added as a way to simulate long-term storage to determine the characteristics of the formulation over time, such as shelf life or stability. Anhydrous pharmaceutical compositions and dosage forms can be prepared using anhydrous or low-moisture ingredients and low-moisture or low-humidity conditions. For example, if there is a large amount of contact with moisture and / or humidity during manufacture, packaging and / or storage, pharmaceutical compositions and dosage forms containing lactose can be prepared as anhydrous, and / or are expected to be stored. Anhydrous pharmaceutical compositions can be prepared and stored so that their anhydrous properties are maintained. Therefore, anhydrous pharmaceutical compositions can be packaged using known materials to prevent exposure to water so that they can be included in suitable preparation kits. Examples of suitable packaging include, but are not limited to, airtight sealed foils, plastics, etc., unit dose containers, blister packs, and strip packs.

[0514] The active ingredient can be combined with a pharmaceutical carrier in a close mixture according to conventional pharmaceutical compounding techniques. The carrier can take a variety of forms, depending on the preparation method required for administration. During the preparation of a pharmaceutical composition for oral dosage forms, any common pharmaceutical medium can be used as a carrier, such as, for example, water, glycol, oil, alcohol, flavoring agent, preservative, colorant, etc. in the case of oral liquid preparations (such as suspensions, solutions and elixirs) or aerosols; or in some embodiments where lactose is not used, carriers such as starch, sugar, microcrystalline cellulose, diluents, granulators, lubricants, binders and disintegrants can be used in the case of oral solid preparations. In some embodiments, the mixture can be blended with lactose, sucrose, starch powder, cellulose esters of alkanoic acid, cellulose alkyl esters, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric acid and sulfuric acid, gelatin, gum arabic, sodium alginate, polyvinyl pyrrolidone and / or polyvinyl alcohol for subsequent preparation. For example, in the case of solid oral preparations, suitable carriers include powders, capsules and tablets. In some embodiments, tablets may be coated by standard aqueous or nonaqueous techniques.

[0515] Non-limiting examples of binders suitable for use in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, corn starch, potato starch, and other starches, gelatin, natural and synthetic gums (such as acacia, sodium alginate, alginic acid, other alginates, powdered tragacanth, guar gum), cellulose and its derivatives (e.g., ethylcellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose), polyvinylpyrrolidone, methylcellulose, pregelatinized starch, hydroxypropyl methylcellulose, microcrystalline cellulose, and mixtures thereof.

[0516] Non-limiting examples of fillers suitable for use in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, talc, calcium carbonate (as granules or powders), microcrystalline cellulose, powdered cellulose, dextrose, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof.

[0517] Disintegrants can be used in pharmaceutical compositions and dosage forms disclosed herein to provide tablets that disintegrate when exposed to an aqueous environment. Too much disintegrant can produce tablets that can disintegrate in a bottle. Too little may not be enough to cause disintegration to occur and may therefore change the rate and degree of release of the active ingredient from the dosage form. Therefore, a sufficient amount of disintegrant (i.e., not too little nor too much to adversely change the release of the active ingredient) can be used to prepare pharmaceutical compositions and dosage forms disclosed herein. The amount of disintegrant can be changed based on the type and mode of administration of the preparation, and may be easily discernible to a person of ordinary skill in the art. For example, in some embodiments, at least one disintegrant of about 0.5 to about 15 total weight percent can be used. In some embodiments, at least one disintegrant of about 1 to about 5 total weight percent can be used in a pharmaceutical composition. Disintegrants that may be used include, but are not limited to, agar, alginic acid, calcium carbonate, microcrystalline cellulose, croscarmellose sodium, crospovidone, polacrilin potassium, sodium starch glycolate, potato or tapioca starch, other starches, pregelatinized starch, other starches, clays, other algins, other celluloses, gums, and mixtures thereof.

[0518] Lubricants that can be used in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerol, sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils (such as peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil), zinc stearate, ethyl oleate, ethyl laurate, agar, syloid silica gel, a condensed aerosol of synthetic silicon dioxide, and mixtures thereof. Lubricants can be optionally added in an amount of less than about 1 percent by weight of the total pharmaceutical composition.

[0519] When aqueous suspensions and / or elixirs are selected for oral administration, the pharmaceutical composition may further comprise at least one additional agent selected from sweeteners, flavoring agents, coloring substances, dyes, emulsifiers, suspending agents and diluents (e.g., water, ethanol, propylene glycol, glycerol, etc.).

[0520] Surfactants that may be included in the pharmaceutical compositions and dosage forms disclosed herein include, but are not limited to, hydrophilic surfactants, lipophilic surfactants, and mixtures thereof. That is, a mixture of hydrophilic surfactants may be used, a mixture of lipophilic surfactants may be used, or a mixture of at least one hydrophilic surfactant and at least one lipophilic surfactant may be used.

[0521] In some embodiments, hydrophilic surfactants have an HLB value of at least about 10, while lipophilic surfactants have an HLB value of less than about 10. An empirical parameter for characterizing the relative hydrophilicity and hydrophobicity of non-ionic amphiphilic compounds is the hydrophilicity-lipophilicity balance ("HLB" value). Surfactants with lower HLB values ​​are more lipophilic or hydrophobic and have better solubility in oil, while surfactants with higher HLB values ​​are more hydrophilic and have better solubility in aqueous solution. Hydrophilic surfactants are generally considered to be those compounds with HLB values ​​greater than about 10, as well as anionic, cationic or zwitterionic compounds (for which the HLB scale is generally not applicable). Similarly, lipophilic (i.e., hydrophobic) surfactants are compounds with HLB values ​​equal to or less than about 10. However, the HLB value of a surfactant is only a rough guide that is generally used to be able to formulate industrial, pharmaceutical and cosmetic emulsions.

[0522] The hydrophilic surfactant may be ionic or nonionic. Suitable ionic surfactants include, but are not limited to, alkylammonium salts; fusidates; fatty acid derivatives of amino acids, oligopeptides and polypeptides; glyceride derivatives of amino acids, oligopeptides and polypeptides; lecithin and hydrogenated lecithin; lysolecithin and hydrogenated lysolecithin; phospholipids and their derivatives; lysolecithin and their derivatives; carnitine fatty acid ester salts; alkyl sulfates; fatty acid salts; sodium docusate; acetylacetylates; mono- and di-acetylated tartaric acid esters of mono- and di-glycerides; succinylated mono- and di-glycerides; citric acid esters of mono- and di-glycerides; and mixtures thereof.

[0523] In the aforementioned group, ionic surfactants include, but are not limited to, lecithin, lysolecithin, phospholipids, lysophospholipids and their derivatives; carnitine fatty acid ester salts; alkyl sulfates; fatty acid salts; docusate sodium; acetylsalicylate; mono- and di-acetylated tartaric acid esters of mono- and di-glycerides; succinylated mono- and di-glycerides; citric acid esters of mono- and di-glycerides; and mixtures thereof.

[0524] Other non-limiting examples of ionic surfactants include ionized forms of lecithin, lysolecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidic acid, phosphatidylserine, lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylglycerol, lysophosphatidic acid, lysophosphatidylserine, PEG-phosphatidylethanolamine, PVP-phosphatidylethanolamine, lactyl esters of fatty acids, stearoyl-2-1 lactylate, stearoyl lactylate, succinylated monoglycerides, monoglycerides, and monoglycerides. tartrates of mono / diglycerides, citric acid esters of mono / diglycerides, cholylsarcosine, caproate, caprylate, caprate, laurate, myristate, palmitate, oleate, ricinoleate, linolenate, linolenate, stearate, lauryl sulfate, teracecyl sulfate, docusate, lauroyl carnitine, palmitoyl carnitine, myristoyl carnitine, and salts and mixtures thereof.

[0525] Non-limiting examples of hydrophilic nonionic surfactants include alkyl glucosides; alkyl maltosides; alkyl glucosides; lauryl macrogolglyceride; polyoxyalkylene alkyl ethers, such as polyethylene glycol alkyl ethers; polyoxyalkylene alkylphenols, such as polyethylene glycol alkylphenols; polyoxyalkylene alkylphenol fatty acid esters, such as polyethylene glycol fatty acid monoesters and polyethylene glycol fatty acid diesters; polyethylene glycol glycerol fatty acid esters; polyglycerol fatty acid esters; polyoxyalkylene sorbitan fatty acid esters, such as polyethylene glycol sorbitan fatty acid esters; hydrophilic transesterification products of polyols with at least one member of glycerides, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols; polyoxyalkylene sterols, derivatives and analogs thereof; polyoxyethylated vitamins and derivatives thereof; polyoxyethylene-polyoxypropylene block copolymers; and mixtures thereof; polyethylene glycol sorbitan fatty acid esters and hydrophilic transesterification products of polyols with at least one member of triglycerides, vegetable oils and hydrogenated vegetable oils. The polyol may be glycerol, ethylene glycol, polyethylene glycol, sorbitol, propylene glycol, pentaerythritol or sugar.

[0526] Other hydrophilic-non-ionic surfactants include, but are not limited to, PEG-10 laurate, PEG-12 laurate, PEG-20 laurate, PEG-32 laurate, PEG-32 dilaurate, PEG-12 oleate, PEG-15 oleate. PEG-20 oleate, PEG-20 dioleate, PEG-32 oleate, PEG-200 oleate, PEG-400 oleate, PEG-15 stearate, PEG-32 distearate, PEG-40 stearate, PEG-100 stearate, PEG-20 dilaurate, PEG-25 triolein, PEG-32 dioleate, PEG-20 laurin, PEG-30 laurin, PEG-20 stearin, PEG-20 olein, PEG-30 olein, PEG-30 laurin, PEG-40 laurin, PEG-40 palm kernel oil, PEG-50 hydrogenated castor oil, PEG-40 castor oil, PEG-35 castor oil, P EG-60 castor oil, PEG-40 hydrogenated castor oil, PEG-60 hydrogenated castor oil, PEG-60 corn oil, PEG-6 capric / caprylic glycerides, PEG-8 capric / caprylic glycerides, polyglyceryl -10 laurate, PEG-30 cholesterol, PEG-25 phytosterols, PEG-30 soybean sterols, PEG-20 trioleate, PEG-40 sorbitan oleate, PEG-80 sorbitan laurate, polysorbate 20, polysorbate 80, POE-9 lauryl ether, POE-23 lauryl ether, POE-10 oleyl ether, POE-20 oleyl ether, POE-20 stearyl ether, tocopherol PEG-100 succinate, PEG-24 cholesterol, polyglyceryl -10 oleate, Tween 40, Tween 60, sucrose monostearate, sucrose monolaurate, sucrose monopalmitate, PEG10-100 nonylphenol series, PEG 15-100 octylphenol series and poloxamer.

[0527] Suitable lipophilic surfactants include, but are not limited to, fatty alcohols; glycerol fatty acid esters; acetylated glycerol fatty acid esters; lower alcohol fatty acid esters; propylene glycol fatty acid esters; sorbitan fatty acid esters; polyethylene glycol sorbitan fatty acid esters; sterols and sterol derivatives; polyoxyethylated sterols and sterol derivatives; polyethylene glycol alkyl ethers; sugar esters; sugar ethers; lactic acid derivatives of mono- and di-glycerides; hydrophobic transesterification products of polyols with at least one member of glycerol esters, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols, vegetable oils, hydrogenated vegetable oils, fatty acids and sterols; oil-soluble vitamins / vitamin derivatives; and mixtures thereof. In this group, non-limiting examples of lipophilic surfactants include glycerol fatty acid esters, propylene glycol fatty acid esters and mixtures thereof, or hydrophobic transesterification products of polyols with at least one member of vegetable oils, hydrogenated vegetable oils and triglycerides.

[0528] In some embodiments, the pharmaceutical compositions and dosage forms disclosed herein may include at least one solubilizing agent to ensure good solubilization and / or dissolution of the compounds disclosed herein and to minimize precipitation of the compounds. This can be used for pharmaceutical compositions for non-oral use, such as pharmaceutical compositions for injection. Solubilizing agents can also be added to increase the solubility of hydrophilic drugs and / or other components (such as surfactants), or to maintain the pharmaceutical composition as a stable or uniform solution or suspension.

[0529] Examples of suitable solubilizers include, but are not limited to, the following: alcohols and polyols such as ethanol, isopropanol, butanol, benzyl alcohol, ethylene glycol, propylene glycol, butanediol and isomers thereof, glycerol, pentaerythritol, sorbitol, mannitol, transcutol, dimethyl isosorbide, polyethylene glycol, polypropylene glycol, polyvinyl alcohol, hydroxypropyl methylcellulose and other cellulose derivatives, cyclodextrins and cyclodextrin derivatives; ethers of polyethylene glycol having an average molecular weight of about 200 to about 6000, such as tetrahydrofuranol PEG ether (glycogen) or methoxy PEG; amides and other nitrogen-containing compounds such as 2-pyrrolidone, 2-piperidone, ε-caprolactam, N-alkylpyrrolidone, N-hydroxyalkylpyrrolidone, N-alkylpiperidone, N-alkylcaprolactam, dimethylacetamide and polyvinylpyrrole esters such as ethyl propionate, tributyl citrate, triethyl acetyl citrate, acetyl tributyl citrate, triethyl citrate, ethyl oleate, ethyl caprylate, ethyl butyrate, glycerol triacetate, propylene glycol monoacetate, propylene glycol diacetate, ε-caprolactone and its isomers, δ-valerolactone and its isomers, β-butyrolactone and its isomers; and other solubilizing agents known in the art such as dimethylacetamide, dimethyl isosorbide, N-methylpyrrolidone, monoetanoin, diethylene glycol monoethyl ether and water.

[0530] Mixtures of solubilizing agents can also be used. Examples include, but are not limited to, triacetin, triethyl citrate, ethyl oleate, ethyl caprylate, dimethylacetamide, N-methyl pyrrolidone, N-hydroxyethyl pyrrolidone, polyvinyl pyrrolidone, hydroxypropyl methylcellulose, hydroxypropyl cyclodextrin, ethanol, polyethylene glycol 200-100, glycogen, transcutol, propylene glycol and dimethyl isosorbide. In some embodiments, the solubilizing agent includes sorbitol, glycerol, triacetin, ethanol, PEG-400, glycogen and propylene glycol.

[0531] The amount of solubilizing agent can change along with composition.The amount of the solubilizing agent provided can be limited to biologically acceptable amount, and it can be easily measured by those skilled in the art.In some cases, it can be advantageous to comprise the amount of the solubilizing agent far exceeding biologically acceptable amount, for example, in order to maximize drug concentration, before pharmaceutical composition is provided to object, conventional techniques such as distillation or evaporation are used to remove excessive solubilizing agent.Therefore, if there is, solubilizing agent can exist in the amount of about 10 % by weight, about 25 % by weight, about 50 % by weight, about 100 % by weight or about 200 % by weight at most of the gross weight based on composition.In some embodiments, solubilizing agent can exist in the amount of about 5%, about 2%, about 1% or even less.In some embodiments, solubilizing agent can exist in the amount of about 1 % by weight to about 100 % by weight such as about 5 % by weight to about 25 % by weight.

[0532] The pharmaceutical composition may further comprise at least one pharmaceutically acceptable excipient. Such excipients include, but are not limited to, detackifiers, defoamers, buffers, polymers, antioxidants, preservatives, chelating agents, viscosity modifiers, tension modifiers, flavoring agents, colorants, oils, odorants, opacifiers, suspending agents, adhesives, fillers, plasticizers, lubricants, and mixtures thereof.

[0533] The limiting examples of preservatives include antioxidants, chelating agents, antimicrobial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives and other preservatives. Exemplary antioxidants include but are not limited to alpha tocopherol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium pyrosulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium pyrosulfite and sodium sulfite. The limiting examples of chelating agents include ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate, disodium edetate, dipotassium edetate, edetic acid, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid and trisodium edetate. Exemplary antimicrobial preservatives include but are not limited to benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerol, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol and thimerosal. Exemplary antifungal preservatives include but are not limited to butylparaben, methylparaben, ethylparaben, propylparaben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate and sorbic acid. Exemplary alcohol preservatives include but are not limited to, ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate and phenylethyl alcohol. Exemplary acidic preservatives include but are not limited to vitamin A, vitamin C, vitamin E, beta carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid and phytic acid. Other preservatives include but are not limited to tocopherol, tocopherol acetate, mesylate deteroxime (deteroxime mesylate), cetrimonium bromide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium bisulfite, sodium pyrosulfite, potassium sulfite, potassium pyrosulfite, Glydant Plus, Phenonip, methyl paraben, Germall 115, Germaben II, Neolone (Neolone), Kathon (Kathon) and Euxyl. In certain embodiments, the preservative can be an antioxidant. In other embodiments, the preservative can be a chelating agent.

[0534] Exemplary oils include, but are not limited to, almond oil, apricot kernel oil, avocado oil, babassu oil, bergamot oil, black currant seed oil, borage oil, juniper oil, chamomile oil, canola oil, coriander, carnauba oil, castor oil, cinnamon oil, cocoa butter, coconut oil, cod liver oil, coffee oil, corn oil, cottonseed oil, emu oil, eucalyptus oil, evening primrose oil, fish oil, linseed oil, citronellol oil, gourd oil, grape seed oil, hazelnut oil, hyssop oil, isopropyl myristate oil, jojoba oil, macadamia nut oil, lavender oil. Oil, lavender oil, lemon oil, litsea cubeba oil, macadamia oil, mallow oil, mango seed oil, meadowfoam seed oil, peppermint oil, nutmeg oil, olive oil, orange oil, orange roughy oil, palm oil, palm kernel oil, peach kernel oil, peanut oil, poppy seed oil, pumpkin seed oil, rapeseed oil, rice bran oil, rosemary oil, safflower oil, sandalwood oil, sasanqua oil, savory oil, sea buckthorn oil, sesame oil, shea butter, silicone oil, soybean oil, sunflower oil, tea tree oil, thistle oil, camellia oil, valerian oil, walnut oil and wheat germ oil. Exemplary oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil and combinations thereof.

[0535] In some embodiments, the composition disclosed herein may be an oil / water formulation. The oil / water emulsion formulation may include at least one emulsifier optionally with at least one fat and / oil. In some embodiments, the composition disclosed herein may include at least one hydrophilic emulsifier optionally with at least one lipophilic emulsifier that can be used as a stabilizer. In some embodiments, both oil and fat can be used. At least one emulsifier optionally with at least one stabilizer can produce at least one emulsifying wax, which can form an emulsified ointment base. The ointment base can form the oily dispersed phase of the cream formulation. Emulsifiers and emulsion stabilizers suitable for disclosed formulations include but are not limited to Tween 60, Span 80, cetearyl alcohol, myristyl alcohol, glyceryl monostearate, sodium lauryl sulfate, glyceryl distearate (alone or with wax), and other materials well known in the art. In some cases, the solubility of the active compound in the oil that may be used in the pharmaceutical emulsion formulation may be low. Straight or branched, mono- or dibasic alkyl esters may aid in dissolution, such as diisoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acid, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate, or a blend of branched esters. These may be used alone or in combination, depending on the desired properties. Alternatively, high melting point lipids such as white soft paraffin and / or liquid paraffin or other mineral oils may be used.

[0536] In addition, acids or bases can be incorporated into the pharmaceutical composition to facilitate processing, enhance stability or for other reasons. Examples of pharmaceutically acceptable bases include amino acids, amino acid esters, ammonium hydroxide, potassium hydroxide, sodium hydroxide, sodium bicarbonate, aluminum hydroxide, calcium carbonate, magnesium hydroxide, magnesium aluminum silicate, synthetic aluminum silicate, hydrotalcite, magnesium aluminum hydroxide, diisopropylethylamine, ethanolamine, ethylenediamine, triethanolamine, triethylamine, triisopropanolamine, trimethylamine and tris (hydroxymethyl) aminomethane (TRIS) and the like. The alkali as the salt of pharmaceutically acceptable acid is also suitable, such as acetic acid, acrylic acid, adipic acid, alginic acid, alkyl sulfonic acid, amino acid, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acid, formic acid, fumaric acid, gluconic acid, hydroquinone sulfonic acid, isoascorbic acid, lactic acid, maleic acid, oxalic acid, p-bromobenzenesulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, uric acid etc. can also be used. The salt of polyacid can be such as sodium phosphate, disodium hydrogen phosphate and sodium dihydrogen phosphate. When the alkali is a salt, the cation can be any suitable and pharmaceutically acceptable cation, such as ammonium, alkali metal, alkaline earth metal etc. Example can include but is not limited to sodium, potassium, lithium, magnesium, calcium and ammonium.

[0537] The non-limiting examples of suitable acid are pharmaceutically acceptable organic or inorganic acids. Suitable examples of inorganic acids include, but are not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, boric acid, phosphoric acid, etc. Suitable examples of organic acids include, but are not limited to, acetic acid, acrylic acid, adipic acid, alginic acid, alkylsulfonic acid, amino acid, ascorbic acid, benzoic acid, boric acid, butyric acid, carbonic acid, citric acid, fatty acid, formic acid, fumaric acid, gluconic acid, hydroquinonesulfonic acid, isoascorbic acid, lactic acid, maleic acid, methanesulfonic acid, oxalic acid, p-bromobenzenesulfonic acid, propionic acid, p-toluenesulfonic acid, salicylic acid, stearic acid, succinic acid, tannic acid, tartaric acid, thioglycolic acid, toluenesulfonic acid, and uric acid, etc.

[0538] In some embodiments, provided herein are pharmaceutical compositions for parenteral administration, containing at least one compound as disclosed herein and at least one pharmaceutically acceptable excipient suitable for parenteral administration. In some embodiments, provided herein are pharmaceutical compositions for parenteral administration, comprising: (i) an effective amount of at least one compound as disclosed herein; optionally (ii) an effective amount of at least one second agent; and (iii) at least one pharmaceutically acceptable excipient suitable for parenteral administration. In some embodiments, the pharmaceutical composition further comprises (iv) an effective amount of at least one third agent.

[0539] Forms that may be incorporated into the disclosed pharmaceutical compositions for administration by injection include aqueous or oily suspensions or emulsions (containing sesame oil, corn oil, cottonseed oil, or peanut oil), as well as elixirs, mannitol, dextrose, or sterile aqueous solutions, and similar pharmaceutical vehicles. Aqueous solutions in saline are also commonly used for injection. Ethanol, glycerol, propylene glycol, liquid polyethylene glycol, benzyl alcohol, etc. (and suitable mixtures thereof), cyclodextrin derivatives, sodium chloride, tragacanth, buffers, and vegetable oils may also be used.

[0540] Aqueous solutions in saline are generally useful for injection. Ethanol, glycerol, propylene glycol, liquid polyethylene glycols, etc. (and suitable mixtures thereof), cyclodextrin derivatives, and vegetable oils may also be used. The desired particle size (in the case of a suspension) may be maintained, for example, by the use of a coating such as lecithin and by the use of a surfactant to maintain appropriate fluidity. Prevention of microbial action may be caused by various antibacterial and antifungal agents (e.g., parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc.).

[0541] In some embodiments, the active ingredient may also be administered by injection in a composition with a suitable carrier (including but not limited to saline, dextrose or water) or with a cyclodextrin (such as Captisol), a co-solvent solubilization (such as propylene glycol) or a micellar solubilization (such as Tween 80).

[0542] Sterile injectable solutions can be prepared by the following steps: as required, the required amount of the compound as disclosed herein is blended with various other ingredients as listed above in an appropriate solvent, and then filtered for sterilization. Typically, dispersions are prepared by incorporating various sterilized active ingredients into a sterile solvent containing a basic dispersion medium and appropriate other ingredients as listed above. In the case of sterile powders for the preparation of sterile injectable solutions, certain preparation methods are vacuum drying and freeze drying techniques, which obtain a powder of the active ingredient plus any other ingredients from a previously sterile filtered solution thereof.

[0543] Sterile injectable preparations can also be sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, such as solutions of 1,3-butanediol. Acceptable vehicles and solvents that can be used are water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile non-volatile oils are generally used as solvents or suspending media. For this purpose, any mild non-volatile oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables.

[0544] The injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating a sterilizing agent in the form of a sterile solid composition that can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use. The injectable composition may contain from about 0.1% to about 5% w / w of a compound as disclosed herein.

[0545] In some embodiments, provided herein are pharmaceutical compositions for topical (e.g., transdermal) administration, comprising at least one compound as disclosed herein and at least one pharmaceutically acceptable excipient suitable for topical administration. In some embodiments, provided herein are pharmaceutical compositions for topical administration, comprising (i) an effective amount of at least one compound disclosed herein; optionally (ii) an effective amount of at least one second agent; and (iii) at least one pharmaceutically acceptable excipient suitable for topical administration. In some embodiments, the pharmaceutically acceptable composition further comprises (iv) an effective amount of at least one third agent.

[0546] The pharmaceutical compositions provided herein can be formulated into preparations in solid, semisolid or liquid form suitable for local / topical application, such as gels, water-soluble jellies, liniments, creams, lotions, suspensions, foams, powders, slurries, ointments, solutions, oils, pastes, suppositories, sprays, emulsions, saline solutions, solutions based on dimethyl sulfoxide (DMSO). Generally, carriers with higher density can provide areas with extended exposure to active ingredients. In contrast, solution formulations can expose active ingredients more directly to selected areas. For example, ointment preparations can have paraffin or water-miscible bases. Alternatively, active ingredients can be configured in creams with water-in-oil cream bases. The aqueous phase of the cream base can include, for example, at least about 30% w / w of polyhydroxy alcohols, such as propylene glycol, butane-1,3-diol, mannitol, sorbitol, glycerol, polyethylene glycol and mixtures thereof.

[0547] Pharmaceutical composition can also comprise suitable solid or gel phase carrier or excipient, it is to increase the permeation of therapeutic molecules through the skin stratum corneum permeation barrier or contribute to the compound of delivering therapeutic molecules through the skin stratum corneum permeation barrier. Many of these penetration enhancing molecules are known to the technicians in the field of topical preparations. Examples of such carriers and excipients include but are not limited to humectants (such as, urea), glycols (such as propylene glycol), alcohols (such as ethanol), fatty acids (such as oleic acid), surfactants (such as isopropyl myristate and sodium lauryl sulfate), pyrrolidone, monolaurin, sulfoxide, terpene (such as, menthol), amine, amide, alkane, alkanol, water, calcium carbonate, calcium phosphate, various sugars, starch, cellulose derivatives, gelatin and polymers such as polyethylene glycol.

[0548] Another exemplary formulation used in the disclosed method adopts a transdermal delivery device ("patch"). The transdermal patch can be used to provide a continuous or discontinuous infusion of a compound as provided herein (with or without another medicament) in a controlled amount. The patch can be a reservoir and a porous membrane type or a solid matrix type. In either case, the active agent can be continuously delivered from the reservoir or microcapsule through a membrane to an active agent permeable adhesive in contact with the skin or mucous membrane of the recipient. If the active agent is absorbed through the skin, a controlled and predetermined active agent flow is applied to the recipient. In the case of microcapsules, an encapsulating agent can also be used as a membrane.

[0549] The construction and use of transdermal patches for the delivery of pharmaceutical agents is well known in the art. See, for example, U.S. Pat. Nos. 5,023,252, 4,992,445, and 5,001,139. ​​Such patches may be constructed for continuous, pulsatile, or on demand delivery of an agent.

[0550] Devices suitable for delivering intradermal pharmaceutically acceptable compositions described herein include short needle devices, such as those described in U.S. Pat. Nos. 4,886,499; 5,190,521; 5,328,483; 5,527,288; 4,270,537; 5,015,235; 5,141,496; and 5,417,662. Intradermal compositions may be administered by devices that limit the length of effective needle penetration into the skin, such as those described in PCT Publication WO 99 / 34850 and functional equivalents thereof. Jet injection devices that deliver liquid vaccines to the dermis via a liquid jet injector and / or via a needle that pierces the stratum corneum and produces a jet that reaches the dermis are suitable. Jet injection devices are described, for example, in U.S. Patent Nos. 5,480,381; 5,599,302; 5,334,144; 5,993,412; 5,649,912; 5,569,189; 5,704,911; 5,383,851; 5,893,397; 5,466,220; 5,339,163; 5,312,335; 5,503,627; 5,064,413; 5,520,639; 4,596,556; 4,790,824; 4,941,880; 4,940,460; and in PCT Publications WO 97 / 37705 and WO 97 / 13537. Ballistic powder / particle delivery devices that use compressed gas to accelerate the vaccine in powder form through the outer layers of the skin to the dermis are suitable. Alternatively or additionally, conventional syringes can be used in the classical mantoux method of intradermal administration.

[0551] Topically administrable preparations may, for example, include about 1% to about 10% (w / w) of the disclosed compound, although the concentration of the compound of formula I may be as high as the solubility limit of the compound in the solvent. In some embodiments, topically administrable preparations may, for example, include about 0.001% to about 10% (w / w) compound, about 1% to about 9% (w / w) compound, such as about 1% to about 8% (w / w), further such as about 1% to about 7% (w / w), further such as about 1% to about 6% (w / w), further such as about 1% to about 5% (w / w), further such as about 1% to about 4% (w / w), further such as about 1% to about 3% (w / w), further such as about 1% to about 2% (w / w) and further such as about 0.1% to about 1% (w / w) compound. In some embodiments, topical preparations include about 0.1 mg to about 150 mg applied one to four times a day, such as once or twice. Preparations for topical administration may also include one or more additional pharmaceutically acceptable excipients described herein.

[0552] In some embodiments, provided herein is a pharmaceutical composition for inhalation administration, comprising at least one compound as disclosed herein and at least one pharmaceutically acceptable excipient suitable for topical administration. In some embodiments, provided herein is a pharmaceutical composition for inhalation administration, comprising: (i) an effective amount of at least one compound disclosed herein; optionally (ii) an effective amount of at least one second agent; and (iii) at least one pharmaceutically acceptable excipient suitable for inhalation administration. In some embodiments, the pharmaceutical composition further comprises: (iv) an effective amount of at least one third agent.

[0553] Pharmaceutical compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, and powders. Liquid or solid pharmaceutical compositions may contain suitable pharmaceutically acceptable excipients as described herein. For example, suitable excipients include, but are not limited to, saline, benzyl alcohol, and fluorocarbons. In some embodiments, the pharmaceutical composition may be administered through the oral or nasal respiratory tract for local or systemic effects. Pharmaceutical compositions in pharmaceutically acceptable solvents may be sprayed using inert gases. Spray solutions may be directly inhaled from a spray device or the spray device may be connected to a mask or intermittent positive pressure respirator. Solutions, suspensions, or powder compositions may be administered orally or nasally, such as from a device that delivers the formulation in an appropriate manner.

[0554] In some embodiments, provided herein is a pharmaceutical composition for ophthalmic administration, comprising at least one compound as disclosed herein and at least one pharmaceutically acceptable excipient suitable for ophthalmic administration. Pharmaceutical compositions suitable for ocular administration may be presented as discrete dosage forms, such as drops or sprays each containing a predetermined amount of active ingredient, solutions or suspensions in aqueous or non-aqueous liquids, oil-in-water emulsions or water-in-oil liquid emulsions. Other forms of administration include intraocular injection, intravitreal injection, local or by using a drug eluting device, microcapsules, implants or microfluidic devices. In some cases, compounds as disclosed herein may be administered together with a carrier or excipient that increases intraocular penetration of the compound, such as colloidal particles having an oily core surrounded by an interfacial membrane and an aqueous emulsion. It is contemplated that all local approaches to the eye may be used, including topical, subconjunctival, periocular, posterior to the eyeball, subfascial, intracameral, intravitreal, intraocular, subretinal, near sclera and urea membrane administration. Systemic or parenteral administration may be feasible, including but not limited to intravenous, subcutaneous and oral delivery. Exemplary administration methods may be intravitreal or subfascial injection of solutions or suspensions, or intravitreal or subfascial placement of bioerodible or bioerodible devices, or by topical ocular administration of solutions or suspensions, or posterior juxtascleral application of gel or cream formulations.

[0555] Eye drops can be prepared by dissolving the active ingredient in a sterile aqueous solution (such as physiological saline, buffer solution, etc.) or by combining a powder composition to be dissolved before use. Other solvents as known in the art can be selected, including but not limited to: balanced salt solution, saline solution, water-soluble polyether (such as polyethylene glycol), polyethylene (such as polyvinyl alcohol and povidone), cellulose derivatives (such as methylcellulose and hydroxypropyl methylcellulose), petroleum derivatives (such as mineral oil and white vaseline), animal fat (such as lanolin), acrylic acid polymers (such as carboxypolyethylene gel), vegetable fat (such as peanut oil) and polysaccharides (such as dextran) and glycosaminoglycans (such as sodium hyaluronate). In some embodiments, additives commonly used in eye drops can be added. Such additives include isotonic agents (such as sodium chloride, etc.), buffers (such as boric acid, sodium monohydrogen phosphate, sodium dihydrogen phosphate, etc.), preservatives (such as benzalkonium chloride, benzethonium chloride, chlorobutanol, etc.), thickeners (such as sugars, such as lactose, mannitol, maltose, etc.; such as hyaluronic acid or its salts, such as sodium hyaluronate, potassium hyaluronate, etc.; such as mucopolysaccharides such as chondroitin sulfate, etc.; such as sodium polyacrylate, carboxyvinyl polymer, cross-linked polyacrylate, polyvinyl alcohol, polyvinyl pyrrolidone, methylcellulose, hydroxypropyl methylcellulose, hydroxyethyl cellulose, carboxymethyl cellulose, hydroxypropyl cellulose or other agents known to those skilled in the art).

[0556] In some cases, the colloidal particles include at least one cationic agent and at least one non-ionic surfactant, such as poloxamer, tyloxapol, polysorbate, polyoxyethylene castor oil derivative, sorbitan ester or polyoxy stearate. In some cases, the cationic agent can be selected from alkylamines, tertiary alkylamines, quaternary ammonium compounds, cationic lipids, amino alcohols, biguanide salts, cationic compounds or mixtures thereof. In some cases, the cationic agent can be a biguanide salt, such as chlorhexidine, polyaminopropyl biguanide, phenformin, alkyl biguanide or mixtures thereof. In some cases, the quaternary ammonium compound can be benzalkonium halide, lauralkonium halide, cetrimonium bromide, hexadecyltrimethylammonium halide, tetradecyltrimethylammonium halide, dodecyltrimethylammonium halide, cetrimonium halide, benzethonium halide, behenalkonium halide, cethylethyldimethylammonium halide, cetylpyridinium halide, benzododecinium halide, chiorallyl methenamine halide, rnyristylalkonium halide, stearalkonium halide, or a mixture of two or more thereof. In some cases, the cationic agent can be benzalkonium chloride, lauralkonium chloride, benzododecinium bromide, benzethonium chloride, hexadecyltrimethylammonium bromide, tridecyltrimethylammonium bromide, dodecyltrimethylammonium bromide, or a mixture of two or more thereof. In some cases, the oil phase can be mineral oil and light mineral oil, medium chain triglycerides (MCT), coconut oil; hydrogenated oils, including hydrogenated cottonseed oil, hydrogenated palm oil, hydrogenated castor oil or hydrogenated soybean oil; polyoxyethylene hydrogenated castor oil derivatives, including poluoxyl-40 hydrogenated castor oil, polyoxyl-60 hydrogenated castor oil or polyoxyl-100 hydrogenated castor oil.

[0557] In some embodiments, the amount of a compound as disclosed herein in a formulation may be from about 0.5% to about 20%, 0.5% to about 10%, or about 1.5% w / w.

[0558] In some embodiments, provided herein is a pharmaceutical composition for controlled release administration, comprising at least one compound as disclosed herein and at least one pharmaceutically acceptable excipient suitable for controlled release administration. In some embodiments, provided herein is a pharmaceutical composition for controlled release administration, comprising: (i) an effective amount of at least one compound disclosed herein; optionally (ii) an effective amount of at least one second agent; and (iii) at least one pharmaceutically acceptable excipient suitable for controlled release administration. In some embodiments, the pharmaceutical composition further comprises: (iv) an effective amount of at least one third agent.

[0559] Active agents such as the compounds provided herein can be administered by controlled release methods or by delivery devices, which are well known to those of ordinary skill in the art. Examples include, but are not limited to, U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123; and 4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; 5,639,480; 5,733,566; 5,739,108; 5,891,474; 5, 922,356; 5,972,891; 5,980,945; 5,993,855; 6,045,830; 6,087,324; 6,113,943; 6,197,350; 6,248,363; 6,264,970; 6,267,981; 6,376,461; 6,419,961; 6,589,548; 6,613,358; 6,699,500, each of which is incorporated herein by reference. Such dosage forms can be used to provide slow or controlled release of one or more active agents using, for example, hydroxypropylmethylcellulose in varying proportions, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or combinations thereof to provide the desired release profile. Suitable controlled release formulations known to those of ordinary skill in the art, including those described herein, can be readily selected for use with the active agents provided herein. Thus, the pharmaceutical compositions provided encompass single unit dosage forms suitable for oral administration, such as, but not limited to, tablets, capsules, caplets, and caplets suitable for controlled release.

[0560] All controlled release medicines have the common goal of improving drug therapy to be superior to drug therapy obtained by their uncontrolled counterparts. In some embodiments, the use of controlled release preparations in medical treatment may be characterized by using the least raw material drug to cure or control diseases, diseases or conditions in the least amount of time. The advantages of controlled release preparations include extended drug activity, reduced dosage frequency and increased subject compliance. In addition, controlled release preparations can be used to affect the onset of action or other characteristics, such as the blood level of the drug, and can therefore affect the occurrence of side (such as unfavorable) effects.

[0561] In some embodiments, the controlled release formulation is designed to initially release an amount of a compound as disclosed herein that can produce a therapeutic effect in a timely manner, and gradually and continuously release other amounts of the compound to maintain that level of therapeutic or preventive effect over an extended period of time. In order to maintain this constant level of compound in the body, the compound should be released from the dosage form at a rate that will replace the amount of drug metabolized and secreted away by the body. Controlled release of the active agent can be stimulated by various conditions including, but not limited to, pH, temperature, enzymes, water, or other physiological conditions or compounds.

[0562] In certain embodiments, the pharmaceutical composition may be administered using intravenous infusion, an injectable osmotic pump, a transdermal patch, a liposome, or other modes of administration. In some embodiments, a pump may be used (see, Sefton, CRC Crit. Ref Biomed. Eng. 14: 201 (1987); Buchwald et al., Surgery 88: 507 (1980); Sandek et al., N. Engl. J. Med. 321: 574 (1989)). In another embodiment, a polymeric material may be used. In yet another embodiment, a controlled release system may be placed at an appropriate site determined by a technician in the subject, i.e., only a portion of the systemic dose is required (see, e.g., Goodson, Medical Applications of Controlled Release, 115-138 (Volume 2, 1984). Other controlled release systems are described in Langer, Science 249:1527-1533 (1990). At least one active agent can be dispersed in a solid inner matrix, such as polymethyl methacrylate, polybutyl methacrylate, plasticized or unplasticized polyvinyl chloride, plasticized nylon, plasticized polyethylene terephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene-vinyl acetate copolymers, silicone rubber, polydimethylsiloxane, silicone carbonate copolymers, hydrophilic polymers (such as hydrogels of esters of acrylic acid and methacrylic acid), collagen, cross-linked polyvinyl alcohol and cross-linked partially hydrolyzed polyvinyl acetate (which is surrounded by an outer polymer membrane, such as polyethylene, polypropylene, ethylene / Propylene copolymers, ethylene / ethyl acrylate copolymers, ethylene / vinyl acetate copolymers, silicone rubber, polydimethylsiloxane, chloroprene rubber, chlorinated polyethylene, polyvinyl chloride, copolymers of vinyl chloride and vinyl acetate, vinylidene chloride, ethylene and propylene, ionomer polyethylene terephthalate, butyl rubber epichlorohydrin rubber, ethylene / vinyl alcohol copolymers, ethylene / vinyl acetate / vinyl alcohol terpolymers and ethylene / vinyloxyethanol copolymers (dissolved in body fluids). Then, in a release rate controlling step, at least one active agent diffuses through the outer polymer film. The percentage of at least one active agent in such parenteral compositions may depend on its specific properties, as well as the needs of the subject.

[0563] The compounds described herein can be delivered in the form of a pharmaceutically acceptable composition, the composition comprising a therapeutically effective amount of at least one compound disclosed herein and / or at least one additional therapeutic agent (such as a chemotherapeutic agent) formulated together with at least one pharmaceutically acceptable excipient. In some embodiments, only the compound provided herein without additional therapeutic agents may be included in the dosage form. In some cases, the compounds described herein and additional therapeutic agents are administered with separate compositions, and may be administered (such as because of different physical and / or chemical properties) by different routes (such as one therapeutic agent may be administered orally, and another may be administered intravenously). In other cases, the compounds described herein and additional therapeutic agents may be administered separately, but only by the same route (such as both are oral or both are intravenous). In other cases, the compounds described herein and additional therapeutic agents may be administered with the same pharmaceutical composition.

[0564] The selected dosage level will depend upon a variety of factors, including the activity of the specific compound employed, the severity of the condition, the route of administration, the time of administration, the rate of excretion or metabolism of the specific compound employed, the rate and extent of adsorption, the duration of the treatment, the administration of other drugs, compounds and / or substances used in combination with the specific compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0565] Dose levels can also be understood by in vitro or in vivo assays, which can be optionally used to help identify the optimal dose range. An effective dose criterion can be extrapolated from a dose response curve derived from an in vitro or animal model system. In addition, after being formulated with an appropriate pharmaceutically acceptable carrier in the desired dosage form, the compositions disclosed herein can be administered to humans or other animals rectally, parenterally, intracisternal, vaginally, intraperitoneally, topically (such as by transdermal patches, powders, ointments or drops), sublingually, buccally (as an oral or nasal spray), etc.

[0566] In general, a suitable daily dose of a compound described herein and / or a chemotherapeutic agent will be that amount of the compound which, in some embodiments, may be the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend on the factors described above. In some embodiments, the dosage of the compounds described herein for use in patients will be in the range of about 0.0001 mg to about 100 mg / day, or about 0.001 mg to about 100 mg / day, or about 0.01 mg to about 100 mg / day, or about 0.1 mg to about 100 mg / day, or about 0.1 mg to about 125 mg / day, or about 0.0001 mg to about 500 mg / day, or about 0.001 mg to about 500 mg / day, or about 0.01 mg to about 1000 mg / day, or about 0.01 mg to about 500 mg / day, or about 0.1 mg to about 500 mg / day, or about 1 mg to about 25 mg / day, or about 1 mg to about 50 mg / day, or about 5 mg to about 40 mg / day. Exemplary dosages may be about 10 to about 30 mg / day. In some embodiments, for 70kg people, suitable dosage will be about 0.05 to about 7g / day, such as about 0.05 to about 2g / day.In some embodiments, daily oral dosage is about 30mg, about 90mg, about 150mg or about 180mg.As used herein, "about" means to modify the value of ± 5%.The actual dosage level of the active ingredient in the pharmaceutical composition described herein can be changed to obtain the amount of active ingredient that effectively realizes the desired therapeutic response for a specific patient, composition and mode of administration and is non-toxic to the patient.In some cases, the dosage level below the lower limit of the aforementioned range may be high enough, and in other cases, a larger dosage can be used without causing any harmful side effects, such as by dividing a larger dosage of magnetic force into several small doses for use all day long.

[0567] In some embodiments, the compound may be administered once a day, once every other day, three times a week, twice a week, once a week, twice every two weeks, or another intermittent schedule. The dosing schedule may include a "drug holiday," i.e., the drug may be administered for two weeks on, one week off, or three weeks on, one week on, or four weeks on, one week off, etc., or may be administered continuously without a drug holiday.

[0568] In some embodiments, a compound as provided herein can be administered at multiple doses. Administration may be about once, twice, three times, four times, five times, six times or more than six times a day. Administration may be about once a month, about once every two weeks, about once a week or about once every other day. In another embodiment, a compound as disclosed herein and another agent are administered together about once a day to about 6 times a day. For example, the compound may be administered once or more a day on a weekly basis (such as every Monday), indefinitely or for a period of several weeks (such as 4-10 weeks). Alternatively, it may be administered daily for a period of several days (such as 2-10 days), and then the compound may not be administered for a period of several days (such as 1-30 days), and this cycle may be repeated indefinitely or for a given number of repetitions, such as 4-10 cycles. As an example, a compound provided herein may be administered once a day for 5 days, then stopped for 9 days, then administered once a day for another 5 days, then stopped for 9 days, and so on, and the cycle may be repeated indefinitely or for a total of 4-10 times. In another embodiment, the administration of the compounds and agents as provided herein lasts for less than about 7 days. In yet another embodiment, the administration lasts for more than about 6, about 10, about 14, about 28 days, about two months, about six months, or about one year. In some cases, continuous administration can be achieved and maintained as long as necessary.

[0569] As long as necessary, the administration of the pharmaceutical composition as disclosed herein can continue. In some embodiments, the medicament as disclosed herein can be administered for more than about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 14, or about 28 days. In some embodiments, the medicament as disclosed herein can be used for less than about 28, about 14, about 7, about 6, about 5, about 4, about 3, about 2, or about 1 day. In some embodiments, the medicament as disclosed herein can be administered on an ongoing basis for a long time, such as for the treatment of chronic effects.

[0570] When used for treatment or known specific disease states or conditions, the effective dose of a compound as disclosed herein may vary according to the specific compound used, the mode of administration, the condition being treated and its severity, and various natural factors associated with the individual being treated. In some embodiments, the effective systemic dose of the compound will generally be in the following range: about 0.01 to about 500 mg compound / patient body weight kg, such as about 0.1 to about 125 mg / kg, and in some cases about 1 to about 25 mg / kg, administered in single doses or multiple doses. The expected daily dose is expected to change with the route of administration. Therefore, parenteral administration will generally be at a level of about 10% to about 20% of the oral administration level. Typically, the compound may be administered to patients in need of such treatment with a daily dose range of about 50 to about 2000 mg / patient. Administration may be daily, weekly (or at other multi-day intervals) or one or more times in an intermittent regimen.

[0571] In some embodiments, the dose of a compound as disclosed herein can be selected from 30, 60, 90, 120, 180, and 240 mg administered orally once daily. Another dosing regimen may include 90 mg administered orally once daily, or a 90 mg dose administered orally daily for 7 days, followed by a 180 mg dose daily. In some embodiments, the compound to be administered is brigatinib Form A.

[0572] Because the compounds described herein can be administered in combination with other treatments (such as additional chemotherapeutic agents, radiation or surgery), the dose of each agent or therapy may be lower than the corresponding dose of a single-dose therapy. The dose of a single-dose therapy may be in the range of, for example, about 0.0001 to about 200 mg, or about 0.001 to about 100 mg, or about 0.01 to about 100 mg, or about 0.1 to about 100 mg, or about 1 to about 50 mg / kg body weight / day.

[0573] When the compounds provided herein are administered in a pharmaceutical composition comprising one or more agents and the one or more agents have a shorter half-life than the compounds provided herein, the unit dosage forms of the agents and compounds provided herein may be adjusted accordingly.

[0574] In some embodiments, kits are provided herein. Kits may include compounds or pharmaceutical compositions as described herein and written materials in suitable packaging, which may include instructions for use, discussion of clinical studies, and lists of side effects, etc. Kits are well suited for delivering solid oral dosage forms such as tablets or capsules. Such kits may also include information, such as scientific literature references, package insert materials, clinical trial results, and / or an overview of these and similar information, which indicates or confirms the activity and / or advantages of the pharmaceutical composition and / or its description of administration, application, side effects, drug interactions, or other information useful to medical care providers. Such information may be based on the results of various studies, such as studies using experimental animals involving in vivo models and studies based on human clinical trials.

[0575] In some embodiments, a memory aid may be provided with the kit, such as in the form of numbers adjacent to the tablets or capsules, whereby the numbers correspond to the days of the regimen on which the tablets or capsules so designated should be ingested. Another example of such a memory aid may be a calendar printed on a card, such as the following "Week 1, Monday, Tuesday... etc....Week 2, Monday, Tuesday..." etc. Other variations of memory aids will be apparent. A "daily dose" may be a single tablet or capsule or several tablets or capsules to be taken on a given day.

[0576] The kit may also contain another medicament. In some embodiments, the compounds and medicaments disclosed herein are provided as separate pharmaceutical compositions in separate containers in separate kits. In some embodiments, the compounds and medicaments disclosed herein are provided as single pharmaceutical compositions in containers in the kit. Suitable packaging and additional available articles (e.g., measuring cups for liquid preparation, foil packaging to minimize exposure to air, etc.) are known in the art and may be included in the kit. In other embodiments, the kit may also include a device for applying an active agent. Examples of such devices include, but are not limited to, syringes, drip bags, patches, and inhalers. The kits described herein may be provided, sold, and / or promoted to health providers, including physicians, nurses, pharmacists, and prescribers, etc. In some embodiments, the kit may also be sold directly to consumers.

[0577] The example of such a kit is a so-called blister pack. Blister packs are well known in the packaging industry and are widely used for packaging drug unit dosage forms (tablets, capsules, etc.). Blister packs are generally composed of a thin sheet of relatively hard material covered by a usually transparent plastic material foil. During the packaging process, a notch is formed in the plastic foil. The notch has the size and shape of the tablet or capsule to be packaged. Next, the tablet or capsule is placed in the notch and the plastic foil is sealed with a thin sheet of relatively hard material on one side of the foil opposite to the direction in which the notch is formed. As a result, the tablet or capsule is sealed in the notch between the plastic foil and the thin sheet. The strength of the thin sheet should be such that a tablet or capsule can be removed from the blister pack by manually applying pressure to the notch, thereby forming an opening at the notch position in the thin sheet. The tablet or capsule can then be removed via the opening.

[0578] The kit may also include a pharmaceutically acceptable solvent for administering one or more active agents. For example, if the active agent is provided in a solid form that must be reconstituted for parenteral administration, the kit may include a sealed container suitable for the solvent, wherein the active agent can be dissolved to form a particle-free sterile solution suitable for parenteral administration. Examples of pharmaceutically acceptable solvents include, but are not limited to: USP Water for Injection; aqueous solvents such as, but not limited to, Sodium Chloride Injection, Ringer's Injection, Dextrose Injection, Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection; water-miscible solvents such as, but not limited to, ethyl alcohol, polyethylene glycol, and polypropylene glycol; and non-aqueous solvents such as, but not limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

[0579] The present disclosure also encompasses anhydrous pharmaceutical compositions and dosage forms comprising active ingredients, since water can promote degradation of some compounds. For example, water (e.g., about 5%) in the pharmaceutical field can be added as a means of simulating long-term storage to determine characteristics such as the shelf life or stability of a preparation over time. Anhydrous pharmaceutical compositions and dosage forms can be prepared using anhydrous or low-moisture ingredients and low-moisture or low-humidity conditions. For example, pharmaceutical compositions and dosage forms containing lactose can be prepared as anhydrous if they are in contact with moisture and / or humidity in large quantities during manufacture, packaging, and / or storage, and / or are expected to be stored. Anhydrous pharmaceutical compositions can be prepared and stored so as to maintain their anhydrous properties. Therefore, anhydrous pharmaceutical compositions can be packaged using known materials to prevent exposure to water so that they can be included in suitable preparation kits. Examples of suitable packaging include, but are not limited to, airtight sealed foils, plastics, etc., unit dose containers, blister packages, and strip packages.

[0580] IV. Treatment Methods

[0581] In some embodiments, a pharmaceutical composition comprising at least one crystalline form of brigatinib can be used to treat cancer by administering a therapeutically effective amount of the pharmaceutical composition to a subject in need thereof. In some embodiments, the cancer is an ALK+-driven cancer. In some embodiments, the cancer is non-small cell lung cancer.

[0582] A "therapeutically effective amount" is an amount effective to detect killing or inhibiting the growth or spread of cancer cells; the size or number of weight; or other measures of the level, stage, progression or severity of cancer. The exact amount required may vary from subject to subject, depending on the race, age and general health of the subject, the severity of the disease, the specific anticancer agent, its mode of administration, combination therapy with other therapies, etc.

[0583] Disclosed herein are compounds having biological properties that make them targets for treating or regulating diseases that may involve kinases, symptoms of such diseases, or the effects of other physiological events mediated by kinases. For example, various compounds as disclosed herein have been shown to inhibit the tyrosine kinase activity of ALK, fak, and c-met, particularly tyrosine kinases believed to mediate the growth, development, and / or metastasis of cancer. It has also been found that various compounds as disclosed herein have effective in vitro activity against cancer cell lines, particularly including karpas 299 cells. Therefore, such compounds are targets for treating cancers (including solid tumors as well as lymphomas and including cancers resistant to other therapies).

[0584] In some embodiments, the cancer is an ALK+-driven cancer. In some embodiments, the cancer is non-small cell lung cancer. In some embodiments, the cancer is ALK-positive NSCLC. In some embodiments, the cancer is locally advanced or metastatic ALK-positive NSCLC. In some embodiments, the cancer / patient has been previously treated with crizotinib or another tyrosine kinase inhibitor. In some embodiments, the cancer / patient has not been previously treated with an ALK inhibitor.

[0585] Such cancers include, but are not limited to, breast cancer, non-small cell lung cancer (NSCLC), neural tumors such as glioblastoma and neuroblastoma; esophageal cancer, soft tissue cancers such as rhabdomyosarcoma, etc.; various forms of lymphoma, such as non-Hodgkin's lymphoma (NHL) known as anaplastic large cell lymphoma (ALCL), various forms of leukemia; and including cancers that are ALK or c-met mediated.

[0586] Anaplastic lymphoma kinase (ALK) is a transcellular membrane receptor tyrosine kinase that belongs to the insulin receptor subfamily. ALK receptor tyrosine kinase (RTK) was initially identified due to its involvement in a subtype of human non-Hodgkin's lymphoma known as anaplastic large cell lymphoma (ALCL). ALK generally has a restricted distribution in mammalian cells and is found to be only at significant levels in the nervous system during embryonic development, indicating a role for ALK in brain development (Duyster, J. et al., Oncogene, 2001, 20, 5623-5637).

[0587] In addition to its role in normal development, expression of full-length normal ALK has been detected in cell lines derived from a variety of tumors, such as neuroblastoma, neuroectodermal tumors (Lamant L. et al., Am. J. Pathol., 2000, 156, 1711-1721; Osajima-Hakomori Y., et al., Am. J. Pathol. 2005, 167, 213-222) and glioblastoma (Powers C. et al., J. Biol. Chem. 2002, 277, 14153-14158; Grzelinski M. et al., Int. J. Cancer, 2005, 117, 942-951; Mentlein, R. et al., J. Neurochem., 2002, 83, 747-753) as well as breast cancer and melanoma lines (Dirk WG. et al., Int. J. Cancer, 2002, 100, 49-56).

[0588] As with other RTKs, translocations affect the ALK gene, resulting in the expression of an oncogenic fusion kinase, the most common of which is NPM-ALK. For example, approximately 60 percent of anaplastic large cell lymphomas (ALCL) are associated with chromosomal mutations that produce a fusion protein consisting of nucleolar phosphatidylcholine (NMP) and the intracellular domain of ALK. (Armitage, JO et al., Cancer: Principle and Practice of Oncology, 6th Edition 2001, 2256-2316; Kutok, JL & Aster JC, J. Clin. Oncol., 2002, 20, 3691-3702; Wan, W. et al., Blood, 2006, 107, 1617-1623. This mutant protein, NPM-ALK, has a constitutively active tyrosine kinase domain, which is responsible for its oncogenic properties by activating downstream effectors (Falini, B et al., Blood, 1999, 94, 3509-3515; Morris, SW et al., Brit. J. Haematol., 2001, 113, 275-295). Experimental data have demonstrated that abnormal expression of constitutively active ALK is directly involved in the pathogenesis of ALCL and that such inhibition of ALK can significantly impede the growth of ALK-positive lymphoma cells (Kuefer, Mu et al., Blood, 1997, 90, 2901 -2910; Bai, RY et al., Exp. Hematol., 2001, 29, 1082-1090; Slupianek, A. et al., Cancer Res., 2001, 61, 2194-2199; Turturro, F. et al., Clin. Cancer. Res., 2002, 8, 240-245). Constitutively activated chimeric ALK has been shown to be present in approximately 60% of inflammatory myofibroblastic tumors (IMT), a slow-growing sarcoma that primarily affects children and young adults (Lawrence, B. et al., Am. J. Pathol., 2000, 157, 377-384). In addition, current reports have described the presence of a variant ALK fusion, TPM4-ALK, in the setting of squamous cell carcinoma (SCC) of the esophagus (Jazzi et al., 2001). fr., et al., World J. Gastroenterol., 2006, 12, 7104-7112; Du X., et al., J. Mol. Med., 2007, 85, 863-875; Aklilu M., Semin. Radiat. Oncol., 2007, 17, 62-69). Thus, ALK is one of the few examples of RTKs involved in neoplasia in both non-hematopoietic and hematopoietic malignancies.Recently, it has been shown that inversion within chromosome 2p leads to the formation of a fusion gene containing a portion of the echinoderm microtubule-associated protein-like 4 (EML4) gene and the anaplastic lymphoma kinase (ALK) gene in non-small cell lung cancer (NSCLC) cells (Soda M., et al., Nature, 2007, 448, 561-567).

[0589] In some embodiments, ALK inhibitors can create durable cures when used as monotherapy or in combination with current chemotherapies for ALCL, IMT, proliferative disorders, glioblastoma, and possibly other solid tumors cited herein, or can be used as monotherapy in a maintenance role to prevent relapse in patients in need of such treatment.

[0590] Compounds as disclosed herein can be administered as part of a treatment regimen in which the compound is the sole active agent, or in combination with one or more other therapeutic agents as part of a combination therapy. When administered as a component of a combination therapy, the therapeutic agents being administered can be formulated as separate compositions that are administered simultaneously or sequentially at different time points (e.g., within 72 hours, 48 ​​hours, or 24 hours of each other), or the therapeutic agents can be formulated together as a single pharmaceutical composition and administered simultaneously.

[0591] Therefore, administration of brigatinib in a form disclosed herein can be combined with at least one additional therapeutic agent known to those skilled in the art for the prevention or treatment of cancer (such as radiotherapy or cytostatics, cytotoxic agents, other anticancer agents, and other drugs) to alleviate the symptoms of cancer or the side effects of any drug. Non-limiting examples of additional therapeutic agents include agents suitable for immunotherapy (such as, for example, PD-1 and PDL-1 inhibitors), anti-angiogenic agents (such as, for example, bevacizumab), and / or chemotherapeutic agents.

[0592] If formulated as a fixed dose, such combination products employ the compounds disclosed herein within an acceptable dosage range. When the combination formulation is appropriate, the compounds disclosed herein may be administered sequentially with other anticancer agents or cytotoxic agents. The compounds disclosed herein may be administered before, simultaneously with, or after the administration of other anticancer agents or cytotoxic agents.

[0593] Currently, standard treatment for primary tumors consists of surgical resection followed by radiation or chemotherapy when appropriate, and is usually administered intravenously (IV). Common chemotherapy regimens consist of DNA alkylating agents, DNA intercalators, CDK inhibitors, or microtubule poisons. The chemotherapy doses used are just below the maximum tolerated dose, and therefore dose-limiting toxicities typically include nausea, vomiting, diarrhea, hair loss, neutropenia, etc.

[0594] There are a large number of antineoplastic agents available for commercial use, clinical evaluation and preclinical development, which can be selected for the treatment of cancer by combination drug chemotherapy. And there are several major categories of such antineoplastic agents, namely antibiotic-type agents, alkylating agents, antimetabolites, antihormonal agents, immune agents, interferon-type agents and a class of miscellaneous agents (miscellaneous agents).

[0595] A first class of antineoplastic agents that may be used in combination with the compounds disclosed herein includes antimetabolite / thymidylate synthase inhibitor antineoplastic agents. Suitable antimetabolite antineoplastic drugs can be selected from, but are not limited to, 5-FU-fibrinogen, acanthoic acid, aminothiadiazole, buquina sodium, carmofur, CibaGeigy CGP-30694, cyclopentylcytosine, cytarabine phosphate stearate, cytarabine conjugate, Lilly DATHF, Merrel Dow DDFC, dezaguanine, dideoxycytidine, dideoxyguanosine, 3,4-dihydroxybenzohydroxamic acid (didox), Yoshitomi DMDC, doxifluridine, Wellcome EHNA, Merck & Co., EX-015, fazarabine, floxuridine, fludarabine phosphate, 5-fluorouracil, N-(21-furanyl)fluorouracil, Daiichi Seiyaku FO-152, isopropylpyrrolizine, Lilly LY-188011, Lilly LY-264618, methobenzaprim, methotrexate, Wellcome MZPES, norspermidine, NCI NSC-127716, NCI NSC-264880, NCI NSC-39661, NCI NSC-612567, Warner-Lambert PALA, pentostatin, pirtrexine, plicamycin, Asahi Chemical PL-AC, Takeda TAC788, thioguanine, thiazofurine, Erbamont TIF, trimetrexate, tyrosine kinase inhibitors, Taiho UFT, and uricytin.

[0596] A second class of antineoplastic agents that can be used in combination with the compounds as disclosed herein consists of alkylating antineoplastic agents. Suitable alkylating antineoplastic drugs can be selected from, but are not limited to, Shionogi 254-S, aldophosphamide analogs, hexamethylmelamine, anaximone, Boehringer Mannheim BBR-2207, betabucil, buducin, Wakunaga CA-102, carboplatin, carmustine, Chinoin-139, Chinoin-153, chlorambucil, cisplatin, cyclophosphamide, American Cyanamid CL-286558, Sanofi CY-233, cyplatate, Degussa D 384, Sumimoto DACHP (Myr) 2, diphenylspiromustine, platinum cell inhibitors, Erba distamycin derivatives, Chugai DWA-2114R, ITI E09, emustine, Erbamont FCE-24517, estramustine sodium phosphate, fotemustine, Unimed GM, Chinoin GYKI-17230, heptylsulfamide, ifosfamide, iproplatin, lomustine, mafosfamide, mitolactolide, Nippon Kayaku NK-121, NCINSC-264395, NCI NSC-342215, oxaliplatin, Upjohn PCNU, prednimustine, Proter PTT-119, ranimustine, semustine, SmithKline SK&F-101772, Yakult Honsha SN-22, spiromustine, TanabeSeiyaku TA-077, tauromustine, temozolomide, teroxirone, tetraplatin, and trimethoprim-sulfamethoxazole.

[0597] The third class of antineoplastic agents that can be used in combination with the compounds disclosed herein include antibiotic antineoplastic agents. Suitable antibiotic antineoplastic agents can be selected from, but are not limited to, Taiho 4181-A, aclarubicin, actinomycin D, actinoplanone, Erbamont ADR-456, aeroplysinin derivatives, Ajinomoto AN II, Ajinomoto AN3, Nippon Soda anisomycin, anthracycline, azinomycin-A, bisucaberin, Bristol-Myers BL-6859, Bristol-Myers BMY-25067, Bristol-Myers BNY-25551, Bristol-Myers BNY-26605IBristolMyers BNY-27557, Bristol-Myers BMY-28438, bleomycin sulfate, bryostatin-1, Taiho C-1027, calichemycin, chromothiocin, dactinomycin, daunorubicin, Kyowa Hakko DC-102, Kyowa Hakko DC-79, Kyowa Hakko DC-88A, Kyowa Hakko, DC89-Al, Kyowa Hakko DC92-B, anthracycline B, Shionogi DOB-41, doxorubicin, doxorubicin-fibrinogen, esacycline-A, epirubicin, erbstatin, esorubicin, esperamicin-Al, esperamicin-Alb, Erbamont FCE21954, Fujisawa FK-973, fostricin, Fujisawa FR-900482, balanoposthicin, gregadine-A, grecomycin, herbimycin, idarubicin, edarubicin, total mycin, kinasapurpurin, KyowaHakko KM-5539, Kirin Brewery KRN-8602, Kyowa Hakko KT-5432, Kyowa Hakko KT-5594, Kyowa Hakko KT-6149, American Cyanamid LL-D49194, Meiji Seika ME 2303, menolipin, mitomycin, mitoxantrone, SmithKline M-TAG, neomycin, Nippon Kayaku NK-313, NipponKayaku NKT-01, SRI InternationalNSC-357704, lysozyme, oxamycin, peclomycin, pyridin, pirarubicin, porothramycin, pyridanicin A, Tobishi RA-I, rapamycin, lisocin, rhodorubicin, sibamicin, svenmycin, Sumitomo SM5887, Snow Brand SN-706, Snow Brand SN-07, sorangicin-A, sparsemycin, SS Pharmaceutical SS-21020, SS Pharmaceutical SS-7313B, SS Pharmaceutical SS-9816B, stilbene B, Taiho 4181-2, talithromycin, TakedaTAN-868A, terpentecin, thrazine, tricrozarin A, UpjohnU-73975, Kyowa Hakko UCN-10028A, Fujisawa WF-3405, Yoshitomi Y-25024, and Zorubicin.

[0598] A fourth class of antineoplastic agents that can be used in combination with the compounds disclosed herein includes miscellaneous antineoplastic agents such as tubulin interacting agents, topoisomerase II inhibitors, topoisomerase I inhibitors and hormonal agents selected from, but not limited to, xcarotene, X-difluoromethyl-arginine, acitretin, Biotec AD-5, Kyorin AHC-52, chishanine, aminonafide, amphetamine, amsacrine, Angiostat, ankinomycin, anti-neoplastic A10, antineoplastic A2, antineoplastic A3, antineoplastic A5, antineoplastic AS2-1F Henkel APD, glycine aphidicolin, asparaginase, Avarol, yancitracin, batracylin, benfluran, benzterpene, Ipsen-Beaufour BIM-23015, bisantrene, BrisoMyers BNY-40481, Vestar boron-10, bromoifosfamide, Wellcome BW-502, Wellcome BW-773, caramelamide, carmezol hydrochloride, Ajinomoto CDAF, chlorothioquinolone, Chemes CHX-2053, Chemex CHX-100, Warner-Lambert CI-921, Warner-Lambert CI-937, Warner-Lambert CI-941, Warner-Lambert CI958, clafluanid, clafadenone, ICN compound 1259, ICN compound 4711, Contracan, Yakult Honsha CPT-11, clinato, curaderm, cytochalasin B. cytarabine, cyclosporine, Merz D-609, DABIS maleate, dacarbazine, datertinib, mescitin-B, dihematoporphyrin ester, dihydrolanperone, dinaline, distamycin, Toyo Pharmar DM-341, Toyo Pharmar DM-75, Daiichi Seiyaku DN-9693, docetaxel elipribine, elliptinium acetate, Tsumura EPMTC, ibosetone, ergotamine, etoposide, etretinate, fenretinide, Fujisawa FR-57704t gallium nitrate, daphne daphne, Chugai GLA-43, Glaxo GR-63178, maitake polysaccharide NMF5N, hexadecylphosphocholine, Green Cross HO-221, homoharringtonine, hydroxyurea, BTG ICRF-187, imofosine, isoglutamine, isotretinoin, Otsuka JI-36, Ramot K-477, OtsuakK-76COONa, Kureha Chemical K-AM, MECT Corp KI-8110, American Cyanamid L-623, leukomodulin, lonidamine, Lundbeck LU 1121 Lilly LY-186641, NCI (US) MAP, Marisin, Merrel Dow MDL-27048, Medco MEDR-340, mebaron, merocyanine derivatives, methylanilinoacridine, Molecular Genetics MGI136, minactivin, mitonadine, mitolaxanthine, mopidal, momiamine, Zenyaku Kogyo MST-16, N-(retinoyl) amino acids, Nisshin Flour Milling N-021, N-acetylated-dehydroalanine, naphthoxazolone, Taisho NCU-190, nocodazole derivatives, Normosang, NCI NSC-145813, NCI NSC-361456, NCI NSC-604782, NCI NSC-95580, ocreotide, Ono ONO-112, quinocin, Akzo Org-10172, paclitaxel, pancratistatin, palzeptin, WarnerLambert PD-111707, Warner-Lambert PD-115934, Warner-Lambert PD-131141, Pierre Fabre PE-1001, ICRT peptide D, pyrroloanthrone, polyhematoporphyrin, polyprotonic acid, Efamol porphyrin, promazine, procarbazine, proglumide, Invitron protease ligase I, Tobishi RA-700, razoxane, Sapporo Breweries RBS, Restrictin-P, Retilipin, Retinoic acid, Rhone-Poulenc RP-49532, Rhone-Poulenc RP-56976, SmithKline SK&F-104864, Sumitomo SM-108, Kuraray SMANCS, SeaPharm SP10094, Furoglossol, Spirocyclopropane derivatives, Spirogermaniumamine, Unimed, SS Pharmaceutical SS-554, Stypoldione, SuntorySUN 0237, Suntory SUN 2071, Superoxide dismutase, Toyama T-506, Toyama T-680, Taxol, Teijin TEI-0303, Teniposide, Tyleratin, Eastman KodakTJB-29, tocotrienol, topotecan, Topostin, Teijin TT82, Kyowa Hakko UCN-01, Kyowa Hakko UCN-1028, ukrain, Eastman Kodak USB-006, vinblastine sulfate, vincristine, vindesine, vinstrexamide, vinorelbine, vinstrol, vinorelbine, withanolide, and Yamanouchi YM.

[0599] Alternatively, the present compounds can also be used in co-therapy with other antineoplastic agents, such as acetomorphan, aclarubicin, aldesleukin, alemtuzumab, alitretinoin, hexamethylmelamine, amifostine, aminolevulinic acid, amrubicin, amsacrine, anagrelide, anastrozole, ANCER, anacetamipril, ARGLABIN, arsenic trioxide, BAM 002 (Novelos), bexarotene, bicalutamide, bromuridine, capecitabine, cymoleukin, cetrorelix, cladribine, clotrimazole, cytarabine phosphodiesterone, DA3030 (Dong-A), daclizumab, denileukin (diftitox), deslorelin, dexrazoxane, delazosine, docetaxel, docosyl, doxorubicin, doxorubicin, bromocriptine, carmustine, cytarabine, fluorouracil, HIT diclofenac, Interferon α, daunorubicin, doxorubicin, tretinoin, edilfosine, edrecolomab, eflornithine, ethiprof, epirubicin, epoetin β, etoposide phosphate, exemestane, exixulin, fadroxil, filgrastim, finasteride, fludarabine phosphate, formestane, fotemustine, gallium nitrate, gemcitabine, gemtuzumab ozogamicin, gemiracil / otrazil / tegafur combination, glucocortin, goserelin, heptaplatin, human chorionic gonadotropin (chorionic gonadotropin) gonadotropin), human fetal alpha-fetoglobulin, ibandronate, idarubicin, (imiquimod, interferon alpha, natural, interferon alpha, interferon alpha-2, interferon alpha-2a, interferon alpha-2b, interferon alpha-NI, interferon alpha-n3, interferon alfacon1, natural, interferon alpha, interferon beta, interferon beta-la, interferon beta-lb, interferon gamma, natural interferon gamma-la, interferon gamma-lb, interleukin-I beta, iobenzylguanidine, irinotecan, irisogladine, lanreotide, LC 9018 (Yakult), leflunomide, levofloxacin, lentinan sulfate, letrozole, interferon alpha, leuprolide, levamisole + fluorouracil, lirazo, lomoplatin, lonidamine, lovastatin, masoprofen, melarsoprol, metoclopramide, mifepristone, miltefosine, milistostatin, mismatched double-stranded RNA, mitoguanidine, dibromodulanol, mitoxantrone, morasporastatin, nafarelin, naloxone + pentazocine, nartograstim, nedaplatin, nilutamide, noscapine, new erythropoiesis stimulating protein, NSC 631570 Octreotide, opreleukin, osatetron, oxaliplatin, paclitaxel, pamidronate, pegaspargase, peginterferon α-2b, sodium polysulfide, pentahistine, sapellose, pirarubicin, rabbit anti-thymocyte polyclonal antibody, peginterferon α-2a, porphyrin, raloxifene, raltitrexed, rasburicase, etidronate rhenium Re186, RII vitamine, rituximab, romotide, samarium (153Sm) lexidronan, sargramostim, sizoran, sobuzosen, sonarmin, strontium-89 chloride, suramin, tasonamine, tazarotene, tegafur, temoporfin, temozolomide, tenipoxib, tetrachloride decaoxide, thalidomide, thymofasin, thyrotropin alfa, topotecan, toremifene, tositumomab-iodine 131, trastuzumab, treoxazole, tretinoin, trilostane, trimetrexate, triptorelin, natural tumor necrosis factor alpha, ubenimex, bladder cancer vaccine, Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin, vinorelbine, VIRULIZIN, nitrosamine, timastatin, timarubicin, or zoledronic acid; abarelix; AE 941 (Aeterna), amoxicillin, antisense oligonucleotides, bcl-2 (Genta), APC 8015 (Dendreon), cetuximab, decitabine, desaminoglucose, diazocone, EL 532 (Elan), EM 800 (Endorecherche), eniluracil, etanidazole, fenretinide I filgrastim SD01 (Amgen), fulvestrant, galocibine, gastrin 17 immunogen, HLA-B7 gene therapy (Vical), granulocyte macrophage colony stimulating factor, histamine dihydrochloride, tentan, isobetaine, ilomastat, IM 862 (Cytran), interleukin ipramoxifene, LDI 200 (Milkhaus), lelipasin, lintuzumab, CA125 MAb (Biomira), cancer MAb (Japan Pharmaceutical Development), HER-2 and Fc MAb (Medarex), idiotype 105AD7 MAb (CRC Technology), idiotype CEAMAb (Trilex), LYM iodine 131 MAb (Techniclone), polymorphic epithelial mucin-yttrium 90 MAb (Antisoma), marimastat, menolipir, mitumomab, motexafin, gadolinium, MX 6 (Galderma), nelarabine, nolatrexed, P 30 protein, pegvisomant, pemetrexed, porfibrinostat, prinomastat, RL 0903 (Shire), rubitecan, satraplatin, sodium phenylacetate, fospartate, SRL 172 (SR Pharma), SU 5416 (SUGEN) ySU 6668 (SUGEN), TA 077 (Tanabe), tetrathiomolybdate, thaliblastine, thrombopoietin, tin ethyl ester of rhodamine, tirapazamine, cancer vaccine (Biomira), melanoma vaccine (New York University), melanoma vaccine (Sloan KetteringInstitute), melanoma tumor lysate vaccine (New York Medical College), viral melanoma cell lysate vaccine (Royal Newcastle Hospital), or valspodar.

[0600] V. Synthesis of Brigatinib Form A

[0601] The following representative synthesis of Brigatinib Form A contains additional information, exemplification, and guidance that may be applicable to the practice of the present invention in its various embodiments and equivalent forms thereof.

[0602] The examples are intended to help illustrate the present invention and are not intended nor should they be construed as limiting the scope of the present invention. Indeed, various modifications of the present invention and many other embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art after reviewing this document (including the examples that follow) and referring to the optional and patent literature cited herein.

[0603] The contents of those cited references are incorporated herein by reference to help illustrate the state of the art. In addition, for purposes of the present invention, chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th edition, inside cover. In addition, general principles of organic chemistry as well as specific functional moieties and reactivities are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999 and "Organic Chemistry", Morrison & Boyd (3rd edition), both of which are incorporated herein by reference in their entirety.

[0604] Step 1: (2-aminophenyl)dimethylphosphine oxide

[0605]

[0606] A mixture of 2-iodoaniline (86 g, 0.393 mol, 1.0 equivalent), dimethylphosphine oxide (36.4 g, 0.466 mol, 1.19 equivalent), potassium phosphate (92.4 g, 0.423 mol, 1.1 equivalent), palladium acetate (II) (4.56 g, 0.02 mol, 0.05 equivalent) and Xantphos (11.6 g, 0.02 mol, 0.05 equivalent) in DMF (700 mL) was stirred at about 120 ° C for about 6 h. The color of the mixture became dark brown. After cooling to room temperature, diatomaceous earth (30 g) was added to the mixture. The mixture was then filtered and the filter pad was rinsed with EtOAc (2 x 250 mL). The filtrate was then concentrated in vacuo to obtain a residue.

[0607] A second batch of (2-aminophenyl)dimethylphosphine oxide was synthesized as above on the same scale and the residues from both batches were combined and subjected to purification as discussed below.

[0608] EtOAc (1L) was added to the combined residue, and the resulting mixture was stirred at room temperature for about 1h. The mixture was filtered and the collected residue was washed with EtOAc (2x 250mL). The combined filtrate was dried over sodium sulfate, filtered and concentrated in vacuo to obtain an oil. The obtained oil was dissolved in a mixture of water / concentrated hydrochloric acid (1.2L / 300mL) at room temperature under stirring, and stirred for 30min. The resulting mixture was filtered, and the collected residue was washed with aqueous hydrochloric acid solution (10%, 300mL). The combined aqueous filtrate was washed with EtOAc (2x 1L washings, then 500mL washings). The water layer was cooled in an ice bath (less than 10°C internal mixture temperature), and the pH of the solution was adjusted to about 12 (as measured by pH paper) by adding aqueous sodium hydroxide solution (30% w / w), while maintaining the internal solution temperature less than 20°C during the entire addition. The resulting solution was extracted with IPA / DCM (1 / 3 v / v, 4 x 1 L), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to give a viscous oil which crystallized upon standing at room temperature. The resulting solid was triturated with EtOAc / heptane (1 / 10 v / v, 2 x 150 mL) to give (2-aminophenyl)dimethylphosphine oxide as a light brown solid.

[0609] Step 2: (2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide

[0610]

[0611] 2,4,5-trichloropyrimidine (54.2 g, 0.296 mol, 1.0 equiv), (2-aminophenyl) dimethyl-phosphine oxide (50.0 g, 0.296 mole, 1.0 equiv), potassium carbonate (49.1 g, 0.355 mol, 1.2 equiv) and tetrabutylammonium hydrogen sulfate (10.2 g, 0.03 mole, 0.1 equiv) were combined in DMF (1050 mL) and heated at 65 ° C for about 8.0-8.5 h. During the heating process, an off-white suspension formed. After cooling, the mixture was cooled to room temperature and filtered. The collected solid was rinsed with DMF (2x50 mL), and the combined filtrate was concentrated in vacuo. The resulting residue was dissolved in EtOAc (1.3 L) and water (350 mL). The aqueous layer was separated and extracted with EtOAc (2x 250 mL). The combined organic layers were washed with brine (20% w / w, 500 mL), dried over sodium sulfate and concentrated in vacuo to give (2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide as an off-white solid.

[0612] Alternative Synthesis of (2-((2,5-Dichloropyrimidin-4-yl)amino)phenyl)-dimethylphosphine oxide

[0613] (2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide can be synthesized according to the previous procedure using the conditions in Table 28.

[0614] Table 28. Reaction conditions for the synthesis of (2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide

[0615]

[0616]

[0617] Step 3: 1-(1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)-4-methylpiperazine

[0618]

[0619] A mixture of 5-fluoro-2-nitroanisole (85.6 g, 0.5 mol, 1.0 equiv), 1-methyl-4-(piperidin-4-yl)piperazine (91.7 g, 0.5 mol, 1.0 equiv) and potassium carbonate (138.5 g, 1.0 mol, 2.0 equiv) in MeCN (500 mL) was stirred at reflux for about 13 h. After cooling to room temperature, DCM (1 L) was added to the mixture and the resulting mixture was filtered. The collected residue was washed with DCM (500 mL). The combined filtrate was washed with water (400 mL) and brine (20% w / w, 300 mL), dried over sodium sulfate, filtered and concentrated in vacuo to give 1-(1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)-4-methylpiperazine as a yellow solid.

[0620] Step 4: 2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline

[0621]

[0622] A mixture of 1-(1-(3-methoxy-4-nitrophenyl)piperidin-4-yl)-4-methylpiperazine (78 g, 0.233 mol) and Pd / C (10% load, 50% wet, 4 g, about 2.5 wt-%) in EtOH (800 mL) was stirred under a hydrogen atmosphere (about 20 psi) for about 2.5 h. Subsequently, the mixture was filtered through a pad of celite (50 g), and the celite pad was rinsed with EtOH (2 x 50 mL).

[0623] The combined filtrates were concentrated in vacuo to give 2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline as a purple solid.

[0624] Step 5: (2-((5-chloro-2-((2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide

[0625]

[0626] A mixture of (2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)-dimethylphosphine oxide (55 g, 0.174 mol, 1.0 eq), 2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline (74.2 g, 0.244 mol, 1.4 eq) and HCl in EtOH (2.5 M, 175 mL) in 2-methoxyethanol (750 mL) was stirred at 120 °C for about 6 h. After cooling to room temperature, the mixture was concentrated in vacuo and the resulting residue was dissolved in water (400 mL) and washed with EtOAc (500 mL). Aqueous sodium hydroxide solution (20% w / w) was added to the aqueous layer until the pH was about 12 (as determined by pH paper). The aqueous layer was extracted with DCM (3 x 500 mL) and the combined organic layers were concentrated in vacuo. The residue was triturated sequentially with EtOAc / MeOH (9 / 1 v / v, 250 mL) and EtOAc / heptane (1 / 2 v / v, 300 mL) at room temperature for about 1 h, then filtered to give a light-colored solid (Batch A).

[0627] Another batch of (2-((5-chloro-2-((2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethyl-phosphine oxide was prepared in 2-methoxyethanol (650 mL) using (2-((2,5-dichloropyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide (50.8 g, 0.161 mol, 1.0 eq), 2-methoxy-4-(4-(4-methylpiperazin-1-yl)piperidin-1-yl)aniline (68.4 g, 0.225 mol, 1.4 eq) and HCl in EtOH (2.5 M, 160 mL). After work-up as described previously, a solid was obtained (Batch B).

[0628] Two batches (batch A and batch B) were combined and ground with MeOH / EtOAc (1% v / v, 500mL) and MeOH / EtOAc (2.5% v / v, 500mL) at room temperature for about 30min, then filtered. The separated solid was then ground with hot EtOAc (500mL) for 15 minutes, then cooled to room temperature, and then filtered. The separated solid was then ground in hot MeOH / EtOAc (2% v / v, 500mL) for 15 minutes, then cooled to room temperature and filtered. The separated solid was then ground at room temperature in DCM (750mL). The resulting solution was filtered and the collected solid was vacuum dried to obtain (2-((5-chloro-2-((2-methoxy-4-(4-(4-methylpiperazine-1-yl)piperidin-1-yl)phenyl)amino)pyrimidin-4-yl)amino)phenyl)dimethylphosphine oxide as a beige solid. 127g, 65% yield. 1H NMR: see Table 2. ESI-MS m / s: 584.2 [M+H] + .

[0629] VI. Pharmaceutical Composition Examples

[0630] Representative pharmaceutical compositions and dosage forms of the compounds as disclosed herein for therapeutic use or prophylactic use in humans (the active ingredient is referred to as the "compound") may be as follows:

[0631] (a) Tablet I mg / tablet

[0632] Compounds ..................................100

[0633] Anhydrous lactose............................182.75

[0634] Croscarmellose sodium...................12.0

[0635] Corn starch paste (5% w / v paste)............2.25

[0636] Magnesium stearate............................3.0

[0637] (b) Tablet II mg / tablet

[0638] Compounds ..................................50

[0639] Anhydrous lactose............................223.75

[0640] Croscarmellose sodium...................6.0

[0641] Corn starch............................15.0

[0642] Polyvinylpyrrolidone (5% w / v paste)...2.25

[0643] Magnesium stearate............................3.0

[0644] (c) Tablet III mg / tablet

[0645] Compounds..............................1.0

[0646] Anhydrous lactose............................93.25

[0647] Croscarmellose sodium...................4.0

[0648] Corn starch paste (5% w / v paste)............0.75

[0649] Magnesium stearate............................1.0-76

[0650] (d) Capsules mg / capsule

[0651] Compounds ..................................10

[0652] Anhydrous lactose............................488.5

[0653] Magnesium..................................1.5

[0654] (e) Injection I (50mg / mL)

[0655] Compound..............................5.0%w / v

[0656] 1M sodium hydroxide solution......................15.0% v / v

[0657] 0.1M hydrochloric acid (to adjust pH to 7.6)

[0658] Polyethylene glycol 400.........................4.5%w / v

[0659] Water for injection up to 100%

[0660] (f) Injection II (10mg / mL)

[0661] Compound..............................1.0%w / v

[0662] Sodium phosphate BP............................3.6%w / v

[0663] O.1M sodium hydroxide solution....................15.0%v / v

[0664] Water for injection up to 100%

[0665] (g) Injection III (1 mg / mL, buffered to pH 6)

[0666] Compound..............................0.1%w / v

[0667] Sodium phosphate BP............................2.26%w / v

[0668] Citric acid..............................0.38% w / v

[0669] Polyethylene glycol 400.........................3.5% w / v

[0670] Water for injection up to 100%

[0671] (h) Aerosol I mg / mL

[0672] Compound ..................................10.0

[0673] Sorbitan trioleate................13.5

[0674] Trichlorofluoromethane.......................910.0

[0675] Dichlorodifluoromethane........................490.0

[0676] (i) Aerosol II mg / mL

[0677] Compound .................................0.2

[0678] Sorbitan trioleate................0.27

[0679] Trichlorofluoromethane.......................70.0

[0680] Dichlorodifluoromethane........................280.0

[0681] Dichlorotetrafluoroethane........................1094.0

[0682] (j) Aerosol III mg / mL

[0683] Compounds ..................................2.5

[0684] Sorbitan trioleate................3.38

[0685] Trichlorofluoromethane.......................67.5

[0686] Dichlorodifluoromethane........................1086.0

[0687] Dichlorotetrafluoroethane.......................191.6

[0688] (k) Aerosol IV mg / mL

[0689] Compounds ..................................2.5

[0690] Soy lecithin....................2.7

[0691] Trichlorofluoromethane.......................67.5

[0692] Dichlorodifluoromethane........................1086.0

[0693] Dichlorotetrafluoroethane.......................191.6

[0694] (l) Ointment Unit / mL

[0695] Compound..............................40mg

[0696] Ethanol................................300μL

[0697] Water..................................300μL

[0698] 1-Dodecylazacycloheptanone................50μL

[0699] Propylene glycol .................................. to 1 mL

[0700] These preparations can be prepared using conventional procedures well known in the pharmaceutical field. If desired, tablets (a)-(c) can be enteric coated by conventional means, for example, to provide a coating of cellulose acetate phthalate. In certain embodiments, tablets suitable for oral administration contain about 30 mg, about 90 mg, about 150 mg, or about 180 mg of substantially pure form A of brigatinib, together with one or more pharmaceutically acceptable excipients, such as described herein. As used herein, "about" means a value modified by ±5%. Aerosol formulations (h)-(k) can be used in conjunction with a standard metered dose aerosol dispenser, and the suspending agents sorbitan trioleate and soy lecithin can be replaced by alternative suspending agents such as sorbitan monooleate, sorbitan sesquioleate, polysorbate 80, polyglycerol oleate, or oleic acid.

[0701] VII. Kinase Inhibition

[0702] The compounds described herein are screened for kinase inhibitory activity as follows. Suitable kinases for use in the following protocols include, but are not limited to, ALK, Jak2, b-Raf, c-Met, Tie-2, FLT3, Abl, Lck, Lyn, Src, Fyn, Syk, Zap-70, Itk, Tec, Btk, EGFR, ErbB2, Kdr, FLT1, Tek, InsR, and AKT.

[0703] Kinases are expressed as kinase domains or full-length constructs fused to glutathione S-transferase (GST) or polyhistidine-tagged fusion proteins in E. coli or baculovirus-High Five expression systems. They are purified to near homogeneity by affinity chromatography as previously described (Lehr et al., 1996; Gish et al., 1995). In some cases, kinases are co-expressed or mixed with purified or partially purified regulatory polypeptides to measure activity.

[0704] Kinase activity and inhibition can be determined by established protocols (see, e.g., Braunwalder et al., 1996). In such cases, 33 The transfer of PO4 from ATP to the synthetic substrate poly(Glu,Tyr) 4:1 or poly(Arg,Ser) 3:1 attached to the bioactive surface of a microtiter plate is used as a measure of enzyme activity. After an incubation period, the amount of transferred phosphate is measured by first washing the plate with 0.5% phosphoric acid, adding liquid scintillant, and then counting in a liquid scintillation detector. 33 The concentration of compound that reduces P by 50% determines the IC 50 .

[0705] Other methods based on the transfer of phosphate groups to peptide or polypeptide substrates containing tyrosine, serine, threonine or histidine alone, in combination with each other or with other amino acids, in solution or immobilized (i.e., solid phase) form may also be used.

[0706] For example, scintillation proximity, fluorescence polarization, or homogenous time-resolved fluorescence can also be used to detect the transfer of phosphate to the peptide or polypeptide. Alternatively, antibody-based methods can be used to measure kinase activity, where antibodies or polypeptides can be used as reagents to detect phosphorylated target polypeptides.

[0707] For further background information on such assays, see, e.g., Braunwalder et al., 1996, Anal. Biochem. 234(1):23; Cleaveland et al., 1990, Anal Biochem. 190(2):249; Gish et al. (1995). Protein Eng. 8(6):609; Kolb et al. (1998). Drug Discov. Toda V. 3:333; Lehr et al. (1996). Gene 169(2):27527-87; Seethala et al. (1998). Anal Biochem. 255(2):257; Wu et al. (2000).

[0708] The inhibition of ALK tyrosine kinase activity can be demonstrated using known methods. For example, in one method, the ability of the test compound to inhibit the kinase activity of ALK expressed by baculovirus can be tested using a modification of the ELISA protocol reported for trkA in Angeles, TS et al., Anal. Biochem. 1996, 236, 49-55, which is incorporated herein by reference. As reported in Rotin, D. et al., EMBO J. 1992, 11, 559-567 (which is incorporated herein by reference), the phosphorylation of the phospholipase C-γ (PLC-γ) substrate produced as a fusion protein with glutathione S-transferase (GST) can be detected with europium-labeled anti-phosphotyrosine antibodies and measured by time-resolved fluorescence (TRF). In this assay, 96-well plates were coated with 100 μL / well of 10 μg / mL substrate (phospholipase C-γ in tris-buffered saline (TBS). Then, 20 nM HEPES (pH 7.2, 1 μM ATP (K m The assay mixture (total volume = 100 μL / well) consisting of 50 nM MnCl2, 5 nM MnCl2, 0.1% BSA, 2.5% DMSO and various concentrations of the test compound is added to the assay plate. The reaction is initiated by adding the enzyme (30 ng / mL ALK) and allowed to proceed at 37°C for 15 minutes. The detection of the phosphorylated product can be performed by adding 100 μL / well of Eu-N1 labeled PT66 antibody (Perkim Elmer #AD0041). The incubation at 37°C is then performed for one hour, followed by the addition of 100 μL of enhancement solution (e.g., Wallac #1244-105). The plate is gently agitated, and the fluorescence of the resulting solution can be measured after thirty minutes (e.g., using an EnVision 2100 (or 2102) multi-label plate reader from Perkin Elmer).

[0709] Data analysis can then be performed. 10 Plotting to calculate IC 50 value.

[0710] Inhibition of ALK tyrosine kinase can also be measured using the recombinant kinase domain of ALK, similar to the VEDG-R kinase assay described in J. Wood et al., Cancer Res 2000, 60, 2178-2189. In vitro enzyme assays using GST-ALK protein tyrosine kinase can be performed in 96-well plates as filter binding assays in 20 mM Tris.HCl, pH 7.5, 3 mM MgCl2, 10 mM MnCl2, 1 nM DTT, 0.1 μCi / assay (= 30 μL) [γ- 33P]-ATP, 2 μM ATP, 3 μg / mL poly (Glu, tyr 4: 1) poly-EY (sigma P-0275), 1% DMSO, 25ng ALK enzyme. The assay can be incubated for 10 min at ambient temperature. The reaction can be terminated by adding 50 μL of 125mM EDTA, and the reaction mixture can be transferred to a MAIP Multiscreen plate (Millipore, Bedford, MA) previously wetted with methanol and rehydrated with water for 5 minutes. After washing (0.5% H3PO4), the plate can be counted in a liquid scintillation counter. IC is calculated by linear regression analysis of percent inhibition 50 value.

[0711] It has also been demonstrated that certain compounds disclosed herein have cytotoxic or growth inhibitory effects on tumors and other cancer cell lines, and therefore certain compounds disclosed herein can be used to treat cancer and other cell proliferation diseases. The anti-tumor activity of the compound is determined using in vivo and in vitro assays known to those skilled in the art. Typically, a preliminary screening of compounds is performed in a cell assay to determine candidate anticancer drugs. Compounds that can then be identified as having antiproliferative activity in such cell-based assays can then be assayed for anti-tumor activity and toxicity in whole organisms. In general, cell-based screening is faster and more economical to perform than assays using whole organisms. As disclosed herein, the terms "anti-tumor" and "anti-cancer" activity can be used interchangeably.

[0712] Cell-based methods for measuring antiproliferative activity are well known and can be used for comparative characterization of compounds as disclosed herein. Typically, cell proliferation and cell viability assays are designed to provide detectable signals when cells are metabolically active. The antiproliferative activity of a compound can be tested by measuring any observable decrease in cellular metabolic activity after cells are exposed to the compound. Common methods include, for example, measuring membrane integrity (as a measure of cell viability) (e.g., using trypan blue exclusion) or measuring DNA synthesis (e.g., by measuring the incorporation of BrdU or 3H-thymidine).

[0713] Some methods for measuring cell proliferation use reagents that are converted to detectable compounds during cell proliferation. Such reagents are tetrazolium salts and include, but are not limited to, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; Sigma-Aldrich, St. Louis, MO), MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium), XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-anilinocarboxamide), INT, NBT, and NTV (Bernas et al. Biochim Biophys Acta 1451(1):73-81, 1999). The more commonly used assay utilizing tetrazolium salts detects cell proliferation by detecting the product of the enzymatic conversion of the tetrazolium salt to a blue formazan derivative, which can be readily and rapidly detected spectrophotometrically (Mosman. J. Immunol. Methods. 65:55-63, 1983).

[0714] Other methods for determining cell proliferation involve incubating cells in a given growth medium (with or without the compound to be determined). The growth conditions of various prokaryotic and eukaryotic cells are well known to those of ordinary skill in the art (Ausubel et al. Current Protocols in Molecular Biology. Wiley and Sons. 1999; Bonifacino et al. Current Protocols in Cell Biology. Wiley and Sons. 1999, both of which are incorporated herein by reference). To detect cell proliferation, a tetrazolium salt is added to the incubated cultured cells so that it is enzymatically converted into a detectable product by living cells. The cells are processed and the optical density of the cells is determined to measure the amount of the formazan derivative. In addition, commercially available kits including reagents and protocols can be purchased, for example, from Promega Corporation (Madison, WI), Sigma-Aldrich (St. Louis, MO) and Trevigen (Gaithersburg, MD).

[0715] In addition, a variety of cell types can be used to screen the anti-proliferative activity of compounds, including the following cell lines, in particular: COLO 205 (colon cancer), DLD-1 (colon cancer), HCT-15 (colon cancer), HT29 (colon cancer), HEP G2 (liver cancer), K-562 (leukemia), A549 (lung cancer), NCI-H249 (lung cancer), MCF7 (breast cancer), MDA-MB-231 (breast cancer), SAOS-2 (osteosarcoma), OVCAR-3 (ovarian cancer), PANC-1 (pancreatic cancer), DU-145 (prostate cancer), PC-3 (prostate cancer), ACHN (renal cancer), CAKI-1 (renal cancer), MG-63 (sarcoma).

[0716] Although cell lines may preferably be mammalian, lower eukaryotic cells such as yeast may also be used to screen compounds. Mammalian cell lines are derived from humans, rats, mice, rabbits, monkeys, hamsters, and guinea pigs because cells from these organisms are well studied and characterized. However, other cells may also be used.

[0717] Suitable mammalian cell lines are usually derived from tumors. For example, the following tumor cell types can be the cell source of cultured cells: melanoma, myeloid leukemia, lung cancer, breast cancer, ovarian cancer, colon cancer, kidney cancer, prostate cancer, pancreatic cancer and testicular cancer), cardiomyocytes, endothelial cells, epithelial cells, lymphocytes (T-cells and B cells), mast cells, eosinophils, vascular intimal cells, hepatocytes, leukocytes (including mononuclear leukocytes), stem cells (such as hematopoietic stem cells, nerves, skin, lungs, kidneys, liver and muscle stem cells (for screening differentiation and dedifferentiation factors)), osteoclasts, chondrocytes and other connective tissue cells, keratinocytes, melanocytes, hepatocytes, kidney cells and adipocytes. Non-limiting examples of mammalian cell lines widely used by researchers include HeLa, NIH / 3T3, HT1080, CHO, COS-1, 293T, WI-38 and CV1 / EBNA-1.

[0718] Other cell assays based on reporter gene detection of metabolically active cells can be used. Non-limiting examples of reporter gene expression systems include green fluorescent protein (GFP) and luciferase. As an example of using GFP to screen potential anti-tumor drugs, Sandman et al. (Chem Biol. 6:541-51; incorporated herein by reference) used HeLa cells containing inducible variants of GFP to detect compounds that inhibit GFP expression and thus inhibit cell proliferation.

[0719] An example of a cell-based assay is shown below. The cell line that can be used in the assay is Ba / F3, a murine Pro-B cell line, which is expressed in the expression vector pClneo encoding NPM-ALK.TM (Promega Corp., Madison WI) stably transfected and subsequently secreted G418-resistant cells. Untransfected Ba / F3 cells depend on IL-3 for cell survival. In contrast, Ba / F3 cells expressing NPM-ALK (i.e., Ba / F3-NPM-ALK) can proliferate in the absence of IL-3 because they obtain proliferation signals through NPM-ALK kinase. Its assumed inhibitor of NPM-ALK kinase eliminates growth signals and produces anti-proliferative activity. However, the anti-proliferative activity of inhibitors of NPM-ALK kinase can be overcome by adding IL-3 that provides growth signals through NPM-ALK independent mechanisms. For similar cell systems using FLT3 kinase, see E. Weisberg et al. Cancer cell, 2002, 1, 433-443. The inhibitory activity of compounds disclosed herein can be determined as follows: BaF3-NPM-ALK cells (15,000 / microtiter plate well) can be transferred to 96-well microtiter plates. A series of concentrations (dilution series) of the test compound (dissolved in DMSO) are then added in such a way that the final concentration of DMSO does not exceed 1% (v / v). After addition, the plates can be incubated for two days, during which control cultures without the test compound can undergo two cell division cycles. TM The growth of BaF3-NPM-ALK cells was measured by staining (T Idziorek et al., J. Immunol. Methods 1995, 185, 249-258). Then, 25 μL of lysis buffer consisting of 20 mM sodium citrate, pH 4.0, 26.8 nM sodium chloride, 0.4% NP40, 20 mM EDTA and 20 mM was added to each well. Cell lysis was completed within 60 minutes at room temperature, and the total amount of Yopro bound to DNA was determined by measurement using, for example, a CytoFluor II 96-well reader (PerSeptive Biosystems). IC 50 It can be determined by computer-aided system using the following formula:

[0720] IC 50 =[(ABS 测试 -ABS 开始 ) / (ABS 对照 -ABS 开始 )]x100

[0721] Among them, ABS is adsorption. IC in this type of experiment 50 Values ​​are given as the concentration of the test compound in question that produces a cell count 50% lower than the cell count obtained using the control in the absence of inhibitor.

[0722] The antiproliferative effect of the compounds disclosed herein can also be determined in the human KARPAS-299 lymphoma cell line by immunoblotting as described by WG Dirks et al., Int. J. Cancer 2002, 100, 49-56, using the method described above for the BaF3-NPM-ALK cell line.

[0723] In another embodiment, the antiproliferative activity can be determined using the KARPAS-299 lymphoma cell line according to the following procedure: the compounds disclosed herein are incubated with the cells for 3 days, and the number of viable cells in each well is indirectly measured using the MTS tetrazolium assay (Promega). This test is a colorimetric method for determining the number of viable cells by measuring the metabolic activity of viable cells. For example, the detection of the product of the enzymatic conversion of tetrazolium salts to blue formazan derivatives is achieved by measuring the absorbance at 490nm using a plate reader. 40 μL of MTS reagent is added to all wells except the edge wells, and the plate is then returned to the incubator at 37°C for 2 hours. The Wallac Victor plate reader is then used at 490nm to measure the absorbance at 490nm. 2 The absorbance in each well was measured using a plate reader. IC was calculated by determining the concentration of compound required to reduce the MTS signal by 50% by comparison to the baseline (DMSO control) which was 0% inhibition using Microsoft XLfit software in the best fit curve. 50 .

[0724] The anti-tumor activity of the compound identified as having anti-cell proliferation activity by such cell assays can then be tested in all organisms (such as mammalian species). Well-characterized mammalian systems for studying cancer include rodents, such as rats and mice. Typically, the target tumor is transplanted into mice with reduced ability to strengthen the immune response to the tumor to reduce the possibility of rejection. Such mice include, for example, nude mice (without thymus) and SCID (heavy comprehensive immunodeficiency) mice. Other transgenic mice such as mice containing proto-oncogenes can be used in this assay (see, for example, USP 4,736,866 and USP 5,175,383). For a review and discussion of anti-tumor drug testing using rodent models, see Kerbel (Cancer Metastasis Rev. 17: 301-304, 1998-99).

[0725] Typically, the target tumor is implanted subcutaneously in the test organism. The organism containing the tumor is treated with a dose of the candidate anti-tumor compound. The size of the tumor is measured regularly to determine the effect of the test compound on the tumor. Some tumor types are implanted in sites other than subcutaneous sites (such as intraperitoneal sites) and survival is measured as an end point. The parameters to be determined by conventional screening include different tumor models, various tumors and drug routes, as well as dosage and regimen. For a review of using mice to detect anti-tumor compounds, see Corbett et al. (Invest New Drugs. 15: 207-218, 1997; incorporated herein by reference).

[0726] The compounds disclosed herein have inhibitory activity against wild-type or mutant (particularly clinically relevant mutant) kinases, particularly kinases such as ALK, Met, Jak2, bRaf, EGFR, Tie-2, FLT3 or another target kinase, with an IC 50 A value of 1 μM or less (as determined using any scientifically acceptable kinase inhibition assay), such as its IC 50 500 nM or better, and also such as IC 50 A value of 250 nM or better; or

[0727] · Has inhibitory activity against a given kinase, with an IC 50 IC values ​​for other target kinases 50 Value at least 100-fold lower; or

[0728] Inhibitory activity against ALK, Met, Jak2, or B-Raf, with IC of 1 μM or better for each 50 value; or

[0729] Cytotoxic or growth inhibitory effects on cancer cell lines (such as Ba / F3 NPM-ALK, Ba / F3 EML4-ALK, Karpas 299 and / or SU-DHL-1 cells) maintained in vitro or in animal studies using scientifically acceptable cancer cell xenograft models, as determined by comparative studies, that are at least as potent as, or at least twice as potent as, or at least 10 times as potent as, known ALK inhibitors (such as NVP-TAE684 and PF2341066, in particular).

[0730] The compounds disclosed herein were found to be effective in inhibiting multiple important kinase targets. When tested, for example, as kinase (ALK) inhibitors, the compounds exhibited IC 50Less than 100 nM, and in many cases less than 10 nM and in many cases less than 1 nM. Some compounds are single digit nanomolar inhibitors of a panel of kinases including kinases such as ALK, FER, FLT3, FES / FPS, FAK / PTK2, BRK, and others.

Claims

1. Crystalline brigatinib.

2. Substantially pure crystalline brigatinib.

3. Crystalline Form A of Brigatinib.

4. The crystalline form A according to claim 3, which has an x-ray powder diffraction pattern comprising at least two peaks expressed in degrees 2θ selected from 9.6, 17.2, 19.4, 20.1, 23.1 and 27.

7.

5. A method for preparing Form A of brigatinib, comprising raising the temperature of crystallization to 60°C and slowly adding a NaOH solution.

6. Crystalline Form B of Brigatinib.

7. The crystalline form B according to claim 6, which has an x-ray powder diffraction pattern comprising at least two peaks expressed in degrees 2θ selected from 11.5, 14.5, 16.9, 19.2 and 23.

2.

8. Crystalline Form C of Brigatinib.

9. The crystalline form C of claim 8, which has an x-ray powder diffraction pattern comprising at least two peaks expressed in degrees 2θ selected from 5.4, 14.9, 15.9, 17.3, 19.2 and 23.

9.

10. Crystalline Form D of Brigatinib.

11. The crystalline form D of claim 10, which has an x-ray powder diffraction pattern comprising at least two peaks expressed in degrees 2θ selected from 9.7, 11.1, 17.4, 18.9 and 23.

7.

12. Crystalline Form E of Brigatinib.

13. The crystalline form E of claim 12, which has an x-ray powder diffraction pattern comprising at least two peaks expressed in degrees 2θ selected from 9.1, 10.2, 15.8, 19.2 and 23.

9.

14. Crystalline Form F of Brigatinib.

15. The crystalline form F of claim 14, which has an x-ray powder diffraction pattern comprising at least two peaks expressed in degrees 2θ selected from 9.8, 17.0, 19.4, 20.3 and 27.

1.

16. Crystalline Form G of Brigatinib.

17. The crystalline form G of claim 16, which has an x-ray powder diffraction pattern comprising at least two peaks expressed in degrees 2θ selected from 8.3, 9.7, 12.9, 15.8, 18.1, 20.7, 22.8 and 26.

8.

18. Crystalline Form H of Brigatinib.

19. The crystalline form H of claim 18, which has an x-ray powder diffraction pattern comprising at least two peaks expressed in degrees 2θ selected from 4.2, 5.2, 8.4, 10.9, 12.7 and 21.

3.

20. A pharmaceutical composition comprising at least one crystalline form of brigatinib and at least one component selected from a pharmaceutically acceptable carrier, a pharmaceutically acceptable solvent and a pharmaceutically acceptable excipient.

21. The pharmaceutical composition of claim 20, wherein the at least one crystalline form of brigatinib is selected from Form A, Form B, Form C, Form D, Form E, Form F, Form G, and Form H.

22. Use of a crystalline form of brigatinib in the preparation of a medicament for treating non-small cell lung cancer.

23. The use of claim 22, wherein the crystalline form of brigatinib is selected from the group consisting of Form A, Form B, Form C, Form D, Form E, Form F, Form G and Form H.

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