BR112017020945B1 - SESQUI-MALEATE CRYSTALLINE SALTS OF A B-RAF KINASE INHIBITOR, METHOD AND PROCESS OF PREPARATION AND USES THEREOF - Google Patents

Abstract

MALEATE SALTS OF A B-RAF KINASE INHIBITOR, CRYSTALLINE FORMS, METHODS OF PREPARATION AND USES THEREOF. The present invention relates to 5 - ((1R, 1aS, 6bR) -1- (6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b-dihydro- 1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one (Compound 1), in particular the sesqui-maleate salt and their crystalline forms, methods of preparation, pharmaceutical compositions and therapeutic uses for the treatment of diseases or disorders mediated by BRAF or other kinases.

Description

TECHNICAL FIELD

[0001] The present invention relates to maleate salts of a B-RAF kinase inhibitor, i.e. 5 - ((1R, 1aS, 6bR) -1- (6- (trifluoromethyl)-1H-benzo [d] imidazol-2-yl)-1a,6b-dihydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one (hence hereinafter referred to as Compound 1), especially Compound 1 sesqui-maleate, and crystalline (polymorph) forms of the sesqui-maleates, and methods of preparation, and uses thereof.

[0002] BACKGROUND OF THE INVENTION

[0003] B-RAF is a kinase that is part of the key RAF-MEKERK MAPK pathway for cell proliferation and survival. B-RAF mutations have been discovered in more than 7% of human cancers, including melanoma (43%) (see H. Davies et al., Nature, 417 (2002), 949-54; DR English et al., Cancer Epidemiol Biomarkers Prev, 17 (2008), 1774-80; GV Long, et al., Lancet Oncol., 13 (2012), 1087-95), thyroid (27%) (see Y. Cohen, J Natl Cancer Inst, 95 (2003), 625-7; ETKimura, et al., Cancer Res, 63 (2003), 1454-7), colorectal (14%) (see H. Davies et al., Nature, 417 (2002), 949 -54; DR English, et al., Cancer Epidemiol Biomarkers Prev, 17 (2008), 1774-80; S. Ogino, et al., Gut, 58 (2009), 90-6; CPVaughn, Cancer Chromosomes Genes, 50 (2011), 307-12), ovary (15%) (see H. Davies, et al., Nature, 417 (2002), 949-54; SE-Russell, J Pathol, 203 (2004), 617-9 ) and lung (2%) (see MSBrose, et al., Cancer Res, 62 (2002), 69977000). More than 90% of B-RAF mutations found in melanoma are a substitution of glutamic acid for valine at the 600th amino acid of the B-RAF protein chain (V600E), resulting in constitutive activation.

[0004] Clinical experience with B-RAF inhibitors such as vemurafenib and dabrafenib in patients with melanoma has been shown to be effective, which validated the concept of targeting tumors depending on B-RAF and MAPK signaling. Selective inhibition in melanoma patients harboring the B-RAF V600E mutation in their tumors has produced impressive objective response rates and prolongation of progression-free survival. However, the first generation of B-RAF inhibitors, including vemurafenib and da-brafenib, have a number of limitations, for example, 1) development of keratoacanthomas or cutaneous squamous cell carcinoma due to the paradoxical increase of the MAPK signal through the induction of B-RAF / C-RAF heterodimer in the context of mutant or activated RAS; and 2) limited clinical activity outside melanoma with B-RAF V600E mutations (eg, colorectal). Therefore, a second-generation inhibitor that can improve in these areas is highly desirable.

[0005] O 5 - ((1R, 1aS, 6bR) -1- (6- (trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b-dihydro-1H-cycloprop [ b ] benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one (free base, Compound 1) has been disclosed as a second generation B-RAF inhibitor, which has demonstrated potent inhibitory activity against the RAF family of serine/threonine kinases. See patent document WO 2013/097224 A1.

[0006]

[0007] Compound 1

[0008] Compound 1 is a molecularly targeted therapeutic agent for the treatment of cancers with aberrations in the RAF-MEK-ERK mitogen-activated protein kinase (MAPK) pathway, including B-RAF mutations and K-RAS / N-RAS mutations , both as monotherapy and in combination with other cancer therapies. However, the relatively poor oral absorption of the free base of Compound 1 makes it unsuitable for drug development.

[0009] It was also found that the free base of Compound 1 was originally obtained as an amorphous solid based on the XRPD results of Figure 1 and that the free base of Compound 1 is substantially insoluble in water with <LOQ to 0.001 mg/mL , and is slightly hygroscopic with 2.2% water gain from 0 to 80% RH.

[0010] Furthermore, the synthesis of Compound 1 was inefficient. The various factors, for example, the requirement of the chiral HPLC column for the preparation of optically pure isomers and the tendency of the free base of Compound 1 in amorphous form to prevent the expulsion of impurities from the process, make the large scale preparation a challenge. and the purification of Compound 1.

[0011] Practically, it is difficult to predict with confidence that the salts of a particular compound are stable and suitable for pharmaceutical processing. It is even more difficult to predict whether or not a given particular will form various crystalline solid state forms and to predict the physical properties of these crystalline solid state forms.

[0012] Therefore, there is a great need for some forms of Compound 1 that have much better bioavailability and chemical and physical stability during the formulation and storage of this drug, as well as a great need for a process suitable for large-scale preparation of Compound 1 with good quality and reproducibility.

[0013] SUMMARY OF THE INVENTION

[0014] The present patent application describes an invention to solve the above challenges and need by providing stable salts of Compound 1 and its crystalline forms.

[0015] In the first aspect, there is provided herein a salt of 5-(((1R, 1aS, 6bR)-1-(6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b -dihydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one ("Compound 1"), which is selected from Compound 1 hydrochloride, methanesulfonate, 2-hydroxyethanesulfonate, L-tartrate, maleate and oxalate. In one embodiment, the salt is pharmaceutically acceptable. In another embodiment, the salt is in a solid state. In a preferred embodiment, the salt is in a crystalline form. The salts of the present application are in solid form and show different crystalline forms from the free base of Compound 1, indicating that they are potential salt candidates. The salts in crystalline form in the present application have superior physical properties suitable for pharmaceutical formulations and can be manufactured on large commercial scales with high quality and good reproducibility.

[0016] Among the hydrochloride, methanesulfonate, 2-hydroxyethanesulfonate, L-tartrate, maleate and oxalate crystalline forms of Compound 1, the inventors of the present invention have unexpectedly found that Compound 1 can form maleate salts, in particular sesqui salt -maleate, in crystalline forms. In particular, it has surprisingly been found that the maleate salt of Compound 1 (hereinafter referred to sometimes as "Maleate Salt of Compound 1" or "Mixed Compound 1") can exist in various crystal line forms (polymorphs), what are referred to herein as Crystalline Forms A*, A**, A, B, C, D, E, F, G, H, I, J, K, L, and M. The maleate salt of Compound 1 and forms thereof crystalline forms, specifically crystalline Form A* and A, possess superior properties, such as improved solubility and stability, particularly long-term chemical/physical stability, to the other salts, which makes them suitable candidates for formulation and clinical applications. More particularly, the improved solubility and/or long-term chemical/physical stability of Forms A*, A, and the other forms of the maleate salts of Compound 1 provide these forms with rapid dissolution in vivo and in vitro, and thus increase bioavailability. compared to the free base.

[0017] In the second aspect, there is provided below a compound of formula (I), which is a maleate salt of 5-(((R, 1aS, 6bR)-1-(6-(trifluoromethyl)-1H-benzo[ d]imidazol-2-yl)-1a,6b-dihydro-1H-cyclopropa[b]benzofuran-5-11)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H) -ona :

[0018]

[0019] where n is a number from about 0.3 to about 1.5.

[0020] In one embodiment, in the compound of formula (I), n is a number selected from the group consisting of 0.5 ± 0.05, 1.0 ± 0.1 and 1.5 ± 0.2 .

[0021] In another embodiment, in the compound of formula (I), n is 0.5, 1.0 or 1.5.

[0022] In another preferred embodiment, the compound of formula (I) is in crystalline form.

[0023] In another preferred embodiment, n is 1.5, and the compound is a crystalline sesqui-maleate salt of 5-((1R, 1aS, 6bR)-1-(6-(trifluoromethyl)-1H-benzoyl [d]imidazol-2-yl)-1a,6b-dihydro-1H-cyclopropa [b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2 (1H )-one of formula (II):

[0024]

[0025] In one embodiment, the compound of formula (II) is in crystalline Form A*, which is characterized by an X-ray powder diffraction pattern comprising seven or more diffraction peaks with independently selected 2θ angular values from the group consisting of: 6.3 ± 0.2, 8.9 ± 0.2, 9.4 ± 0.2, 11.2 ± 0.2, 12.6 ± 0.2, 13.4 ± 0.2, 17.9 ± 0.2, 18.6 ± 0.2, 18.8 ± 0.2, 19.3 ± 0 .2, 20.1 ± 0.2 , 20.7 ± 0.2, 21.2 ± 0.2, 21.8 ± 0.2, 22.4 ± 0.2, 22.6 ± 0.2, 23.3 ± 0.2, 23.8 ± 0.2, 24.7 ± 0.2, 25.6 ± 0.2, 26.1 ± 0.2, 27.4 ± 0.2, 28.3 ± 0.2, 28.6 ± 0 .2, 29.0 ± 0.2, 29.4 ± 0.2 and 30.4 ± 0.2 degrees.

[0026] In another embodiment, the compound of formula (II) is a single crystal in Crystalline Form A**.

[0027] In another embodiment, the compound of formula (II) is in crystalline Form A, which is characterized by an X-ray powder diffraction pattern comprising three or more diffraction peaks with angular values 2θ selected independently of the group consisting of: 8.3 ± 0.2, 11.2 ± 0.2, 17.9 ± 0.2, 18.4 ± 0.2, 18.6 ± 0.2, 19.3 ± 0.2, 20.8 ± 0.2 and 22.5 ± 0.2 degrees.

[0028] In another embodiment, the compound of formula (I) is in crystalline Form B, which is characterized by a powder X-ray diffraction pattern comprising three or more diffraction peaks with angular values 2θ selected independently of the group consisting of: 11.1 ± 0.2, 15.8 ± 0.2, 17.7 ± 0.2, 18.4 ± 0.2, 19.6 ± 0.2, 22.3 ± 0, 2, 23.1 ± 0.2 and 28.8 ± 0.2 degrees.

[0029] In another embodiment, the compound of formula (I) is in crystalline Form C, which is characterized by an X-ray powder diffraction pattern comprising three or more diffraction peaks having angular values 2θ selected independently of the group consisting of: 3.1 ± 0.2, 8.8 ± 0.2, 11.2 ± 0.2, 17.8 ± 0.2, 18.5 ± 0.2, 19.3 ± 0, 2, 20.1 ± 0.2, 20.7 ± 0.2, 21.9 ± 0.2 and 22.4 ± 0.2 degrees.

[0030] In another embodiment, the compound of formula (I) is in crystalline Form D, which is characterized by a powder X-ray diffraction pattern comprising three or more diffraction peaks with angular values 2θ selected independently of the group consisting of: 8.9 ± 0.2, 14.9 ± 0.2, 16.7 ± 0.2, 17.8 ± 0.2, 19.9 ± 0.2, 20.4 ± 0, 2, 20.9 ± 0.2 and 26.9 ± 0.2 degrees.

[0031] In another embodiment, the compound of formula (I) is in crystalline Form F, which is characterized by an X-ray powder diffraction pattern comprising three or more diffraction peaks with angular values 2θ selected independently of the group consisting of: 12.9 ± 0.2, 17.0 ± 0.2, 18.5 ± 0.2, 19.4 ± 0.2, 20.5 ± 0.2, 22.5 ± 0, 2 and 24.1 ± 0.2 degrees.

[0032] In another embodiment, the compound of formula (I) is in crystalline Form G, which is characterized by a powder X-ray diffraction pattern comprising seven or more diffraction peaks with angular values 2θ selected independently from the group consisting of: 3.4 ± 0.2, 5.6 ± 0.2, 7.0 ± 0.2, 10.3 ± 0.2, 10.9 ± 0.2, 11.7 ± 0.2 , 12.4 ± 0.2, 13.1 ± 0.2, 14.0 ± 0.2, 14.9 ± 0.2, 16.4 ± 0.2, 17.4 ± 0.2, 18 .6 ± 0.2, 19.3 ± 0.2, 20.1 ± 0.2, 21.0 ± 0.2, 21.9 ± 0.2, 23.6 ± 0.2, 24.2 ± 0.2, 25.6 ± 0.2 and 26.4 ± 0.2 degrees.

[0033] In another embodiment, the compound of formula (I) is in crystalline Form H, which is characterized by an X-ray powder diffraction pattern comprising seven or more diffraction peaks with angular values 2θ selected independently from the group consisting of: 6.3 ± 0.2, 9.0 ± 0.2, 10.1 ± 0.2, 11.2 ± 0.2, 12.7 ± 0.2, 14.5 ± 0.2, 16.1 ± 0.2, 16.6 ± 0.2, 17.9 ± 0.2, 18.1 ± 0.2, 18.5 ± 0.2, 19.0 ± 0.2 , 20.1 ± 0.2, 21.9 ± 0.2, 22.4 ± 0.2, 23.9 ± 0.2, 25.1 ± 0.2, 26.2 ± 0.2 and 28 .7 ± 0.2 degrees.

[0034] In another embodiment, the compound of formula (I) is in crystalline Form I, which is characterized by a powder X-ray diffraction pattern comprising five or more diffraction peaks with angular values 2θ selected independently from the group consisting of: 3.1 ± 0.2, 5.5 ± 0.2, 6.8 ± 0.2, 10.8 ± 0.2, 11.5 ± 0.2, 13.7 ± 0.2 , 16.1 ± 0.2, 16.3 ± 0.2, 17.8 ± 0.2, 19.8 ± 0.2, 21.5 ± 0.2, 23.8 ± 0.2, 24 .4 ± 0.2 and 28.3 ± 0.2 degrees.

[0035] In another embodiment, the compound of formula (I) is in crystalline Form J, which is characterized by a powder X-ray diffraction pattern comprising seven or more diffraction peaks with angular values 2θ selected independently from the group consisting of: 5.5 ± 0.2, 8.2 ± 0.2, 10.9 ± 0.2, 11.3 ± 0.2, 13.6 ± 0.2, 14.8 ± 0.2 , 15.8 ± 0.2, 17.4 ± 0.2, 18.0 ± 0.2, 19.1 ± 0.2, 19.8 ± 0.2, 20.0 ± 0.2, 20 .4 ± 0.2, 21.1 ± 0.2, 21.9 ± 0.2, 22.6 ± 0.2, 23.4 ± 0.2, 24.1 ± 0.2, 25.0 ± 0.2, 26.1 ± 0.2, 26.9 ± 0.2, 27.3 ± 0.2, 28.4 ± 0.2, 29.1 ± 0.2, 33.1 ± 0 .2 and 35.9 ± 0.2 degrees.

[0036] In another embodiment, the compound of formula (I) is in crystalline Form K, which is characterized by an X-ray powder diffraction pattern comprising seven or more diffraction peaks with angular values 2θ selected independently from the group consisting of: 3.1 ± 0.2, 8.9 ± 0.2, 9.3 ± 0.2, 11.2 ± 0.2, 16.7 ± 0.2, 17.9 ± 0.2 , 18.6 ± 0.2, 18.8 ± 0.2, 19.4 ± 0.2, 20.2 ± 0.2, 21.9 ± 0.2, 22.4 ± 0.2, 23 .4 ± 0.2, 23.9 ± 0.2, 24.6 ± 0.2, 26.2 ± 0.2, 27.4 ± 0.2, 28.5 ± 0.2, 29.4 ± 0.2 and 30.4 ± 0.2 degrees.

[0037] In another embodiment, the compound of formula (I) is in crystalline Form L, which is characterized by an X-ray powder diffraction pattern comprising diffraction peaks having the following 2θ angular values: 9.7 ± 0.2 and 14.1 ± 0.2 degrees.

[0038] In another embodiment, the compound of formula (I) is in Crystalline Form M, which is characterized by a powder X-ray diffraction pattern comprising seven or more diffraction peaks with selected 2θ angular values regardless of the group consisting of: 5.0 ± 0.2, 9.3 ± 0.2, 11.3 ± 0.2, 14.9 ± 0.2, 15.9 ± 0.2, 17.4 ± 0.2, 18.0 ± 0.2, 18.7 ± 0.2, 19.4 ± 0.2, 20.2 ± 0.2, 22.1 ± 0.2, 23.4 ± 0, 2 , 24.3 ± 0.2, 25.4 ± 0.2, 26.5 ± 0.2, 27.5 ± 0.2, 28.5 ± 0.2 and 29.3 ± 0.2 degrees .

[0039] In some embodiment, in the compound of formula (I), n is 1, i.e. Compound 1 maleate (1:1). In another embodiment, there is provided herein, crystalline Form N of Compound 1 maleate (1:1), which is characterized by a power X-ray diffraction pattern comprising seven or more diffraction peaks with angular values 2θ independently selected from the group consisting of: 3.30 ± 0.2, 6.61 ± 0.2, 9.88 ± 0.2, 11.73 ± 0.2, 13.14 ± 0.2, 15 .23 ± 0.2, 16.56 ± 0.2, 17.94 ± 0.2, 18.72 ± 0.2, 19.34 ± 0.2, 19.93 ± 0.2, 20.76 ±0.2, 22.04±0.2, 22.95±0.2, 23.86±0.2, 25.19±0.2, 26.61±0.2, 28.36±0.2, 30.13±0.2, 31.36±0.2, 33.49±0. 2 and 37.22 ± 0.2.

[0040] In another embodiment, the compound of formula (I) is in a crystalline form substantially characterized by an X-ray powder diffraction pattern selected from the group consisting of Figure 2, Figure 16, Figure 17, Figure 18 , figure 19, figure 20, figure 21, figure 22, figure 23, figure 24, figure 25, figure 26, figure 27, figure 28 and figure 33.

[0041] In some embodiment, the Crystalline Form of compound 1 maleate (1:1) is a chemically stable crystalline form substantially characterized by an X-ray powder diffraction pattern as shown in Figure 33.

[0042] In another aspect, provided herein is a method for preparing a crystalline form of 5 - (((1R, 1aS, 6bR) -1- (6- (trifluoromethyl) - 1H-benzo[d]imidazole-2 -yl)-1a,6b-dihydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one of general formula (I), comprising - one of the following procedures:

[0043] (a) dissolution of the free base or a salt other than the maleate salt of 5 - ((1R, 1aS, 6bR) -1- (6- (trifluoromethyl)-1H-benzo[d]imidazol-2-yl) -1a,6b-dihydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one in a solvent or mixture of solvent to form a solution or suspension; mixing the resulting solution or suspension with maleic acid to form a mixture; and precipitation of 5 - ((1R, 1aS, 6bR) -1- (6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b-dihydro-1H-cyclopropa [ b] benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one in a target crystalline form;

[0044] (b) dissolving or suspending a sesqui-maleate salt of 5-((1R, 1aS, 6bR)-1-(6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a , 6b-dihydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one in a solvent or solvent mixture; and precipitation of 5 - ((1R, 1aS, 6bR) -1- (6- (trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b-dihydro-1H-cyclopropa [ b] benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one in a target crystalline form;

[0045] (c) storage of a sesqui-maleate salt of 5-(((1R, 1aS, 6bR)-1-(6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b-dihydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one for an extended period to obtain a crystalline form target;

[0046] (d) heating a crystalline 5-(((1R, 1aS, 6bR)-1-(6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a sesqui-maleate salt or amorphous, 6b-dihydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one at elevated temperature and salt cooling to obtain a target crystalline form; It is

[0047] (e) exposure of a maleate salt of 5 - (((1R, 1aS, 6bR) -1- (6- (trifluoromethyl) -1H-benzo[d]imidazol-2-yl) -1a, 6b- crystalline or amorphous dihydro -1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one to a solvent vapor to obtain a crystalline form target.

[0048] In one embodiment of this aspect, procedure (a) or (b) further comprises one or more procedures independently selected from heating, filtration to remove undissolved impurities, solvent distillation, addition of a counter-solvent or solvent mixing, adding crystal granules, adding precipitation inducing agent(s), cooling, precipitating and filtering to collect the crystalline product.

[0049] In another embodiment of this aspect, procedure (a) or (b), wherein the solvent or solvent mixture is selected from the group consisting of water, lower alkyl alcohols, ketones, ethers, esters, lower aliphatic carboxylic acids, lower aliphatic nitriles, optionally halogenated aromatic solvents and combinations thereof.

[0050] In another embodiment of this aspect, in procedure (a) or (b) the solvent is isopropanol, ethanol, methanol, acetone, THF, 1,4-dioxane, acetic acid, acetonitrile, water, or a combination thereof same.

[0051] In another embodiment of this aspect, in procedure (a) the free base is an isolated and purified free base, an isolated but unpurified free base, or a crude reaction product containing the free base.

[0052] In another embodiment of this aspect, in procedure (c) the extended period is at least three days, at least one week, or at least two weeks.

[0053] In another embodiment of this aspect, in procedure (d) the elevated temperature is at least 40 °C, at least 60 °C, at least 80 °C, or at least 100 °C, but less than the temperature decomposition of the sesqui-maleate salt.

[0054] In another embodiment of this aspect, in procedure (e) the vapor is an acetic acid vapor.

[0055] In another embodiment of this aspect, the method is selected from:

[0056] 1) procedure (a) or (b) using isopropanol-water (v/v>60/40) as solvent to produce Crystalline Form A*;

[0057] 2) procedure (a) or (b) using acetone as solvent to form Crystalline Form A**;

[0058] 3) procedure (a) or (b) using an IPA-water mixture (v: v = 4: 1) as solvent to produce Crystalline Form A;

[0059] 4) procedure (a) or (b) using 1,4-dioxane as solvent to produce Crystalline Form B;

[0060] 5) the procedure (a) or (b) using ethanol as the solvent to form Crystalline Form C;

[0061] 6) the procedure (a) or (b) using methanol as solvent to form Crystalline Form D;

[0062] 7) procedure (a) or (b) using a mixture of acetonitrile-water (v: v = 1: 1) as solvent to produce crystalline Form F;

[0063] 8) the procedure (a) or (b) using an acetic acid-water mixture as a solvent to produce Crystalline Form G;

[0064] 9) procedure (a) or (b) using tetrahydrofuran (THF) as solvent to produce Crystal Form H;

[0065] 10) procedure (a) or (b) using an IPA-water mixture (v: v = 3: 1) as solvent to produce Crystalline Form I;

[0066] 11) the procedure (c) of storing Crystal Form D under an ambient temperature for two weeks to produce Crystal Form K;

[0067] 12) the procedure (c) of storing Crystalline Form J under room temperature for two weeks to produce Crystalline Form M;

[0068] 13) the procedure (d) of heating Crystalline Form G to 140 °C and then cooling to room temperature to produce Crystalline Form L; It is

[0069] 14) the procedure (e) of letting Crystalline Form A A interact with acetic acid vapor to produce Crystalline Form J.

[0070] In one embodiment, there is provided herein a process for preparing a crystalline form of the sesqui-maleate salt of Compound 1 (hereinafter, sometimes referred to as "Sesqui-maleate salt of Compound 1" or "Sesqui-maleate of Compound 1") comprising mixing under a temperature below the reflux temperature, for example mixing under about 50 °C a mixture of Compound 1 and maleic acid in a mixed solvent of i-PrOH and H2O, or mixing at a temperature below the reflux temperature, for example mixing under about 50 °C Compound 1 with a mixture or a suspension or a solution of maleic acid in a mixed solvent of i-PrOH and H2O, or mixing under a temperature below the temperature reflux, for example, mixing under about 50oC a mixture or a suspension or a solution of Compound 1 in a mixed solvent of i-PrOH and H2O with maleic acid, in which the amount of i-PrOH is greater than 40% in volume in terms of the total volume of i -PrOH and water, preferably 60vol%, and most preferably 90vol%. In some preferred embodiments, the aforementioned mixed solvent is replaced by i-PrOH. In another embodiment, the process further comprises adding some crystal granules to the resulting mixture after cooling to room temperature, and then allowing the mixture to stand for a certain period of time, such as 12 hours, 24 hours, 2, 3 or 4 days or 1 week, 2 weeks.

[0071] In another embodiment, there is provided herein a process for preparing a crystalline form of Compound 1 maleate (1:1) comprising mixing a crystalline form of the sesqui-maleate of Compound 1 with methanol. In another embodiment, the above mixing is carried out under agitation. In another embodiment, the maleate crystalline form of Compound 1 (1:1) is Form N as described herein. In another embodiment, the crystalline form of Compound 1 maleate is any one selected from the group consisting of Crystalline Forms A*, A**, A, B, C, D, F, G, H, I, J, K, L and M. In another embodiment, the above mixture is formed for approximately one day, or 2 days, or 3 days, or 4 days, 5 days, or 6 days, or one week, or 2 weeks. This conversion of Sesqui-Maleate Salt of Compound 1 (1:1.5) to Maleate Salt (1:1) provides an economical process for the manufacture and/or further purification of the active pharmaceutical ingredient of Compound 1 with very simple procedure, by for example, simply stirring Sesqui-Maleate of compound 1, for example any Crystalline Form A*, A** and A in methanol under room temperature or elevated temperature. This conversion has been found to give rise to a reduced crystalline particle size for the resulting crystalline substance (i.e., Crystalline Form of compound 1 maleate (1:1), which may in turn increase the rate of dissolution to the pharmaceutical product and further enhance the chemical purity of the medicine. In this regard, the conversion is expected to further simplify the pharmaceutical manufacturing process, for example, without using micronization and roller compaction process.

[0072] In another aspect, there is provided herein a pharmaceutical composition comprising a compound of formula (I) or formula (II) according to any one of the embodiments described herein and a pharmaceutically acceptable carrier.

[0073] In one embodiment, the pharmaceutical composition is suitable for oral administration.

[0074] In another embodiment, the pharmaceutical composition is in tablet or capsule form.

[0075] In another embodiment, the unit dosage of the tablet or capsule is 5-80 mg.

[0076] In another embodiment, the weight percentage of the compound in the pharmaceutical composition is 1-99%.

[0077] In another aspect, there is provided herein a method of treating or pre-ventilating a disease or disorder in a subject, such as a human, comprising administering to said subject a therapeutically effective amount of a compound of formula ( I) or formula (II) according to any one of the embodiments described herein or a pharmaceutical composition comprising a compound of formula (I) or formula (II).

[0078] In one embodiment, the disease or disorder is a cancer selected from the group consisting of brain cancer, lung cancer, kidney cancer, bone cancer, liver cancer, bladder cancer, breast cancer, head and neck cancer, ovarian cancer, melanoma, skin cancer, adrenal cancer, cervical cancer, colorectal cancer, lymphoma or thyroid tumors and their complications.

[0079] In another embodiment, the disease is selected from the group consisting of BRAF, NRAS and KRAS mutant brain cancer, lung cancer, kidney cancer, bone cancer, liver cancer, bladder cancer, breast cancer , head and neck cancer, ovarian cancer, melanoma, skin cancer, adrenal cancer, cervical cancer, colorectal cancer, lymphoma or thyroid tumors and their complications.

[0080] In another embodiment, the administered dosage of the tablet is 1-100 mg/day and the administration frequency is from one to three times a day.

[0081] In another embodiment, the administered dosage of the tablet is 5-50 mg/day and the administration frequency is from one to three times a day.

[0082] In another embodiment, the administered dosage of the tablet is 10-40 mg/day, and the frequency of administration is once a day.

[0083] In another embodiment, the compound is 5-((1R, 1aS, 6bR)-1-(6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b-di -hydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one in a crystalline form, selected from the group consisting of Forms Crystals A*A**,A,B,C,D,F,G,H,I,J,K,L,M,N.

[0084] In another aspect, there is provided the use of a combination of formula (I) or formula (II) according to any one of the embodiments described herein in the manufacture of a medicament for the treatment of an associated disease or disorder BRAF, NRAS and KRAS activities.

[0085] In a preferred embodiment, the disease is a cancer.

[0086] In another preferred embodiment, the compound is 5 - (((1R, 1aS, 6bR) -1- (6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b -dihydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one in a crystalline form, selected from the group consisting in A*, A**, A, B, C, D, F, G, H, I, J, K, L, M and N crystalline forms. These and other aspects of the present invention will be better appreciated in light of the following drawing, detailed description and claims.

[0087] BRIEF DESCRIPTION OF THE DRAWINGS

[0088] Figure 1 shows an X-ray diffraction pattern of Compound 1 in amorphous form.

[0089] Figure 2 shows an X-ray diffraction pattern of Crystalline Form A* of Sesqui-Maleate Compound 1 (crystallization from isopropanol/water).

[0090] Figure 3 shows an X-ray diffraction pattern of the hydrochloride salt of Compound 1 (1:1).

[0091] Figure 4 shows an X-ray diffraction pattern of the methanesulfonate salt of Compound 1 (1:1).

[0092] Figure 5 shows an X-ray diffraction pattern of the 2-hydroxyethanesulfonate salt of Compound 1 (1:1).

[0093] Figure 6 shows an X-ray diffraction pattern of the L-tartarate salt of Compound 1 (1: 1)

[0094] Figure 7 shows an X-ray diffraction pattern of Compound 1 oxalate (1:1).

[0095] Figure 8 shows an overlay of X-ray diffraction patterns of solids obtained from Compound 1 with phosphoric acid (1: 1), toluenesulfonic acid (1: 1), ethanedisulfonic acid (1: 1), acid sulfuric (1: 1), API and API suspension control.

[0096] Figure 9 shows the superposition of X-ray diffraction patterns of solids obtained from Compound 1 with L-lactic acid (1: 1), L-maleic acid (1: 1), malonic acid (1: 1: 1), nicotinamide (1:1), saccharin (1:1), succinic acid (1:1), API and API suspension control.

[0097] Figure 10 shows the superimposition of X-ray diffraction patterns of solids obtained from Compound 1 with benzoic acid (1: 1), citric acid (1: 1), fumaric acid (1: 1), acid gentisico (1:1), glycolic acid (1:1), suspension control API and API.

[0098] Figure 11 shows a superimposition of X-ray diffraction patterns of the solid obtained from Compound 1 with nicotinic acid (1: 1), nicotinic acid suspension control, API suspension control and API.

[0099] Figure 12 shows the superimposition of X-ray diffraction patterns of the solid obtained from Compound 1 with adipic acid (1: 1), adipic acid suspension control, API suspension control and API.

[0100] Figure 13 shows the absolute structure of Single Crystalline Form A ** of Sesqui-Maleate of Compound 1 (single crystals obtained by crystallization from acetone).

[0101] Figure 14 illustrates hydrogen bonds of Form A ** single crystalline Sesqui-Maleate of Compound 1.

[0102] Figure 15 shows a crystal packaging of form A ** single crystalline ** Sesqui-Maleate Compound 1.

[0103] Figure 16 shows a theoretical XRPD pattern of single Crystalline Form A** of Compound 1 Sesqui-Maleate calculated using the MERCURY software.

[0104] Figure 17 shows an X-ray diffraction pattern of crystalline Form A of Sesqui-Maleate of Compound 1 (obtained by recrystallization from isopropanol / water).

[0105] Figure 18 shows an X-ray diffraction pattern of crystalline Form B of Compound 1 maleate.

[0106] Figure 19 shows an X-ray diffraction pattern of crystalline Form C of Compound 1 maleate.

[0107] Figure 20 shows an X-ray diffraction pattern of crystalline Form D of Compound 1 maleate.

[0108] Figure 21 shows an X-ray diffraction pattern of crystalline Form F of Compound 1 maleate.

[0109] Figure 22 shows an X-ray diffraction pattern of crystalline Form G of Compound 1 maleate.

[0110] Figure 23 shows an X-ray diffraction pattern of the crystalline H form of Compound 1 maleate.

[0111] Figure 24 shows an X-ray diffraction pattern of crystalline Form I of Compound 1 maleate.

[0112] Figure 25 shows an X-ray diffraction pattern of crystalline Form J of Compound 1 maleate.

[0113] Figure 26 shows an X-ray diffraction pattern of crystalline Form K maleate Compound 1.

[0114] Figure 27 shows an X-ray diffraction pattern of crystalline Form L maleate Compound 1.

[0115] Figure 28 shows an X-ray diffraction pattern of crystalline Form M of Compound 1 maleate.

[0116] Figure 29 shows a 1H-NMR spectrum of Crystalline Form A* of Sesqui-Maleate of Compound 1.

[0117] Figure 30 shows a 13C NMR spectrum of Sesqui-Maleate Crystalline Form A* of Compound 1.

[0118] Figure 31 shows hygroscopicity (ie, moisture absorption) of Crystalline Form A * of Sesqui-Maleate of Compound 1 by DVS.

[0119] Figure 32 shows a 1H-NMR spectrum of Crystalline Form N of the Maleate Salt of Compound 1 (1:1).

[0120] Figure 33 shows an X-ray diffraction pattern of crystalline Form N of Compound 1 maleate (1: 1).

[0121] Figure 34 shows a 1H-NMR spectrum of Sesqui-Maleate Crystalline Form A of Compound 1.

[0122] Figure 35 shows a 1H-NMR spectrum of Crystalline Form B of Compound 1 maleate (1:1.2).

[0123] Figure 36 shows a 1H-NMR spectrum of Crystalline Form C of Compound 1 maleate (1: 1.3).

[0124] Figure 37 shows a 1H-NMR spectrum of Crystalline Form D of Compound 1 maleate (1:1).

[0125] Figure 38 shows a 1H-NMR spectrum of Crystalline Form F of Compound 1 maleate (1:0.5).

[0126] Figure 39 shows a 1H-NMR spectrum of Crystalline Form G of Compound 1 maleate (1:0.5).

[0127] Figure 40 shows a 1H-NMR spectrum of Crystalline Form H of Compound 1 maleate (1: 1).

[0128] Figure 41 shows a 1H-NMR spectrum of Crystalline Form I maleate of Compound 1 (1:0.5).

[0129] Figure 42 shows a 1H-NMR spectrum of crystalline Form J maleate Compound 1 (1: 1).

[0130] Figure 43 shows a 1H-NMR spectrum of Crystalline Form K Compound 1 maleate (1:1).

[0131] Fig. 44 shows a 1H-NMR spectrum of Crystalline L-Form Compound 1 maleate (1:0.3).

[0132] Figure 45 shows a 1H-NMR spectrum of Crystalline Form M of Compound 1 maleate (1:1.3).

[0133] DETAILED DESCRIPTION OF THE INVENTION

[0134] In the course of preparing the salts of the present invention, many salt-forming agents commonly used in the pharmaceutical industry were investigated. In particular, the salt forming agents investigated in the present invention include the following 25 acids or salt forming agents selected from the group consisting of hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, glycolic acid, l-lactic acid, fumaric acid, l-tartaric acid, citric acid, l-maleic acid, succinic acid, hippuric acid, maleic acid, adipic acid, benzoic acid, gentisic acid, malonic acid, ethanedisulfonic acid, toluenesulfonic acid, oxalic acid, nicotinic acid, nicotinamide and saccharin.

[0135] However, the reaction of Compound 1 with only six salts showed the formation of salts. In particular, solid forms of Compound 1 salts with six acids, including hydrochloric acid (Figure 3), methanesulfonic acid (Figure 4), 2-hydroxyethanesulfonic acid (Figure 5), L-tartaric acid (Figure 6), maleic acid ( figure 2, figure 17, etc.) and oxalic acid (figure 7) showed a different crystalline form of free base, API control and solid acid control. In contrast to the above six crystalline salts of the present application, the other salts did not crystallize.

[0136] In one aspect, there is provided Sesqui Maleate Salt Compound 1, which is prepared by the process described herein in bulk quantities. As shown in Figure 2, the Sesqui-Maleate Salt of Compound 1 is in a crystalline form (herein referred to as Crystalline Form A*) and its X-ray powder diffraction spectra typically have the following peak diffraction angles (where " spacing" is shown as the "D value " in figure 2)

[0137] Table 1. X-Ray Diffraction Pattern of Compound 1 Sesqui-Maleate (Crystalline Form A*)

[0138] The Sesqui-Maleate Crystalline Form A* of Compound 1 is a very stable crystalline form. After micronization to obtain well-distributed fine particles with an average particle size (D90) of approximately 1-10 microns, it can be readily formulated into a drug for clinical use.

[0139] In another embodiment, a Sesqui-Maleate Compound 1 Sesqui-Maleate Crystalline Form A** is provided herein. The structure of Sesqui-Maleate Compound 1 Crystalline Form A** was determined using a dataset from diffraction collected from a single cultured crystal by slow cooling in acetone. Crystal data and structure refinement are listed in Table 2.

[0140] Table 2: Single crystal data and structure refinement of Sesqui-Maleate Compound 1 (Crystalline Form A **).

[0141] The structure of a single crystal of the maleate of Compound 1, shown in Figure 13, confirms a sesqui-maleate salt, in which the imidazole nitrogen atom of the free base molecule is protonated. The absolute configurations of C1(S), C2(R), and C3(R) were determined with high probability as indicated by the absolute structure parameter of 0.1 (5). The hydrogen bonding interactions are demonstrated in Figure 14. The structure demonstrates a two-dimensional structure in the bc plane. The free base molecules of compound 1 and maleate anions are bonded by intermolecular interaction (N3 ... N3 2.797A) and hydrogen bonding (N2-H2 ... O4 2.760A) along the b-axis. The strand structures are then bonded by maleate anions through intermolecular interactions (N3...O7 3.008A and N4...O7 2.715A) and hydrogen bonding (N1-H1...O8 2.659A) along the c axis to form a two-dimensional structure. The crystal package of the sesqui-maleate salt of Compound 1 is shown in figure 15. The theoretical PXRD pattern of the single crystalline form A** of Sesqui-Maleate compound 1 calculated using the MERCURY software is shown in figure 16.

[0142] In another embodiment, crystalline Form A of the Sesqui-Maleate of Compound 1 is provided herein. As shown in Figure 17, the X-ray powder diffraction spectra of crystalline Form A typically have the following diffraction angles peak (where "spacing" is shown as the "d-value" in Figure 17):

[0143] Table 3. X-Ray Diffraction Pattern of Compound 1 Sesqui-Maleate (Crystalline Form A).

[0144] Peak # Diffraction angle (2 theta) Spacing Relative to voltage.

[0145] In another embodiment, crystalline Form B maleate of Compound 1 is provided herein. As illustrated in Figure 18, X-ray powder diffraction spectra of crystalline Form B typically exhibit the following peak diffraction angles (where "spacing" is shown as the "d value" in figure 18):

[0146] Table 4. X-ray diffraction pattern of Compound 1 maleate (Crystalline Form B)

[0147] In another embodiment, crystalline Form C of Compound 1 maleate is provided herein. As illustrated in Figure 19, X-ray powder diffraction spectra of crystalline form C typically exhibit the following peak diffraction angles (where "spacing" is shown as the "d value" in Figure 19):

[0148] Table 5. X-ray diffraction pattern of compound 1 maleate (crystalline form C)

[0149] In another embodiment, crystalline Form D maleate of Compound 1 is provided herein. As illustrated in Figure 20, X-ray powder diffraction spectra of crystalline Form D typically exhibit the following diffraction angles of peak (where the "spacing" is shown as the "d value" in figure 20):

[0150] Table 6. X-ray diffraction pattern of compound 1 maleate (crystalline D form)

[0151] In another embodiment, crystalline Form F of Compound 1 maleate is provided herein. As illustrated in Figure 21, X-ray powder diffraction spectra of crystalline form F typically exhibit the following peak diffraction angles ( where "spacing" is shown as the "d value" in figure 21):

[0152] Table 7. X-ray diffraction pattern of compound 1 maleate (crystalline F form)

[0153] In another embodiment, crystalline Form G maleate of Compound 1 is provided herein. As shown in Figure 22, X-ray powder diffraction spectra of crystalline Form G typically exhibit the following peak diffraction angles (where the "spacing" is shown as the "d value" in figure 22):

[0154] Table 8. X-ray diffraction pattern of compound 1 maleate (crystalline G form)

[0155] In another embodiment, crystalline Form H of Compound 1 maleate is provided herein. As shown in Figure 23, X-ray powder diffraction spectra of crystalline Form H typically exhibit the following peak diffraction angles (where "spacing" is shown as the "d value" in Figure 23):

[0156] Table 9. X-ray diffraction pattern of compound 1 maleate (Crystalline Form H)

[0157] In another embodiment, crystalline Form I maleate of Compound 1 is provided herein. As shown in Figure 24, X-ray powder diffraction spectra of crystalline Form I typically exhibit the following peak diffraction angles ( where the "spacing" is shown as the "d value" in figure 24):

[0158] Table 10. X-ray diffraction pattern of Compound 1 maleate (Crystalline Form I)

[0159] In another embodiment, crystalline Form J of Compound 1 maleate is provided herein. As illustrated in Figure 25, X-ray powder diffraction spectra of crystalline Form J typically exhibit the following peak diffraction angles (where the "spacing" is shown as the "d value" in figure 25):

[0160] Table 11. X-ray diffraction pattern of compound 1 maleate (Crystalline Form J)

[0161] In another embodiment, crystalline Form K of Compound 1 maleate is provided herein. As illustrated in Figure 26, X-ray powder diffraction spectra of crystalline Form K typically exhibit the following peak diffraction angles (where "spacing" is shown as the "d value" in figure 26):

[0162] Table 12. X-ray diffraction pattern of compound 1 maleate (Crystalline Form K)

[0163] In another embodiment, crystalline Form L maleate of Compound 1 is provided herein. As shown in Figure 27, X-ray powder diffraction spectra of crystalline form L typically show the following diffraction angles peak (where "spacing" is shown as the "d value" in figure 27):

[0164] Table 13. X-ray diffraction pattern of Compound 1 maleate (Crystalline Form L)

[0165] In another embodiment, crystalline Form M of the maleate of Compound 1 is provided herein. As shown in Figure 28, the X-ray powder diffraction spectra of crystalline Form M typically show the following peak diffraction angles (where "spacing" is shown as the "d value" in figure 28):

[0166] Table 14. X-ray diffraction pattern of compound 1 maleate (Crystalline form M)

[0167] In another embodiment, there is provided herein the Crystalline Form of the maleate of compound 1 (1:1). As shown in figure 33, X-ray powder diffraction spectra of crystalline Form N T typically show the following peak diffraction angles (where the "spacing" is shown as the "D value" in figure 33):

[0168] Table 15. X-Ray Diffraction Pattern of Compound 1 maleate (1: 1) (Crystalline Form N)

[0169] For Crystalline Forms A *, A **, A, B, C, D, F, G, H, I, J, K, L, M and N described above, only the main peaks (i.e., characteristic, significant, unique and/or reproducible maximum peaks) are summarized; additional peaks can be obtained from the diffraction spectra by conventional methods. The main peaks described above can be reproduced within the margin of error (+ or -2 in the last decimal place, or + or - 0.2 in the indicated value).

[0170] In another aspect, a method for preparing the compound of formula (I) or formula (II) is provided herein.

[0171] In one embodiment, there is provided herein a Sesqui-Maleate Crystalline Form A* of Compound 1 prepared or purified according to the procedures described in Scheme 1. The new synthetic methods and crystallization/recrystallization processes described herein surpass many issues associated with previously reported processes, such as preparing the key chiral intermediate with >99% optical purity, and provides many advantages over existing processes. Notably, the methods described herein are especially suited for the reproducible commercial scale manufacture of the Sesqui-Maleate Salt of Compound 1 in high quality and good yields.

[0172] Scheme 1

[0001] Sesqui-maleate salt crystalline forms of compound 1 can be prepared by the following general method: Sesqui-maleate salt crystalline form A* of 1 lb -1 is heated with a solvent until completely dissolved. After filtration, cooling, crystallization, filtration and drying, the corresponding different crystalline forms are obtained. An example of the crystallization process for preparing Crystalline Form A of the sesqui-maleate Salt of Compound 1 is described in Example 3 (below). The crystallization described above can be carried out in a single solvent, a mixture of organic solvents, or a mixture of water and organic solvents. Suitable organic solvents for crystallization may be selected from, but not limited to, lower alkyl alcohols, ketones, ethers, esters, halogenated hydrocarbons, alkanes, halogenated benzene, aliphatic nitrile and other aromatic solvents. Preferred solvents include, for example, isopropanol, ethyl acetate, water, N,N-dimethylformamide, methanol, ethanol, acetone, acetonitrile and mixtures thereof.

[0173] The free base of Compound 1 was originally obtained as an amorphous solid, which is slightly hygroscopic with 2.2% water gain from 0 to 80% RH. No shape transformation was observed after dynamic vapor sorption (DVS). It exhibited a non-birefringence phenomenon with an irregular shape of the polarized light microscope.

[0174] Described herein are the hydrochloride, methanesulfonate, 2-hydroxy-ethanesulfonate, L-tartrate, maleate and oxalate salts of Compound 1, wherein the maleate salt, in particular the sesqui-maleate salt, has surprisingly been found to possess the properties desired characteristics of a drug candidate. For example, Compound 1 did not appear to form a crystalline tartrate salt, and the oxalate salt has poor crystallinity. Although the 2-hydroxyethanesulfonate, mesylate, and HCl salts have relatively good solubility in water, the HCl salt is chemically unstable with significant degradation after one week at 60 °C and 40 °C / 75% RH and 2- hydroxyethanesulfonate and mesylate are both hygroscopic with about 4% weight gain from 0 to 80% RH. The 2-hydroxyethanesulfonate and mesylate salts also showed degradation after one week at 60 °C and 40 °C / 75% RH, and therefore are not expected to have prolonged chemical stability for a long time. On the other hand, the sesqui-maleate salt (1:1.5 free base/maleic acid) has good crystallinity as demonstrated by the XRPD patterns disclosed here, is slightly soluble in water, and showed no degradation after one week at 60° C and 40 °C / 75% RH, indicating possible long-term stability; therefore, the sesqui-maleate salt was chosen for further development.

[0175] If desirable or necessary, the purity of the sesqui-maleate salt of compound 1 could be further improved by converting the sesqui-maleate salt into the maleate salt (1:1 free base/acid ratio) via a process solution methanol suspension. The formation of 1:1 maleate crystalline salt caused efficient expulsion of impurities. The amorphous free base form can also be manufactured in high purity by treating the maleate salt 1:1 with a base and then solvent extraction. Crystalline sesqui-maleate salt with much higher purity was prepared by treating the free base with maleic acid (Scheme 2)

[0176] Scheme 2

[0177] The term "lower alkyl alcohols" herein includes C1-C8, preferably C1-C6, more preferably C1-C8, straight chain or branched alkyl alcohols. Specific examples include, but are not limited to, methanol, ethanol, isopropanol, and butanol.

[0178] The term "approximately", as used herein, unless otherwise indicated, denotes that a number (e.g., temperature, pH, volume, etc.) can vary within ± 10%, preferably within of ± 5%.

[0179] The crystallization of the crystalline forms of the present invention can also be carried out in an appropriate solvent system that contains at least one solvent by evaporating the solvent, cooling and / or by adding anti-solvents (solvents that are less able to solubilize the Compound 1 Sesqui-Maleate, including but not limited to those described herein) to achieve supersaturation in the solvent system.

[0180] Crystallization can be done with or without crystal granules, which is described in the present invention.

[0181] The individual crystalline forms provided by the present invention develop under specific conditions depending on the thermodynamic properties and the particular balance of the crystallization process. Therefore, a person skilled in the art will know that the crystals formed are a consequence of the kinetic and thermodynamic properties of the crystallization process. Under certain conditions (e.g., solvent, temperature, pressure, and compound concentration), a particular crystalline form may be more stable than another crystalline form (or, in fact, more stable than any other crystalline form). However, the relatively low thermodynamic stability of particular crystals can have advantageous kinetic stability. Additional factors other than kinetics, such as time, impurity distribution, agitation, and the presence or absence of crystal granules, etc., can also affect the crystalline form.

[0182] In another aspect, pharmaceutical compositions each containing an effective amount of Compound 1 maleate in any of the Crystalline Forms A*, A**, A, B, C, D, F described above are provided herein. G, H, I, J, K, L, M and N, in particular, Sesqui-Maleate Crystalline Form A* of Compound 1 and a pharmaceutically acceptable carrier. The active compound(s) may be 1-99% (by weight), preferably 1-70% (by weight), or more preferably 1-50% (by weight), or more preferably, 5-40% (by weight) of the composition.

[0183] The pharmaceutical compositions can be administered orally in forms such as capsules, tablets, pills, powders, extended release injection in the form of a sterile solution, suspension or emulsion; through a form of local treatment such as paste, cream or ointment; or through a rectal form, such as suppositories. The pharmaceutical compositions can be in unit dosage form which is suitable for precise dosing applications. In addition, pharmaceutical compositions can include other active ingredients.

[0184] Suitable pharmaceutical carriers include water, various organic solvents, and various inert diluents or fillers. If necessary, the pharmaceutical compositions can contain various additives, such as spices, adhesives and excipients. For oral administration, tablets and capsules may contain various excipients such as citric acid, a variety of disintegrating agents such as starch, alginic acids and certain silicates, and a variety of properties such as sucrose, gelatin and acacia. Additionally, lubricants containing magnesium stearate and talc fillers are commonly used in tabletting. The same types of solid components can also be used to formulate soft and hard gelatin capsules. When an aqueous suspension is required for oral gavage, the active compound can be mixed with a variety of sweetening or flavoring agents, pigments or color combinations. If necessary, a variety of emulsifiers can be employed or suspensions prepared; Diluents such as water, ethanol, propylene glycol, glycerin or a combination thereof may be used.

[0185] The pharmaceutical compositions described above are preferably administered orally.

[0186] The pharmaceutical compositions described above are preferably in capsule or tablet form.

[0187] In another aspect, there is provided herein the use of the compounds of the present application (i.e., the maleate of Compound 1 and any of the Crystalline Forms A*, A**, A, B, C, D described above F, G, H, I, J, K, L, M and N) in the manufacture of medicaments that are useful for treating cancers sensitive to inhibition of Braf kinase or other kinases such as EGFR, VEGFR, EPHA, EPHB, or the like.

[0188] In one embodiment, there is provided herein the use of the compounds of the present application (i.e., the maleate of Compound 1 and any of the Crystalline Forms A*, A**, A, B, C, D described above F, G , H, I, J, K, L, M and N) in the manufacture of medicaments that are useful for the treatment or prevention of mammalian pancreatitis, kidney disease, cancer, angiogenesis or angiogenesis-related diseases.

[0189] The maleate of compound 1 and any of the Crystalline Forms A *, A **, A, B, C, D, F, G, H, I, J, K, L, M and N or Compound maleate 1 (1:1) of the Crystalline Form N of the present application can be used to treat or prevent selected diseases of, but not limited to, tumor angiogenesis, chronic inflammatory disease such as rheumatoid arthritis, atherosclerosis, skin diseases such as psoriasis and scleroderma, diabetes-induced skin diseases, diabetic retinopathy, premature retinopathy, age-related degenerative spots, hemangioma, glioma, internal Kaposi tumor, ovarian cancer, breast cancer, lung cancer, pancreatic cancer, lymphatic cancer , prostate, colon and skin tumors and their complications. Among the mammals mentioned here, humans are preferred.

[0190] The target diseases for the above-described treatment methods are preferably selected from B-RAF, NRAS and K-RAS mutant tumors, such as B-RAF or NRAS or K-RAS non-small cell lung cancer , colorectal cancer, endometrial cancer, colon cancer, esophageal cancer, gastric cancer, colon cancer, rectal cancer, breast cancer, ovarian cancer, melanoma, skin cancer, adrenal cancer, cervical cancer, lymphoma , thyroid tumors and their complications.

[0191] The methods described above can be applied in combination with any chemical therapy (eg MEK inhibitors), biological therapy or radiation therapy.

[0192] The dosage of the active ingredient or compound when administered will be determined by the individual needs of the patient to be treated, by schedule of administration, severity of the disease or malignancy, dosage schedule, as well as the evaluation and judgment of the designated physician. However, based on the active compound, the preferred effective dosage range may be approximately 0.01-120 mg per day per kilogram of body weight; or more preferably 0.1-10 mg per day per kilogram of body weight in single or separate doses. In some cases, it is more suitable to apply the lower levels of the dosage ranges described above, while in other cases the higher doses can be used without causing harmful side effects.

[0193] Another aspect of the present application is the provision of Sesqui-Maleate Compound 1 for clinical applications. In particular, the present invention relates to clinical treatment with Compound 1 maleate with the following treatment options for cancer patients: the dosage of Compound 1 maleate and/or Crystalline form A*, A**, A, B, C, D, F, G, H, I, J, K, L, M and N can be 1-100 mg / day, the frequency of administration being 1 to 3 times a day; a preferred dosage is 5-50 mg/day, with the frequency of administration being 1 to 3 times a day; a more preferred dosage is 5-60 mg/day, the frequency of administration being 1 hour/day; an even more preferred dosage is 10 to 50 mg/day, the frequency of administration being 1 time per day.

[0194] The following synthetic methods, specific examples, and efficacy tests further describe some aspects of the present invention. They should not limit or restrict the scope of the present invention in any way.

[0195] EXAMPLES

[0196] The examples below are intended to be exemplary and efforts have been made to ensure accuracy with respect to the numbers used (eg, range, quantities, temperature, etc.), but some errors and experimental deviations may occur. Unless otherwise noted, temperature is in degrees centigrade. Reagents were purchased from commercial suppliers such as Sigma-Aldrich, Alfa Aesar or TCI, and were used without further purification unless otherwise noted.

[0197] Unless otherwise indicated, the reactions shown below were carried out under a positive pressure of nitrogen or argon or with a drying tube in anhydrous solvents; the reaction flasks were equipped with rubber stoppers for the introduction of substrates and reagents through the syringe; and the utensil has been oven dried and/or dried by application of heat.

[0198] Unless otherwise noted, column chromatography purification was conducted on a Biotage system (Manufacturer: Dyax Corporation) having a silica gel column or on a silica SepPak cartridge (Waters), or was conducted on a Teledyne Isco Combiflash purification system using prepackaged silica gel cartridges.

[0199] 1H NMR and 13 C NMR spectra were recorded on a Varian instrument at 400 MHz using DMSO-d6 as solvent.

[0200] X-ray intensity data from a colorless plateless crystal were measured at 173 (2) K using a Bruker APEX-II CCD diffractometer (radiation Cu K,, À = 1.54178 Â) . The microscopic image of polarized light was captured at room temperature.

[0201] In the following examples, the abbreviations below were used:

[0202] AcOH acetic acid

[0203] ACN Acetonitrile

[0204] The pharmaceutical ingredient API Active, hereinafter referred to as Compound 1

[0205] [Aqueous water]

[0206] Saturated aqueous solution of saturated sodium chloride

[0207] Bn Benzila

[0208] BnBr benzyl bromide

[0209] CH2Cl2 Dichloromethane

[0210] DMF N, N-Dimethylformamide

[0211] Dppf 1.1 "-bis(diphenylphosphine) ferrocene

[0212] [0212] DBU 1,8-diazabicyclo [5.4.0] undec-7-ene

[0213] DIEA or DIPEA N, N-diisopropylethylamine

[0214] DMAP 4-N, N-dimethylaminopyridine

[0215] DMF N, N-dimethylformamide

[0216] DMSO Dimethylsulfoxide

[0217] EtOAc ethyl acetate

[0218] EtOH Ethanol

[0219] Et2O or ether Diethyl ether

[0220] G grams

[0221] [0221] h or time

[0222] HATU O-(7-Azabenzotriazol-1-yl)-N, N, N', N'-tetramethyluronium hexafluorophosphate

[0223] HCl Hydrochloric acid

[0224] High performance HPLC liquid chromatography

[0225] IPA 2-propanol

[0226] i-PrOH Isopropyl alcohol

[0227] Mg milligrams

[0228] [0228] mL of milliliters

[0229] Millimole Millimol

[0230] [0230] MeCN Acetonitrile

[0231] MeOH methanol

[0232] [0232] min minutes

[0233] MS or MS mass spectrum

[0234] Sodium Na2SO4 sulfate

[0235] PE petroleum ether

[0236] [0236] Polyphosphoric acid PPA

[0237] [R] Rt retention time

[0238] Rt or rt Ambient temperature

[0239] [0239] TBAF tetrabutyl ammonium fluoride

[0240] TBSCl tert-butyldimethylsilyl chloride

[0241] TFA Trifluoroacetic acid

[0242] THF Tetrahydrofuran

[0243] TLC Thin Layer Chromatography

[0244] [0244] TMSCl Trimethylsilyl chloride

[0245] μL Microliters

[0246] [0246] Example 1: Formation of Compound 1 Salts with Various Salt Forming Agents

[0247] Example 1A: Preparation of the free base of Compound 1 and Crystalline Form A * of the Sesqui-Maleate of Compound 1

[0248] Step 1: Synthesis of Intermediate 1.

[0249] Intermediary 1

[0250] To a stirred solution of EtONa (154 kg) in DMF (989 kg) was added EtSH (68.6 kg) at an internal temperature < 35 ° C under nitrogen protection. The mixture was stirred for 60 ~ 90 min at internal temperature <35 °C. 5-Methoxybenzofuran (58.75 kg) in DMF (55.0 kg) was added. The mixture was heated to 110-130 °C, stirred for 45 hours and then concentrated under vacuum under 90 °C. After the mixture was cooled to 10 ~ 20 °C, HCl was added dropwise 2N (1326 kg), followed by addition of EtOAc (531 kg) and H2O2 (129 kg) at internal temperature <35°C. The mixture was stirred for 30 ~ 60 min. After separating the organic layer, the aqueous phase was extracted with EtOAc. The combined organic phase was washed twice with saturated brine and then the solvent was evaporated to dryness. MeOH and a solution of NaOH (44.5 kg) in water (185 kg) were added dropwise to the residue below 40 °C. The mixture was stirred for 5-7 hours at 30 ~ 40 °C. active carbon (74 kg) with water (77 kg). The mixture was stirred for 4-6 hours at 30~40 °C and filtered; and the filter cake was washed with MeOH and water. DCM was placed in the filtrate and the pH was adjusted to 1 with 35% aq. HCl below 40 °C. The aqueous phase was extracted with DCM, and the organic phase was washed with 25% NaCl and concentrated below 40 °C. The residue was used in the next step directly. 1H NMR (400 MHz, DMSO-d6) δ 9.14 (s, 1H), 7.86 (d, J = 2.0 Hz, 1H), 7.36 (d, J = 8.8 Hz, 1H), 6.94 (d, J = 2.4 Hz, 1H), 6.79 (dd, J = 2.0, 0.9 Hz, 1H), 6.74 (dd, J = 8.8 , 2.4Hz, 1H) ppm. MS: M / e 135 (M + 1) +.

[0251] Step 2: Intermediate 2 Synthesis

[0252] Intermediary 2

[0253] To a stirred solution of benzofuran-5-ol (Intermediate 1, 33.1 kg) and Et3N (50.8 kg) in DCM (155 kg) was added dropwise a solution of TMSCl (30.4 kg ) in DCM (50 kg) at -5 to 0°C. The mixture was warmed to 0 ~ 10 °C and stirred at the temperature for 2 hours (IPC found INT-1 / INT-2 = 37.4%). The mixture was cooled to between -5 and 0 °C and a solution of TMSCl (10.6 kg) in DCM (8 kg) was added dropwise, and then the mixture was warmed to 0 ~ 10 °C and stirred at room temperature for 1 h. The mixture was concentrated below 40 °C, and n-heptane was added to the mixture. The mixture was stirred for 20-30 min and filtered, and the filter cake was washed with n-heptane. The solvent was distilled off the filtrate to obtain crude intermediate 2 (INT-2%: 62.7%, KF: 0.01%). To a stirred solution of crude intermediate 2 above and Et3N (8.6 kg) in DCM (149 kg) was added dropwise a solution of TMSCl (9.0 kg) in DCM (10 kg) at -5 to 0 °C. The mixture was warmed to 0 ~ 10 °C and stirred at room temperature for 1 h (TLC showed that the reaction was over). The reaction mixture was concentrated below 40°C, and to the mixture was added n-heptane. The mixture was stirred for 20 to 30 minutes and then filtered, and the filter cake was washed with n-heptane. The solvent was distilled off the filtrate to obtain Intermediate 2 (41.5 kg, INT-2%: 98.1%) as a colorless oil. 1H NMR (400 MHz, DMSO-d6) δ 7.69 (d, J = 2.0 Hz, 1H), 7.21 (d, J = 8.8 Hz, 1H), 6.84 (d, J = 2.5 Hz, 1H), 6.61 (d , J = 2.0 Hz, 1H), 6.56 (dd, J = 8.8, 2.5 Hz, 1H), 0.00 ( s, 9H) ppm.

[0254] Step 3: Intermediate 3 Synthesis

[0255] Intermediary 3

[0256] Copper triflate (I) triphate (2:1 complex with toluene, 0.41 kg) and (S, S)-Vans binder (0.552 kg) were stirred in DCM (160 kg) at room temperature under N2 atmosphere for 1 -2 hrs. Intermediate 2 (37.0 kg) was added followed by slow addition of ethyl diazoetanoate (58 kg) in DCM (450 kg) at 20 ~ 30 °C. The reaction was stirred for 0.5 ~ 1 h at 20 ~ 30oC (CPI: INT-2 / INT-3<0.2%, residual N2CHCO2Et: 0.05% <1.0%). A disodium EDTA solution (0.05 mol/L, 150 kg) was added to the reaction mixture for 40~50 min at 20~30oC in three times. The organic phase was washed with 25% aqueous NaCl at 20~30oC twice and concentrated below 30°C. The residue was distilled under reduced pressure and the crude Intermediate 3 (36.26 kg, 84.5%) was collected at 120 ~ 144°C. The crude compound incorporated the endo-enantiomer which could be removed in the next step. 1H NMR (400 MHz, DMSO-d6) δ 6.79 (d, J = 2.4 Hz, 1H), 6.59 (d, J = 8.4 Hz, 1H), 6.42 (dd, J = 8.4, 2.4 Hz, 1H), 4.95 (dd, J = 5.4, 1.0 Hz, 1H), 3.08 (dd, J = 5.4, 3.2 Hz , 1H), 1.02 (dd, J = 3.1, 1.2 Hz, 1H), 0.00 (s, 9H) ppm.

[0257] Steps 4 and 5: synthesis of intermediate 5 and intermediate 6

[0258]

[0259] Intermediate 5 Intermediate 6

[0260] To a solution of Intermediate 4 (36.3 kg) in MeOH (108 kg) was added HCl / MeOH solution (5M, 0.11 kg) at 20 ~ 30 °C and the mixture was stirred for 2 -3 hours (IPC: L/M: 0.5%, chiral purity 90.0%). Et3N (0.22 kg) was added dropwise at 20 ~ 30 °C. The mixture was concentrated and the residue was diluted with n-heptane / EtOAc (4: 1) and then concentrated. After adjusting the temperature to 10 ~ 20 °C and stirring for 2 ~ 4 hours at 10 ~ 20 °C, the mixture was filtered to give a wet product (Intermediate 5: 94.0%, chiral purity: 90.5% ). 1H NMR (400 MHz, DMSO-d6) δ 9.05 (s, 1H), 6.89 (d, J = 2.4 Hz, 1H), 6.72 (d, J = 8.4 Hz, 1H), 6.55 (dd, J = 8.4, 2.4 Hz, 1H) dd, J = 5.4, 1.0 Hz, 1H), 3.27 (dd, J = 5.4, 3.0 Hz, 1H), 1.19-1.17 (m, 1H) ppm. The crude product was suspended with n-heptane/EtOAc (20:1) three times to give a pale yellow solid, which was dried for 12 ~ 16 hours at 40 ~ 50 °C to yield 16.55 kg of product (Intermediate 6: 98.6%, chiral purity: 99.3%).

[0261] Steps 6 and 7: synthesis of intermediate 7 and intermediate 8

[0262] Intermediate 7 Intermediate 8

[0263] To a solution of Intermediate 6 (14 kg) and 5-fluoro-3,4-dihydro-1,8-naphthyridin-2(1H)-one (SM2, 11.2 kg) in DMF (66 kg) Cs2CO3 (26 kg) was added at 40 ~ 60 °C, and the mixture was heated to 110 ~ 120 °C and stirred for 3 hours at 110 ~ 120 °C. The reaction pH was adjusted to 6 with acetic acid ( 12.0 kg) at 25 ~ 35 °C. Water (520 kg) was added and the mixture was stirred for 1 to 2 hours. After filtration, the solid was slurried with EA (78kg) to obtain a wet product (the purity: (Intermediate 7 + Intermediate 8)%: 98%). An aqueous sodium hydroxide solution (125 kg, 2 M) was added to a stirred solution of the wet product in THF (240 kg) and stirred for 2 ~ 3 hours at 20 ~ 30 °C (IPC: INT-7 / INT -8: 0.9%). The mixture was adjusted to pH 4 ~ 5 with 4N HCl (37 kg) at 20 ~ 30 °C and then stirred for 0.5 ~ 1 h. The mixture was concentrated below 50 °C and a solid precipitated out of solution. After filtration, the wet product was resuspended in THF at 35 ~ 45 °C for 1 to 2 hours and then filtered. The resulting wet product was dried for 40 hours at 45 ~ 65 °C to provide the title compound Intermediate 8 (18.95 kg: chemical purity 99%, chiral purity 100%). 1H NMR (400 MHz, DMSO-d6) δ 12.59 (s, 1H), 10.43 (s, 1H), 7.92 (d, J = 5.8 Hz, 1H), 7.29 ( d, J = 2.4 Hz, 1H), 6.97 (d, J = 8.8 Hz, 1H), 6.93 (dd, J = 8.8, 2.4 Hz, 1H), 6, 21 (d, J = 5.8 Hz, 1H), 5.21 (dd, J = 5.4, 1.0 Hz, 1H), 3.27 - 3.25 (m, 1H), 2.89 (t, J = 7.8 Hz, 2H), 2.51 (d, J = 8.8 Hz, 2H), 1.19 (dd, J = 3.0, 1.0 Hz, 1H) ppm. MS: M / e 339 (M + 1) +.

[0264] Step 8: Intermediate 9 Synthesis

[0265] Intermediary 9

[0266] A solution of Intermediate 8 (13.3kg), DIPEA (16kg) and HATU (18.1kg) in DMF (167kg) was added dropwise to a mixture of 4-(trifluoromethyl)benzene-1,2 -diamine (SM3, 7.6 kg) in DMF (74 kg) at 0 ~ 15 °C. The mixture was stirred at 20 ~ 25 °C for 4 ~ 6hrs (IPC: INT-9 / INT-9: not detected ). Active carbon (5.3 kg) in DMF (7.5 kg) was added to the reaction mixture, stirred for 2 ~ 4 hours at 40 ~ 45 °C, and then filtered. Water (846 kg) was added dropwise to the filtrate at 15 ~ 30°C and a solid precipitated out of solution when stirred for 1 to 2 hours. The precipitate was collected and slurried in EtOH at 20~30°C for 2-4 hours. After filtration, the wet product was dried for 37 hours at 45 ~ 60 °C to obtain the title compound Intermediate 9 (17.60 kg: 95.5%).

[0267] Step 9: Synthesis of free base of Compound 1

[0268]

[0269] Compound 1 free base

[0270] A solution of Intermediate 9 (17 kg) and water (1.5 kg) in AcOH (360 kg) was stirred at 65-70°C for 20 hours (IPC: R/S < 1.0%). The mixture was concentrated to dryness below 55 °C and active carbonate (17 kg) with MeOH (32 kg) was added to the residue. The mixture was stirred for 1 h at about 50 °C. After filtration, the filtration was concentrated to remove the solvent at a temperature below 45 °C. EA (160 kg) and water (330 kg) were added to the residue, followed by an aqueous solution of NaOH (2 mol / L) to pH 8-9 at 20-30oC. The organic layer was separated and the aqueous phase was extracted with EA. The combined organic phase was washed with water twice, concentrated to dryness to give Compound 1 as the free base. 1H-NMR (600 MHz, DMSO-d6) δ 12.84 (s, 1H), 10.47 (s, 1H), 7.98 (d, J = 5.8 Hz, 1H), 7.86 ( d, J = 1.2 Hz, 1H) 7.69 (d, J = 2.6 Hz, 1H), 7.08 (d, J = 8.7 Hz, 1H), 7.02 (dd, J = 8.7 , 2.6 Hz, 1H), 6.29 (d, J = 5.8 Hz, 1H), 5.43 (dd, J = 5.4, 1.2 Hz, 1H), 3 .55 (dd, J = 5.3, 3.3 Hz, 1H), 2.95 (t , J = 7.7 Hz, 2H), 2.55 (t, J = 7.7 Hz, 2H) , 1.97 (d, J = 1.3 Hz, 1H) ppm.

[0271] Step 10: synthesis of Sesqui-Maleate of compound 1

[0272] Compound 1 Sesqui-Maleate

[0273] IPA (83 kg) was added to the residue from step 9. Maleic acid (5 kg) in water (29 kg) was added to the mixture and stirred for 4 hours at about 50 °C and then cooled to 35oC and stirred for 12 hours at that temperature. The resulting solid was filtered, dried at 40 ~ 60oC and micronized in a micronizer to provide a white powder (Sesqui-Maleate Salt of Compound 1, 8.36 kg) with particle sizes of D90 = 4.1μm, D10 = 1.5μm, D50 = 2.4μm. The X-ray powder diffraction pattern method was used to characterize the structure of Crystalline Form A*, see Figure 2. 1H-NMR spectra for Crystalline Form A* of the Sesqui-Maleate of Compound 1 are shown in Figure 29 13 C NMR spectra for Sesqui-Maleate Crystalline Form A* of compound 1 are shown in Figure 30.

[0274] The solubility study showed a much better aqueous solubility (0.020 mg / mL) of Crystalline Form A * of the Sesqui-Maleate of Compound 1 obtained, than the (<LOQ at 0.001 mg / mL) of the free base of Compound 1.

[0275] Sesqui-Maleate Crystalline Form A* of Compound 1 was found to be non-hygroscopic with only 0.1801% absorption of water exposed to 95% RH as shown in figure 31.

[0276] Example 1B: Preparation of Compound 1 salts with other salt-forming agents

[0277] About 150 mg of Compound 1 free base were dissolved in a 40 mL glass vial with 2 mL of IPA individually. A solution of appropriate salt forming agents in IPA (Salt forming agents were pre-dissolved in IPA, for salt forming agents that could not be dissolved, suspensions were heated to dissolve) according to the molar ratio listed in Table 16, was slowly titrated into the free base solution, respectively, on the magnetic stirrer, and then under stirring at room temperature for 24 hours to precipitate the solid. If no solids were obtained, an anti-solvent (eg heptane) would be added slowly into the solution to obtain precipitation. The centrifugal solid was determined by XRPD to determine if the new crystalline form was obtained and then dried under vacuum at 40 °C overnight for further characterization. API and pure solid acid suspended in IPA were used as API control and pure solid salt forming agent control to distinguish salt / co-crystal formation and polymorphism of API or solid salt forming agents.

[0278] In addition to the formation of crystalline salts with maleic acid, solids of crystalline forms were also found from the following five acids, including hydrochloric acid (figure 3), methanesulfonic acid (figure 4), 2-hydroxyethanesulfonic acid (figure 5) , L-tartaric acid (figure 6, the true tartrate salt may not be formed and is just the polymorph of the free base, since no chemical change to tartrate was detected) and oxalic acid (figure 7), which presented different crystal forms of free base, API control and solid acid control. Among the crystalline salts formed, 2-hydroxyethanesulfonate and maleate have better crystallinity than the other four (hydrochloric acid, methanesulfonic acid, L-tartaric acid and oxalic acetic acid). The solids obtained from other salt forming agents were all amorphous in form (figure 8, figure 9, figure 10) or the same form with pure solid salt forming agents (figure 11 and figure 12), suggesting free forms or amorphous salts or obtaining the mixture of salt forming agents with free form.

[0279] Table 16. Summary of Compound 1 salt formation with various salt-forming agents

[0280] Example 2: Preparation of Sesqui-Maleate Single Crystal Form A ** of compound 1

[0281] A single crystal growth screening was performed under 94 different conditions, varying solvent, temperature and recrystallization methods, from which single crystals suitable for structure determination were obtained by slow cooling in acetone.

[0282] Example 3: Preparation of Sesqui-Maleate Crystalline Form A of Compound 1

[0283] The free base of Compound 1 (145 g) was combined with 63.5 g of maleic acid (1.8 equivalents) in 1750 mL of i-PrOH / H 2 O (V: V = 4: 1). The mixture was refluxed until all solids dissolved. The clear solution was cooled to room temperature, some granules of crystals were added, and then the mixture was allowed to stand for 24 hours. White solids were precipitated and filtered. The filter cake was washed with about 500 ml of i-PrOH / H 2 O (V:V = 4:1) and dried under high vacuum at 50°C for 48 hours to give the title product (112 g) as crystalline crystals. A powder X-ray diffraction pattern method was used to characterize the structure of Crystalline Form A; see figure 17. The TGA result indicated a weight loss of 0.3 wt% up to 120°C. The DSC result showed a melting endotherm at 194.2°C (onset temperature). The 1H-NMR spectrum showed a stoichiometric ratio of 1 / 1.5 free base / maleic acid (see figure 34).

[0284] Example 1A is directed to the mass production of the crystalline form of the Sesqui-Maleate of Compound 1 (i.e. designated as Form A*) and Example 3 is directed to the laboratory preparation of the crystalline form of the Compound 1 Sesqui-Maleate (ie, designated as Form A). X-ray powder diffraction pattern, 1H-NMR spectra and other techniques show the consistency between the two crystalline forms of Crystalline Form A* and Crystalline Form A.

[0285] Example 4: Preparation of Crystalline Form B of Compound 1 maleate

[0286] Crystalline Form B of Compound 1 maleate was prepared by cooling a 1,4-dioxane solution of Crystalline Form A of Sesqui-Maleate from 60 °C to 5 °C at a rate of 0.1 ° C / min for crystallization. Procedures: Weigh 20.0 mg of Form A solids into a 3 mL glass vial and add 1 mL of 1,4-dioxane to the vial; stir the mixture at 50 °C magnetically with a speed of 800 RPM; Sample filtration after equilibration at 60 °C for 2 hours using 0.45 µm nylon membrane; and cooling the filtrate at a rate of 0.1°C/min from 60°C to 5°C. The sample was stored at 5°C prior to isolation. A powder X-ray diffraction pattern method was used to characterize the structure of Crystalline Form B; see Figure 18. The TGA result indicated a weight loss of 15.1% by weight up to 150°C (containing molecules of 1,4-dioxane crystal solvate/hydrate). The DSC result showed multiple overlapping endotherms (181.4 and 189.6 °C) before decomposition. The 1H-NMR spectrum showed a stoichiometric ratio of about 1 / 1.2 free base / maleic acid (see figure 35).

[0287] Example 5: Preparation of Crystalline Form C of Compound 1 maleate

[0288] Crystalline Compound 1 maleate Form C was prepared by cooling an ethanol solution of Compound 1 from 60 °C to 5 °C at a rate of 0.1 °C / min for crystallization. Procedures: Weigh 21.3 mg of Form A solids into a 3 mL glass vial and add 1 mL of ethanol to the vial; stir the mixture at 50 °C magnetically with a speed of 800 RPM; filter the sample after equilibration at 60 °C for 2 hours using a 0.45 μm nylon membrane; and cooling the filtrate at a rate of 0.1°C/min from 60°C to 5°C. The sample was stored at 5°C prior to isolation. A powder X-ray diffraction pattern method was used to characterize the structure of Crystalline Form C; see Figure 19. The TGA result indicated a weight loss of 7.0 wt% up to 180 °C (containing molecules of crystal hydrate). The DSC result showed four endotherms (107.5, 162.7, 179.3 and 196.0 °C) before decomposition. The 1H-NMR spectrum showed a stoichiometric ratio of about 1 / 1.3 free base / maleic acid (see figure 36).

[0289] Example 6: Preparation of crystalline Form D of Compound 1 maleate

[0290] Compound 1 Maleate Crystalline Form D was prepared by slowly evaporating the MeOH solution of Compound 1 Sesqui-Maleate Crystalline Form A in the presence of ionic liquid [dmin]CF3COO at RT. Procedures: Weigh 150.2 mg of Form A solids into a 20 mL glass vial and add 10.2 mL of MeOH to the vial; Filter the mixture to obtain a saturated Stoke's solution using a 0.45 µm nylon membrane; add 2.8 mg ionic [dmin]CF3COO to the 1 mL MeOH stock solution; and stirring under TA to induce precipitation. A powder X-ray diffraction pattern method was used to characterize the structure of Crystalline Form D; see Figure 20. The TGA result indicated a weight loss of 6.3 wt% up to 120 °C (containing molecules of crystal hydrate). The DSC result showed two endotherms (75.8 and 161.4 °C) before decomposition. The 1H-NMR spectrum showed a stoichiometric ratio of about 1/1 free base / maleic acid (see figure 37).

[0291] Example 7: Preparation of crystalline Form F F of Compound 1 maleate

[0292] Compound 1 Maleate Crystalline Form F was prepared by crystallizing Compound 1 Sesqui-Maleate Crystalline Form A from an ACN/H2O solution (1:1, v/v). Procedures: Weigh 18.9 mg of Form A solids into a 3 mL glass vial and add 1.5 mL of ACN/H2O (1:1, v/v); and add about 2 mg of polymer blend (polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), polyvinyl chloride (PVC), polyvinyl acetate (PVAC), hypromellose (HPMC) and methylcellulose (MC) to a mass ratio of 1:1:1:1:1:1) into the suspension and stir at RT to cause precipitation.

[0293] An X-ray powder diffraction pattern method was used to characterize the structure of Crystalline Form F; see Figure 21. The TGA result indicated a weight loss of 3.2 wt% up to 130 °C (containing molecules of ACN solvate / hydrate). The DSC result showed three endotherms (62.4, 156.9 and 169.3 °C) before decomposition. The 1H-NMR spectrum showed a stoichiometric ratio of about 1 / 0.5 free base / maleic acid (see figure 38).

[0294] Example 8: Preparation of Crystalline Form G of Compound 1 maleate

[0295] Crystalline Form G of the maleate of Compound 1 was pre-prepared by adding water to the acetic acid solution of Crystalline Form A of the Sesqui-Maleate of Compound 1. [0297] Procedures: weigh 20.2 mg of solids of Form A in a 20 mL glass vial; add 1.0 mL of acetic acid to the vial and shake at RT to obtain a clear solution; and add 5 mL of water into the stepwise solution to precipitate the ingestion.

[0296] An X-ray powder diffraction pattern method was used to characterize the structure of Crystalline Form G; see figure 22. The TGA result indicated a weight loss of 11.0 wt% up to 120°C (containing molecules of crystal hydrate). The DSC result showed four endotherms (84.3, 126.9, 139.4 and 187.3 °C) before decomposition. The 1H-NMR spectrum showed a stoichiometric ratio of about 1 / 0.5 free base / maleic acid (see figure 39).

[0297] Example 9: Preparation of Crystalline Form H of Compound 1 maleate

[0298] Crystalline Form H of Compound 1 maleate was prepared by suspending Crystalline Form A of Compound 1 Sesqui-Maleate in THF at RT. Procedures: Weigh 17.7 mg of Form A solids into a 1.5 mL flask and add 0.3 mL of THF to the flask to obtain a suspension; and stir the mixture with RT magnetically at a speed of 800 RPM for 3 days. A powder X-ray diffraction pattern method was used to characterize the structure of Crystalline Form H; see Figure 23. The TGA result indicated a weight loss of 13.0 wt% up to 150°C (containing molecules of THF crystal solvate / hydrate). The DSC result showed 2 endotherms (124.5 and 178.0 °C) before decomposition. The 1H-NMR spectrum showed a stoichiometric ratio of about 1/1 free base / maleic acid (see figure 40).

[0299] Example 10: Preparation of Crystalline Form I of Compound 1 maleate

[0300] Compound 1 Maleate Crystalline Form I was prepared by slowly evaporating an IPA/H2O (3:1, v/v) solution of Compound 1 Sesqui-Maleate Crystalline Form A at RT. Procedures: Weigh 13.8 mg of Form A solids into a 3 ml glass vial and add 1.5 ml of IPA/H2O (3:1, v/v) into the vial to obtain a clear solution; Evaporate solvent from solution at RT to induce precipitation. A powder X-ray diffraction pattern method was used to characterize the structure of Crystalline Form I; see Figure 24. The TGA result indicated a two-cycle weight loss of 20.7 wt% at 180 °C (containing molecules of crystal IPA solvate / hydrate). The DSC result showed four overlapping endotherms (85.8, 115.1 and 138.2 °C) before decomposition. The 1H-NMR spectrum showed a stoichiometric ratio of about 1 / 0.5 free base / maleic acid (see figure 41).

[0301] Example 11: Preparation of Crystalline Form J of Compound 1 maleate

[0302] Crystalline Form J of Compound 1 maleate was prepared through the interaction between the solids of Crystalline Form A of Compound 1 maleate and acetic acid vapor. Procedures: Weigh 14.2 mg of Form A solvents into a 3 mL glass vial; seal the 3 mL vial in a 20 mL glass vial containing 2 mL of acetic acid; and keep the system under AT for 8 days, allowing the steam to interact with solids. A powder X-ray diffraction pattern method was used to characterize the structure of Crystalline Form J; see figure 25. The TGA result indicated a weight loss of 19.3% by weight up to 130°C (contained molecules of acetic acid solvate / acetic acid hydrate). The DSC result showed three endotherms (102.9, 155.7 and 187.7 °C) before decomposition. The 1H-NMR spectrum showed a stoichiometric ratio of about 1/1 free base / maleic acid (see figure 42).

[0303] Example 12: Preparation of Crystalline Form K of Compound 1 maleate

[0304] Compound 1 maleate crystalline Form K was obtained after storage of Compound 1 maleate crystalline Form D under ambient conditions for two weeks. A powder X-ray diffraction pattern method was used to characterize the structure of Crystalline Form K; see Figure 26. The TGA result indicated a weight loss of 4.2% by weight up to 150°C (containing molecules of crystal hydrate). The DSC result showed two endotherms (59.6 and 163.5 °C) before melting at 186.6 °C (maximum temperature). The 1H-NMR spectrum showed a stoichiometric ratio of about 1/1 free base / maleic acid (see Figure 43).

[0305] Example 13: Preparation of the Crystalline Form L of the Maleate of Compound 1

[0306] Compound 1 Maleate Crystalline Form L was obtained by heating Compound 1 Maleate Crystalline Form G to 140 °C and cooling under RT. A powder X-ray diffraction pattern method was used to characterize the structure of Crystalline Form L; see Figure 27. The TGA result indicated a weight loss of 3.9% by weight up to 120°C (containing molecules of crystal hydrate). The DSC result showed an endotherm (141.1 °C, onset temperature) before melting at 183.9 °C (maximum temperature). The 1H-NMR spectrum showed a stoichiometric ratio of about 1 / 0.3 free base / maleic acid (see figure 44).

[0307] Example 14: Preparation of Crystalline Form M of Compound 1 maleate

[0308] Crystalline Form M of Compound 1 Maleate was obtained after storing Crystalline Form J of Compound 1 maleate under ambient conditions for two weeks. A powder X-ray diffraction pattern method was used to characterize the structure of Crystalline Form L; see figure 28. The TGA result indicated a weight loss of 1.4 wt% up to 140 °C (containing molecules of crystalline acetic acid hydrate / hydrate). The DSC result showed multiple endotherms (123.0, 156.5, 171.9, 176.1 and 195.3 °C). The 1H-NMR spectrum showed a stanchiometric ratio of about 1 / 1.3 free base / maleic acid (see figure 45).

[0309] Example 15: Scan of conditions for the formation of Compound 1 maleate and sesqui-maleate

[0310] Condition 1: Compound 1 reacted with maleic acid in EtOAc.

[0311] To a stirred solution of Compound 1 (47.8 mg) in EtOAc (5 mL) was added a solution of maleic acid in EtOAc (0.2 mol/L, v mL) at room temperature. The mixture was stirred for 30 min. The solid was filtered and dried under high vacuum to give the title salt. Surprisingly, when 0.5, 0.9, 1.8 or 3.0 eq. of maleic acid was used, a crystalline Sesqui-Maleate salt of Compound 1 (1:1.5) was obtained under all conditions.

[0312] Condition 2: Compound 1 was reacted with maleic acid in EtOH.

[0313] Procedure: A mixture of Compound 1 and maleic acid in EtOH was refluxed for 20 minutes until all solids dissolved. The solution was cooled to room temperature and stood for 3 hours. A white solid precipitated, which was filtered and dried under high vacuum to provide the title compound. It was surprisingly found that when one equivalent of maleic acid was used, a crystalline maleate salt of Compound 1 was obtained (1:1); but when the amount of maleic acid was increased to 1.3, 1.5 or 3.5 eq., all conditions provided a crystalline Sesqui-Maleate salt of Compound 1 (1:1.5) as the product.

[0314] Condition 3: Compound 1 reacted with maleic acid in MeOH.

[0315] Procedure: A mixture of Compound 1 and maleic acid in MeOH was refluxed for 20 min until all solids dissolved. The solution was cooled to room temperature and allowed to stand for 3 hours. A white solid precipitated out, which was filtered and dried under high vacuum to provide a maleate salt of Compound 1 (1:1). Surprisingly, using MeOH as solvent, when 1.0, 1.5 or 3.5 eq. of maleic acid, the product was consistently a powdered Compound 1 maleate salt (1:1).

[0316] Condition 4: Compound 1 reacted with maleic acid in i-PrOH / water.

[0317] Process: A mixture of Compound 1 (145 g, 0.3 mol) and maleic acid (63 g, 0.54 mol) in a mixed solvent (i-PrOH: H2O = 4: 1 by volume, 1, 7 L) was refluxed until all solid dissolved. The mixture was allowed to cool to room temperature, some granules of crystals were added to the solution and the mixture was allowed to stand for 24 hours. A white crystal precipitated and was filtered. The filter cake was washed with 500 mL of the mixed solvent, dried under high vacuum at 50 °C for 48 hours to give Pa-1 sesqui-maleate salt (112 g, 56.7%) as crystalline needles. Interestingly, when 40% by volume of i-PrOH was used as solvent, crystalline hemi-maleate salt of compound 1 (1:0.5) was obtained; whereas when 60vol% i-PrOH, 90vol% i-PrOH or 100% i-PrOH were used as solvent, all conditions provided a crystalline (1:1.5) sesqui-maleate salt of Compound 1.

[0318] Condition 5: compound 1 reacted with maleic acid in other solvents.

[0319] When other solvents (THF, acetone, DME, 1,4-dioxane) were used, according to similar procedures described above, the product obtained was consistently a Sesqui-Maleate salt of Compound 1 (1: 1.5 ).

[0320] Condition 6: Conversion of the Sesqui-Maleate Salt of Compound 1 (1: 1.5) into the Maleate Salt (1: 1).

[0321] A residue of Sesqui-Maleate of Compound 1 (1.0 g, 1.53 mmol) was suspended in MeOH (20 mL) and the mixture was stirred at room temperature for 1 day. The white solid was filtered. The filter cake was washed with MeOH (10 mL) and dried under infrared lamp at 50°C for 16 hours to provide maleate salt of Compound 1 (1:1) (820 mg, 90%) as a white crystalline solid . 1H NMR (400 MHz, dmso) δ 10.50 (s, 1H), 7.97 (d, J = 5.6 Hz, 1H), 7.86 (s, 1H), 7.68 (d, J = 8.4 Hz, 1H), 7.47 (dd J = 8.4, 1.2 Hz, 1H), 7.38 (d, J = 2.4 Hz, 1H), 7.08 (d , J = 8.8 Hz, 1H), 7.01 (dd, J = 8.8, 2.4 Hz, 1H), 6.31 - 6.24 (m, 3H), 5.43 (dd, J = 5.2, 1.2 Hz, 1H), 3.55 (dd, J = 5.2, 3.2 Hz, 1H), 2.94 (t, J = 7.6 Hz, 2H), 2.54 ( t, J = 7.6 Hz, 2H), 1.97 (dd, J = 3.2, 1.2 Hz, 1H) (detailed spectrum see figure 32).

[0322] An X-ray powder diffraction pattern study was used to determine the structure of the white crystalline solid resulting under Condition 6 and named Crystalline Form N (1: 1) of Compound 1 (figure 33)

[0323] Condition 7: Conversion of Maleate Salt of Compound 1 (1: 1) into Sesqui-Maleate Salt.

[0324] A mixture of maleate salt of Compound 1 (1: 1) (4.3 g) and maleic acid (0.67 g, 0.8 eq of maleic acid) in a mixture of i-PrOH / H2O (4 : 1, v/v, 50 mL) was refluxed for 1 hour. A small amount of solid remained. An additional 5 mL of the mixed solvent was added and the resulting mixture was refluxed for 30 minutes, and all solids dissolved. The mixture was allowed to cool to room temperature and was then allowed to stand for 16 hours. A white crystal precipitated out and was filtered. The filter cake was washed with 20 mL of the mixed solvent, dried under an infrared lamp for 24 hours to provide the Sesqui-Maleate salt of Compound 1 (3.2 g, 68%) as crystalline needles.

[0325] Example 16

[0326] Expulsion of impurities from compound 1 API: 28.5 kg of crystallized sesqui-maleate (chemical purity = 98.82%) was stirred with methanol at 45-50 °C for 4-5 h. The suspended mixture was then cooled to 20-30 °C for 3-4 h, filtered to give a wet cake (30.3 Kg, chemical purity = 99.76%). The wet cake was mixed with ethyl acetate and water, treated with aqueous sodium carbonate until pH = 7-8 and then washed with water. Compound 1 free base (16.2 kg, chemical purity = 99.81%) in ethyl acetate was obtained. Ethyl acetate was replaced by i-propanol by distillation. The free base in i-PrOH was stirred with an aqueous maleic acid solution and crystal granules of the sesquimaleate. The mixture was stirred at 40-50°C for a few hours and at 20-30°C for a few hours. The resulting solid was filtered to provide the 2nd crystallized sesqui-maleate salt (19.6 Kg, Chiral Purity = 99.82%, Optical Purity = 100%)

[0327] Example 17: Stability Study of Salts or Crystalline Forms of Compound 1

[0328] Example 17A: Short-Term Stability Study

[0329] A short-term stability study was conducted to evaluate the HCl salt, the mesylate salt, the 2-hydroxyetensulphonate salt and the maleate salt.

[0330] Weigh about 2 mg (for XRPD: 10 mg) of each of the above salts into a 40 mL glass vial separately. The free base form was used as a control. Closed samples were covered and sealed with parafilm, while open samples were covered with aluminum foil with a pinhole hole. All samples were stored in the stability chamber or in the dry oven under the corresponding conditions (60 °C and 40 °C / 75% RH) for 1 week.

[0331] The HCl salt appeared to be light yellow powder in both conditions after 1 week, the appearance of 2-hydroxyethanesulfonate mesylate and maleate showed no change under visual observation. XRPD results showed that no crystalline form transformation was detected for all salt samples.

[0332] The chemical stability results are compiled in Table 17. Variable degradation was detected with increasing TRS (HCl salt > 2-hydroxyetensulphonate > mesylate > maleate > free base). In contrast, the temperature stress condition may have a greater impact than the moisture stress condition for Compound 1 salt formation, since transmission Raman spectroscopy (TRS) of salt samples increased more than 60 ° C.

[0333] Table 17. Summary of the short-term stability study Crystalline form A* of the compound Sesqui-Maleate of compound 1 0.08 0.17 0.10

[0334] Example 17B: Long-Term Stability Study

[0335] Long-term stability studies of Crystalline Form A * of Sesqui-Maleate of Compound 1 showed that there was no significant change in chemical purity that occurred when stored at 25 ° C ° C / 60% RH for up to 18 months (Assay w/w: T0 = 99.1% and T18 = 98.8%) and at 40°C / 75% RH condition for up to 6 months (Assay w/w: T0 = 99.1% and T6 = 98.9%). The results further showed that Crystalline Form A* of Compound 1 Sesqui-Maleate still maintained 98.5% purity after the 24 month stability study at 25°C / 75% RH. Furthermore, no crystal shape and optical purity changes were observed when stored at 25 °C / 60% RH for up to 18 months and at 40 °C / 75% RH for up to 6 months.

[0336] Efficacy Tests

[0337] Test 1: Inhibition and Selectivity of Kinases by Sesqui-Maleate of Compound 1 (Crystalline Form A * tested)

[0338] Methods:

[0339] RaF kinase enzymatic assays:

[0340] Crystalline Form A * of Compound 1 sesqui-maleate was tested against recombinant B-Raf (V600E) (PV3849), C-Raf (Y340D / Y341D) (PV3805) and wild-type Braf (PV3848) from of Life Technologies in kinase activity assays based on homogeneous fluorescission methodology (HTRF) (Cisbio Bioassays). Assays were performed in a reaction mixture containing RAF kinases, ATP at KM concentration, GST-labeled MeK1 (K97R) and Crystalline Form A* of Compound 1 Sesqui-Maleate or DMSO in buffer containing 25 mM Tris pH 7.4 , 10 mM MgCl2, 0.5 mM EGTA, 0.5 mM Na3VO4, 5 mM beta-glycerophosphate, 0.01% Triton X-100, 2.5 mM DTT, and 0.1% BSA. The kinase was incubated with Sesqui-Maleate Compound 1 Crystalline Form A* for 1 hour at room temperature (RT) and the reaction was initiated by the addition of ATP and GST-MEK1 (K97R mutated full-length protein, recombinant protein purified from the bacterial expression system). After the RT reaction for 1 hour, the stop/detection solution was added according to the manufacturer's instructions (Bioisights Cisbio). The stop/detection solution contained anti-phosphorus MEK1/2 conjugated rabbit polyclonal antibody (Ser217/221) and D2-conjugated anti-GST monoclonal antibody in buffer containing 25 mM Tris pH 7.4, 400 mM KF, 50 mM EDTA , 0.1% BSA and 0.01% Triton X100. TR-FRET signals (ratio of fluorescence emission at 665 nm over emission at 620 nm with excitation wavelengths of 337 nm) were measured in a PHERAstar FS plate reader (BMG Labtech) after 1.5 hours of incubation. MEK1 phosphorylation led to binding of the anti-phospho-MEK1/2 antibody to the GST-MEK1 protein which placed the donor fluorophore (Eu3+cryptate) in close proximity to the d2 receptor on the antiGST antibody, thus resulting in a high degree of of fluorescence resonance energy transfer from the donor fluorophore (at 620 nm) to the acceptor fluorophore (at 665 nm). The IC50 for Crystalline Form A* of Sesqui-Maleate Compound 1 was derived by adapting the dose-response % inhibition data to a four-parameter logistic model using Graphpad Prism software.

[0341] Kinase assays for EGFR, EPHA2, PDGFR-beta, TXK, FLT3, VEGFR2, RET, BTK, ITK, TEC and SRC

[0342] Sesqui-Maleate Crystalline Form A* of Compound 1 was tested against recombinant EGFR / EPHA2 / BTK (from Carna Biosciences), ITK / TEC / SRC / PDGFR-beta / TXK / FLT3 / VEGFR2 / RET (from Life Tech-nologies) and HER2 (aa676-1255, recombinant protein purified from insect expression system) in the Bioisays Cisbio HTRF KinEASE-TK assay according to the manufacturer's instructions. The assay was performed in a reaction mixture containing kinases, ATP at KM concentration, biotinylated peptide substrate (61TK0BLC, Cisbio Bioassys) and Crystalline Form A* of Compound 1 Sesqui-Maleate. The kinase was incubated with Crystalline Form A* of Compound 1 Sesqui-Maleate at RT for 1 h or 2 h (for EGFR) and the reaction was initiated by the addition of ATP and substrate. After the reaction, the stop/detection solution was added according to the manufacturer's instructions. The detection/stop solution contained Eu + cryptate anti-phosphotyrosine conjugated antibody (61T66kLB, CisBio Bioassys) and Streptavi-din-XL665 (610SAXLB, CisBio Bioassys) in buffer containing 25 mM Tris pH 7.4, 400 mM KF, EDTA 50 mM, 0.01% BSA, 0.01% Triton X-100. TR-FRET signals (ratio of fluorescence emission at 665 nm over emission at 620 nm with excitation at 337 nm wavelength) were recorded in a PHERAstar FS plate reader (BMG Labtech) after 1 hour of incubation. Phosphorylation of the biotinylated peptide substrate led to the binding of the Eu+-conjugated anti-phosphotyrosine antibody and the streptavidad-XL665 to the biotinylated peptide. The proximity of the donor fluorophore (Eu + cryptate) and acceptor (XL665) led to a high degree of fluorescence resonance energy transfer from the donor fluorophore (at 620 nm) to the acceptor fluorophore (at 665 nm). The IC50 for Sesqui-Maleate Crystalline Form A* of Compound 1 was derived by fitting the %-dose-response inhibition data to a four-parameter logistic model by Gra-phpad Prism software.

[0343] Results:

[0344] Compound 1 Sesqui-Maleate (Crystalline Form A*) is a potent and reversible inhibitor of mutant B-RAF enzymes V600E, wild-type B-RAF and C-RAF. Furthermore, among the following kinases that have been tested so far, the potent Sesqui-Maleate compound inhibits EGFR, EPHA2, PDGFR-beta, FLT3, VEGFR2 and RET, has weak inhibition for HER2, SRC and TXK and no inhibition for BTK, ITK and TEC. Many of these kinases that Compound 1 Sesqui-Maleate Inhibitor have been identified as potential targets for anti-cancer therapy and as such may enhance the potential efficacy as well as the potential applicability of Compound 1 Sesqui-Maleate to a wide range of tumors

[0345] Table 18. Enzyme Inhibition Activities of Crystalline Form A* of Sesqui-Maleate of Compound 1

[0346] Test 2: Inhibition of in vitro growth of human tumor cell lines by Sesqui-Maleate Compound 1 (Crystalline Form A * tested)

[0347] Methods:

[0348] Cell lines A375, Sk-Mel-28, HT29, Colo205, A431 and HCC827 were purchased from the American Type Culture Collection. All cell lines used in the cell assays were cultured in the designated medium supplemented with 10% fetal bovine serum (FBS, Thermo Scientifico), 100 units/ml penicillin (Gibco), 0.1 mg/ml streptomycin (Gibco) and in a humidified environment of 37°C with 5% CO2. Cell lines were reintegrated from frozen stocks established within three passages of the original cells purchased and passaged no more than 30 times.

[0349] Cellular phospho-ERK and phospho-EGFR were measured using a TR-FRET-based method. Cells were seeded at 3 x 10 4 per well of a 96-well plate and allowed to rest for 16 hours. The growth medium was then replaced with 100 µL of serum-free medium. The cells were then treated with a 10 point titre of the compound. After 1 h of compound treatment, 50 µL of lysis buffer (Cisbio) was added to each well. The plates were then incubated at room temperature under agitation for 30 min. A total of 16 µL of cell lysate from each well of a 96-well plate was transferred to a small volume white 384-well plate. The lysate from each well was incubated with 2 µl of anti-ERK or anti-EGFR (Cisbium) antifreeze and anti-ERK (Cisbium) labeled with 2 µl of anti-ERK antibody (Cisbium) and 2 µl of anti-phospho-ERK labeled with D2 (receptor) or anti-phospho-EGFR Antibody (Cisbio) for 2 h at room temperature. FRET signals were measured using a PHERAstar FS reader (BMG Labtech). IC50 values for ERK or EGFR phosphorylation were determined using GraphPad Prism software.

[0350] The growth inhibitory activity of compounds on a panel of melanoma, colon, breast, and lung cancer cells was determined using the CellTiter-Glo luminescent cell viability assay (Promega). The number of cells seeded per well of a 96-well plate was optimized for each cell line to ensure logarithmic growth over the 3-day treatment period. Cells were allowed to attach for 16 hours and then treated with a 10-point dilution series in duplicate. After a 3-day exposure to the compound, a volume of CellTiter-Glo reagent equal to the volume of cell culture medium present in each well was added. The mixture was placed on an orbital shaker for 2 minutes to allow cell lysing, followed by a 10 minute incubation at room temperature to allow luminescent signal development and stabilization. The luminescent signal was measured using the PHERAstar FS reader (BMG Labtech). EC50 values for cell viability were determined with GraphPad PrismResults software:

[0351] Cellular assays confirmed that Sesqui-Maleate from Compound 1 inhibits a number of direct signaling intermediates downstream of RAFs. For example, Compound 1 Sesqui-Maleate inhibited ERK phosphorylation with an IC50 of 32 and 77 nM in A375 eSek-Mel-28 melanoma cells driven by mutant B-RAF V600E. It also inhibits ERK phosphorylation with an IC50 of 103 and 53 nM in HT29 and Colo205 colorectal cancer cells reared by B-RAF mutant V600E. Furthermore, Compound 1 Sesqui-Maleate inhibits EGFR phosphorylation with an IC50 of 385 and 161 nM in EGFR overexpressing A431 mutant HCC827 lung cancer cell lines and mutant EGFR. Compound 1 Sesqui-Maleate was evaluated for its anti-proliferation activity in a panel of cancer cell lines. It showed antiproliferative activity against a number of cell lines harboring B-RAF cell mutations (Table 19).

[0352] Table 19. Crystalline Form A* of Compound 1 Sesqui-Maleate inhibited BRAF V600E driven cell proliferation.

[0353] Test 3: In Vivo Pharmacology of Crystalline Form A* of Compound 1 Sesqui-Maleate

[0354] Tumor transplantation methods:

[0355] Human cancer cells were inoculated subcutaneously into the right axillary region of BALB/C mice to generate tumor xenografts. On the day of implantation, the cell culture medium was replaced with fresh medium. Three hours later, the medium was removed and the cells were harvested and resuspended in cold PBS (4 °C) before inoculation. The right axillary region of each mouse was cleaned with 70% ethanol prior to cell inoculation. Each animal was injected subcutaneously with cells in 200 µl of cell suspension into the right front flank through a 26 gauge needle. After implantation, tumor volumes were measured twice a week in two dimensions using calipers. Tumor volume was calculated using the formula: V = 0.5 x (a x b2) where a and b are the long and short tumor diameters, respectively.

[0356] For in vivo efficacy studies, when the average tumor size reached 100 ~ 200 mm 3 , animals were assigned into the desired number of groups with 6-10 mice per group using a stratified randomization procedure. The groups consisted of a control group (no drug treatment) and different dose levels of Crystalline Form A* from the Compound 1 Sesqui-Maleate treatment groups (ranged from 1-20 mg/kg). All doses were based on free base weight. Drugs were given by oral administration once (qd) or twice (throw) daily; Dosing frequency depended on the individual case for tumor growth. Treatments were administered by oral gavage (p.o.) in a volume of 10 ml/kg body weight. Body weight was assessed immediately before dosing and volume dosing was adjusted accordingly. Individual body weights and tumor volumes were determined twice a week, with mice monitored daily for clinical signs of toxicity during the study period. Mice were euthanized using carbon dioxide once the tumor volume reached □ 2000 mm3, the tumor was ulcerated, or the body weight loss exceeded 20%. All group data were analyzed using the t-test. If the p value was <0.05, it was considered statistically significant.

[0357] Tumor growth inhibition (TGI) was calculated using the following formula:

[0358] Treated t = tumor volume treated at time t

[0359] Treated t0 = volume of tumor treated at time 0

[0360] placebo t = placebo tumor volume at time t

[0361] placebo t0 = placebo tumor volume at time 0

[0362] For the PD study, when the average tumor size reaches 140900 mm3, animals were assigned to the desired number of groups with 4 mice per group using a stratified randomization procedure. The groups consisted of a control group (no drug treatment) and different dose levels of Crystalline Form A* from the Compound 1 Sesqui-Maleate treatment groups (ranged from 1 to 20 mg/kg). All doses were based on free base weight. Drugs were administered by oral gavage once (qd) or twice (throw) daily. All treatments were administered by oral gavage (p.o.) in a volume of 10 ml/kg body weight. Body weight was assessed immediately before dosing and volume dosing was adjusted accordingly. Mice were euthanized using carbon dioxide at the desired time points after administration. Tumor tissues were dissected immediately after euthanization and snap-frozen in the tube pre-filled with PM beads using liquid nitrogen and stored at -80 °C prior to p-ERK testing. Fifteen microliters of complete lysis buffer were added to tumors frozen with MP beads. Tumor tissue homogenization was performed in the MP homogenization unit and the lysates were then centrifuged at 13,000 rpm for 10 min at 4 °C to remove insoluble material. Two micrograms of sodium protein were used to measure the phosphorylated ERK1/2 level by the AlphaScreen® SureFire® p-ERK1/2 assay (PerkinElmer).

[0363] Results:

[0364] Compound 1 Sesqui-Maleate (Crystalline Form A*) has potent kinase inhibitory activity against B-RAF V600E, antiproliferative activity in cell-based assays, and antitumor activity in xenograft models. Sesqui Maleate Compound 1 (Crystalline Form A*) marked antitumor activity against A375 human melanoma (B-RAF V600E mutation), LOX melanoma (B-RAF V600E mutation), Colo205 colorectal carcinoma (B-RAF V600E mutation), Carcinoma HT29 colorectal carcinoma (B-RAF V600E mutation), WiDr colorectal carcinoma (B-RAF V600E mutation), HCC827 lung carcinoma mutations (EGFR mutation), and A431 squamous cell carcinoma (EGFR deletion) xenografts in mice. In addition, Compound 1 Sesqui-Maleate (Form A* from Crystal Line) showed remarkable antitumor activity in Calu-6 human lung adenocarcinoma tumors (K-RAS mutation) when combined with docetaxel or the MEK inhibitor, selumetinib (AZD6244).

[0365] Oral administration of Crystalline Form A* of Sesqui Maleate compound 1 resulted in time-dependent and dose-dependent inhibition of ERK1/2 phosphorylation in A375 xenografts in mice. Inhibition of p-ERK levels in tumor tissues correlates well with plasma and tumor drug concentrations of Compound 1. In addition, Sesqui-Maleate (Crystalline Form A) and the MeK inhibitor AZD6244 of Compound 1 demonstrated a strong synergistic effect on p-ERK inhibition in Calu-6 K-RAS mutant non-small cell adenocarcinoma xenograft models.

[0366] Test 5: Toxicology of Sesqui-Maleate Compound 1 (Crystalline Form A*)

[0367] A comprehensive non-clinical toxicity study program, including 28-day GLP studies in rats and dogs and several investigative studies, was conducted to evaluate the preclinical safety of Sesqui- Compound 1 Maleate (Crystalline Form A*). These studies took into account available regulatory guidance for the preclinical development of anticancer drugs. In these studies, Compound 1 Sesqui-Maleate demonstrated a favorable toxicology pharmacology and safety profile. The results of pharmacological safety tests showed that Compound 1 Sesqui-Maleate (Crystalline Form A*) had no effect on respiration, blood pressure and circulatory function, and no effect on autonomic nervous system or central nervous system activity. The results of additional toxicity tests did not show teratogenic, mutagenic or reproductive toxicity.

[0368] Test 6: Pharmacokinetics of Crystalline Form A * of Sesqui Maleate of Compound 1

[0369] The fully validated LC-MS/MS method has been well utilized for the pharmacokinetic (PK) studies of Crystalline Form A* of Compound 1 Sesqui-Maleate in Sprague-Dawley rats and Beagle dogs following single-dose and multiple-administrations .

[0370] Crystalline Form A* of Sesqui-Maleate Compound 1 has high oral bioavailability in both rats (51% to 102%) and dogs (41% to 82%). Elimination half-lives ranged from 4.4 to 9.4 hours in rats and from 3.3 to 4.9 hours in dogs following oral administration.

[0371] The kinetics were linear over the dose range of 0.5 to 15 mg/kg in rats and 1.5 to 15 mg/kg in dogs. After multiple administration, a small accumulation (~2-fold) was observed in rats. This small accumulation was statistically significant in female rats but not in male rats. No accumulation was observed after multiple administration in dogs.

[0372] Test 7: ADME of Crystalline Form A * of Compound Sesqui-Maleate 1

[0373] Compound 1 was widely distributed in various tissues, but was low in brain tissue, indicating that the drug does not readily cross the blood-brain barrier. No significant induction effect was found on P450 enzymes in rat liver, no inhibitory activity was found on drug metabolism enzymes for Compound 1 Sesqui-Maleate, with the exception of weak inhibition on CYP2C8 (IC50 = 1.03 µM ) and CYP2C19 (IC50 = 1.96 µM). CYP3A was the major CYP isoform responsible for Compound 1 metabolism while CYP1A2, CYP2C8, CYP2C9, CYP2C19 and CYP2D6 contribute to Compound 1 metabolism to a lesser extent.

[0374] Test 8: Pharmacokinetic Comparison of Crystalline Form A* of Sesqui Maleate Compound 1 and Free Base.

[0375] Drugs and reagents: Sesqui-Maleate crystalline form A * of compound 1 with particle sizes of D90 = 4.1um, D10 = 1.5um, D50 = 2.4um after micronization. The material content (purity) was not less than 98.0%.

[0376] Experimental animals: Beagle dogs, male and female.

[0377] Pharmaceutical preparation: Weigh the appropriate amount of each substance and disperse in 0.5% sodium carboxymethylcellulose (for the free base of Compound 1) or 0.5% methylcellulose solution (for Sesqui Maleate of Compound 1). Prepare a suspension in the desired concentration of Compound 1. All doses and concentrations of Compound 1 were calculated on a free base basis in this study.

[0378] Administration and sampling: the dosing solutions were freshly prepared before dose administration. Actual body weights and actual volume injected are recorded accordingly. Dogs were fasted overnight and allowed to eat four hours after administration. Each suspension was administered orally to dogs at a dose ranging from 0.5 to 15 mg/kg. Blood samples (~1.0 mL) will be collected pre-dose and at different times up to 36 hours post-dose via cephalic vein plexus. Whole blood will be processed by centrifugation and plasma samples will be collected and kept in the freezer prior to analysis. Plasma samples were processed for protein precipitation. Compound 1 concentrations in plasma samples were determined using a validated tandem mass liquid chromatography (LC-MS/MS) method. Plasma concentration-time data were analyzed using a non-compartmental model using Pharsight Win-Nonlin.

[0379] Table 20. The Cmax and area under the concentration-time curve for Compound 1 in free bases and salts.

[0380] The above experiment showed that the Cmax (ng/mL) and AUC0-inf (ng.h/mL) of Crystalline Form A* of Sesqui-Maleate Compound 1 were as high as approximately 2-3 times that of free base form. Therefore, Crystalline Form A* of Sesqui-Maleate of Compound 1 has significantly better relative bioavailability than the free base of Compound 1 (amorphous form).

[0381] Test 9: clinical trials

[0382] Using Crystalline Form A* of Compound 1 Sesqui-Maleate to prepare capsules, a Phase I clinical safety study was conducted in 25 subjects administered single doses of 5, 10, 20, 30, 40, and 50 mg . Results showed that single doses of 5-50 mg were safe and well tolerated. Compound 1 treatment caused partial responses in BRAF V600E melanoma patients, partial responses in BRAF V600E PTC patients, and some antitumor activities in BRAF V600E CRC patients. 16 KRAS cancer patients were also recruited and treated, 3 PR, and at least 12 patients survived more than 2.2 months (a reported mean PFS for untreated KRAS cancer patients, see Oncotarget, 5, 19, 2014). These preliminary data demonstrated that Compound 1 Sesqui-Maleate (Crystalline Form A*) was effective in the treatment of Braf and K-ras mutant cancers.

[0383] The foregoing examples and description of certain embodiments are to be taken as illustrating, rather than limiting, the present invention as defined by the claims. As will be readily appreciated, numerous variations and combinations of the features set out above can be used without departing from the present invention as set out in the claims. All such variations are intended to be included within the scope of the present invention. All cited references are hereby incorporated by reference in their entirety.

[0384] It should be understood that if any prior art publication r is referred to herein, such reference does not constitute an admission that the publication forms part of common general knowledge in the prior art in any country.

[0385] In the claims that follow and in the foregoing description of the invention, except where the context requires otherwise due to express language or necessary implication, the word "comprises" or variations such as "comprises" or "comprising" is used in an inclusive sense, i.e. to specify the presence of the indicated features, but not to preclude the presence or addition of additional features in various embodiments of the invention.

[0386] The descriptions of all publications, patents, patent applications and published patent applications referred to herein by an identifying citation are hereby incorporated by reference in their entirety.

Claims (17)

1. 5-(((1R, 1aS, 6bR)-1-(6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b-di) sesqui-maleate crystalline salt -hydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one characterized by having the structure of formula (II) : wherein said crystalline salt is selected from: (a) crystalline form A* characterized by an X-ray powder diffraction pattern comprising seven or more diffraction peaks having 2θ angle values independently selected from the group of: 6.3 ±0.2, 8.9±0.2, 9.4±0.2, 11.2±0.2, 12.6±0.2, 13.4±0.2, 17.9±0 .2, 18.6±0.2, 18.8±0.2, 19.3±0.2, 20.1±0.2, 20.7±0.2, 21.2±0.2 , 21.8±0.2, 22.4±0.2, 22.6±0.2, 23.3±0.2, 23.8±0.2, 24.7±0.2, 25 .6±0.2, 26.1±0.2, 27.4±0.2, 28.3±0.2, 28.6±0.2, 29.0±0.2, 29.4±0 ,2 and 30.4±0.2 degrees; or (b) crystalline form A with characterization by an X-ray powder diffraction pattern comprising three or more diffraction peaks having 2θ angle values independently selected from the group of: 8.3±0.2, 11.2±0 .2, 17.9±0.2, 18.4±0.2, 18.6±0.2, 19.3±0.2, 20.8±0.2 and 22.5±0.2 degrees. 2. Crystalline salt according to claim 1, wherein said crystalline salt is crystalline form A* is characterized by an X-ray powder diffraction pattern comprising seven or more diffraction peaks having 2θ angle values independently selected from the group consisting of: 6.3±0.2, 8.9±0.2, 9.4±0.2, 11.2±0.2, 12.6±0.2, 13.4±0.2, 17.9±0.2, 18.6±0.2, 18.8±0.2, 19.3±0.2, 20.1±0.2, 20.7±0.2 . 29.0 ±0.2, 29.4±0.2, and 30.4±0.2 degrees. 3. Crystalline salt according to claim 2, characterized in that it has an X-ray powder diffraction pattern of Figure 2. A crystalline salt according to claim 1, wherein said crystalline salt is crystalline form A characterized by an X-ray powder diffraction pattern comprising three or more diffraction peaks having 2θ angle values independently selected from the group consisting of : 8.3±0.2, 11.2±0.2, 17.9±0.2, 18.4±0.2, 18.6±0.2, 19.3±0.2, 20.8±0.2, and 22.5±0.2 degrees. 5. Crystalline salt according to claim 4, characterized in that it has an X-ray powder diffraction pattern of Figure 17. 6. Method for preparing the crystalline salt as defined in claim 1, characterized in that it comprises any of the following procedures: (a) dissolving the free base or a different salt of 5 - (((1R, 1aS, 6bR) -1 - (6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a,6b-dihydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-di- hydro-1,8-naphthyridin-2(1H)-one in a solvent or mixture of solvents to form a solution or suspension; mixing the resulting solution or suspension with maleic acid to form a mixture; and precipitation of 5-(((1R, 1aS, 6bR)-1-(6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b-dihydro maleate salt -1H-cyclopropa[b]benzofuran -5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one in a target crystalline form; or (b) dissolving or suspending 5-(((1R, 1aS, 6bR)-1-(6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b-dihydro-1H-cyclopropa [benzazolan-5-yl)oxy salt)-3,4-dihydro-1,8-naphthyridin-2(1H)-one in a solvent or in a mixture of solvents; and precipitating 5-(((1R, 1aS, 6bR)-1-(6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b-dihydro-1H maleate salt -cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one in a target crystalline form. 7. Method according to claim 6, characterized in that said procedure (a) or (b) further comprises one or more independently selected procedures of heating, filtration to remove undissolved impurities, solvent distillation, addition of a counter -solvent or mixture of solvents, addition of crystal seeds, addition of precipitation inducing agent(s), cooling, precipitation and filtration to collect the crystalline product. 8. Method according to claim 6 or 7, characterized in that it comprises said procedure (a) or (b), wherein the solvent or mixture of solvents is selected from the group consisting of water, lower alkyl alcohols, ketones, ethers, esters, lower aliphatic carboxylic acids, lower aliphatic nitriles, optionally halogenated aromatic solvents and combinations thereof. Method according to claim 6 or 7, characterized in that the solvent is isopropanol, ethanol, methanol, acetone, THF, 1,4-dioxane, acetic acid, acetonitrile, water or a combination thereof. 10. Method according to claim 6, characterized in that it comprises said procedure (d), wherein said elevated temperature is at least 40°C, at least 60°C, at least 80°C, or at least 100° C, but below the decomposition temperature of the sesqui-maleate salt. Method according to claim 6, characterized in that: 1) said procedure (a) or (b) comprises the use of isopropanol-water (v / v > 60/40) as solvent to produce the crystalline form A *; or 2) said procedure (a) or (b) comprises the use of acetone as a solvent to produce crystalline form A**; 3) said procedure (a) or (b) comprising the use of an IPA-water mixture (v / v = 4:1) as a solvent to produce crystalline form A. 12. Use of crystalline salt as defined in any one of claims 1 to 5, characterized in that it is intended for the production of a drug for the treatment or prevention of a cancer selected from the group consisting of brain cancer, lung cancer, kidney cancer, bone cancer, liver cancer, bladder cancer, breast cancer, head and neck cancer, ovarian cancer, melanoma, skin cancer, adrenal cancer, cervical cancer, colorectal cancer, lymphoma or thyroid tumors and their complications. 13. Use according to claim 12, characterized in that the cancer is selected from the group consisting of BRAF, NRAS and KRAS mutant brain cancer, lung cancer, kidney cancer, bone cancer, liver cancer, bladder cancer, breast cancer, head and neck cancer, ovarian cancer, melanoma, skin cancer, adrenal cancer, cervical cancer, colorectal cancer, lymphoma or thyroid tumors and their complications. 14. Process for the preparation of a sesqui-maleate of 5 - (((1R, 1aS, 6bR) -1- (6- (trifluoromethyl) -1H-benzo[d]imidazol-2-yl)-1a, 6b-di crystalline -hydro-1H-cyclopropa [b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one crystalline, characterized in that it comprises: a) mixture a about 50 °C of a mixture of 5 - (((1R, 1aS, 6bR) -1- (6-(trifluoromethyl)-1H-benzo[d]imidazol-2-yl)-1a, 6b-dihydro -1H -cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one and maleic acid in a mixed solvent of i-PrOH and H2O, or b) mixing at about 50 °C 5 - (((1R, 1aS, 6bR) -1- (6- (trifluoromethyl) - 1H-benzo[d]imidazol-2-yl)-1a, 6b-di- hydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthyridin-2(1H)-one with a mixture or a suspension or a solution of maleic acid in a mixed solvent of i-PrOH and H2O, or c) mixing at about 50 °C a mixture or a suspension or a solution of 5 - (((1R, 1aS, 6bR) -1- (6- (trifluoromethyl) - 1H-benzo[d]imidazol-2-yl)-1a,6b-dihydro-1H-cyclopropa[b]benzofuran-5-yl)oxy)-3,4-dihydro-1,8-naphthy- ridin-2(1H)-one in a mixed solvent of i-PrOH and H2O with maleic acid, wherein in each of (a), (b) and (c), the amount of i-PrOH is greater than 40% by volume in terms of the total volume of i-PrOH and water. Process according to claim 14, characterized in that in each of (a), (b) and (c), the amount of i-PrOH is greater than 60% by volume in terms of the total volume of i -PrOH and water. Process according to claim 14, characterized in that the mixed solvent is replaced by i-PrOH. Process according to any one of claims 14 to 16, characterized in that it comprises adding some crystal seeds to the resulting mixture after cooling to room temperature, and then letting the mixture rest for a certain period of time.