HK1059085B - Salts of a isothiazole-4-carboxamide and their use as anti-hyperproliferation agents - Google Patents

Description

Salts of isothiazole-4-carboxamides and their use as anti-hyperproliferative agents

Background

The present invention relates to salts of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide having the formula.

Formula I

The free base form of the compound of formula I is described in co-pending U.S. Ser. No. 09/316,837, filed on 21/5/1999, and the disclosure of which is hereby incorporated by reference in its entirety. The foregoing application is assigned with the present application. The free base of formula I is useful for the treatment of hyperproliferative diseases, such as cancer.

The present invention provides hydrochloride, hydrobromide, hemi-citrate, acetate, p-toluenesulfonate, L-tartrate, hemi-succinate and methanesulfonate salts of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide.

The invention also relates to a process for the production of hydrochloride, hydrobromide, hemi-citrate, acetate, p-toluenesulfonate, L-tartrate, hemi-succinate and methanesulfonate salts of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide. The invention also relates to pharmaceutical compositions containing the hydrochloride, hydrobromide, hemi-citrate, acetate, p-toluenesulfonate, L-tartrate, hemi-succinate and methanesulfonate salts of the compounds of formula I. The salts of the invention are useful for the treatment of hyperproliferative diseases, such as cancer, in mammals, particularly in humans. The invention also relates to methods of treating hyperproliferative diseases by administering the salts of formula I.

Brief description of the drawings

FIG. 1 is an X-ray powder diffraction pattern of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide hydrochloride salt which was prepared and isolated according to the procedures described in example 2 of the present invention.

FIG. 2 is an X-ray powder diffraction spectrum of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide hydrobromide salt prepared and isolated according to the procedure described in example 3 of this invention.

FIG. 3 is an X-ray powder diffraction pattern of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide hemi-citrate salt prepared and isolated according to the procedures outlined in example 4 of the present invention.

FIG. 4 is an X-ray powder diffraction pattern of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide acetate prepared and isolated according to the procedures described in example 5 of the present invention.

FIG. 5 is an X-ray powder diffraction pattern of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide p-toluenesulfonate salt, which was prepared and isolated as described in example 6 of the present invention.

FIG. 6 is an X-ray powder diffraction pattern of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide L-tartrate, prepared and isolated according to the procedures described in example 7 of the present invention.

FIG. 7 is an X-ray powder diffraction pattern of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide hemisuccinate salt prepared and isolated as described in example 8 of the present invention and identified as hemisuccinate type A.

FIG. 8 is an X-ray powder diffraction pattern of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide hemisuccinate salt prepared and isolated as described in example 9 of the present invention and identified as the type B hemisuccinate salt.

FIG. 9 is an X-ray powder diffraction pattern of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide mesylate salt prepared and isolated as described in example 10 of the present invention.

In the X-ray powder diffraction spectra of FIGS. 1-9, the horizontal axis is the diffraction 2-theta angle and the vertical axis is the diffraction intensity, in Cps.

Summary of the invention

The present invention relates to hydrochloride, hydrobromide, hemi-citrate, acetate, p-toluenesulfonate, L-tartrate, hemi-succinate and methanesulfonate salts of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide having the formula:

formula I

The invention also relates to a process for the preparation of hydrochloride, hydrobromide, hemi-citrate, acetate, p-toluenesulfonate, L-tartrate, hemi-succinate and methanesulfonate of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide which comprises combining the free base with one of the above salts in a suitable organic solvent.

The hydrochloride, hydrobromide, hemi-citrate, acetate, p-toluenesulfonate, L-tartrate, hemi-succinate and methanesulfonate salts of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide are characterized by X-ray powder diffraction.

The X-ray powder diffraction patterns of the crystalline hydrochloride, hydrobromide, hemi-citrate, acetate, p-toluenesulfonate, L-tartrate, hemi-succinate (forms a and B) and methanesulfonate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide are substantially the same as the X-ray powder diffraction patterns shown in figures 1-9, respectively. However, it is known that the obtained X-ray powder diffraction spectrum has measurement errors depending on the measurement conditions. In particular, it is generally known that the intensity of an X-ray powder diffraction spectrum may fluctuate depending on the measurement conditions. Thus, it should be understood that the salt of the present invention should not be limited to crystals having an X-ray powder diffraction spectrum identical to that of FIGS. 1 to 8, and any crystals having an X-ray powder diffraction spectrum substantially identical to that of the above-described X-ray powder diffraction spectrum are within the scope of the present invention. Substantial identity of the X-ray powder diffraction spectra can be readily determined by those skilled in the art of X-ray powder diffraction studies.

In general, the measurement error of the diffraction angle in the conventional X-ray powder diffraction method is about 5% or less, and such a measurement error degree should be considered to be related to the diffraction angle. In addition, it should be understood that the intensity may fluctuate depending on the test conditions.

The hydrochloride salt of the compound of formula I is characterized by crystals having a high intensity diffraction peak [% relative intensity ] at 2-theta diffraction angles of: 8.623[90.7], 12.121[38.9], 17.298[95.2], 23397[44.7], 23.944[51.7], 24.119[62.7], 24.873[55.7], 25.948[100] and 28.821[39.6 ]. The hydrochloride salt of the present invention has an X-ray powder diffraction pattern substantially the same as that shown in FIG. 1.

Characteristic 2-theta (2 theta) values and Relative Intensity (RI) percentages of the hydrochloride diffraction spectrum of the compound of formula I are shown in Table 1.

TABLE 1

2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)
										6.225	10.5	17.298	95.2	23.397	44.7	28.821	39.6	33.407	10.4
8.623	90.7	17.868	17.6	23.944	51.7	29.438	19.0	33.778	15.3
										12.121	38.9	18.712	17.1	24.119	62.7	30.543	15.3	34.920	10.5
12.522	16.8	18.880	14.2	24.873	55.7	31.144	8.7	35.273	9.4
										12.873	6.0	19.549	13.8	25.948	100.0	31.757	8.7	36.321	13.6
14.206	4.7	20.552	8.5	27.216	12.6	32.348	14.6	38.409	15.5
										15.951	5.1	21.896	22.2	28.146	6.0	32.640	11.4	39.500	9.9
16.736	12.4

A hydrobromide salt of a compound of formula I characterized by crystals having a high intensity diffraction peak [% relative intensity ] at 2-theta diffraction angles of: 8.687[100.0], 12.264[35.9], 17.374[42.3], 23.711[24.0], 24.335[20.7] and 25.769[34.3 ]. The X-ray powder diffraction spectrum of the hydrobromide of the present invention was substantially the same as that shown in FIG. 2.

Characteristic 2-theta (2 theta) values and relative intensities (%) of the hydrobromide diffraction spectrum of the compound of formula I are shown in table 2.

TABLE 2

2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)
										3.156	2.5	17.374	42.3	23.188	5.6	28.528	6.1	33.256	3.7
4.615	2.4	17.767	5.6	23.711	24.0	28.916	9.4	33.897	7.9
										6.331	9.3	18.185	3.2	24.335	20.7	29.418	7.7	34.628	2.8
8.687	100.0	18.913	16.6	25.435	9.1	30.266	4.6	34.999	3.3
										12.264	35.9	19.528	9.5	25.769	34.3	31.561	3.9	35.432	6.1
12.890	2.2	20.286	2.5	26.940	4.1	32.082	3.4	36.006	4.3
										13.445	1.4	20.581	2.9	27.345	7.0	32.638	5.3	37.361	3.4
14.140	3.4	21.874	13.6	28.160	5.7	32.925	4.4	38.224	4.8
										16.063	4.0

The hemi-citrate salt of a compound of formula I is characterized by crystals having a high intensity diffraction peak [% relative intensity ] at 2-theta diffraction angles of: 4.306[79.9], 16.317[100.0], 20.988[32.7], 21.476[30.9], 22.643[48.7], 23.384[76.9], 24.891[76.0], 27.573[47.9] and 27.840[32.3 ]. The X-ray powder diffraction spectrum of the hemi-citrate salt of the present invention is substantially the same as the X-ray powder diffraction spectrum shown in figure 3.

Characteristic 2-theta (2 theta) values and relative intensities (%) of the hemi-citrate diffraction spectra of the compounds of formula I are shown in table 3.

TABLE 3

2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)
										3.201	14.3	13.766	12.4	18.693	15.8	24.217	28.8	29.630	17.4
4.306	79.9	14.086	7.0	19.344	23.4	24.891	76.0	31.251	14.6
										6.429	7.0	14.710	9.5	20.394	16.4	25.320	20.4	31.848	14.2
8.620	6.0	15.297	16.0	20.988	32.7	25.948	28.0	32.235	11.8
										9.589	5.6	16.317	100.0	21.476	30.9	26.370	25.7	34.147	11.0
10.583	6.8	17.309	14.4	21.994	27.3	27.573	47.9	35.878	16.2
										11.449	20.9	17.572	16.5	22.643	48.7	27.840	32.3	37.337	12.3
12.300	8.7	18.258	13.7	23.384	76.9	28.609	19.6

The acetate salt of the compound of formula I is characterized by crystals having a high intensity diffraction peak [% relative intensity ] at 2-theta diffraction angles of: 6.096[21.7], 12.183[21.4], 17.451[33.3], 18.288[100.0], 22.441[57.7], 23.086[19.9] and 24.439[20.7 ]. The X-ray powder diffraction spectrum of the acetate of the invention is basically the same as that shown in figure 4.

Characteristic 2-theta (2 theta) values and relative intensities (%) of the acetate diffraction spectrum of the compound of formula I are shown in table 4.

TABLE 4

2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)
										6.096	21.7	16.793	4.5	21.346	8.9	27.930	5.8	32.271	4.1
8.625	2.8	17.121	12.8	22.441	57.7	28.820	10.0	33.127	5.1
										11.840	2.9	17.451	33.3	23.086	19.9	29.648	6.1	35.030	3.4
12.183	21.4	17.920	8.7	24.038	7.5	30.634	3.3	36.445	3.2
										14.836	4.2	18.288	100.0	24.439	20.7	31.112	3.2	37.830	3.0
15.264	9.2	20.088	3.6	24.760	11.3	31.951	2.9	39.478	2.5
										15.824	5.0	20.458	11.3	25.861	5.0

The p-toluenesulfonate salt of the compound of formula I is characterized in that its crystals have a high intensity diffraction peak [% relative intensity ] at 2-theta diffraction angle of: 20.446[100.0], 20.760[74.0], 22.092[81.7], 22.371[70.8], 23.190[65.2] and 26.239[61.5 ]. The X-ray powder diffraction spectrum of the p-toluenesulfonate of the present invention is substantially the same as the X-ray powder diffraction spectrum shown in FIG. 5.

Characteristic 2-theta (2 theta) values and relative intensities (%) of the p-toluenesulfonate diffraction spectrum of the compound of the formula I are shown in Table 5.

TABLE 5

2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)
										6.817	50.3	13.373	53.9	18.174	21.3	23.190	65.2	28.167	34.5
7.515	28.0	14.337	18.3	18.976	40.4	24.110	30.5	29.672	19.6
										7.822	22.3	15.001	22.2	19.739	36.7	25.471	40.2	31.038	19.3
11.157	15.0	15.601	21.4	20.446	100.0	25.932	50.4	31.586	21.2
										12.205	24.5	15.297	14.0	20.760	74.0	26.239	61.5	35.357	19.6
12.800	30.0	16.943	32.0	22.092	81.7	27.355	48.8	36.800	16.4
										13.047	43.6	17.362	23.5	22.371	70.8	27.833	39.0

The L-tartrate salt of the compound of formula I is characterized by crystals having a high intensity diffraction peak [% relative intensity ] at 2-theta diffraction angles of: 4.061[82.9], 20.821[85.6], 21.634[100.0], 22.179[94.0] and 25.858[95.1 ]. The X-ray powder diffraction spectrum of the L-tartrate salt of the present invention is substantially the same as the X-ray powder diffraction spectrum shown in FIG. 6.

Characteristic 2-theta (2 theta) values and relative intensities (%) of the L-tartrate diffraction spectra of the compound of formula I are shown in table 6.

TABLE 6

2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)
										4.061	82.9	14.631	27.2	20.010	53.0	24.788	53.8	32.465	36.1
6.678	11.8	15.428	22.7	20.334	58.7	25.081	60.9	33.442	33.7
										8.057	29.4	16.143	31.2	20.821	85.6	25.858	95.1	34.090	34.6
9.383	8.7	16.853	65.3	21.634	100.0	26.803	59.6	34.642	26.8
										10.647	8.3	17.338	56.2	22.179	94.0	28.386	34.3	35.635	33.7
11.711	60.2	18.400	97.0	22.730	73.8	29.067	34.1	36.073	28.5
										12.075	27.4	18.639	98.2	23.477	77.1	29.844	30.5	36.771	24.5
12.868	33.8	18.994	52.4	24.257	67.8	31.309	39.4	38.080	22.6
										13.320	22.7	19.722	42.9

Crystals of hemisuccinic anhydride of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide were found to have hygroscopic properties at 90% humidity. Two crystalline forms of the hemisuccinate salt crystal of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide were identified. The crystalline hemisuccinate salt form A of the compound of formula I was found to be 0.6% w/w hygroscopic at 30 ℃ and 90% RH. Crystalline form B hemisuccinate salt of the compound of formula I was found to be 1.5% w/w hygroscopic at 30 ℃ and 90% RH. Hemisuccinate form B was converted to hemisuccinate form a in refluxing ethanol over 24 hours. The X-ray powder diffraction spectra of the hemisuccinate salts of the invention, type A and type B, are substantially the same as the X-ray diffraction spectra of figures 7 and 8, respectively.

The hemisuccinate salt form A of the compound of formula I is characterized by a crystal having a high intensity diffraction peak [% relative intensity ] at 2-theta diffraction angles of: 4.634[100.0], 16.735[67.2], 22.179[60.8] and 25.002[70.3 ].

The hemisuccinate salt form B of the compound of formula I is characterized by a crystal having a high intensity diffraction peak [% relative intensity ] at 2-theta diffraction angles of: 6.714[ 31.0], 15.272[100.0], 19.197[59.3], 19.457[50.0], 24.487[99.0] and 24.802[79.1 ].

Characteristic 2-theta (2 theta) values and relative intensities (%) of diffraction spectra of the type a and type B hemisuccinates of the compound of formula I are shown in tables 7 and 8, respectively.

TABLE 7

2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)
										4.634	100.0	13.683	13.8	17.760	20.6	22.866	38.7	27.609	23.7
6.149	15.5	14.440	13.5	18.679	18.4	23.255	39.5	30.106	14.3
										9.843	6.2	14.896	21.9	19.421	30.2	24.079	44.6	30.797	13.7
11.392	11.3	15.996	14.4	20.586	29.2	25.002	70.3	37.769	11.4
										11.937	19.2	16.735	67.2	22.179	60.8	26.549	26.8

TABLE 8

2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)
										6.714	31.0	15.272	100.0	19.197	59.3	24.487	99.0	29.732	14.1
8.666	7.3	15.813	19.8	19.457	50.0	24.802	79.1	30.796	17.5
										11.092	8.6	16.551	17.5	20.597	17.2	25.640	15.9	33.484	9.4
11.696	21.9	16.875	26.4	21.160	28.8	26.641	20.1	34.594	11.4
										12.008	15.9	17.365	12.6	21.648	21.7	27.090	15.1	37.212	15.5
12.630	7.3	17.986	8.6	22.988	15.0	27.843	21.5	37.905	8.9
										13.466	17.2	18.710	26.4	23.568	18.6	28.552	19.7	39.023	8.9
13.774	13.9

The mesylate salt of the compound of formula I is characterized by crystals having a high intensity diffraction peak [% relative intensity ] at 2-theta diffraction angles of: 4.417[99.3], 17.288[45.5], 20.828[39.6], 21.677[43.5], 22.148[68.3], 25.427[100.0] and 27.006[37.5 ]. The X-ray powder diffraction spectrum of the mesylate of the invention is substantially the same as the X-ray powder diffraction spectrum shown in figure 9.

Characteristic 2-theta (2 theta) values and relative intensities (%) of the mesylate salt diffraction spectrum of the compound of formula I are shown in table 9.

TABLE 9

2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)	2θ	RI(％)
										4.417	99.3	15.619	9.1	21.677	43.5	27.006	37.5	32.337	9.0
8.806	7.2	17.288	45.5	22.148	68.3	28.757	8.8	32.940	7.8
										11.664	7.0	17.654	17.2	23.231	14.1	29.564	6.7	33.796	11.8
12.267	18.7	17.993	15.9	23.966	28.8	30.560	6.2	34.550	8.7
										12.610	8.3	18.728	9.8	24.602	30.2	31.173	9.7	35.308	7.6
13.224	5.4	20.358	15.0	25.427	100.0	31.722	13.3	36.883	9.1
										14.915	27.1	20.828	39.6	26.226	16.1

The X-ray powder diffractogram is but one of many methods of characterizing the atomic arrangement that constitutes 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide. Other methods well known in the art, such as single X-ray crystallography, can be used to identify the salts of formula I above.

It has surprisingly been found that the acetate, hydrochloride, hydrobromide, hemisuccinate and mesylate salts of the compound of formula I have a high degree of crystallinity, i.e. are substantially free of amorphous material. The advantage of these salts is that they provide more reproducible dosing results. The hydrochloride, hydrobromide and hemisuccinate salts of the compounds of formula I are substantially hydroscopic stable, which reduces potential problems associated with weight changes of the active ingredient during capsule or tablet manufacture. The hydrochloride and hydrobromide salts of the compounds of formula I have the further advantage that, after standing, they have a lower tendency to form concentrated aqueous solutions which can form viscous mixtures. Furthermore, the hydrobromide salt of the compound of formula I has a mild sedative effect at low to moderate doses. The mesylate salt of the compound of formula I has fast dynamic water solubility, which simplifies administration of the aqueous solution and makes it suitable for injectable dosage forms. In addition, the enhanced solubility of the mesylate salt of the compound of formula I may facilitate timely dissolution of the solid dosage form.

The compounds of formula I have greater kinetic solubilities for the p-toluenesulfonate, L-tartrate and hemi-citrate salts than for the free base or hydrochloride salt. In addition, the hygroscopicity of the p-toluenesulfonate, L-tartrate and hemi-citrate salts of the compound of formula I is less than that of the methanesulfonate salt of the compound of formula I. Thus, the p-toluenesulfonate, L-tartrate and hemi-citrate salts of the compounds of formula I are more stable in air and can be used without deliquescence.

The invention also relates to a pharmaceutical composition for treating hyperproliferative diseases in mammals, which comprises a therapeutically effective amount of salt of the compound of formula I or hydrate thereof and a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutical composition is for the treatment of cancer, such as brain, lung, squamous cell, bladder, stomach, pancreas, breast, head, neck, kidney, prostate, colorectal, esophageal, gynecological (e.g., ovarian), or thyroid cancer. In another embodiment, the pharmaceutical composition is for use in treating a non-cancerous hyperproliferative disease, such as benign hyperplasia of the skin (e.g., psoriasis) or of the prostate (e.g., Benign Prostatic Hypertrophy (BPH)).

The invention also relates to a pharmaceutical composition for treating pancreatitis or kidney diseases (including proliferative glomerulonephritis and diabetic kidney disease) in a mammal, comprising a therapeutically effective amount of a salt of a compound of formula I, or a hydrate thereof, and a pharmaceutically acceptable carrier.

The invention also relates to a pharmaceutical composition for preventing blast implantation in a mammal, comprising a therapeutically effective amount of a salt of a compound of formula I or a hydrate thereof, and a pharmaceutically acceptable carrier.

The invention also relates to a pharmaceutical composition for treating diseases related to angiogenesis or vasculogenesis in mammals, which comprises a therapeutically effective amount of the salt of the compound shown in the formula I or the hydrate thereof and a pharmaceutically acceptable carrier. In one embodiment, the pharmaceutical composition is for the treatment of a disease selected from tumor angiogenesis, chronic inflammatory diseases such as rheumatoid arthritis, atherosclerosis, skin diseases such as psoriasis, excema and scleroderma, diabetes, diabetic retinopathy, retinopathy of prematurity, age-related macular degeneration retinopathy, hemangioma, glioma, melanoma, kaposi's sarcoma and ovarian, breast, lung, pancreatic, prostate, colon and epidermoid cancers.

The present invention also relates to a method of treating a hyperproliferative disorder in a mammal comprising administering to said mammal a therapeutically effective amount of a salt of a compound of formula I or a hydrate thereof. In one embodiment, the method relates to the treatment of cancer, such as brain, squamous cell, bladder, stomach, pancreas, breast, head, neck, esophagus, prostate, colorectal, lung, kidney, gynecological (such as ovarian) or thyroid cancer. In another embodiment, the method relates to the treatment of a non-cancerous hyperproliferative disease, such as benign hyperplasia of the skin (e.g., psoriasis) or of the prostate (e.g., Benign Prostatic Hypertrophy (BPH)).

The present invention also relates to a method of treating a hyperproliferative disorder in a mammal comprising administering to said mammal a therapeutically effective amount of a salt of a compound of formula I or a hydrate thereof in combination with an anti-neoplastic agent selected from the group consisting of mitotic inhibitors, alkylating agents, antimetabolites, intercalating antibiotics, growth factor a preparations, cell cycle inhibitors, enzymes, topoisomerase inhibitors, biological response modifiers, anti-hormones, and anti-androgens.

The present invention also relates to a method for treating pancreatitis or kidney disease in a mammal which comprises administering to said mammal a therapeutically effective amount of a salt of a compound of formula I, or a hydrate thereof.

The invention also relates to a method of preventing blast implantation in a mammal comprising administering to said mammal a therapeutically effective amount of a salt of a compound of formula I or a hydrate thereof.

The present invention also relates to a method of treating a disease associated with angiogenesis or vasculogenesis in a mammal, the method comprising administering to the mammal an effective amount of a salt of a compound of formula I or a hydrate thereof. In one embodiment, the method is for treating a disease selected from tumor angiogenesis, chronic inflammatory diseases such as rheumatoid arthritis, atherosclerosis, skin diseases such as psoriasis, excema and scleroderma, diabetes, diabetic retinopathy, retinopathy of prematurity, macular degeneration retinopathy, hemangioma, glioma, melanoma, kaposi's sarcoma, and ovarian, breast, lung, pancreatic, prostate, colon, and epidermoid cancers.

Furthermore, the compounds of the present invention may be used as contraceptives in mammals. In a preferred embodiment, the compounds of the invention are useful for preventing conception in a female mammal.

Patients that can be treated according to the methods of the invention with a salt of a compound of formula I or a hydrate of said compound include, for example, patients who have been diagnosed with psoriasis, BPH, lung cancer, bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer or cancer of the anal region, stomach cancer, colon cancer, breast cancer, gynecological tumors (e.g., uterine sarcoma, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer or vulvar cancer), hodgkin's disease, esophageal cancer, small bowel cancer, cancer of the endocrine system (e.g., thyroid, parathyroid or adrenal cancer), soft tissue sarcoma, urinary tract cancer, penile cancer, prostate cancer, chronic or acute leukemia, solid tumors of childhood, lymphocytic lymphomas, bladder cancer, cancer of the kidney or ureter (e.g., renal cell carcinoma, renal pelvis cancer), or tumors of the central nervous system (e.g., primary CNS lymphoma, CNS lymphoma, Spinal axis tumors, brain stem glioma or pituitary adenoma).

The invention also relates to a pharmaceutical composition for the treatment of infections in a mammal, including a human, which infections are such as malaria or hepatitis delta, facilitated by farnesyl protein transferase which comprises a salt of a compound of formula I, a prodrug or solvate thereof, in an amount effective to treat abnormal cell growth, as described above, and a pharmaceutically acceptable carrier.

As used herein, "abnormal cell growth" generally refers to cell growth that is not constrained by normal regulatory mechanisms (e.g., loss of contact inhibition), unless otherwise indicated. This includes (1) abnormal growth of tumor cells (tumors) that express the activated Ras oncogene; (2) oncogene mutation in another gene, resulting in abnormal growth of tumor cells in which Ras protein is activated; (3) abnormal growth of benign and malignant cells of other proliferative diseases, in which aberrant Ras is activated; and (4) abnormal growth of tumor cells proliferating by farnesyl protein transferase.

The term "treating," as used herein, unless otherwise indicated, generally refers to reversing, alleviating, inhibiting the progression of, or preventing the disease in which it is used, or one or more symptoms of the disease. The term "treatment" as used herein, unless otherwise indicated, refers to the act of "treating" as defined immediately above.

Detailed description of the invention

The present invention relates to hydrochloride, hydrobromide, hemi-citrate, acetate, p-toluenesulfonate, L-tartrate, hemi-succinate and methanesulfonate salts of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide.

The invention also relates to a process for the preparation of hydrochloride, hydrobromide, hemi-citrate, acetate, p-toluenesulfonate, L-tartrate, hemi-succinate and methanesulfonate salts of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide. The salts of the invention are useful for the treatment of hyperproliferative diseases in mammals, such as cancer in mammals, especially humans, and for pharmaceutical compositions containing these compounds.

The salts of the compounds of formula I are characterized by X-ray powder diffraction. The X-ray powder diffraction patterns of the hydrochloride, hydrobromide, hemi-citrate, acetate, p-toluenesulfonate, L-tartrate, hemi-succinate (form a), hemi-succinate (form B) and methanesulfonate salts of the compound of formula I are substantially the same as those of figures 1-9.

The hydrochloride salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide of the present invention is characterized by a crystalline X-ray powder diffraction spectrum having a high intensity diffraction peak at diffraction angle (2-theta) of about: 8.623, 12.121, 17.298, 23.397, 23.944, 24.119, 24.873, 25.948, and 28.821.

The hydrobromide salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide according to the present invention is characterized by having a crystalline X-ray powder diffraction spectrum with high intensity diffraction peaks at diffraction angles (2-theta) of about: 8.687, 12.264, 17.374, 23.711, 24.335 and 25.769.

The hemi-citrate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide of the present invention is characterized by a crystalline X-ray powder diffraction spectrum having a high intensity diffraction peak at diffraction angle (2-theta) of about: 4.306, 16.317, 20.988, 21.476, 22.643, 23.384, 24.891, 27.573, and 27.840.

The acetic acid salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide according to the present invention is characterized by a crystalline X-ray powder diffraction spectrum having a high intensity diffraction peak at diffraction angle (2-theta) of about: 6.096, 12.183, 17.451, 18.288, 22.441, 23.086, and 24.439.

The p-toluenesulfonate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide of the present invention is characterized by a crystalline X-ray powder diffraction spectrum having a high intensity diffraction peak at diffraction angle (2-theta) of about: 20.446, 20.760, 22.092, 22.371, 23.190 and 26.239.

The L-tartrate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide of the present invention is characterized by a crystalline X-ray powder diffraction spectrum having a high intensity diffraction peak at diffraction angle (2-theta) of about: 4.061, 20.821, 21.634, 22.179 and 25.858.

The hemisuccinate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide of the present invention (form a) is characterized by a crystalline X-ray powder diffraction spectrum having a high intensity diffraction peak at diffraction angle (2-theta) of about: 4.634, 16.735, 22.179 and 25.002. At a relative humidity of 90%, the type a hemisuccinate absorbs 0.6% of the water.

The hemisuccinate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide of the present invention (form B) is characterized by a crystalline X-ray powder diffraction spectrum having a high intensity diffraction peak at diffraction angle (2-theta) of about: 6.714, 15.272, 19.197, 19.457, 24.487 and 24.802. At a relative humidity of 90%, the type B hemisuccinate absorbs 1.5% of the water.

The mesylate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide of the present invention is characterized by a crystalline X-ray powder diffraction spectrum having a high intensity diffraction peak at diffraction angle (2-theta) of about: 4.417, 17.288, 20.828, 21.677, 22.148, 25.427, and 27.006.

The in vitro activity of a compound of formula I to inhibit the KDR/VEGF receptor can be determined by the following method.

The ability of a compound of the invention to inhibit tyrosine kinase activity may be determined using a recombinase in an assay for determining the inhibition of the exogenous substrate polyGluTyr (PGT, Sigma)^TM4: 1) the ability to phosphorylate. The kinase domain of the human KDR/VEGF receptor (amino acids 805-1350) was expressed in Sf9 insect cells as glutathione S-transferase (GST) -fusion protein using a baculovirus expression system. This protein was purified from lysates of these cells using a glutathione agarose affinity column. Assays for this enzyme were performed in 96-well plates (0.625. mu.g PGT/well) embedded with PGT substrate. Test compounds were diluted in dimethyl sulfoxide (DMSO) and then added to the PGT plate to give a final DMSO concentration of 1.6% (v/v) in this assay. In phosphorylation buffer (50mM hepes, pH7.3, 125mM NaCl, 24mM MgCl)₂) Diluting the recombinant enzyme. This reaction was initiated by adding ATP to a final concentration of 10. mu.M. Incubate at room temperature for 30 minutes while shaking, then withdraw the reaction and buffer with a washThe plates were washed with solution (PBS-containing 0.1% Tween-20). The amount of phosphorylated PGT was determined by incubation with an HRP-coupled (HRP is horseradish peroxidase) PY-54 antibody (Transduction Labs), development with TMB peroxidase (TMB is 3, 3 ', 5, 5' -tetramethylbenzidine), and staining with 450nM BioRad^TMThe reaction was measured by a Microplate reader. The inhibition of the kinase enzyme activity is determined by the test compound as the absorbance decreases, and the concentration of the compound required to inhibit the signal to 50% is reported as the IC of this test compound₅₀The value is obtained.

To determine the ability of this compound to inhibit the activity of KDR tyrosine kinase on the full-length protein present in the pre-and post-cellular sequences, Porcine Aortic Endothelial (PAE) cells transfected with human KDR (Waltenberger et al, J.biol.chem.269: 26988, 1994) were used. Cells were cultured on plates, allowed to attach to 96-well plates in the same medium (Ham's F12) containing 10% v/v FBS (fetal bovine serum), and then serum-depleted medium (0.1% v/v FBS) containing 0.1% (v/v) Bovine Serum Albumin (BSA) was added and cultured for 16-24 hours. Serum-depleted medium (0.1% v/v FBS) (BSA-free) was added to the cells just prior to compound administration. Test compounds dissolved in DMSO were diluted in this medium (final DMSO concentration 0.5% (v/v)). At the end of the 2 hour incubation, VEGF was added₁₆₅(50ng/ml final concentration) was added to the medium and incubated for 8 minutes. The cells were washed and lysed in 50. mu.lysis buffer containing 20mM Tris-HCL (pH8), 150mM NaCl, 1% v/v NP40, 2mM NaVO₄500 μ M EDTA, 1mM PMSF and 1 plate/25 ml EDTA free complete  Protease Inhibitor Table, Roche. Then in PBS/1mM NaVO₄The cell lysate was diluted to a final volume of 150. mu.l. The extent of KDR phosphorylation was determined using ELISA. Prior to addition of anti-flk-1C-20 antibody (0.5. mu.g/well, Santa Cruz), the reactive coat-anti-rabbit plates (Pierce) were blocked with Superlock buffer (Pierce). All unbound antibody was washed off the plate before adding 100 μ l of cell lysate. After incubation of this lysate with flk-1 antibody for 2 hours, KDR junctions were determined by development with the HRP-conjugated PY-54 antibody and TMB as described aboveA synthetic phosphotyrosine. The ability of this compound to inhibit VEGF-stimulated autophosphorylation to 50% (compared to VEGF-stimulated controls) was reported as the IC of this test compound₅₀The value is obtained.

By means of these compound pairs³Incorporation of H-thymidine into HUVE cells (human umbilical vein endothelial cells, Clonetics)^TM) And their ability to inhibit human endothelial cell division is determined. This assay has been described in detail in the literature (Waltenberger J et al J.biol.chem.269: 26988, 1994; Cao Y et al J.biol.chem.271: 3154, 1996). Briefly, culture 10 was performed on collagen-embedded 24-well plates⁴Cells, to which they are attached. The cells were re-nourished in serum-free medium and 24 hours later with different concentrations of compounds (prepared in DMSO, final DMSO concentration in this assay is 0.2% v/v) and 2-30ng/ml VEGF₁₆₅And (6) processing. During the last 3 hours of 24 hours compound treatment, with³These cells were pulse labeled with H thymidine (NEN, 1. mu. Ci/well). The medium was then removed and the cells were rinsed thoroughly with ice cold Hank balanced salt solution and then 2 times with ice cold trichloroacetic acid (10% v/v). These cells were lysed by adding 0.2ml of 0.1N NaOH and the lysate was transferred to scintillation vials. The wells were then rinsed with 0.2ml 0.1N HCl and the rinses were transferred to the scintillation vials. Determination by scintillation counting³The extent of H thymidine incorporation. The ability of this compound to inhibit incorporation to 50% (compared to the control (VEGF treated with DMSO vehicle only)) is reported as the IC of this test compound₅₀The value is obtained.

The in vivo activity of the compound of formula I is determined by the amount of inhibition of tumor growth of the test compound compared to the control group. According to the Corbett T.H. and the like in the development of transplantable colon cancer used for chemotherapy test mice, the tumor induction relationship and the attention to the structure of carcinogenic substances are realized,Cancer Res.35, 2434-2439(1975) and Corbett, T.H. et al in "murine models of colon cancer for experimental treatment",Cancer Chemother.Rep.(Part 2)5, 169-186(1975), with minor modifications, different compounds were determinedThe tumor growth inhibitory effect of (1). 1X 10 suspended in 0.10ml of RPMI 1640 by subcutaneous injection⁶Tumor cells (human MDA-MB-468 breast or human HN5 head and neck cancer cells) were cultured in log phase, inducing tumors in the left flank. After a sufficient period of time, after palpable tumors (2-3 mm in diameter), the active compound is administered either intraperitoneally (ip) or orally (po), by dissolving in DMSO, usually at a concentration of 50-100mg/mL, and then diluting 1: 9 in saline, or 1: 9 in a solution containing 0.1% Pluronic^TMP105 in 0.9% saline) were treated to test animals (athymic mice) 2 times a day (i.e. 1 time every 12 hours) for 5 consecutive days. To determine the antitumor effect, 2 diameters of the tumor cross were measured in mm with a vernier caliper and the tumor weight was determined by the formula (length x [ width ])]²) The tumor size (mg) was calculated according to the method of Geran, R.l. et al, "report on experiments screening chemical drugs and natural products against animal tumors and other biological systems", third edition, cancer Chemotherm. The results are expressed as percent inhibition according to the formula: inhibition ratio (%) (TuW)_Control-TuW_{Test of})/TuW_ControlX 100%. The dose/response effect of this tumor implantation at the flank site was reproducible for various chemotherapeutic drugs and this measurement (tumor diameter) method was a reliable method to assess the tumor growth rate.

Administration of the compounds of the present invention (hereinafter "active compounds") can be accomplished by a variety of methods that deliver the compounds to the site of action. These methods include oral, intraduodenal, parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion), topical and rectal administration.

The amount of active compound administered will be determined according to the subject being treated, the severity of the disease, the rate of administration and the judgment of the prescribing physician. However, an effective dose is about 0.001 to 100mg/kg body weight/day, preferably about 1 to 35 mg/kg/day, in single or divided doses. For a 70kg human, the dosage should be about 0.05 to 7 g/day, preferably about 0.2 to 2.5 g/day. In some instances, dosage levels below the lower limit of the aforesaid dosage range may not be sufficient, while in other instances larger doses may be employed without causing any harmful side effects, provided that such larger doses are first divided into several small doses for administration throughout the day.

The active compound may be applied as a monotherapy or may include one or more other anti-tumour agents, for example those selected from: mitotic inhibitors, such as vinblastine; alkylating agents, such as cis-platinum ammonia, carboplatin, and cyclophosphamide; antimetabolites, for example 5-fluorouracil, cytarabine and hydroxyurea, or a preferred antimetabolite, for example as disclosed in European patent application No.239362, such as N- (5- [ N- (3, 4-dihydro-2-methyl-4-oxoquinazolin-6-ylmethyl) -N-methylamino]-2-thenoyl) -L-glutamic acid; a growth factor inhibitor; a cell cycle inhibitor; intercalating antibiotics, such as doxorubicin and bleomycin; enzymes, such as interferon; and anti-hormones, e.g. anti-estrogens, such as Nolvadex^TM(tamoxifen), or, for example, antiandrogens, such as Casodex^TM(4 '-cyano-3- (4-fluorophenylsulfonyl) -2-hydroxy-2-methyl-3' - (trifluoromethyl) -propionylanilide). Such combination therapy can be achieved by the simultaneous, sequential or separate administration of the individual therapeutic ingredients.

For example, the pharmaceutical composition may be in a form suitable for oral administration, such as tablets, capsules, pills, powders, sustained release formulations, solutions, suspensions; may be in a form suitable for parenteral injection, such as a sterile solution, suspension or emulsion; may be in a form suitable for topical administration, such as an ointment or cream; or may be in a form suitable for rectal administration, such as a suppository. The pharmaceutical composition may be in unit dosage form suitable for single administration of precise dosages. The pharmaceutical composition will include a conventional pharmaceutical carrier or excipient, and a compound of the invention as an active ingredient. In addition, it may include other pharmaceutical or pharmaceutical ingredients, carriers, adjuvants, etc.

Typical parenteral administration forms include solutions or suspensions of the active compounds in sterile aqueous solutions, for example, aqueous propylene glycol or glucose solutions. Such dosage forms may be suitably buffered with a buffer, if desired.

Suitable pharmaceutical carriers include inert diluents or fillers, water and various organic solvents. Such pharmaceutical compositions may contain additional components, such as flavoring agents, binders, excipients, and the like, if desired. Thus for oral administration, tablets containing various excipients, such as citric acid, may be employed together with various disintegrants such as starch, alginic acid and certain complex silicates, together with binding agents such as sucrose, gelatin and acacia. Additionally, lubricating agents, such as magnesium stearate, sodium lauryl sulfate, and talc, are often used for tableting. Solid compositions of a similar type may also be employed in soft and hard-filled capsules. Thus, preferred starting materials include lactose and high molecular weight polyethylene glycols. When aqueous suspensions or elixirs are desired for oral administration, the active compound may be combined with various sweetening or flavoring agents, coloring matter or dyes, and, if desired, emulsifying or suspending agents, together with a diluent such as water, ethanol, propylene glycol, glycerol, or combinations thereof.

Methods for preparing various pharmaceutical compositions containing a particular amount of an active compound are well known or will be apparent to those skilled in the art. For example, see《Remington′s Pharmaceutical Sciences》Mack publishing company, Easter, Pa., 15 th edition (1975).

The following examples and formulations further illustrate and exemplify the compounds of the present invention, as well as methods for preparing such compounds. It will be clear that the scope of the examples and formulations which follow are not intended to limit the scope of the invention in any way.

Diffraction spectra of figures 1-9 were recorded using a Siemens θ/2 θ powder diffractometer, configured as follows: a 40 bit autosampler, a fixed slit goniometer, a sealed tube copper (Cu) X-ray source (wavelength 1: 1.54056, wavelength 2: 1.54439), and a Kevex solid state detector. Tube power: 40-mA x 50-kV, or as appropriate. Slit: 1X 0.6mm (source, anti-scatter and detector slits, respectively). Step length: 0.04 degree in 2T. The time of each step: for 1 second. Scanning is started: 3 degrees in 2T. And (3) terminating the scanning: 2T medium 40 degrees.

Example 1

Free base of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid

The free base of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid was prepared according to the procedure described in example 30 of U.S. serial No. 09/316837, filed on 21/5/1999, which disclosure is hereby incorporated herein by reference in its entirety. And Mp: 208 Deg.C (DSC). Characteristic X-ray powder diffraction peaks (2-theta, [% relative intensity ]): 9.314[100.0], 11.356[44.8], 15.897[49.6], 22.059[84.5], 22.520[63.3], 22.726[70.0], 23.927[67.6], 24.307[60.5], 25.310[64.8] and 26.551[86.6 ].

Example 2

Hydrochloride salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrazin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide

Reacting 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido]-isothiazole-4-carboxylic acid amide (500mg, 0.939mmol) was dissolved in refluxing EtOH (20mL), allowed to cool to room temperature, and treated with HCl (0.94mL of 1.0M Et₂Solution O) while swirling the flask. Then, the mixture was gently shaken while heating at 50 ℃ for 3 hours and kept at room temperature for 3 days. The solid was filtered and dried under high vacuum to give a white solid (468mg, 0.823mmol, 82%). Melting point: 230 Deg.C (DSC). Moisture absorption: 1% by weight at room temperature under 90% Relative Humidity (RH). Characteristic X-ray powder diffraction peaks (2-theta, [% relative intensity)])：8.623[90.7]、12.121[38.9]、17.298[95.2]、23.397[44.7]、23.944[51.7]、24.119[62.7]、24.873[55.7]、25.948[100]And 28.821[39.6]]。

Example 3

Hydrobromide salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid

Hydrobromic acid (1.0mL of a 47-49% aqueous 8.9M solution) was added to 4mL of MeOH in a graduated cylinder, which was then filled with MeOH to 8.9 mL. In addition, 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido]-isothiazole-4-carboxylic acid (500mg, 0.939mmol) dissolved in CH₂Cl₂(10mL) and MeOH (4mL) and treated with HBr (1.0mL of the above solution). This solution was then placed in a flask containing Et₂O in a diffusion cell. After 16 hours, a solid appeared. With fresh Et₂Replacement of Et therein by O₂O, and diffusion was continued overnight. A white solid was obtained (529mg, 0.863mmol, 86%). Melting point: 201.0 Deg.C (DSC). Moisture absorption: 0.1% at 87% relative humidity RH. Characteristic X-ray powder diffraction peaks (2-theta, [% relative intensity)])：8.687[100.0]、12.264[35.9]、17.374[42.3]、23.711[24.0]、24.335[20.7]And 25.769[34.3]]。

Example 4

Hemi-citrate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid

3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid (532mg, 1.00mmol), citric acid (96mg, 0.50mmol) and MeOH (8mL) were combined in a septum-capped 16mL vial at 75 deg.C and heated, shaken and allowed to stir for 24 h. The mixture was cooled to room temperature and filtered. The solid was rinsed with MeOH and air was continuously passed through the solid for drying. A white solid was obtained (530mg, 0.843mmol, 84%). Melting point: 201.7 Deg.C (DSC). Moisture absorption: 0.43% at 87% relative humidity RH. Characteristic X-ray powder diffraction peaks (2-theta, [% relative intensity ]): 4.306[79.9], 16.317[100.0], 20.988[32.7], 21.476[30.9], 22.643[48.7], 23.384[76.9], 24.891[76.0], 27.573[47.9] and 27.840[32.3 ].

Example 5

Acetate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid

3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] at 75 deg.C]Isothiazole-4-carboxylic acid (532mg, 1.00mmol), acetic acid (57 μ L, 1.0mmol) and MeOH (3mL) were combined in a 8mL vial with septum cap and heated, shaken and allowed to stand for 24 h. The mixture was cooled to room temperature and placed in the flask containing Et₂O in a box. After 5 hours, the reaction mixture was poured off, washed with MeOH and then Et₂The solid was washed with O to obtain large crystals. Drying is carried out by briefly passing air through the solid. A white solid was obtained (330mg, 0.557mmol, 56%). Melting point: 175 Deg.C (DSC). Characteristic X-ray powder diffraction peaks (2-theta, [% relative intensity)])：6.096[21.7]、12.183[21.4]、17.451[33.3]，18.288[100.0]，22.441[57.7]，23.086[19.9]And 24.439[20.7]]。

Example 6

Para-toluenesulfonate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid

Reacting 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido]-isothiazole-4-carboxylic acid (532mg, 1.00mmol), p-toluenesulfonic acid monohydrate (179mg, 1.00mmol), MeOH (10mL) and CH₂Cl₂(1ml) are mixed and filtered to remove small amounts of fine particles and use additional CH₂Cl₂(3ml) rinse thoroughly. This solution was added to additional CH₂Cl₂(4ml) and MeOH (1ml) and placed in a flask containing Et₂O overnight in diffusion cells. Without any crystal formation, pentane then replaces Et₂O, overnight. With Et₂O washes the solid and dries by continuously passing air through the solid. Obtaining a white pigmentA colored solid (572mg, 0.812mmol, 81%). Melting point: 140 and 174 ℃ (DSC). Moisture absorption: 0.9% at 87% relative humidity RH. Characteristic X-ray powder diffraction peaks (2-theta, [% relative intensity)])：20.446[100.0]、20.760[74.0]、22.092[81.7]、22.371[70.8]、23.190[65.2]、26.239[61.5]。

Example 7

L-tartrate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid

3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid (532mg, 1.00mmol), L-tartaric acid (150mg, 1.0mmol) and MeOH (8mL) were mixed and heated in a 16mL vial with septum cap at 75 deg.C and shaken for 24 h. The mixture was cooled to room temperature and filtered. The solid was rinsed with MeOH and dried by continuously passing air through the solid. A white solid was obtained (617mg, 0.904mmol, 90%). Melting point: 206 ℃ (DSC). Moisture absorption: 0.3% at 100% relative humidity RH. Characteristic X-ray powder diffraction peaks (2-theta, [% relative intensity ]): 4.061[82.9], 20.821[85.6], 21.634[100.0], 22.179[94.0], 25.858[95.1 ].

Example 8

Hemisuccinate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid (type A)

3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid (532mg, 1.00mmol), succinic acid (59mg, 0.50mmol) and MeOH (8mL) were combined in a septum cap 16mL vial at 75 deg.C, heated, and shaken for 24 h. The mixture was cooled to room temperature and filtered. The solid was rinsed with MeOH and dried by continuously passing air through the solid. A white solid was obtained (500mg, 0.845mmol, 85%). Type a melting point: 216 Deg.C (DSC). Moisture absorption: 0.6% at 90% relative humidity RH. Characteristic X-ray powder diffraction peaks (2-theta, [% relative intensity ]): type A: 4.634[100.0], 16.735[67.2], 22.179[60.8] and 25.002[70.3 ].

Example 9

Hemisuccinate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidino-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid (type B)

3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid (100mg) was dissolved with heating in 6mL EtOH: MeOH (2: 1). Succinic acid (11.1mg, 0.5eq) dissolved in EtOH was added to the previous solution. The mixture was cooled to room temperature and stirred for 20 minutes. The solid was filtered, rinsed with MeOH, and dried by continuously passing air through the solid. A white solid was obtained (80mg, 70%). Type B hygroscopicity: 1.5% at 90% relative humidity RH. Characteristic X-ray powder diffraction peaks (2-theta, [% relative intensity ]): 6.714[ 31.0], 15.272[100.0], 19.197[59.3], 19.457[50.0], 24.487[99.0] and 24.802[79.1 ].

Example 10

Methanesulfonic acid salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid

Reacting 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido]-isothiazole-4-carboxylic acid (10.9g, 20.5mmol) was dissolved in MeOH (150mL) and cooled to 0 ℃. In a liquid separation bottle, H is put₃CSO₃H (1.33mL) was added to 0 ℃ MeOH (15 mL). This acid solution was added dropwise to the amine starting solution over 10 minutes. The solution was warmed to room temperature, filtered to remove minor solid impurities, and treated with Et₂O (1L) was diluted and stirred for 1 hour. The mixture was further diluted with hexane (500mL) and cooled to 0 ℃ while continuing to stir. After standing overnight at 0 deg.C, the crystals were filtered, washed with hexane and dried by suction to give a white solid (11.1g, 17.7mmol, 86%). Characteristic X-ray powder diffraction peaks (2-theta, [% relative intensity)])：4.417[99.3]、17.288[45.5]、20.828[39.6]、21.677[43.5]、22.148[68.3]、25.427[100.0]And 27.006[37.5]]。

Claims (20)

1. A hydrobromide salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide.

2. The salt according to claim 1, wherein the salt has an X-ray diffraction spectrum at 2-theta substantially the same as: 8.687, 12.264, 17.374, 23.711, 24.335, or 25.769.

3. A hemi-citrate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide.

4. A salt according to claim 3, wherein the salt has an X-ray diffraction spectrum at 2-theta substantially the same as: 4.306, 16.317, 20.988, 21.476, 22.643, 23.384, 24.891, 27.573, or 27.840.

5. An acetate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide.

6. The salt according to claim 5, wherein the salt has an X-ray diffraction spectrum at 2-theta substantially the same as: 6.096, 12.183, 17.451, 18.288, 22.441, 23.086, or 24.439.

7. A p-toluenesulfonate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide.

8. The salt according to claim 7, wherein the salt has an X-ray diffraction spectrum at 2-theta substantially the same as: 20.446, 20.760, 22.092, 22.371, 23.190, or 26.239.

9. An L-tartrate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide.

10. The salt according to claim 9, wherein the salt has an X-ray diffraction spectrum at 2-theta substantially the same as: 4.061, 20.821, 21.634, 22.179, or 25.858.

11. A hemisuccinate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide which is form A.

12. The salt according to claim 11, wherein the salt has an X-ray diffraction spectrum at 2-theta substantially the same as: 4.634, 16.735, 22.179, or 25.002.

13. A hemisuccinate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide which is form B.

14. The salt according to claim 13, wherein the salt has an X-ray diffraction spectrum at 2-theta substantially the same as: 6.714, 15.272, 19.197, 19.457, 24.487, or 24.802.

15. Use of a compound of claim 1, 3, 5, 7, 9, 11, or 13 for the manufacture of a medicament for treating a hyperproliferative disorder in a mammal.

16. The use of claim 15, wherein the use is for the treatment of a cancer selected from brain, squamous cell, bladder, stomach, pancreas, breast, head, neck, esophagus, prostate, colorectal, lung, kidney, ovarian, gynecological and thyroid cancer.

17. A pharmaceutical composition comprising an amount of a compound of claim 1, 3, 5, 7, 9, 11, or 13 effective to treat a hyperproliferative disorder in a mammal and a pharmaceutically acceptable carrier.

18. The pharmaceutical composition of claim 17, wherein the hyperproliferative disease is a cancer selected from the group consisting of brain, lung, squamous cell, bladder, stomach, pancreas, breast, head, neck, kidney, ovary, prostate, colorectal, esophagus, gynecological, and thyroid cancers.

19. A process for preparing a hydrobromide salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide comprising reacting hydrobromic acid with 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide.

20. A process for preparing a hemi-citrate salt of 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide comprising reacting citric acid with 3- (4-bromo-2, 6-difluoro-benzyloxy) -5- [3- (4-pyrrolidin-1-yl-butyl) -ureido ] -isothiazole-4-carboxylic acid amide.