TW201425298A - 4-(4-(3-(4- chloro-3-(trifluoromethyl) phenyl) ureido)-3- fluorophenoxy)-2- (N-1', 1', 1'-d3-methyl) picolinamide salt, solvate thereof, preparation method and uses - Google Patents

Abstract

Disclosed are a 4-(4-(3-(4- chloro-3-(trifluoromethyl) phenyl) ureido)-3- fluorophenoxy)-2- (N-1', 1', 1'-d3-methyl) picolinamide salt, a solvate thereof and a preparation method thereof. The 4-(4-(3-(4- chloro-3-(trifluoromethyl) phenyl) ureido)-3- fluorophenoxy)-2- (N-1', 1', 1'-d3-methyl) picolinamide salt or its solvate can be used to prepare a pharmaceutical composition for inhibiting the phosphokinase activity.

Description

4-(4-(3-(4-chloro-3-(trifluoromethyl)phenyl) guanidine)-3-fluoro- Salt of phenoxy)-2-(N-1',1',1'-tridemethyl)pyridiniumamine, solvate thereof, preparation method thereof and use thereof

The present invention relates to a fluorine-containing deuterated diphenylurea compound, in particular to a 4-(4-(3-(4-chloro-3-(trifluoromethyl)phenyl)guanidine)- A salt of 3-fluoro-phenoxy)-2-(N-1',1',1'-tridemethyl)pyridiniumamine, a solvate thereof, a process for the preparation thereof, and use thereof.

The known ω-diphenylurea derivatives are compounds of c-RAF kinase activity. A class of ω-carboxyaryl substituted diphenylureas, and their use in the treatment of cancer and related diseases are disclosed, for example, in WO 2000/042012.

Omega-diphenyl urea compounds such as Sorafenib were first discovered to be inhibitors of c-RAF kinase, and subsequent studies have found that it also inhibits MEK and ERK signaling pathways, vascular endothelial growth factor-2 (VEGFR-2), vascular endothelial growth factor-3 (VEGFR-3), and platelet-derived growth factor-β (PDGFR-β) tyrosine kinase activity (Curr Pharm Des 2002; 8: 2255-2257), It is therefore called a multi-kinase inhibitor and has a dual anti-tumor effect.

Sorafenib, trade name Nexavar, is a new oral multi-kinase inhibitor developed by Bayer and ONXY, due to its superior performance in a phase III clinical study of advanced kidney cancer. In December 2005, it was quickly approved by the FDA for the treatment of advanced renal cell carcinoma. It was launched in China in November 2006. However, Sorafenib has various side effects such as high blood pressure, weight loss, and rash.

BAY 73-4506 is a 3-fluoro-based derivative of sorafenib and is also targeted as an inhibitor of multikinases in tumors and their blood vessels. In particular, BAY 73-4506 is a potent inhibitor of raf kinase, p38 kinase, platelet growth factor receptor (PDGFR) kinase, and vascular endothelial growth factor receptor (VEGFR) kinases 2 and 3. Inhibition of these specific kinases has been closely related to the prevention or treatment of osteoporosis, inflammatory diseases, proliferative diseases, angiogenic diseases including cancer (Dumas et al, patent PCT Publication WO 2005009961 A2, Hedbom S et al, Journal of Clinical Oncology , 25, (Suppl. 18): Abs, 3593). The compound is currently undergoing renal cell carcinoma (ClinicalTrials.gov Web Site 2008, May 05), solid tumor (J. Clin. Oncol., 2008, 26 (15, Suppl.): Abst 2558), and liver cancer (ClinicalTrials.gov Web Clinical evaluation of Site 2010, May 17) and metastatic colon cancer (ClinicalTrials.gov Web Site 2010, May 17). BAY 73-4506 is undergoing preclinical evaluation of the treatment of multiple myeloma (Blood, 2008, 112(11): Abst 2766).

However, there is still a need in the art to develop compounds having inhibitory activity against raf kinase, or better pharmacodynamic properties, and processes thereof.

It is an object of the present invention to provide a novel class of compounds having raf kinase inhibitory activity and better pharmacodynamic properties and uses thereof.

Another object of the present invention is to provide a series of synthetic methods for deuterated ω-diphenylurea and intermediates thereof, thereby meeting the standards in the pharmaceutical industry production, and improving operability and safety.

In a first aspect of the invention, there is provided an intermediate of formula B2.

In a second aspect of the invention, there is provided a process for the preparation of a compound of the formula:

The method comprises: (a) reacting a compound of formula B2 with a compound of formula V in an inert solvent and in the presence of a base to form the compound;

Wherein X is Cl, Br, I, or OSO ₂ CF ₃ ; alternatively, the method comprises: (b) reacting a compound of formula B' with CD ₃ NH ₂ or CD ₃ NH ₂ ‧ HCl in an inert solvent, Forming the compound;

Wherein R is H, a C ₁ -C ₈ linear or branched alkyl group, or an aryl group; or the method comprises: (c) 4-chloro-3-trifluoromethylbenzene in an inert solvent The isocyanate (VIII) is reacted with a compound of formula 5 to form the compound;

Alternatively, the method comprising: (d) in an inert solvent, in CH ₂ Cl ₂ and CDI exist, compounds of formula 5 with a compound of formula 6, the compound formed.

Alternatively, the process comprises: (e) reacting 3-fluoro-4-amino-phenol with a formula V in an inert solvent to form a compound of formula B1; reacting the compound of formula B with formula W in an inert solvent To form a compound of formula M; and reacting a compound of formula M with 4-chloro-3-trifluoromethylaniline in an inert solvent to form the compound

Wherein R is hydrogen, methyl, nitro and chloro; the inert solvent is selected from the group consisting of chloroform, dichloromethane, tetrahydrofuran, 1,4-dioxane, toluene, pyridine, DMF, DMSO, or a mixture thereof a solvent; the base is selected from the group consisting of pyridine, potassium carbonate, sodium carbonate, sodium hydrogencarbonate, potassium hydrogencarbonate, triethylamine, N , N -diethylisopropylamine, N , N -dimethylamine pyridine, N -A Kemorphin, or a combination thereof; or, the method comprises: (f) reacting 4-chloro-3-trifluoromethylaniline with a formula W in an inert solvent to form a compound of formula B; in an inert solvent The compound of formula B is reacted with a compound of formula B to form the compound.

Wherein R is hydrogen, methyl, nitro and chloro; the inert solvent is selected from the group consisting of chloroform, dichloromethane, tetrahydrofuran, 1,4-dioxane, toluene, pyridine, DMF, DMSO, or a mixture thereof a solvent; the base is selected from the group consisting of pyridine, potassium carbonate, sodium carbonate, sodium hydrogencarbonate, potassium hydrogencarbonate, triethylamine, N, N -diethylisopropylamine, N , N -dimethylamine pyridine, N -A Kemorphin, or a combination thereof.

In another preferred embodiment, the compound of the formula B2 is prepared by the following method: 3-fluoro-4-aminophenol and 4-chloro-3-trifluoromethylphenylisocyanate are carried out in an inert solvent. Condensation to form a compound of formula B2:

In another preferred embodiment, the inert solvent includes, but is not limited to, dichloromethane, dichloroethane, acetonitrile, n-hexane, toluene, tetrahydrofuran, N , N -dimethylformamide, N. , N -dimethylacetamide, N -methylpyrrolidone, dimethylhydrazine, or a combination thereof.

In another preferred embodiment, the base is selected from the group consisting of potassium t-butoxide, sodium hydride, potassium hydride, potassium carbonate, cesium carbonate, potassium phosphate, potassium hydroxide, sodium hydroxide or a combination.

In another preferred embodiment, the method (a) further comprises: carrying out the reaction in the presence of a catalyst selected from the group consisting of a combination of cuprous iodide and valine acid, or a combination of cuprous iodide and picolinic acid.

In another preferred embodiment, each reaction is carried out at a temperature of from 0 to 200 °C.

In another preferred embodiment, the reaction time is 0.2-72 hours, The preferred area is 0.5-64 hours, more preferably 1-48 hours.

In a third aspect of the invention, there is provided a process for the preparation of 4-chloropyridin-2-( N -1',1',1'-tridecylidenemethyl)guanamine comprising the steps of:

(a1) reacting methyl 4-chloro-2-picolinate with 1,1,1-tris-methylamine or a salt thereof (such as a hydrochloride) under an alkaline condition in an inert solvent, thereby Forming 4-chloro-pyridin-2-(N-1',1',1'-tridemethyl) decylamine; or

(a2) 4-chloro-2-picolinic acid is reacted with 1,1,1-tridemethylamine or a salt thereof (e.g., hydrochloride) in an inert solvent to form 4-chloro-pyridine-2 -( N -1',1',1'-tridemethyl)carbamamine; or

(a3) reacting 4-chloro-pyridine-2-carboxamidine with 1,1,1-tridecylmethylamine in an inert solvent to form 4-chloro-pyridine-2-( N -1', 1',1'-triterpene methyl)formamidine

In another preferred embodiment, the inert solvent includes tetrahydrofuran, ethanol, methanol, water, or a mixed solvent thereof.

In another preferred embodiment, in the steps (a1) and (a2), the reaction temperature is -10 ° C to reflux temperature, preferably -4 ° C to 60 ° C, more preferably 5-50 °C.

In another preferred embodiment, in the steps (a1) and (a2), the reaction time is from 0.5 to 72 hours, preferably from 1 to 64 hours, more preferably from 2 to 48 hours.

In another preferred embodiment, in the step (a1), the alkaline condition is that potassium carbonate, sodium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide or a combination thereof is present in the reaction system.

In a fourth aspect of the invention, there is provided a process for the preparation of a compound of formula B2 comprising the steps of: 3-fluoro-4-aminophenol and 4-chloro-3-trifluoromethylphenylisocyanate in an inert solvent Condensation is carried out to form a compound of formula B2:

In a fifth aspect of the invention, there is provided the use of an intermediate according to the first aspect of the invention, which is used for the preparation of deuterated diphenylurea or as a starting material for the preparation of deuterated diphenylurea.

In another preferred embodiment, the deuterated diphenyl urea comprises 4-(4-(3-(4-chloro-3-(trifluoromethyl)phenyl) guanidine)-3-fluoro-benzene Oxy)-2-( N -1',1',1'-tridemethyl)pyridiniumamine (CM4309); 4-(4-(3-(4-chloro-3-(trifluoromethyl)) (phenyl) guanidinium)-3-fluoro-phenoxy)-2-( N -1',1',1'-trimethylaminomethylmercapto)pyridine-1-oxide; -(4-(3-(4-chloro-3-(trifluoromethyl)phenyl) guanidine)-3-fluoro-phenoxy)-2-( N -1',1',1'- Triterpene methyl) pyridinium p-toluenesulfonate (CM4309‧TsOH); 4-(4-(3-(4-chloro-3-(trifluoromethyl)phenyl)indole)-3- Fluoro-phenoxy)-2-( N -1',1',1'-tridemethylmethyl)pyridinium hydrochloride; or 4-(4-(3-(4-chloro-3-) (trifluoromethyl)phenyl)carbazide)-3-fluoro-phenoxy)-2-( N -1',1',1'-tridemethyl)pyridinium methanesulfonate Things.

In a sixth aspect of the invention, there is provided an intermediate of formula V,

Wherein X is Cl, Br, I, or OSO ₂ CF ₃ .

In another preferred embodiment, X is Cl and the structural formula is .

In a seventh aspect of the invention, there is provided the use of the intermediate of the sixth aspect of the invention, which is used for the preparation of deuterated diphenylurea or as a starting material for the preparation of deuterated diphenylurea.

In an eighth aspect of the invention, there is provided the use of a compound of the formula (CM4309) for the preparation of a pharmaceutical composition for inhibiting a phosphokinase, such as raf kinase.

In another preferred embodiment, the pharmaceutical composition is for treating and preventing a disease: cancer, cardiovascular disease, inflammation, immune disease, kidney disease, angiogenesis, or prostate disease.

In another preferred embodiment, the cancer includes (but is not limited to ): non-small cell lung cancer, uterine cancer, rectal cancer, brain cancer, head cancer, neck cancer, bladder cancer, prostate cancer, breast cancer, solid tumor, kidney cancer, blood cancer, liver cancer, stomach cancer, or pancreatic cancer.

In a ninth aspect of the invention, there is provided a method of treatment comprising the step of administering a CM4309 compound of the invention, or a crystalline form, a pharmaceutically acceptable salt, hydrate or solvate thereof, to a subject in need of treatment, or A pharmaceutical composition comprising the compound is administered to inhibit a phosphokinase (such as raf kinase). Preferably, the disease comprises cancer, cardiovascular disease, inflammation, immune disease, kidney disease, angiogenesis, or prostate disease.

It is to be understood that within the scope of the present invention, the various technical features of the present invention and the technical features specifically described hereinafter (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here.

Figure 1 shows the inhibition of CM4306, CM4308 and CM4309 on human hepatocellular carcinoma SMMC-7721 nude mouse transplantation model (Mean+SEM). In the figure, "treatment" means that the treatment time is 14 days. "control" means a blank control.

The present inventors have unexpectedly discovered that the fluorine-containing deuterated omega-diphenylurea of the present invention and pharmaceutically acceptable salts thereof have not been deuterated Compared with compounds, the compounds have significantly better pharmacokinetic and/or pharmacodynamic properties, and thus are more suitable as compounds for inhibiting raf kinase, and thus more suitable for the preparation of drugs for treating cancer and related diseases. Further, the inventors have found that a novel high-purity diphenylurea compound can be produced efficiently and simply by using a novel intermediate of the formula V or the formula B2. The present invention has been completed on this basis.

definition

As used herein, "halogen" refers to F, Cl, Br, and I. More preferably, the halogen atom is selected from the group consisting of F, Cl and Br.

As used herein, "alkyl" includes straight or branched alkyl groups. Preferred alkyl groups are C ₁ -C ₄ alkyl groups such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl and the like.

As used herein, "deuterated" means that one or more hydrogens in a compound or group are replaced by deuterium. Deuterated can be monosubstituted, disubstituted, polysubstituted or fully substituted. The terms "one or more deuterated" are used interchangeably with "one or more deuterated".

In another preferred embodiment, the cerium isotope content of cerium at the cerium substitution site is greater than the natural strontium isotope content (0.015%), more preferably greater than 50%, more preferably greater than 75%, and even more preferably greater than 95%, more preferably The ground is greater than 97%, more preferably greater than 99%, and even more preferably greater than 99.5%.

In another preferred embodiment, the compound of formula (I) contains at least one deuterium atom, more preferably three deuterium atoms, more preferably five deuterium atoms.

As used herein, the term "compound CM4306" refers to the compound 4-(4-(3-(4-chloro-3-(trifluoromethyl)phenyl)indole)-phenoxy) -N -methylpyridinium amine.

As used herein, the term "compound CM4308" refers to the compound 4- (4- (3- (4-chloro-3- (trifluoromethyl) phenyl) acyl urea) -3-fluoro - phenoxy) - N - Methyl pyridinamine.

As used herein, the term "compound CM4309" refers to the compound 4-(4-(3-(4-chloro-3-(trifluoromethyl)phenyl)guanidine)-3-fluoro-phenoxy)-2- ( N -1',1',1'-tridemethyl)pyridinium.

As used herein, the term "TsOH" means p-toluenesulfonic acid. CM4309‧TsOH represents the p-toluenesulfonate of compound CM4309.

Deuterated diphenylurea

Preferred deuterated diphenyl urea compounds of the invention have the structure of formula (I):

Wherein X is N or N ⁺ -O ^- ; R ¹ is halogen (such as F, Cl or Br), one or more deuterated or fully deuterated C ₁ -C ₄ alkyl groups; R ² is not Deuterated, one or more deuterated or fully deuterated C ₁ -C ₄ alkyl groups, or a partially or fully halogen-substituted C ₁ -C ₄ alkyl group; R ³ , R ⁴ , R ⁵ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R ¹⁴ are each hydrogen, deuterium, or halogen (such as F, Cl, or Br); R ⁶ is hydrogen, deuterium, or one or more deuterated Or fully deuterated C ₁ -C ₄ alkyl; R ⁷ is hydrogen, deuterium, or one or more deuterated or fully deuterated C ₁ -C ₄ alkyl; with the proviso that R ² , R ³ And at least one of R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ or R ¹⁴ is deuterated C ₁ -C ₄ alkyl or fluorene.

In another preferred embodiment, the cerium isotope content of the cerium at the cerium substitution site is at least greater than the natural strontium isotope content (0.015%), preferably greater than 30%, more preferably greater than 50%, and even more preferably greater than 75%. Preferably, the ground is greater than 95%, more preferably greater than 99%.

In another preferred embodiment, all elements other than H (such as N, C, O, F, etc.) in the compound of formula (I) are all or substantially (>99 wt%) being the most naturally occurring element, For example ¹⁴ N, ¹² C, ¹⁶ O and ¹⁹ F.

In another preferred embodiment, the compound of formula (I) contains at least one deuterium atom, more preferably three deuterium atoms, more preferably five deuterium atoms.

In another preferred embodiment, R ^{1 is} selected from halogen; more preferably chlorine.

In another preferred embodiment, R ² is trifluoromethyl.

In another preferred embodiment, R ⁶ or R ⁷ are each independently selected from hydrogen, deuterium, deuterated methyl, or deuterated ethyl; more preferably, selected from monomethyl, dimethyl, Triterpene methyl, monoethylidene, dieththylethyl, tridecylethyl, tetradecylethyl, or pentadecylethyl.

In another preferred embodiment, R ⁶ or R ⁷ are each independently selected from hydrogen, methyl or trimethylmethyl.

In another preferred embodiment, R ³ , R ⁴ or R ⁵ are each independently selected from hydrogen or hydrazine.

In another preferred embodiment, R ⁸ , R ⁹ , R ¹⁰ or R ¹¹ are each independently selected from hydrogen or hydrazine.

In another preferred embodiment, R ¹² , R ¹³ or R ¹⁴ are each independently selected from hydrogen or hydrazine.

In another preferred embodiment, the compound is a preferred compound selected from the group consisting of N- (4-chloro-3-(trifluoromethyl)phenyl) -N '-(4-(2-( N - 1',1',1'-trimethylaminomethylmercapto)-4-pyridyloxy)phenyl)urea;

4-(4-(3-(4-chloro-3-(trifluoromethyl)phenyl)ureido)phenoxy)-2-( N -1 ',1',1'-trimethylmethyl Aminomethylmercapto)pyridine-1-oxide;

4-(4-(3-(4-Chloro-3-(trifluoromethyl)phenyl) guanidine)-3-fluoro-phenoxy)-2-( N -1',1',1' - triterpene methyl) pyridinamine (CM4309);

4-(4-(3-(4-Chloro-3-(trifluoromethyl)phenyl) guanidine)-3-fluoro-phenoxy)-2-( N -1',1',1' - trimethylaminomethylmercapto)pyridine-1-oxide;

Intermediate

As used herein, the term "intermediate of the invention" refers to a compound of formula V, a compound of formula B' (particularly a compound of formula B2).

Other elements other than H (such as N, C, O, etc.) in the above formula are all or substantially (>99 wt%) being the most abundant naturally occurring elements, such as ¹⁴ N, ¹² C and ¹⁶ O.

Active ingredient

The term "compound of the invention" as used herein refers to a compound of formula (I). The term also encompasses various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of formula (I).

A particularly preferred compound is CM4309 or a pharmaceutically acceptable salt thereof.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt of the compound of the invention formed with an acid or base suitable for use as a medicament. Pharmaceutically acceptable salts include inorganic and organic salts. A preferred class of salts are the salts of the compounds of the invention with acids. Suitable acids for forming salts include, but are not limited to, mineral acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, An organic acid such as maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, benzene methanesulfonic acid or benzenesulfonic acid; and an acidic amino acid such as aspartic acid or glutamic acid.

Preparation

The preparation of the structural compound of the formula (I) of the present invention and the novel intermediate will be described more specifically below, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention may also be conveniently prepared by combining various synthetic methods described in the specification or known in the art, and such combinations are readily made by those skilled in the art to which the present invention pertains.

The preparation of the undeuterated ω-diphenylurea and its physiologically compatible salts used in the present invention are known. The preparation of the corresponding deuterated ω-diphenylurea can be carried out by the same route using the corresponding deuterated starting compound as a starting material. For example, the compound of the formula (I) of the present invention can be produced by the production method described in WO 2000/042012, except that the raw material used for deuteration in the reaction replaces the non-deuterated raw material.

Usually, in the preparation scheme, each reaction is usually carried out in an inert solvent at room temperature to reflux temperature (e.g., 0 ° C to 80 ° C, preferably 0 ° C to 50 ° C). The reaction time is usually from 0.1 to 60 hours, preferably from 0.5 to 48 hours.

Taking compound CM4309 as an example, a preferred preparation process is as follows:

As shown in Scheme 1, 3-fluoro-4-amino-phenol (Compound I) and 4-chloro-3-trifluoromethylaniline (Compound II) in N , N '-carbonyldiimidazole, phosgene or The reaction gave triphosgene to give 1-(4-chloro-3-trifluoromethyl-phenyl)-3-(2-fluoro-4-hydroxy-phenyl)-urea (Compound III). Methyl picolinate (compound IV) and deuterated methylamine or deuterated methylamine hydrochloride under the action of a base (such as sodium carbonate, potassium carbonate, sodium hydroxide, triethylamine, pyridine, etc.) or directly mixed reaction Pyridine-2-( N -1',1',1'-tridemethyl)carbamamine (Compound V) was obtained. Compound III and Compound V are in a base (such as potassium t-butoxide, sodium hydride, potassium hydride, potassium carbonate, cesium carbonate, potassium phosphate, potassium hydroxide, sodium hydroxide) or an optional catalyst (such as cuprous iodide and Compound CM-4309 was obtained by the action of valine acid, or cuprous iodide and picolinic acid. The above reaction is carried out in an inert solvent such as dichloromethane, dichloroethane, acetonitrile, n-hexane, toluene, tetrahydrofuran, N,N-dimethylformamide, dimethyl hydrazine, etc., at a temperature of 0-200 ° C. get on.

Taking compound CM4309 as an example, another preferred preparation process is as follows:

As shown in Scheme 2, the picolinate (compound VI) and 3-fluoro-4-amino-phenol (compound I) are in the base (such as potassium t-butoxide, sodium hydride, potassium hydride, potassium carbonate, cesium carbonate). , potassium phosphate, potassium hydroxide, sodium hydroxide) or an optional catalyst (such as cuprous iodide and valine, or cuprous iodide and pyridine Under the action of formic acid, an amine (compound VII) is obtained. Further, urea (Compound IX) is obtained by reacting Compound II with N,N'-carbonyldiimidazole, phosgene or triphosgene or with 4-chloro-3-trifluoromethyl-phenylisocyanate (Compound VIII). Compound IX and deuterated methylamine or deuterated methylamine hydrochloride are reacted by a base (for example, sodium carbonate, potassium carbonate, sodium hydroxide, triethylamine, pyridine, etc.) or directly to obtain a compound CM4309. The above reaction is carried out in an inert solvent such as dichloromethane, dichloroethane, acetonitrile, n-hexane, toluene, tetrahydrofuran, N,N-dimethylformamide, dimethyl hydrazine, etc., at a temperature of 0-200 ° C. get on.

Taking compound CM4309 as an example, another preferred preparation process is as follows:

As shown in Scheme 3, 2-fluoro-4-methoxyaniline (Compound X) and Compound II are under N , N '-carbonyldiimidazole, phosgene or triphosgene, or 4-chloro-3- Trifluoromethyl-phenylisocyanate (Compound VIII) is reacted to give urea (Compound XI). 1-(4-Chloro-3-(trifluoromethyl)phenyl)-3-(2-fluoro-4-hydroxyphenyl)urea (Compound III) is obtained by various demethylation methods known in the art. Compound III and Compound V are then reacted to give compound CM4309 using the same procedure as described in Scheme 1, or various synthetic methods known in the art. The above reaction is carried out in an inert solvent such as dichloromethane, dichloroethane, acetonitrile, n-hexane, toluene, tetrahydrofuran, N , N -dimethylformamide, dimethyl hydrazine, etc. at a temperature of 0-200 ° C. get on.

Taking compound CM4309 as an example, another preferred preparation process is as follows:

As shown in Scheme 4, compound V and the compound 3-fluoro-4-amino-phenol are reacted in an inert solvent under basic conditions to give 4-(4-amino-3-fluoro-phenoxy)-pyridine. -2-( N -1',1',1'-trideuteromethyl)decylamine (Compound B1). The compound of formula B is reacted with formula W in an inert solvent to form a compound of formula M; and the compound of formula M is reacted with 4-chloro-3-trifluoromethylaniline in an inert solvent to provide compound CM4309 . The above reaction is carried out in an inert solvent such as dichloromethane, dichloroethane, acetonitrile, n-hexane, toluene, tetrahydrofuran, N , N -dimethylformamide, dimethylhydrazine, pyridine, etc., temperature 0-200 Perform at °C.

Taking compound CM4309 as an example, another preferred preparation process is as follows:

As shown in Scheme 5, 4-chloro-3-trifluoromethylaniline is reacted with Formula W under basic conditions to form a compound of Formula B; under basic conditions, the B1 compound is reacted with a compound of Formula B. The reaction was carried out to thereby give the compound CM4309. The above reaction is carried out in an inert solvent such as dichloromethane, dichloroethane, acetonitrile, n-hexane, toluene, tetrahydrofuran, N , N -dimethylformamide, dimethylhydrazine, pyridine, etc., temperature 0-200 Perform at °C.

Taking compound CM4309 as an example, a particularly preferred preparation process is as follows:

Deuterated can be introduced by deuterated methylamine.

The deuterated methylamine or its hydrochloride can be obtained by the following reaction. Nitromethane in alkali (sodium hydride, potassium hydride, deuterated sodium hydroxide, deuterated hydration Potassium, potassium carbonate, etc., or under a phase transfer catalyst, react with hydrophobic water to obtain deuterated nitromethane, and if necessary, repeat the above experiment to obtain high purity deuterated nitromethane. Deuterated nitromethane reduction, such as zinc powder, magnesium powder, iron or nickel, to obtain deuterated methylamine or its hydrochloride.

Further, deuterated methylamine or its hydrochloride can be obtained by the following reaction.

The key intermediate 3 can also be synthesized from deuterated methanol by the following method.

The specific synthesis method thereof is described in detail in Example 1.

The main advantages of the invention include:

(1) The compound of the present invention has excellent inhibitory properties against a phosphate kinase such as raf kinase.

(2) By using the intermediate of the formula V, the formula B or the formula B2 of the present invention, various deuterated diphenylureas can be prepared conveniently, efficiently, and with high purity.

(3) The reaction conditions are milder and the operation process is safer.

The invention is further illustrated below in conjunction with specific embodiments. It is to be understood that the examples are not intended to limit the scope of the invention. The experimental methods in the following examples which do not specify the specific conditions are usually in accordance with conventional conditions or according to the conditions recommended by the manufacturer. Parts and percentages are parts by weight and percentage by weight unless otherwise stated.

Example 1: N- (4-Chloro-3-(trifluoromethyl)phenyl) -N '-(4-(2-( N -1',1',1'-trimethylamino) Mercapto)-4-pyridyloxy)phenyl)urea (compound CM4307)

Process one

Preparation of 1:4-chloro-pyridin-2-( N -1',1',1'-tridemethyl)carbamamine (3)

In a 250 m L single-necked round bottom flask equipped with an exhaust gas treatment device, add hydrazine chloride (60 mL), maintain the temperature between 40-50 ° C, slowly add anhydrous DMF (2 mL), and add dropwise After completion, stirring was continued for 10 minutes, and nicotinic acid (20 g, 162.6 mmol) was added portionwise thereto over 20 minutes, and the color of the solution gradually changed from green to light purple. The temperature was raised to 72 ° C and stirred under reflux for 16 hours to produce a large solid precipitate. It was cooled to room temperature, diluted with toluene (100 mL), concentrated to dryness, then diluted with toluene and concentrated to dryness. Filtration and washing with toluene gave a pale yellow 4-chloro-pyridin-2-carboyl chloride solid. This solid was slowly added to a saturated solution of deuterated methylamine in tetrahydrofuran under ice cooling, maintaining the temperature below 5 ° C and stirring was continued for 5 hours. Was concentrated, ethyl acetate was added, the precipitated white solid was filtered off, the filtrate was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to dryness to give a pale yellow 4-chloro - pyridin -2- (N -1 ', 1' , 1'-tridemethyl)carbamamine (3) (20.68 g), yield 73%.

¹ H NMR (CDCl ₃ , 300 MHz): δ 8.37 (d, 1H), 8.13 (s, 1H), 7.96 (br, 1H), 7.37 (d, 1H).

Preparation of 2: 4-(4-Aminephenoxy)-2-pyridine-( N -1',1',1'-tridemethyl)carbamamine (5)

To 100 mL of dry anhydrous DMF, p-aminophenol (9.54 g, 0.087 mol) and potassium butoxide (10.3 g, 0.092 mol) were sequentially added, and the solution became dark brown, and stirred at room temperature for 2 hours, and then added thereto. 4-Chloro-pyridin-2-( N -1',1',1'-tridemethyl)methaneamine (3) (13.68 g, 0.079 mol), anhydrous potassium carbonate (6.5 g, 0.0467 mol) After the temperature of the reaction solution was raised to 80 ° C, stirring was continued overnight. After the reaction was completed by TLC, the mixture was cooled to room temperature, and the mixture was poured into a mixed solution of ethyl acetate (150 mL) and saturated brine (150 mL). (100 mL × 3), the combined extracts were washed with saturated water (100 mL × 3), dried over anhydrous sodium sulfate and evaporated to give pale yellow 4-(4-aminophenoxy)-2-pyridine-( N -1) ',1',1'-triterpene methyl) formamidine (18.00 g), yield 92%.

_{^{1 H NMR (CDCl 3, 300MHz}} ): δ8.32 (d, 1H), 7.99 (br, 1H), 7.66 (s, 1H), 6.91-6.85 (m, 3H), 6.69 (m, 2H), 3.70 (br, s, 2H).

3: N- (4-chloro-3-(trifluoromethyl)phenyl) -N '-(4-(2-( N -1',1',1'-trimethylaminocarbamidine) Of benzyl-4-pyridyloxy)phenyl)urea (CM4307)

To a solution of 120 mL of dichloromethane was added 5-amino-2-chloro-trifluoromethylbenzene (15.39 g, 78.69 mol), N , N '-carbonyldiimidazole (CDI) (13.55 g, 83.6 mmol), room temperature After stirring for 16 hours, 4-(4-aminophenoxy)-2-pyridine-( N -1',1',1'-tridecylidenemethyl)carbenamide (18 g, was slowly added dropwise thereto. A solution of 73 mmol) in dichloromethane (180 mL) was stirred at room temperature for 18 h. After the TLC detection reaction was completed, a part of the dichloromethane solvent was rotated to about 100 mL, and left at room temperature for several hours. A large amount of white solid was precipitated, suction filtered, and the solid was washed with a large amount of dichloromethane. After the filtrate was concentrated to remove a portion of the solvent, a portion of solid was precipitated, and the solids were combined and washed again with a large portion of dichloromethane to give N- (4-chloro-3-(trifluoromethyl)phenyl) -N as a white powder. '-(4-(2-( N -1',1',1'-trimethylaminocarbamoyl)-4-pyridyloxy)phenyl)urea CM4307 pure product (20.04g), produced The rate is 58%.

¹ H NMR (CD ₃ OD, 300 MHz): δ 8.48 (d, 1H), 8.00 (d, 1H), 7.55 (m, 5H), 7.12 (d, 1H), 7.08 (s, 2H), ESI- HRMS m/z: C ₂₁ H ₁₃ D ₃ ClF ₃ N ₄ O ₃ , calcd. 467.11, found 490.07 [M+Na] ⁺ .

Alternatively, the compound CM4307 can be dissolved in dichloromethane and reacted with peroxybenzoic acid to give the corresponding oxidation product: 4-(4-(3-(4-chloro-3-(trifluoromethyl)benzene) Urinyl)phenoxy)-2-( N -1',1',1'-trimethylaminomethylmercapto)pyridine-1-oxide.

Example 2

Synthesis of CM4309 based on 4-chloro-pyridine-2-( N -1',1',1'-tridemethyl) decylamine (Intermediate A2)

Preparation of 1:4-chloro-pyridin-2-( N -1',1',1'-tridemethyl)guanamine (Intermediate A2)

method 1:

Methyl 4-chloro-2-picolinate (50 g, 291 mmol, 1 eq) was placed in a three-necked flask containing 250 mL of tetrahydrofuran, and stirred, and 1,1,1-tridecylmethylamine hydrochloride (31 g, 437 mmol) was added. Anhydrous potassium carbonate (80 g, 583 mmol, 2 eq) was stirred at room temperature for 20 hr then water (250 mL) and EtOAc. The aqueous phase was extracted with EtOAc (EtOAc)EtOAc.

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.64 (dd, J = 2 Hz, 5.2 Hz, 1H), 7.97 (d, J = 1.6 Hz, 1H), 8.54 (d, J = 5.2 Hz, 1H) , 8.74 (br, 1H).

MS (ESI, m/z) Calcd. for C ₇ H ₄ D ₃ ClN ₂ O: 173, found: 174 [M+H] ⁺ .

Method 2:

4-Chloro-2-picolinic acid (200 mg, 1.27 mmol, 1 eq) was dissolved in N , N -dimethylformamide (DMF, 5 mL), and 1,1,1-tridemethylamine hydrochloride (179 mg, 2.54 mmol, 2 eq), 2-(7-azobenzotriazole) -N , N , N ', N '-tetramethyluron hexafluorophosphate (HATU, 965 mg, 2.54 mmol, 2 eq , N , N -diisopropylethylamine (DIEA, 492 mg, 3.81 mmol, 3 eq), and the mixture was stirred at room temperature for 16 hrs, water (20 mL), ethyl acetate (30 mL) The organic layer was dried over anhydrous sodium sulfate (MgSO4).

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.64 (dd, J = 2 Hz, 5.2 Hz, 1H), 7.97 (d, J = 1.6 Hz, 1H), 8.54 (d, J = 5.2 Hz, 1H) , 8.74 (br, 1H).

MS (ESI, m/z) Calcd. for C ₇ H ₄ D ₃ ClN ₂ O: 173, found: 174 [M+H] ⁺ .

2: 4-(4-Amino-3-fluoro-phenoxy)-pyridine-2-( N -1',1',1'- Preparation of triterpene methyl) guanamine B1

Potassium tert-butoxide (15 g, 130 mmol, 1.3 eq) was placed in a solution containing N,N-dimethylacetamide (DMA, 50 mL), and 3-Fluoro was slowly added dropwise at 0-5 ° C. A solution of 4-amino-phenol (16 g, 127 mmol, 1.3 eq) in DMA (50 mL), stirred at room temperature for 20 min, warmed to 100 ° C and slowly added dropwise 4-chloro-pyridine-2-( N -1 ',1',1'-triterpenemethyl)guanamine A2 (17 g, 97 mmol, 1 eq) of DMA (50 mL), after stirring, stirring was continued for 0.5 hour, cooled to room temperature, and acetic acid was added to the reaction mixture. Ethyl acetate (1.5 L) was diluted and stirred for 0.5 hr, and the organic salt was removed by filtration, washed with water (500 mL × 3), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure, and ethanol (100 mL) was added to the crude and stirred for 2 hr. Filtration to room temperature gave the title compound as a brown solid, 20 g, mp.

¹ H NMR (CD ₃ OD-d ₄ , 400 MHz): δ 6.74-6.77 (m, 1H), 6.87 (dd, J = 2.4 Hz, 11.6 Hz, 1H), 6.93 (t, J = 10 Hz, 1H), 7.02 (dd, J = 2.8 Hz, 6 Hz, 1H), 7.54 (d, J = 2.4 Hz, 1H), 8.44 (d, J = 6 Hz, 1H).

MS (ESI, m / z) calcd.for Cl 3 H 9 D 3 ClN 3 O 2: 264, found: 265 [M + H] +.

3: 4-(4-(3-(4-Chloro-3-(trifluoromethyl)phenyl))urea)-3-fluoro-phenoxy)-2-( N -1',1', Preparation of 1'-triterpene methyl)pyridiniumamine (CM4309)

method 1:

4-Chloro-3-trifluoromethylphenylisocyanate (13 g, 58 mmol, 1.1 eq) was placed in a single-necked flask containing dichloromethane (70 mL) and 4-(4-amine a solution of benzyl-3-fluoro-phenoxy)-pyridine-2-( N -1',1',1'-tridemethyl)guanamine (14 g, 53 mmol, 1 eq) in dichloromethane (350 mL) The mixture was slowly added dropwise to the above solution, and the mixture was stirred at room temperature for 20 hr, and then filtered and washed with dichloromethane (20 mL × 2) to give a pale brown solid 13 g, purity 98%, yield 50%.

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.06-7.10 (m, 1H), 7.19 (dd, J = 2.4 Hz, 5.6 Hz, 1H), 7.35 (dd, J = 2.8 Hz, 12 Hz, 1H) , 7.43 (d, J = 2.4 Hz, 1H), 7.63 (m, 2H), 8.14 (br, 1H), 8.17 (t, J = 8.8 Hz, 1H), 8.53 (d, J = 5.6 Hz, 1H) , 8.75 (d, J = 1.6 Hz, 1H), 8.78 (br, 1H), 9.54 (br, 1H).

MS (ESI, m/z) calcd. for C ₂₁ H ₁₂ D ₃ ClF ₄ N ₄ O ₃ : 485, found: 486 [M+H] ⁺ .

Method 2:

4-Chloro-3-trifluoromethyl-phenylamine (300 mg, 1.53 mmol, 1.05 eq) was dissolved in dichloromethane (5 mL) and N , N -carbonyldiimidazole (CDI, 265 mg, 1.64 mmol, 1.12 eq) and stirred at room temperature for 16 hours, 4-(4-amine-3-fluoro-phenoxy)-2-pyridine-( N -1',1',1'-triterpene The guanamine (386 mg, 1.46 mmol, 1 eq) was added to the reaction mixture, and the mixture was stirred at room temperature for 20 hr. The solvent was evaporated under reduced pressure. EtOAc (EtOAc) Filtration to room temperature afforded CM4309 as s.

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.06-7.10 (m, 1H), 7.19 (dd, J = 2.4 Hz, 5.6 Hz, 1H), 7.35 (dd, J = 2.8 Hz, 12 Hz, 1H) , 7.43 (d, J = 2.4 Hz, 1H), 7.63 (m, 2H), 8.14 (br, 1H), 8.17 (t, J = 8.8 Hz, 1H), 8.53 (d, J = 5.6 Hz, 1H) , 8.75 (d, J = 1.6 Hz, 1H), 8.78 (br, 1H), 9.54 (br, 1H).

MS (ESI, m/z) calcd. for C ₂₁ H ₁₂ D ₃ ClF ₄ N ₄ O ₃ : 485, found: 486 [M+H] ⁺ .

Example 3

Synthesis of CM4309 based on 4-chloro-pyridine-2-( N -1',1',1'-tridemethyl) decylamine (Intermediate A2)

Preparation of 1-(4-chloro-3-trifluoromethyl-phenyl)-3-(2-fluoro-4-hydroxy-phenyl)-urea B2

3-Fluoro-4-amino-phenol (500 mg, 3.93 mmol, 1 eq) was dissolved in N , N -methylformamide (3 mL) at room temperature and 4-chloro-3-trifluoro A solution of methyl phenyl isocyanate (917 mg, 4.13 mmol, 1.05 eq) in dichloromethane (3 mL) was evaporated. The organic layer was washed with EtOAc EtOAc (EtOAc m. The compound was 1.2 g as a brown solid, purity 98%, yield 89%.

_{^{1 H NMR (DMSO-d 6}} , 400MHz): δ 6.56-6.59 (m, 1H), 6.64 (dd, J = 2.4Hz, 12.4Hz, 1H), 7.59-7.63 (m, 3H), 8.11 (br, 1H), 8.28 (br, 1H), 9.34 (br, 1H), 9.69 (br, 1H).

MS (ESI, m/z) calcd. for C ₁₄ H ₉ ClF ₄ N ₂ O ₂ : 348, Found: 349 [M+H] ⁺ .

2: 4-(4-(3-(4-Chloro-3-(trifluoromethyl)phenyl))urea)-3-fluoro-phenoxy)-2-( N -1',1', Preparation of 1'-triterpene methyl)pyridiniumamine (CM4309)

Potassium tert-butoxide (906 mg, 8.09 mmol, 1.4 eq) was dissolved in N , N -methylacetamide (DMA, 5 mL), and 1-(4-chloro-3-3) was slowly added dropwise at 0 °C. A solution of fluoromethyl-phenyl)-3-(2-fluoro-4-hydroxy-phenyl)-urea (2.8 g, 8.09 mmol, 1.4 eq) in DMA (5 mL) was added to the above solution and stirred at room temperature After 20 minutes, the temperature was raised to 100 ° C, and a solution of 4-chloro- N -deuterated methylpyridine-2-carboxamide (1 g, 5.78 mmol, 1 eq) in DMA (5 mL) was added dropwise. The mixture was diluted with ethyl acetate (200 mL) and stirred for 0.5 hour. The inorganic salt was removed by filtration, washed with water (5mL × 3), dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. Ether: ethyl acetate = 4:1) 520 mg of CM4 s.

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.06-7.10 (m, 1H), 7.19 (dd, J = 2.4 Hz, 5.6 Hz, 1H), 7.35 (dd, J = 2.8 Hz, 12 Hz, 1H) , 7.43 (d, J = 2.4 Hz, 1H), 7.63 (m, 2H), 8.14 (br, 1H), 8.17 (t, J = 8.8 Hz, 1H), 8.53 (d, J = 5.6 Hz, 1H) , 8.75 (d, J = 1.6 Hz, 1H), 8.78 (br, 1H), 9.54 (br, 1H).

MS (ESI, m/z) calcd. for C ₂₁ H ₁₂ D ₃ ClF ₄ N ₄ O ₃ : 485, found: 486 [M+H] ⁺ .

Example 4

Synthesis of CM4309 based on 4-chloro-pyridine-2-( N -1',1',1'-tridemethyl) decylamine (Intermediate A2)

Preparation of 1:(4-chloro-3-trifluoromethyl-phenyl)-carbamic acid phenyl ester B3

4-Chloro-3-trifluoromethylaniline (4.9 g, 25 mmol, 1 eq) was added to dichloromethane (50 mL) and phenyl chloroformate (3.9 g, 25 mmol, 1 eq) was slowly added dropwise at room temperature. A solution of methyl chloride (10 mL) was continued and the reaction was stirred for 1 hour. Diluted hydrochloric acid (1 N, 50 mL) was added and washed with water (40 mL×3) and layered. The organic phase was dried over anhydrous sodium

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.25-7.30 (m, 3H), 7.45 (t, J = 8 Hz, 2H), 7.67-7.78 (m, 2H), 8.07 (d, J = 2 Hz, 1H), 10.73 (br, 1H).

MS (ESI, m/z) Cald. for C ₁₄ H ₉ ClF ₃ NO ₂ : 315, Found: 316 [M+H] ⁺ .

2:4-[4-({[4-chloro-3-(trifluoromethyl)phenyl]methyl)amino)-3-fluorophenoxy] -N -deuterated methylpyridine-2 -Preparation of methotrexate CM4309

(4-Chloro-3-trifluoromethyl-phenyl)-carbamic acid phenyl ester (0.95 g, 3.0 mmol, 1 eq) was dissolved in pyridine (10 mL) at room temperature and 4-(4-) Amine-3-fluoro-phenoxy)-2-pyridine-( N -1',1',1'-tridecylmethyl)decylamine (0.79 g, 3.0 mmol, 1 eq), warmed to 85 ° C and stirred Reaction for 2 hours. The solvent was evaporated under reduced pressure. EtOAc (EtOAc)EtOAc. The ester (3 mL) was stirred for 1 hr.

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.06-7.10 (m, 1H), 7.19 (dd, J = 2.4 Hz, 5.6 Hz, 1H), 7.35 (dd, J = 2.8 Hz, 12 Hz, 1H) , 7.43 (d, J = 2.4 Hz, 1H), 7.63 (m, 2H), 8.14 (br, 1H), 8.17 (t, J = 8.8 Hz, 1H), 8.53 (d, J = 5.6 Hz, 1H) , 8.75 (d, J = 1.6 Hz, 1H), 8.78 (br, 1H), 9.54 (br, 1H).

MS (ESI, m/z) calcd. for C ₂₁ H ₁₂ D ₃ ClF ₄ N ₄ O ₃ : 485, found: 486 [M+H] ⁺ .

Example 5

Synthesis of CM4309 based on 4-chloro-pyridine-2-( N -1',1',1'-tridemethyl) decylamine (Intermediate A2)

Preparation of [2-fluoro-4(2-deuterated methylformamido-pyridin-4-yloxy)-phenyl]-carbamic acid phenyl ester B4

2.60 g of 4-(4-Amine-3-fluoro-phenoxy)-2-pyridine-( N -1',1',1'-tridecylmethyl)decylamine, 10 mmol, 1 eq) Dichloromethane (15 mL), pyridine (1.58 g, 20 mmol, 2 eq) was added, and phenyl chloroformate (1.88 g, 12 mmol, 1.2 eq) was dissolved in dichloromethane (15 mL). The reaction was stirred at room temperature for 1 hour. It was washed with dilute hydrochloric acid (1 N, 30 mL), brine (10 mL×2) and layered. The organic phase was dried over anhydrous sodium

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.10-7.13 (m, 1H), 7.21. (dd, J = 2.8 Hz, 5.6 Hz, 1H), 7.26 (t, J = 6.4 Hz, 3H), 7.36 (dd, J = 2.8 Hz, 11.6 Hz, 1H), 7.43-7.47 (m, 3H), 7.80 (t, J = 8.4 Hz, 1H), 8.55 (d, J = 5.2 Hz, 1H), 8.79 (br , 1H), 10.10 (br, 1H).

MS (ESI, m/z) calcd. for C ₂₀ H ₁₃ D ₃ FN ₃ O ₄ : 384, Found: 385 [M+H] ⁺ .

2:4-[4-({[4-chloro-3-(trifluoromethyl)phenyl]methyl)amino)-3-fluorophenoxy] -N -deuterated methylpyridine-2 -Preparation of methotrexate CM4309

[2-Fluoro-4(2-deuterated methylformamido-pyridin-4-yloxy)-phenyl]-carbamic acid phenyl ester (1.92 g, 5 mmol, 1 eq) Dissolved in pyridine (10 mL), added 4-chloro-3-trifluoromethylaniline (0.98 g, 5 mmol, 1 eq). The solvent was evaporated under reduced pressure. EtOAc (EtOAc m. (5 mL) and stirred at 50 ° C for 30 min.

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.06-7.10 (m, 1H), 7.19 (dd, J = 2.4 Hz, 5.6 Hz, 1H), 7.35 (dd, J = 2.8 Hz, 12 Hz, 1H) , 7.43 (d, J = 2.4 Hz, 1H), 7.63 (m, 2H), 8.14 (br, 1H), 8.17 (t, J = 8.8 Hz, 1H), 8.53 (d, J = 5.6 Hz, 1H) , 8.75 (d, J = 1.6 Hz, 1H), 8.78 (br, 1H), 9.54 (br, 1H).

MS (ESI, m/z) calcd. for C ₂₁ H ₁₂ D ₃ ClF ₄ N ₄ O ₃ : 485, found: 486 [M+H] ⁺ .

Example 6

Synthesis of CM4309 based on intermediate B7

4-{4-[3-(4-Chloro-3-trifluoromethyl-phenyl)-urea]-3-fluoro-phenoxy}-pyridine-2-carboxylic acid (intermediate B7) as the key intermediate Body route

Preparation of 1:4-chloro-pyridine-2-carboxylic acid tert-butyl ester A7

4-Chloro-pyridine-2-carboxylic acid (10.5 g, 66.64 mmol) was suspended in hydrazine chloride (40 mL), heated to reflux at 80 ° C, and N,N-dimethylformamide (0.2 mL) was added dropwise. The reaction was refluxed for 2 hours, and dimethyl hydrazine was removed under reduced pressure to give pale yellow chlorobenzene, and dichloromethane (60mL). The above-prepared dichloromethane solution was added dropwise to a mixed solution of a third butanol (25 mL), pyridine (20 mL) and dichloromethane (80 mL) at -40 °C. The reaction solution was warmed to 50 ° C and stirred for 16 hours. The solvent was removed under reduced pressure, and ethyl acetate (150 mL) was evaporated and evaporated. The solvent was removed under reduced pressure and dried title crystals crystals crystals

_{^{1 H NMR (DMSO-d 6}} , 400MHz): δ 1.56 (s, 9H), 7.80 (dd, J = 2.4Hz, 5.2Hz, 1H), 8.02 (d, J = 2Hz, 1H), 8.69 (d, J = 5.2 Hz, 1H).

MS (ESI, m/z) calcd. for C ₁₀ H ₁₂ ClNO ₂ : 213, Found: 158 (M-Bu ^t + H) ⁺ .

Preparation of 2: 4-(4-Amino-3-fluoro-phenoxy)-pyridine-2-carboxylic acid tert-butyl ester B5

Potassium tert-butoxide (5 g, 23.47 mmol, 1.4 eq) was dissolved in N,N-dimethylacetamide (DMA, 12 mL), and 3-fluoro-4-aminophenol was slowly added dropwise at 0 °C. A solution of 4.2 g, 32.86 mmol, 1.4 eq) in DMA (15 mL) was stirred at room temperature for 30 min. The temperature was raised to 100 ° C, and a DMA solution (18 mL) of 4-chloro-pyridine-2-carboxylic acid tert-butyl ester was slowly added dropwise, stirred for 1 hour, cooled to room temperature, ethyl acetate (500 mL) was added, and the insoluble matter was removed by filtration. The organic layer was washed with EtOAc (EtOAc m. , purity 98%, yield 20%.

¹ H NMR (CD ₃ OD-d ₄ , 400 MHz): δ 1.61 (s, 9H), 6.78-6.80 (m, 1H), 6.89-6.97 (m, 2H), 7.07 (dd, J = 2.4 Hz, 5.6 Hz, 1H), 7.55 (d, J = 2.4 Hz, 1H), 8.48 (d, J = 5.6 Hz, 1H).

MS (ESI, m/z) calcd. for C ₁₆ H ₁₇ FN ₂ O ₃ : 304, found: 249 [M-Bu ^t + H] ⁺ .

Preparation of 3:4-{4-[3-(4-chloro-3-trifluoromethyl-phenyl)-urea]-3-fluoro-phenoxy}-pyridine-2-carboxylic acid tert-butyl ester B6

4-Chloro-3-trifluoromethylphenylisocyanate (1.06 g, 4.77 mmol, 1.05 eq) was dissolved in dichloromethane (10 mL) at room temperature and 4-(4-amino-3- A solution of butyl-phenoxy)-pyridine-2-carboxylic acid tert-butyl ester (1.38 g, 4.54 mmol, 1 eq) in dichloromethane (50 mL) was slowly added dropwise to the above solution Stir at room temperature for 16 hours. The solvent was evaporated under reduced pressure. EtOAcjjjjjjjjjjj

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 1.54 (s, 9H), 7.08 (dd, J = 2 Hz, 8.8 Hz, 1H), 7.18 (dd, J = 2.8 Hz, 5.6 Hz, 1H), 7.34 (dd, J = 2.4 Hz, 3.6 Hz, 1H), 7.46 (d, J = 2.4 Hz, 1H), 7.61-7.66 (m, 2H), 8.13-8.19 (m, 2H), 8.58 (d, J = 5.6 Hz, 1H), 8.76 (d, J = 1.6 Hz, 1H), 9.53 (br, 1H).

MS (ESI, m / z) calcd.for C 24 H 20 ClF 4 N 3 O 4: 525, found: 526 [M + H] +.

Preparation of 4:4-{4-[3-(4-chloro-3-trifluoromethyl-phenyl)-urea]-3-fluoro-phenoxy}-pyridine-2-carboxylic acid B7

4-{4-[3-(4-Chloro-3-trifluoromethyl-phenyl)-urea]-3-fluoro-phenoxy}-pyridine-2-carboxylic acid tert-butyl at room temperature The ester (1.8 g, 3.43 mmol) was dissolved in dichloromethane (15 mL), trifluoroacetic acid (15 mL) and triethyl decane (0.2 mL) were added, and the mixture was warmed to 50 ° C and stirred for 16 hours. The solvent was removed under reduced br~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ , purity 85%, yield 92%.

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.13 (dd, J = 1.2 Hz, 9.2 Hz, 1H), 7.38-7.41 (m, 2H), 7.61-7.66 (m, 3H), 8.15 (s, 1H), 8.22 (t, J = 8.8 Hz, 1H), 8.67 (d, J = 5.6 Hz, 1H), 9.06-9.08 (m, 1H), 10.21-10.28 (m, 1H), 11.96 (br, 1H) ).

MS (ESI, m/z) calcd. for C ₂₀ H ₁₂ ClF ₄ N ₃ O ₄ : 469, Found: 468 [MH] ^- .

5:4-[4-({[4-chloro-3-(trifluoromethyl)phenyl]methyl)amino)-3-fluorophenoxy] -N -deuterated methylpyridine-2 -Preparation of methotrexate CM4309

method 1:

4-{4-[3-(4-Chloro-3-trifluoromethyl-phenyl)-urea]-3-fluoro-phenoxy}-pyridine-2-carboxylic acid (1.48 g) at room temperature , 3.15 mmol, 1 eq) was dissolved in N , N -dimethylformamide (20 mL), and 1,1,1-tridemethylamine hydrochloride (0.45 g, 6.29 mmol, 2 eq), 2- 7-Azobenzotriazole)- N , N , N ', N '-tetramethylurea hexafluorophosphate (HATU, 2.40 g, 6.29 mmol, 2 eq) with N , N -diisopropyl B The amine (DIEA, 1.20 g, 9.44 mmol, 3 eq) was stirred at room temperature for 16 h. Water (50 mL) was added dropwise to the reaction mixture, and the mixture was stirred for 0.5 hr. EtOAc (EtOAc) Filtration and removal of the solvent under reduced pressure afforded EtOAc EtOAc EtOAc EtOAc EtOAc Compound

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.06-7.10 (m, 1H), 7.19 (dd, J = 2.4 Hz, 5.6 Hz, 1H), 7.35 (dd, J = 2.8 Hz, 12 Hz, 1H) , 7.43 (d, J = 2.4 Hz, 1H), 7.63 (m, 2H), 8.14 (br, 1H), 8.17 (t, J = 8.8 Hz, 1H), 8.53 (d, J = 5.6 Hz, 1H) , 8.75 (d, J = 1.6 Hz, 1H), 8.78 (br, 1H), 9.54 (br, 1H).

MS (ESI, m/z) calcd. for C ₂₁ H ₁₂ D ₃ ClF ₄ N ₄ O ₃ : 485, found: 486 [M+H] ⁺ .

Method 2:

4-{4-[3-(4-Chloro-3-trifluoromethyl-phenyl)-urea]-phenoxy}-pyridine-2-carboxylic acid (1 g, 2.13 mmol) was suspended in methanol (20 mL) Concentrated sulfuric acid (5 mL) was added at room temperature and refluxed for 3 hours. The solvent was removed under reduced pressure and purified by column chromatography (dichloromethane:methanol = 10:1) to afford 4-{4-[3-(4- Methyl chloro-3-trifluoromethyl-phenyl)-ureido]-phenoxy}-pyridine-2-carboxylate B8 (0.83 g), purity 97%, yield 80%.

MS (ESI, m/z) Calcd. for C ₂₁ H ₁₄ ClF ₄ N ₃ O ₄ : 483, Found: 484 [M+H] ⁺ .

Methyl 4-{4-[3-(4-chloro-3-trifluoromethyl-phenyl)-urea]-phenoxy}-pyridine-2-carboxylate (0.83 g, 1.71 mmol, 1 eq) Into a three-necked flask containing 15 mL of tetrahydrofuran, and stirred, and added 1,1,1-tridemethylamine hydrochloride (0.24 g, 3.43 mmol, 2 eq), anhydrous potassium carbonate (0.47 g, 3.43 mmol, 2 eq). After stirring the reaction for 20 hours at room temperature, water (10 mL) and methyl t-butyl ether (25 mL) were added, stirred and layered to give an organic phase. The aqueous phase was extracted with EtOAc (EtOAc) (EtOAc)EtOAc.

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.06-7.10 (m, 1H), 7.19 (dd, J = 2.4 Hz, 5.6 Hz, 1H), 7.35 (dd, J = 2.8 Hz, 12 Hz, 1H) , 7.43 (d, J = 2.4 Hz, 1H), 7.63 (m, 2H), 8.14 (br, 1H), 8.17 (t, J = 8.8 Hz, 1H), 8.53 (d, J = 5.6 Hz, 1H) , 8.75 (d, J = 1.6 Hz, 1H), 8.78 (br, 1H), 9.54 (br, 1H).

MS (ESI, m/z) calcd. for C ₂₁ H ₁₂ D ₃ ClF ₄ N ₄ O ₃ : 485, found: 486 [M+H] ⁺ .

Example 7

Preparation of oxide BO of CM4309

4-(4-(3-(4-Chloro-3-(trifluoromethyl)phenyl))urea)-3-fluoro-phenoxy)-2-( N -1' at room temperature , 1',1'-trideuteromethyl)pyridiniumamine (0.5 g, 1.03 mmol, 1 eq) was dissolved in dichloromethane (10 mL) and tetrahydrofuran (10 mL), and m-chloroperoxybenzoic acid (0.54 g, 3.09 mmol, 3 eq), the mixture was heated to 50 ° C and stirred for 20 hours. The solvent was removed under reduced pressure, and ethyl acetate (50 mL) was evaporated and evaporated. The organic phase was dried over anhydrous sodium sulfate.

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 7.10 (dd, J = 1.2 Hz, 8.8 Hz, 1H), 7.32-7.38 (m, 2H), 7.61-7.63 (m, 2H), 7.89-7.91 ( m,1H), 8.13-8.17 (m, 2H), 8.42 (d, J = 6.8 Hz, 1H), 8.76 (d, J = 1.6 Hz, 1H), 9.53 (br, 1H), 11.35 (br, 1H) ).

MS (ESI, m/z) calcd. for C ₂₁ H ₁₂ D ₃ ClF ₄ N ₄ O ₄ : 501, found: 502 [M+H] ⁺ .

Example 8

Preparation of CM4309 salt

Preparation of 1:CM4309 hydrochloride (4309‧HCl)

4-(4-(3-(4-Chloro-3-(trifluoromethyl)phenyl) guanidine)-3-fluoro-phenoxy)-2-(N-1',1',1 '-Trisylmethyl)pyridiniumamine (2.5 g, 5.16 mmol, 1 eq) was suspended in ethanol (50 mL), heated to reflux until the solution was clarified, and filtered to remove insolubles. (37%, 6.39 mmol, 0.6 mL, 1.24 eq), the clear liquid was kept for about 0.5 minutes, the oil bath was turned off and allowed to cool to room temperature, stirred for 1 hour, filtered to give insoluble material, and dried under vacuum at 50 ° C for 20 hours to give the title compound. , light gray solid 2.56 g, yield: 95%.

_{^{1 H NMR (DMSO-d 6}} , 400MHz): δ 7.08-7.10 (m, 1H), 7.24 (dd, J = 3.2Hz, 5.6Hz, 1H), 7.36 (dd, J = 2.8Hz, 11.6Hz, 1H ), 7.52 (d, J = 2.4 Hz, 1H), 7.64 (s, 2H), 8.14 (s, 1H), 8.18 (t, J = 9.2 Hz, 1H), 8.56 (d, J = 5.6 Hz, 1H) ), 8.89 (br, 1H), 8.97 (br, 1H), 10.05 (br, 1H), 10.23 (br, 1H).

Melting point: 240 ° C - 241.5 ° C

Elemental analysis Theoretical value: C, 48.29%; H, 3.67%; N, 10.73%. Found: C, 48.33%; H, 3.20%; N, 10.93%.

2: CM4309 methanesulfonic acid ethanolate (4309‧MeSO ₃ H‧CH ₃ CH ₂ Preparation of OH)

4-(4-(3-(4-Chloro-3-(trifluoromethyl)phenyl))urea)-3-fluoro-phenoxy)-2-( N -1',1',1 '-Trisylmethyl)pyridiniumamine (2.1 g, 4.33 mmol, 1 eq) was suspended in ethanol (50 mL), heated to reflux until the solution was clarified, and filtered to remove insolubles. Acid (85%, 8.66 mmol, 0.6 mL, 2 eq), the clear liquid was kept for about 0.5 minutes, the oil bath was turned off and cooled to room temperature, stirred for 1 hour, filtered to give insoluble material, and dried under vacuum at 50 ° C for 20 hours to give the title compound. It was 2.25 g of a pale red solid with a yield of 89%.

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 1.05 (t, J = 6.8 Hz, 3H), 2.48 (s, 3H), 3.44 (q, J = 6.8 Hz, 2H), 7.11 (dd, J = 1.2Hz, 8.4Hz, 1H), 7.31 (dd, J = 2.8Hz, 6Hz, 1H), 7.38 (dd, J = 2.4Hz, 11.6Hz, 1H), 7.61 - 7.65 (m, 3H), 8.14 - 8.19 (m, 2H), 8.60 (d, J = 6 Hz, 1H), 8.90 (t, J = 1.2 Hz, 1H), 8.99 (br, 1H), 9.39 (br, 1H), 9.66 (br, 1H).

Melting point: 157.5 ° C - 158.1 ° C.

3: Preparation of CM4309 p-toluenesulfonate (4309‧TsOH)

4-(4-(3-(4-Chloro-3-(trifluoromethyl)phenyl))urea)-3-fluoro-phenoxy)-2-( N -1',1',1 '-Trisylmethyl)pyridiniumamine (2.16 g, 4.33 mmol, 1 eq) was suspended in ethanol (50 mL), heated to reflux until the solution was clarified, and filtered to remove insolubles. Quickly add p-toluenesulfonic acid (70%, 8.66mmol, 1mL, 2eq), the clear solution is kept for about 0.5 minutes, turn off the oil bath and naturally cool to room temperature, stir for 1 hour, filter to obtain insoluble matter, vacuum dry at 50 °C 20 In an hour, 2.36 g of white crystals were obtained with a yield of 83%.

¹ H NMR (DMSO-d ₆ , 400 MHz): δ 2.30 (s, 3H), 7.09-7.14 (m, 3H), 7.24 (dd, J = 2.4 Hz, 5.2 Hz, 1H), 7.37 (dd, J = 2.4 Hz, 11.6 Hz, 1H), 7.48-7.50 (m, 3H), 7.61-7.66 (m, 2H), 8.15-8.19 (m, 2H), 8.56 (d, J = 5.6 Hz, 1H), 8.79 ( Br, 1H), 8.85 (br, 1H), 9.56 (br, 1H), 10.38 (br, 1H).

Melting point: 240.7 ° C - 241 ° C.

Example 9

For c-Kit, PDGFR-β protein tyrosine kinase molecular layer inhibitory activity

1. Experimental method

In this example, the enzyme-linked immunosorbent Assay (ELISA) was used to determine the molecular level inhibitory activity of the diphenylurea compound against c-Kit and PDGFR-β protein tyrosine kinase.

Compounds determined: CM4306, CM4308 (4-(4-(3-(4-chloro-3-(trifluoromethyl)phenyl))urea)-3-fluoro-phenoxy) -N -methylpyridine Guanamine), CM4309.

Main reagents:

The reaction substrate Poly(Glu, Tyr) _{4:1 was} purchased from Sigma; the anti-phosphotyrosine monobody antibody PY99 was purchased from Santa Cruz; horseradish peroxidase-labeled goat anti-mouse IgG was purchased from Calbiochem ATP, DTT, OPD were purchased from Amresco; ELISA was purchased from Corning; Su11248 was purchased from Merk.

experimental method:

See Roskoski, R., Jr. Sunitinib: a VEGF and PDGF receptor protein kinase and angiogenesis inhibitor. Biochem Biophys Res Commun, 356: 323-328, 2007, specifically including: kinase reaction substrate Poly (Glu, Tyr) _4:1 The ELISA plate was coated by diluting to 20 μg/mL with PBS without potassium ions. The test sample of diphenylurea was added to the well-coated ELISA well (the test sample was first prepared with DMSO to prepare a storage solution of 10 ^-2 M, diluted with the reaction buffer to the desired concentration before use, plus Into the experimental well, the final concentration in the 100 μL reaction system was 10 ^-5 mol/L). A positive control well was also established and the positive control compound Su11248 was added.

ATP solution diluted with the reaction buffer (ATP final concentration: 5 μM) was added, and finally, the test tyrosine kinase diluted with the reaction buffer was added. The total volume of the reaction system was 100 μL. Negative control wells and enzyme-free control wells were also established.

The reaction system was placed in a wet box, shaken at 37 ° C for 1 h, and T-PBS was washed three times after the reaction. The antibody PY99 was added to 100 μL/well and shaken at 37 ° C for 30 min. T-PBS was washed three times. Horseradish peroxidase-labeled goat anti-mouse IgG 100 μL/well was added and shaken at 37 ° C for 30 min. T-PBS was washed three times. Add 100 μL/well of OPD coloring solution and avoid the light reaction at room temperature for 1-10 min. The reaction was stopped by the addition of 2 M H ₂ SO ₄ 50 μL, and the A492 value was measured with a tunable wavelength ELISA reader VERSAmax.

The inhibition rate of the sample is calculated by the following formula:

2, the experimental results

The above experimental results are the mean of the two trials.

3. Evaluation criteria and results evaluation

Under the premise that the inhibitory activity of the positive control compound meets the reference range, the test compound is effective at an experimental concentration of 10 ^-5 mol/L, and the inhibition rate is greater than 50%. The inhibition rate is less than 50%.

The results showed that CM4306, CM4308 and CM4309 inhibited the protein tyrosine kinases c-Kit and PDGFR-β by more than 50%, so they have significant inhibitory activities against c-Kit and PDGFR-β tyrosine kinase at the molecular level. .

Example 10

Pharmacokinetics study of CM4306, CM4308 and CM4309 in rats

3 Male Sprague-Dawley rats weighing about 220g, after fasting overnight, were also given a mixed solution of CM4306, CM4308 and CM4309 (PEG400 as carrier) with 4mg/kg (calculated as alkali). Blood was collected at 0.25, 0.5, 1.0, 2.0, 3.0, 5.0, 7.0, 9.0, 24, 36, 48 and 72 h after administration, and the concentrations of CM4306 and CM4307 in plasma were determined by LC/MS/MS.

The results showed that the t1/2 of CM4309 (23.7h±2.9h) was significantly higher than that of CM4306 (12.5h±2.4h) and CM4308 (11.1h±0.9h).

The AUC (0-∞) of CM4309 (32611ng‧h/mL±7866ng‧h/mL) was significantly higher than that of CM4308 (13371ng‧h/mL±4958ng‧h/mL).

Example 11

In vivo pharmacodynamic study of CM4306, CM4308 and CM4309

Establishment of a tumor nude mouse transplantation model: SMMC-7721 cells in logarithmic growth phase were harvested. After counting, the cells were suspended in 1×PBS, the cell suspension concentration was adjusted to 1.5×10 ⁷ /mL, and the tumor was inoculated subcutaneously in the right side of the nude mouse. Cells, 3 × 10 ⁶ /0.2 mL / only. A total of 45 nude mice were inoculated. When the tumor volume reached 30-130 mm ³ , the animals were randomly grouped, and a total of 34 animals were obtained, so that the tumor difference of each group was less than 10% of the mean, and administration was started. Ten nude mice in the control group and 8 nude mice in each group in the treatment group. CM4306, CM4308 and CM4309 were administered orally in a single dose of 30 mg/kg for 2 weeks. After stopping the drug, the observation was continued until the tumor volume was greater than 2000 mm ³ . The body weight and tumor size were measured twice a week during the test. The compound was dissolved in Cremophor EL/ethanol/water (12.5:12.5:75).

Tumor volume (TV) is calculated as: TV = a × b ² /2. Where a and b represent the length and width of the tumor measurement, respectively. The relative tumor volume (RTV) is calculated as: RTV = Vt / V ₀ . Where V ₀ is the tumor volume at the time of group administration, and Vt is the tumor volume at the time of measurement. The evaluation index of antitumor activity is the relative tumor growth rate T/C (%), and the calculation formula is: T/C (%) = (T _RT V / C _RTV ) × 100%. T _RTV was the treatment group RTV, and C _RTV was the negative control group RTV.

Efficacy evaluation criteria: T / C (%) 40% and statistically analyzed p<0.05 was effective.

After the administration, the T/C (%) of 30 mg/kg of CM4306 was 64.1%, the T/C (%) of CM4308 was 51.0%, and the CM4309 was 27.0%. Among them, the inhibition rate of CM4309 at 30mg/kg was <40%, and the tumor volume was significantly different from the control (p<0.01), which showed significant inhibition of tumor growth. There was no significant difference in tumor volume between CM4306 and CM4308 at the same dose (p>0.05), whereas there was a significant difference between CM4309 and CM4306 (p<0.05) and CM4308 (p<0.01) tumor volume (Fig. 1). .

Examples 12-14 Pharmaceutical Compositions

Wherein the active compounds are prepared in Examples 6-8, respectively. CM4309, CM4309 oxide or methanesulfonic acid ethanolate of CM4309. The above materials were uniformly mixed according to a conventional method, and then filled into ordinary gelatin capsules, each of which obtained 1000 capsules.

All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the In addition, it should be understood that various modifications and changes may be made by those skilled in the art without departing from the scope of the invention.

Claims (10)

a salt of a compound of the formula: Wherein the salt of the compound is a hydrochloride selected from the compound, a p-toluenesulfonate of the compound, or a methanesulfonate of the compound. A method for preparing a salt of a compound of the formula, comprising the steps of: reacting a compound of the formula with an acid, wherein the acid is selected from hydrochloric acid, p-toluenesulfonic acid or methanesulfonic acid; A method for producing a salt of a compound of the formula according to Claim 2, wherein, when the acid is hydrochloric acid, 4-(4-(3-(4-chloro-3-(trifluoromethyl)phenyl) fluorene) Urea)-3-fluoro-phenoxy)-2-(N-1',1',1'-tridecylmethyl)pyridiniumamine is mixed with ethanol and heated to reflux until the solution is clarified, followed by filtration The filtrate was heated to reflux, and then hydrochloric acid was added thereto, and the mixture was cooled to room temperature and filtered to obtain a cake, followed by drying. A method for producing a salt of a compound of the formula according to claim 2, wherein, when the acid is p-toluenesulfonic acid, 4-(4-(3-(4-chloro-3-)trifluoromethyl)benzene is used.醯))urea)-3-fluoro-phenoxy)-2-(N-1',1',1'-tridecylmethyl)pyridiniumamine is mixed with ethanol and heated to reflux until the solution is clear. Filtration treatment was carried out, and the filtrate was heated under reflux. Then, p-toluenesulfonic acid was added, and the mixture was cooled to room temperature and filtered to obtain a cake, followed by drying. a solvate of a compound of the formula: Wherein the solvate of the compound is an methanesulfonic acid ethanolate selected from the compound. A process for the preparation of a solvate of the compound of the formula of claim 5, which comprises the step of: 4-(4-(3-(4-chloro-3-(trifluoromethyl)phenyl) guanidine) -3-fluoro-phenoxy)-2-(N-1',1',1'-tridecylidenemethyl)pyridiniumamine is mixed with ethanol and heated to reflux until the solution is clarified, followed by filtration, and The filtrate was heated to reflux, and then methanesulfonic acid was added thereto, and the mixture was cooled to room temperature and filtered to obtain a cake, followed by drying. 4-(4-(3-(4-Chloro-3-(trifluoromethyl)phenyl) guanidine)-3-fluoro-phenoxy)-2-( N -1 ',1',1 '-Trimethylaminomethylmercapto)pyridine-1-oxide is represented by the formula: Preparation of 4-(4-(3-(4-chloro-3-(trifluoromethyl)phenyl)guanidine)-3-fluoro-phenoxy)-2-(N-1',1', A method of 1'-trimethylaminomethylmercapto)pyridine-1-oxide, comprising the steps of: including 4-(4-(3-(4-chloro-3-)trifluoromethyl) A reaction of a mixture of phenyl)carbazide)-3-fluoro-phenoxy)-2-(N-1 ',1',1'-tridemethyl)pyridiniumamine with an oxidizing agent. a use of a salt of a compound of the formula: A salt of the compound for use in the preparation of a pharmaceutical composition for inhibiting phosphokinase, wherein the salt of the compound is selected from the group consisting of the hydrochloride salt of the compound, the p-toluenesulfonate of the compound, or the mesylate salt of the compound. . Use of a solvate of a compound of the formula: A solvate of the compound is used to prepare a pharmaceutical composition for inhibiting phosphokinase, wherein the solvate of the compound is an methanesulfonic acid ethanolate selected from the compound.