A tyrosine kinase inhibitor and its application

Abstract

The present invention discloses a tyrosine kinase inhibitor and its application. The structure of the tyrosine kinase inhibitor compound or pharmaceutically acceptable salt is shown in formula (I): #imgabs0# wherein R1, R3 and R4 are independently hydrogen, substituted or unsubstituted C1-4 alkyl; the substitution is substituted by at least one of the following groups: halogen, nitro, amino, cyano and phenyl; R2 is independently hydrogen, halogen, nitro, cyano, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1-2 alkoxy; the substitution is substituted by at least one of the following groups: halogen, nitro and phenyl; the tyrosine kinase inhibitor compound or pharmaceutically acceptable salt exists in the form of a pharmaceutical composition during actual application, and the pharmaceutical composition also contains a pharmaceutically acceptable carrier and / or excipient. The tyrosine kinase inhibitor and its application can be used to prevent and treat tyrosine kinase-mediated diseases, including thyroid cancer, renal cell carcinoma, liver cancer, etc., mainly HCC.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and more particularly to a tyrosine kinase inhibitor and its application in preventing and treating tyrosine kinase-mediated diseases, in particular to a tyrosine kinase inhibitor and its application. Background Art

[0002] Liver cancer is the sixth leading cause of cancer worldwide and the fourth most common cause of cancer death. China is a major country for liver cancer. According to statistics, there are about 700,000 new liver cancer patients in the world each year, and China accounts for about half, about 360,000.

[0003] Hepatocellular carcinoma (HCC) accounts for 75%-85% of all primary liver cancers and is a solid tumor with high vascular dependence. During the development of HCC, abnormalities in vascular endothelial growth factor receptor (VEGFR) and platelet-derived growth factor receptor (PDGFR) can upregulate the downstream RAS / MAPK pathway, leading to increased angiogenesis and cell proliferation, which are associated with HCC and metastasis.

[0004] Lenvatinib was approved by the FDA in February 2015 for the treatment of advanced radioactive-iodine refractory differentiated thyroid cancer. In 2018, the FDA confirmed the approval of lenvatinib for the treatment of advanced, inoperable hepatocellular carcinoma. Less than a month later, China's CFDA officially confirmed that lenvatinib was approved for marketing in China for the first-line treatment of advanced liver cancer. Although studies have shown that lenvatinib is more effective than sorafenib, it still has defects.

[0005] When lenvatinib was first developed, it was intended for the treatment of thyroid cancer, not HCC. In vitro studies have shown that it has a better effect on VEGFR, but a weaker effect on PDGFR. It is 10 times more selective for VEGFR2-3 than for FGFR1 and PDGFR.

[0006] In addition, since the free state of lenvatinib (structural formula as formula ①) is difficult to dissolve in various common solvents, when synthesizing its mesylate, acetic acid is selected as the solvent, and then methanesulfonic acid is added to form a salt (refer to Japanese Eisai patent WO2016 / 031841). This process will inevitably lead to the appearance of genotoxic impurities (structural formula as formula ②). When synthesizing lenvatinib, the genotoxic impurities can only be reduced as much as possible, but it is difficult to remove the impurities.

[0007]

[0008] Oral lenvatinib has many adverse reactions, such as high blood pressure, diarrhea, decreased appetite, weight loss, and fatigue. Such a large number of side effects may also be related to the excessive amount of various genotoxic impurities in lenvatinib.

[0009] In view of the above defects, the present invention improves the structure of lenvatinib in the hope that it can exert better efficacy against liver cancer, while reducing genotoxic impurities in the drug and reducing drug side effects; therefore, we propose a tyrosine kinase inhibitor and its application to solve the above problems. Summary of the invention

[0010] The object of the present invention is to provide a tyrosine kinase inhibitor and its application for preventing and treating tyrosine kinase-mediated diseases, including thyroid cancer, renal cell carcinoma, liver cancer, gastric cancer, etc.

[0011] To achieve the above object, the present invention provides the following technical solution: a tyrosine kinase inhibitor, wherein the structure of the tyrosine kinase inhibitor compound or a pharmaceutically acceptable salt thereof is as shown in formula (I):

[0012]

[0013] Wherein, R1, R3 and R4 are independently hydrogen, substituted or unsubstituted C1-4 alkyl; the substitution is substitution by at least one of the following groups: halogen, nitro, amino, cyano and phenyl;

[0014] R2 is independently hydrogen, halogen, nitro, cyano, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1-2 alkoxy; the substitution is substituted by at least one of the following groups: halogen, nitro and phenyl.

[0015] Preferably, the structural formula of the preferred compound of the tyrosine kinase inhibitor or pharmaceutically acceptable salt is as shown in Formula (II) and Formula (III):

[0016]

[0017] Preferably, the salt includes but is not limited to acetate, ascorbate, benzoate, benzenesulfonate, citrate, fumarate, hydrochloride, hydrobromide, maleate, methanesulfonate, nitrate, oxalate, phosphate, succinate and sulfate.

[0018] Another technical solution provided by the present invention is to provide an application of a tyrosine kinase inhibitor, wherein the tyrosine kinase inhibitor compound or pharmaceutically acceptable salt exists in the form of a pharmaceutical composition during actual application, and the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

[0019] Preferably, the tyrosine kinase inhibitor compound or pharmaceutically acceptable salt also includes use in the preparation of a drug for preventing or treating a tyrosine kinase-mediated disease.

[0020] Preferably, the diseases mediated by the tyrosine kinase inhibitor compound or pharmaceutically acceptable salt include thyroid cancer, renal cell carcinoma, liver cancer, etc., especially hepatocellular carcinoma.

[0021] Compared with the prior art, the beneficial effects of the present invention are: the tyrosine kinase inhibitor and its application can be used to prevent and treat tyrosine kinase-mediated diseases, including thyroid cancer, renal cell carcinoma, liver cancer, etc., mainly HCC. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a schematic diagram of the preparation process of compound (II) in the present invention;

[0023] Figure 2 The experimental data analysis chart in the present invention is shown in FIG. Figure 1 ;

[0024] Figure 3 The experimental data analysis chart in the present invention is shown in FIG. Figure 2 ;

[0025] Figure 4 The experimental data analysis chart in the present invention is shown in FIG. Figure 3 ;

[0026] Figure 5 The experimental data analysis chart in the present invention is shown in FIG. Figure 4 ;

[0027] Figure 6 The experimental data analysis chart in the present invention is shown in FIG. Figure 5 . DETAILED DESCRIPTION

[0028] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] The present invention provides a technical solution: a tyrosine kinase inhibitor, wherein the structure of the tyrosine kinase inhibitor compound or a pharmaceutically acceptable salt thereof is as shown in formula (1):

[0030]

[0031] Wherein, R1, R3 and R4 are independently hydrogen, substituted or unsubstituted C1-4 alkyl; the substitution is substitution by at least one of the following groups: halogen, nitro, amino, cyano and phenyl;

[0032] R2 is independently hydrogen, halogen, nitro, cyano, substituted or unsubstituted C1-2 alkyl, substituted or unsubstituted C1-2 alkoxy; the substitution is substituted by at least one of the following groups: halogen, nitro and phenyl.

[0033] In the present invention, the preferred structural formulas of the tyrosine kinase inhibitor or pharmaceutically acceptable salt are as shown in Formula (II) and Formula (III):

[0034]

[0035] In the present invention, the salts include but are not limited to acetate, ascorbate, benzoate, benzenesulfonate, citrate, fumarate, hydrochloride, hydrobromide, maleate, methanesulfonate, nitrate, oxalate, phosphate, succinate and sulfate.

[0036] Another technical solution provided by the present invention is to provide an application of a tyrosine kinase inhibitor, wherein the tyrosine kinase inhibitor compound or pharmaceutically acceptable salt exists in the form of a pharmaceutical composition during actual application, and the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

[0037] The present invention further includes the use of the tyrosine kinase inhibitor compound or pharmaceutically acceptable salt in the preparation of a drug for preventing or treating a tyrosine kinase-mediated disease.

[0038] In the present invention, the diseases mediated by the tyrosine kinase inhibitor compound or pharmaceutically acceptable salt include thyroid cancer, renal cell carcinoma, liver cancer, etc., especially hepatocellular carcinoma.

[0039] Embodiment 1:

[0040] The preparation of compound (II) is as follows Figure 1 As shown in;

[0041] Wherein: the synthesis of compound 3 is;

[0042] 900 ml of DMSO was added to a 3 L reaction bottle, and then 64 g of KOH was added. The reaction bottle was evacuated and replaced with nitrogen once. While maintaining the nitrogen atmosphere, 98.9 g of compound 1 was added to the reaction bottle.

[0043] Rinse the sticky material with 100 ml DMSO. Cool the reaction solution to 20-25°C, add water to the reaction bottle (dropping time 10-15 minutes), (the reaction is slightly exothermic) and control the temperature below 40°C.

[0044] After the water is added, keep stirring until the system changes from reddish brown to dark black (about 0.5-1h), and continue stirring for 10-15 minutes after the system changes color.

[0045] Add 100g of compound 2 to the reaction bottle at one time, heat to 90℃, stir and react for 2 hours, cool to 55℃, add 500ml of acetone and 1000ml of water to the reaction solution in sequence, keep warm at 50 to 55℃ and stir for 30 minutes. Cool to 30-35℃, then cool to 5-10℃ in an ice water bath, keep warm and stir for 30 minutes.

[0046] Filter by suction, wash the filter cake with a mixed solution of 100 ml acetone / 200 ml water, and then wash the filter cake with water until the filtrate is basically colorless; dry under vacuum / air drying at below 90°C until the moisture content is less than 0.2%, to obtain 132.0-138.0 g of off-white to reddish yellow solid (Compound 3), with a yield of 91-95%;

[0047] Submit for inspection. If the single impurity is ≥ 0.2%, it is purified. The purification steps are as follows: add 132g of crude product to 10ml / gTHF, heat and reflux for 2-3h, cool to 10-20℃, filter, rinse the filter cake with a small amount of THF, and dry in vacuum at 60-80℃ to obtain 118.8g of finished product, with a yield of 85-90%.

[0048] Among them: Synthesis of compound 4;

[0049] Weigh 130 g of compound 3 into a 3 L three-necked flask under nitrogen protection, add 1300 ml of DMF, 65.8 g of pyridine, and 5.4 g of water, stir, cool to 0-5°C, add 142.0 g of benzyl chloroformate dropwise into the reaction flask, and react at 0-10°C after the addition is complete.

[0050] About 30 minutes after the addition, solid will precipitate out of the system and the system will become viscous.

[0051] After about 3 hours of reaction, 1300 ml of water was added dropwise to the reaction solution, and the temperature was controlled to be less than or equal to 20°C. (heat was released when water was just added). After the addition was completed, the mixture was stirred for 0.5 hours, and the temperature was controlled to 10-20°C. The mixture was filtered and the filter cake was washed with 520 ml of water.

[0052] Then wash with 520 ml of acetone, and dry in vacuum at 60°C-70°C until the moisture content is less than 0.5%, to obtain 162 g of white or off-white solid (Compound 4), with a yield of about 90%;

[0053] The synthesis of compound (II)

[0054] Weigh 125.7g of adamantane hydrochloride and 92.6g of potassium carbonate into a 3L three-necked flask under nitrogen protection, add 800ml of DMF, heat to 50℃ and stir for 1 hour, add 53g of pyridine, then add 160g of compound 4, and stir at 50℃ for about 2 hours. Add 1600ml of water to the reaction solution, control the temperature to be less than or equal to 20℃, add dropwise, (heat will be released when water is just added), stir for 0.5 hour, control the temperature to 10-20℃, filter, wash the filter cake with 1120ml of water, and then wash with 560ml of ethanol. Dry under vacuum at 60℃-70℃ until the moisture is less than 0.5%, and obtain 157g of white or off-white solid (compound II). The yield is about 90%.

[0055] Purification: Add 800 ml of ethanol to the crude product and reflux for 1 hour, then cool and filter to obtain the pure product.

[0056] MS: m / z(ES):521.2,522.2[M+1].

[0057] 1H NMR:

[0058] (400MHz,d6-DMSO):8.67(d,2H),8.28(d,1H),8.01(s,1H),7.86(s,1H),7.75(s,1H),7.51(s,1H),7.47( d,1H),7.18~7.21(dd,1H),6.87(s,1H),6.52(d,1H),4.03(s,3H),2.03(s,3H),1.96(s,6H),1.64(s,6H).

[0059] Embodiment 2:

[0060] Preparation of compound (III):

[0061] Preparation process: Refer to the synthesis of compound (II) in Example 1; replace adamantane hydrochloride with memantine hydrochloride to obtain compound (III).

[0062] MS: m / z(ES):549.2,550.2[M+1].

[0063] 1H NMR:

[0064] (400MHz,d6-DMSO):8.67(d,2H),8.27(d,1H),8.00(s,1H),7.86(s,1H),7.75(s,1H),7.51(s,1H),7.47(d,1H),7.18~7.21(dd,1H) ,6.90(s,1H),6.51(d,1H),4.03(s,3H),2.10(s,1H),1.78(s,2H),1.60(s,4H),1.35(d,2H),1.28(d,2H),1.12(s,2H),0.83(s,6H).

[0065] Embodiment 3:

[0066] Preparation of compound (II) hydrochloride:

[0067] Weigh 10 g of compound (II), add 300 ml of ethanol to dissolve, add 10 ml of 15% hydrogen chloride ethanol solution dropwise, stir at room temperature for 2 hours, filter, wash the filter cake with ethanol, and dry in vacuo to obtain about 9 g of white solid.

[0068] Embodiment 4:

[0069] Preparation of compound (III) hydrochloride:

[0070] The preparation process can be prepared by referring to Example 3 to obtain the hydrochloride of compound (III).

[0071] Purity experiment 1

[0072] 1. Experimental conditions:

[0073] Detector: UV absorbance meter (measurement wavelength: 252nm);

[0074] Column: WONDASIL C18 SUPERB 5um 4.6MM*250MM;

[0075] Column temperature: constant temperature close to 40°C;

[0076] Mobile phase: As shown in Table 1, use a linear gradient to elute Solution A and Solution B with the following compositions;

[0077] Solution A: water / acetonitrile / 70.0-72.0% analytical grade perchloric acid (990:10:1, V / V / V);

[0078] Solution B: water / acetonitrile / 70.0-72.0% analytical grade perchloric acid (100:900:1, V / V / V);

[0079] Flow rate: 1.0ml / min

[0080]

[0081]

[0082]

Table 1

[0083] 2. Experimental principle: The content of formula ② in the hydrochloride of compound (II) or (III) prepared in Example 3 or 4 and the mesylate of formula ① prepared by the method of patent WO2016 / 031841 was detected by external standard method;

[0084] 3. Experimental results:

[0085] name Content of impurity formula ② (by mass) Compound (II) hydrochloride Not detected Compound (III) hydrochloride Not detected Formula ① Methanesulfonate 78ppm

[0086]

Table 2

[0087] As shown in Table 2, the hydrochlorides of compound (II) and compound (III) did not detect the genotoxic impurity formula ② mentioned above. However, the existing process of lenvatinib mesylate will inevitably produce genotoxic impurities of formula ②;

[0088] Test Example 1: In vitro enzyme inhibition activity test:

[0089] 1. Test principle:

[0090] The chemiluminescence in the assay plate was detected by Envision, and the IC50 value of the compound was used as an indicator to evaluate the inhibitory effect of the compound on tyrosine kinase.

[0091] 2. Test method (taking PDGFRα as an example):

[0092] Dilute enzyme, substrate, ATP, and inhibitors in kinase buffer.

[0093] Add to wells of a 384 low volume plate: 1 μl inhibitor or (5% DMSO), 2 microliters enzyme (2 ng), 2 μl substrate / ATP mix (25 uM ATP, 0.2 μg / μl PolyE4Y1), then incubate at 25°C for 120 minutes.

[0094] Add 5 μl ADP-Glo TM The reagents were incubated at 25°C for 40 minutes.

[0095] Add 10 μl of kinase assay reagent and incubate at 25°C for 30 minutes.

[0096] Luminescence was recorded (integration time 0.5 s). Kinase buffer: 40 mM Tris, pH 7.5; 20 mM magnesium chloride; 0.1 mg / ml bovine serum albumin; 50 μM DTT;

[0097] 3. Test results

[0098]

[0099] [Table 3 Inhibitory effects of the compounds on in vitro enzymes]

[0100] As shown in Table 3, Example compounds (II) and (III) have inhibitory effects on the tyrosine kinases in the test, and their inhibitory effects on PDGFRα kinase are better than those of the control formula ① (lenvatinib), especially compound (II). The specific test data analysis is shown in Figure 2 , Figure 3 , Figure 4 , Figure 5 and Figure 6 , wherein LQTX001-A in the spectrum corresponds to the compound (II) described above, LQTX001-B corresponds to the compound (III) described above, and LQTX001-DZ corresponds to lenvatinib.

[0101] The contents not described in detail in this specification belong to the prior art known to the professional and technical personnel in this field. Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for the technical solutions described in the aforementioned embodiments to be modified, or some of the technical features thereof to be replaced by equivalents, and any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A tyrosine kinase inhibitor, characterized in that: The structure of the tyrosine kinase inhibitor compound or pharmaceutically acceptable salt is shown in formula (1): Wherein, R1, R3 and R4 are hydrogen or unsubstituted C1-4 alkyl; Wherein, R2 is hydrogen, halogen, nitro, cyano, unsubstituted C1-2 alkyl or unsubstituted C1-2 alkoxy.

2. A tyrosine kinase inhibitor according to claim 1, characterized in that: The structural formula of the tyrosine kinase inhibitor or the pharmaceutically acceptable salt thereof is shown in Formula (II) and Formula (III):

3. A tyrosine kinase inhibitor according to claim 1, characterized in that: The salt is acetate, ascorbate, benzoate, benzenesulfonate, citrate, fumarate, hydrochloride, hydrobromide, maleate, methanesulfonate, nitrate, oxalate, phosphate, succinate or sulfate.

4. A pharmaceutical composition containing the tyrosine kinase inhibitor according to claim 1, characterized in that: The pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

5. The use of a tyrosine kinase inhibitor according to claim 1, characterized in that: Use of the tyrosine kinase inhibitor compound or pharmaceutically acceptable salt in preparing drugs for preventing or treating tyrosine kinase-mediated diseases.

6. The use of a tyrosine kinase inhibitor according to claim 5, characterized in that: The disease is thyroid cancer, renal cell carcinoma or liver cancer.

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