WO2013007184A1 - Antineoplastic drug tetrahydronaphthalene amide-group compound and pharmaceutically acceptable salt, preparation method and use thereof - Google Patents

Abstract

The present invention relates to a novel compound tetrahydronaphthalene amide-group compound as represented by general formula I, or a pharmaceutically acceptable salt thereof, serving as an antineoplastic drug. Also provided are a preparation method of the compound, a pharmaceutical composition comprising the compound, a result of exosomatic and endosomatic antineoplastic effect, and a study on acute toxicity. The antineoplastic drug tetrahydronaphthalene amide-group compound of the present invention has better antineoplastic activity and safety, can be used to treat tumors such as leukemia, lung cancer, colon cancer, ovarian cancer and kidney cancer and the like, thus having a wide therapeutic window and a high application value as an antineoplastic agent in the pharmaceutical field.

Description

 Antitumor drug tetralinamide type compound and pharmaceutically acceptable salt thereof, and preparation method and application thereof

 [0001] The present invention relates to the field of medicine, and in particular to a tetrahydronaphthylamine compound which inhibits tumor cell growth and exerts an antitumor effect, and a pharmaceutically acceptable salt thereof, a preparation method and use thereof.

Background technique

 [0002] Imatinib competitively inhibits the binding site of adenosine triphosphate (ATP) to a thymidine kinase (TK) receptor such as KIT, blocks TK phosphorylation, thereby inhibiting signaling, and inhibits KIT associated with kinase activity. Mutations (causing KIT receptor activation) and wild-type KIT. There are three main target sites: Abelson (ABL) protein, KIT protein and platelet-derived growth factor (PDGF) receptor. Imatinib can reduce kinase phosphorylation of GIST-derived cell lines (GIST882) by functionally acquired KIT mutations that are independent of stem cell factor activation, and completely inhibit kinase phosphorylation at concentrations up to 1 μmol/L Chemical.

 [0003] Imatinib, approved by the FDA for the treatment of chronic myeloid leukemia (CML) in May 2001, was approved by the FDA in 2003 for the treatment of gastrointestinal stromal tumors (GIST).

[0004] Its structure is:

Imatinib has significant adverse effects, and most patients experience some adverse reactions during their use, but most are mild to moderate.

 [0005] In the clinical trial of CML, patients who discontinued due to drug-related adverse reactions accounted for only 1% of patients with chronic phase failure of α-interferon therapy, 2% of the accelerated phase, and 5 of the blast phase. %.

 [0006] In the GIST clinical trial, 3.4% discontinued due to drug-related adverse events.

[0007] The adverse effects of CML and GIST patients are similar, with only two exceptions: GIST patients have less bone marrow suppression, and intratumoral hemorrhage is only observed in GIST patients. Among CML and GIST patients, the most common reported drug-related adverse events were mild nausea (50 to 60%), vomiting, diarrhea, abdominal pain, fatigue, myalgia, tendon and erythema. These adverse events are easy. deal with. [0008] In all studies, edema and water retention were reported, with incidences of 47 to 59% and 7 to 13%, respectively, of which severe cases were 1 to 3% and 1 to 2%, respectively. The edema of most patients is characterized by periorbital and lower extremity edema. There have been individual reports of glaucoma, related to water retention. There are also reports of pleural effusion, ascites, pulmonary edema, and rapid weight gain. These events can usually be alleviated by suspension of imatinib, use of diuretics or other supportive care. But individual patients are in serious condition and even life-threatening. One patient with blast crisis died of complicated clinical conditions involving pleural effusion, congestive heart failure, and renal failure.

 [0009] When imatinib is used in combination with high-dose chemotherapeutic agents, transient hepatotoxicity, such as elevated transaminases and hyperbilirubinemia, can occur.

Summary of the invention

 [0010] In order to solve the above problems, an object of the present invention is to provide an antitumor drug tetralinamide which has superior antitumor activity and safety than imatinib and a therapeutic window width, and is pharmaceutically acceptable Salt.

 Another object of the present invention is to provide a process for producing an antitumor drug tetralinamide type compound and a pharmaceutically acceptable salt thereof, and use thereof.

 [0012] In the present invention, in order to change the above-mentioned weakness of imatinib, we structurally modify it to synthesize various tetrahydronaphthalene amide compounds and pharmaceutically acceptable salts thereof, and study their antitumor activities. The result was found to have the following general formula

The compounds represented by [I] and [11] have extremely excellent antitumor activity, stability and safety.

The present invention relates to a compound represented by the general formula [I] and a pharmaceutically acceptable salt thereof:

In the formula:

Is a saturated cyclic amino group selected from the group consisting of piperidinyl, piperazinyl, imidazolidinyl, pyrrolidinyl, azacyclobutane and morpholinyl, or each group may be The ground is replaced by 1, 2, 3 or 4, which is a hydrogen atom, a halogen, an amino group, a cyano group, a C ₆ alkyl group, a d- ₆ hydroxyalkyl group, a d- ₆ halogenated alkyl group or a d- ₆ cyanide. Alkyne.

D is selected from pyridyl, pyrimidinyl, pyrazinyl, triazinyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, isoxazolyl, triazolyl, or pyrazolyl, or each radical optionally substituted with 1, 2, 3, or R ₂ is substituted, the R ₂ is independently selected from halo, cyano, amino, C ^ ₆ alkyl, d- ₆ hydroxyalkyl, d — ₆ haloalkyl, or — ₆ cyanyl.

E is selected from pyridyl, pyrimidinyl, pyrazinyl, triazinyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, isoxazolyl, triazolyl, pyrazolyl, nitrogen Azyl, pyrrolopyridyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl, quin Orolinyl, isoquinolyl, quinazolinyl, piperazinyl or morpholinyl, or each group optionally substituted by 1, 2, or 3 R ₃ , said R _{3 being} independently selected Or halogen, cyano, amino, C^ ₆ alkyl, d- ₆ hydroxyalkyl, d- ₆ haloalkyl, or C^ ₆ cyanoalkyl.

 The above compound represented by the formula [I] and a pharmaceutically acceptable salt thereof, preferably, have the following structural formula [11]:

Its towel _:

R4 and R _{5 are} independently selected from a hydrogen atom, a cyano group, C ^ ₆ alkyl, d- ₆ hydroxyalkyl, d- ₆ haloalkyl, d- ₆ cyanoalkyl;

R ₆ is independently selected from hydrogen atoms, C ^ ₆ alkyl, C ₂ - ₆ hydroxyalkyl, C ₂ - ₆ haloalkyl, d- ₆ cyanoalkyl, acyl and carboxylate;

p is 0, 1, or 2.

 a compound represented by the above formula [II] and a pharmaceutically acceptable salt thereof, preferably, D is a pyrimidinyl group, and E is a pyridyl group, having the formula [III]

The compound represented by the above formula [III] and a pharmaceutically acceptable salt thereof, preferably the compound 1 is: D is a pyrimidinyl group, E is a pyridyl group, R4 and R ₅ are a hydrogen atom, and R ₆ is a methyl group. , p is 1, and its structural formula is as follows: ,

 Compound 1

 a compound represented by the above formula [II] and a pharmaceutically acceptable salt thereof, preferably, D is a pyrimidinyl group, and hydrazine is a pyrimidyl group, having a knot of the formula [IV]

The compound represented by the above formula [IV] and a pharmaceutically acceptable salt thereof, preferably the compound 2 is: D is a pyrimidinyl group, hydrazine is a pyrimidinyl group, and R4 and R ₅ p are 1, and the structural formula is as follows:

 Compound 2

 The above compound and a pharmaceutically acceptable salt thereof are used as an active ingredient in the preparation of a medicament for treating a tumor disease.

 A pharmaceutical composition comprising a compound as described above, and a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

[0023] The use of a medicament for treating a tumor disease includes: leukemia, lung cancer, liver cancer, colon cancer, nerve Cancer, melanoma, ovarian cancer, kidney cancer, prostate cancer, and breast cancer.

 [0024] The beneficial effects of the present invention are: the antitumor drug tetralinamide compound obtained by the invention and the pharmaceutically acceptable salt thereof have better antitumor activity and safety, and can be used for treating leukemia, lung cancer, The application of tumors such as colon cancer, ovarian cancer and kidney cancer has a wide anticancer spectrum and a wide therapeutic window, so it is very useful as an antitumor agent in the medical field.

DRAWINGS

 BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a diagram showing the synthesis mechanism of a compound represented by the formula [I] of the present invention and a pharmaceutically acceptable salt thereof.

2 is a synthetic process route diagram of the compound 1 of the present invention.

3 is a synthetic process route diagram of the compound 2 of the present invention.

detailed description

 The present invention will be more specifically described by the following examples, but the present invention is not limited by these examples.

[0029] Exemplary embodiments of the present invention will be described in detail below. However, these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

 The compounds of the invention also include pharmaceutically acceptable salts. A pharmaceutically acceptable salt refers to a form which converts a basic group in the parent compound into a salt. Pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic groups such as amines (amino) groups.

 The pharmaceutically acceptable salts can be prepared from inorganic or organic acids, which may include, but are not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids may include, but are not limited to, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, almonds Acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like.

 [0032] As used herein, "pharmaceutically acceptable carrier" includes any and all pharmaceutically acceptable solvents, dispersion media, coatings, isotonic and absorption delaying agents, and the like, such media and agents are employed in pharmaceutically active materials, in It is well known in the art that unless any conventional media or agent is incompatible with the active ingredient, its use in a therapeutic composition is contemplated, and the additional active ingredient may be incorporated into the compositions.

 The pharmaceutical composition of the present invention can be used orally, by injection, by spray inhalation, for external use on the skin, for rectal administration, nasal, vaginal, intraperitoneal, or by implantation of a reservoir or a transdermal patch.

 An important aspect of the present invention is that the present invention is useful for treating diseases involving tumors, including: leukemia, lung cancer, colon cancer, ovarian cancer, and renal cancer.

Another aspect of the invention relates to a process for the preparation of a compound of the general formula [I]. As shown in Figure 1, the compound of the general formula [I] can be produced by the following methods and processes. [0036] 6-methoxy-1-tetralinone is refluxed in a 48% HBr solution to give intermediate 1-1, the phenolic hydroxyl group is protected with trifluoromethanesulfonic anhydride to give intermediate 1-2, 1-2 Reaction with CO in DMF, using palladium as a catalyst, such as palladium diacetate, 1,3-bis(phenylphosphine)propane (dppp) or bis(triphenylphosphine)palladium(II) chloride [II] [(PPh ₃ ) ₂ PdCl ₂ ] Intermediates 1-3 are obtained, which are reduced in an alcohol solvent (for example, ethanol) by a reducing agent (for example, sodium borohydride) to give Intermediate 1-4. The hydroxy group of 1-4 is chlorinated under the action of a reagent such as thionyl chloride to give the chloro group of the intermediate 1-5, 1-5 substituted by a cyclic amino group (or cyclic amino group), using triethylamine or Potassium carbonate is used as a base to obtain intermediates 1-6, 1-6 in a solution of trimethylaluminum toluene with 6-methyl-!^-[(4-pyridin-3-yl)pyrimidin-2-yl] The benzene-1,3-diamine is reacted to give a compound of the general formula [I]. It is also possible to hydrolyze from the ester group in 1-6 to give intermediates 1-7, which are reacted with an alcohol or an amino group after formation of an acid chloride under the action of thionyl chloride or in a coupling agent such as EDCI (1-ethyl-3). -(3-dimethylaminopropyl)carbodiimide) or DCC (1,3-dicyclohexylcarbodiimide) to DMAP (4,4-dimethylaminopyridine) or hydrazine (1-hydroxybenzene) And the triazole is directly reacted with an alcohol or an amino group as a catalyst to obtain a compound of the general formula [I].

[0037]

Example 1 Preparation of Compound 1 and Its Pharmaceutically Acceptable Salt

 (i) N-{4-methyl-3-[(4-pyridin-3-yl)pyrimidin-2-amino]phenyl}-5-(4-methylpiperazine small group)-5,6,7 Preparation of 8-tetrahydronaphthalene-2-amide (Compound 1), the structural formula of Compound 1 is as follows:

 Compound 1

 Step A: Synthesis of 6-Hydroxy-1-tetralinone

6-methoxy-1-tetralinone (17.6 g, 100 mmol) was reacted in 100 ml of 48% hydrogen bromide solution at 100 ° C for 24 hours, then cooled to room temperature, and the reaction solution was added to ice water and stirred. A large amount of solid was precipitated, the solid was filtered, washed with 5% aqueous sodium bicarbonate, washed with EtOAc EtOAc EtOAc EtOAc EtOAc. 13.9 g, yield 86.0 o MS (M+l) = 163.14o Step B: Synthesis of 5-family-5,7,8-tetrahydronaphthalene-2-ester of trifluoromethanesulfonate

6-Hydroxy-1-tetralinone (8.1 g, 50 mmol) was dissolved in 50 ml of dichloromethane, and 14.1 g of trifluoromethanesulfonic anhydride diluted in 20 ml of dichloromethane was slowly added, 0~5. Under stirring at 0 °C, 5.05 g of triethylamine diluted with 20 ml of dichloromethane was slowly added dropwise. After about 10 minutes, the mixture was slowly added to room temperature and stirred for 2 hours. Then, the reaction was stopped, and the reaction solution was diluted with 5% hydrogen carbonate. The sodium aqueous solution was washed twice, and brine was washed twice. The organic phase was dried over anhydrous sodium sulfate and filtered and then evaporated. MS (M+1) = 295.21.

Step C: Synthesis of methyl 5-amino-5,6,7,8-tetrahydro-2-carboxylate

Dissolve 5-fluoro-5-carbonyl-5,6,7,8-tetrahydronaphthalene-2-ester (11.8 g, 40 mmol) in 50 mL of methanol, add DMF 5 mL, DIEA (N, N-diisopropylethylamine 5 ml, 4 g of palladium diacetate, 4 g of 1,3-bis(diphenylphosphino)propane, sealed reaction system, vacuumed, added with nitrogen, vacuumed, passed The carbon monoxide balloon was stirred at 65 ° C for 22 hours, and the reaction solution was filtered through Celite. The celite solid was washed with ethanol, and then the filtrate phase was combined, concentrated, dissolved in ethyl acetate (200 ml), and washed with brine. After 200, 200, 100), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column, and 5.3 g of product was isolated under ethyl acetate:hexanehexane = 1:2 mobile phase. The yield was 65%. MS (M+1) = 205.34.

 Step D: Synthesis of 5-hydroxy-5,6,7,8-tetracarboxylic acid methyl ester

Methyl 5-carbonyl-5,6,7,8-tetrahydronaphthalene-2-carboxylate (4.08 g, 20 mmol) was dissolved in 50 mL of ethanol and sodium borohydride (0.76 g, After stirring at room temperature for 3 hours, the reaction mixture was concentrated under reduced pressure. EtOAc (EtOAc) %. MS (M+1) = 207.12.

Step E: Synthesis of methyl 5-chloro-5,6,7,8-tetrahydronaphthalene-2-carboxylate

Methyl 5-hydroxy-5,6,7,8-tetrahydronaphthalene-2-carboxylate (3.09 g, 15 mmol) was dissolved in 20 mL of dichloromethane, slowly at 0-5 ° C A solution of thionyl chloride (7.08 g, 60 mmol) in 10 ml of dichloromethane was added dropwise. After the completion of the dropwise addition, the reaction mixture was allowed to react to room temperature for 6 hours. The reaction mixture was concentrated under reduced pressure. ethyl acetate (50 ml), EtOAc (EtOAc) Product, yield 94.5%. MS (M+1) = 225.21.

Step F: Synthesis of 5-(4-methylpiperazine-1-yl-5,6,7,8-tetrahydronaphthalene-2-carboxylic acid methyl ester

Methyl 5-chloro-5,6,7,8-tetrahydronaphthalene-2-carboxylate (2.41 g, 10 mmol) was dissolved in 20 mL of DMF solvent and potassium carbonate (2.78 g, 20 mmol) , N-methylpiperazine (2 g, 20 mmol), after reacting at 50 ° C for 5 hours, the reaction solution was added to ethyl acetate (80 ml), and the pH was adjusted to neutral with dilute aqueous hydrochloric acid. The ethyl ester phase was washed with saturated brine (80×3), dried over anhydrous sodium sulfate, filtered, filtered, evaporated, evaporated, evaporated.克, yield 73.9 %, MS (M+1) = 289.31.

Step G: Synthesis of 5-(4-methylpiperazin-1-yl)-5,7,8-tetrahydronaphthalene-2-carboxylic acid

Dissolve methyl 5-(4-methylpiperazin-1-yl)-5,6,7,8-tetrahydronaphthalene-2-carboxylate (2.88 g, 10 mmol) in 30 mL of methanol and 20 mL In a solution of tetrahydrofuran, 2N NaOH (10 ml, 20 mmol) was added, and the mixture was reacted at room temperature for 16 hours. After adjusting pH = 4 with 2N aqueous hydrochloric acid solution, the organic solvent was evaporated under reduced pressure, ethyl acetate (60 ml) and saturated brine. Washed with water (60 X 3), the ethyl acetate phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated to dryness. Separate, methanol: dichloromethane: glacial acetic acid = 20: 1: 5 drops gave product 2.16 g, yield 78.8%, MS (M+1) = 275.18.

 Step H: N-{4-methyl-3-[(4-pyridin-3-yl)pyrimidin-2-amino]phenyl}-5-(4-methylpiperazin-1-yl)- Synthesis of 5,6,7,8-tetrahydronaphthalene-2-amide (Compound 1)

method one:

Methyl 5-(4-methylpiperazin-1-yl)-5,6,7,8-tetrahydronaphthalene-2-carboxylate (2.88 g, 10 mmol) and 6-methyl-! -[(4-Pyridin-3-yl)pyrimidin-2-yl]benzene-1,3-diamine (3.32 g, 12 mmol) was suspended in 30 ml of toluene, and 2 M trimethylaluminum toluene solution was added. (5 ml, 10 mmol), the mixture was reacted at 10 CTC for 6 hours, and the solution was cooled. An aqueous solution of sodium potassium tartrate (50 ml) was added with stirring. The solution was extracted with ethyl acetate three times (100, 100, 50 ml). The combined extracts were washed with sodium hydrogen carbonate (100 ml) and brine (100, 100, 50 ml) and dried over anhydrous sodium sulfate. Filtration, the filtrate was concentrated under reduced pressure and purified by silica gel column eluting with 50% ethyl acetate/dichloromethane/1% triethylamine mobile phase, Compound 1, 2.32 g, yield 43.5%.

[0046] Method two:

5-(4-Methylpiperazine small)-5,6,7,8-tetrahydronaphthalene-2-carboxylic acid (1.37 g, 5 mmol), 6-methyl-!^-[(4 -pyridin-3-yl)pyrimidin-2-yl]benzene-1,3-diamine (1.83 g, 5.5 mmol), EDCI (1-ethyl-3-(3-dimethylaminopropyl) carbon two Imine) (1.15 g, 6 mmol), DMAP (4,4-methylaminopyridine) (0.31 g, 2.5 mmol) was added to a mixture of 30 ml of tetrahydrofuran and 10 ml of DMF, and reacted at room temperature for 4 hours. The reaction solution was evaporated under reduced pressure of tetrahydrofuran, and ethyl acetate (50 ml) was added, and washed with brine (3, (60, 60, 30 ml), ethyl acetate phase dried over anhydrous sodium sulfate, filtered, Solvent, silica gel column chromatography was carried out under 50% ethyl acetate/dichloromethane/1% triethylamine mobile phase to give the desired product, Compound 1, 1.65 g, yield 61.8%.

MS (M+1) = 534.21. 1H-NMR (DMSO-J ₆ ppm): δ 9.98 (s, IH); 9.07 (s, IH); 8.67 (s, lH); 8.61 (d, J = 4.8 Hz, IH); 8.54 (d, J = 4.8 Hz, IH); 8.47 (d, J = 9.0 Hz, IH); 7.89 (s, lH); 7.76 (s, lH); 7.71 (d, J = 9.0 Hz, IH); 7.54 (dd, J = 8.0 Hz, 4.8 Hz, IH); 7.37 (d, J = 8.0 Hz, IH); 7.33 (d, J = 4.8 Hz, IH); 7.17 (d, J = 9.0 Hz, IH); 6.97 (d, J = 9.0 Hz, IH); 4.34 (t, J = 9.0 Hz, IH); 2.41-2.83 (m, 10H); 2.05 (s, 3H); 2.00 (s, 3H); 1.92 (m, 2H); 1.76 (m, 2H).

(b) N-{4-methyl-3-[(4-pyridin-3-yl)pyrimidin-2-amino]phenyl}-5-(4-methylpiperazin-1-yl)- Synthesis of 5,6,7,8-tetrahydronaphthalene-2-carboxamide methanesulfonate (cyclopentinib), the structural formula is as follows:

The reaction flask was added to the compound 1 (5.33 g, 10 mmol), 100 ml of anhydrous methanol, methanesulfonic acid (1.01 g, 10.5 mmol), and refluxed at 70 ° C for 1 hour, then added pharmaceutically decolorized charcoal 2 g After refluxing for an additional 1 hour, it was suction filtered, and the filtrate was concentrated under reduced pressure. Filtration and drying to give the product N-{4-methyl-3-[(4-pyridin-3-yl)pyrimidin-2-amino]phenyl}-5-(4-methylpiperazin-1-yl)- 5,6,7,8-tetrahydronaphthalene-2-carboxamide methanesulfonate 5.94 g, yield

94.5%. Melting point: 191~193 °C. Elemental analysis: C ₃₃ H ₃₉ N ₇ 0 ₄ S

Calculated for C, 62.94; H, 6.24; N, 15.57; ???: C 62.82; H 6.44; N 15.31.

 [0049]

Example 2 4-{6-[({4-methyl-3-[(4-Pyridin-3-ylpyrimidin-2-yl)amino]phenyl}amino))]-1,2,3, Preparation of 4-tetrahydronaphthalene small group} piperazine-1-carboxylic acid tert-butyl ester

Its structural formula is as follows:

Step A: Synthesis of 4-[6-(methoxyloxy)-1,2,3,4-tetrahydronaphthalen-1-yl]piperazine-l-carboxylic acid tert-butyl ester

Methyl 5-chloro-5,6,7,8-tetrahydronaphthalene-2-carboxylate (22.4 g, 100 mmol), potassium carbonate (27.8 g, 200 mmol), tert-Butyl-1-carboxylic acid tert-butyl ester (20.5 g, 110 mmol) was added to 300 ml of DMF, stirred at 40 ° C for 5 hours, filtered, and filtrate was added to ethyl acetate (800 ml) with brine. 3 X 800 ml), the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure. The reaction was carried out in a hydrogen bromide solution at 100 ° C for 24 hours, and then cooled to room temperature. The reaction solution was added to ice water and stirred to precipitate a large amount of solid. The solid was filtered and rinsed with 5% aqueous sodium hydrogencarbonate and washed with deionized water until neutral. The crude product was purified by silica gel column eluting with ethyl acetate: hexane = hexanes: MS (M+1) = 375.35.

 Step B: Synthesis of 5-(4-tert-butoxymethyl-piperidin-1-yl)-5,6,7,8-tetrahydronaphthalene-2-carboxylic acid

把 4-[6- (甲氧基羰基 )-1,2,3,4-四氢萘 -1-基]哌嗪 -1 基-羧酸叔丁基酯 (3.74 克， 10 毫摩尔)溶解 到 80毫升甲醇及 20毫升四氢呋喃溶液中， 加入含 2N的 NaOH(10毫升， 20毫摩尔)， 室温 下反应 12小时， 用 2N盐酸水溶液调 PH=5~6后， 减压旋出有机溶剂， 加入乙酸乙酯 100毫 升并用饱和食盐水洗涤 (3 X 100毫升)， 乙酸乙酯相用无水硫酸钠干燥， 过滤， 滤液减压旋出 有机溶剂后硅胶柱层析分离， 甲醇： 二氯甲烷： 冰乙酸 =20: 1： 5 滴的条件下分离得到产物 3.07克， 产率 85.3% ， MS(M+1)=361.27。

Dissolve 4-[6-(methoxycarbonyl)-1,2,3,4-tetrahydronaphthalen-1-yl]piperazine-1 -carboxylic acid tert-butyl ester (3.74 g, 10 mmol) To a solution of 80 ml of methanol and 20 ml of tetrahydrofuran, 2N NaOH (10 ml, 20 mmol) was added, and the reaction was carried out for 12 hours at room temperature. After adjusting the pH to 5 to 6 with 2N aqueous hydrochloric acid, the organic solvent was evaporated under reduced pressure. Add 100 ml of ethyl acetate and wash with saturated brine (3×100 ml). The ethyl acetate phase is dried over anhydrous sodium sulfate and filtered and evaporated. : glacial acetic acid = 20: 1: 5 The product was isolated under the conditions of 3.07 g, yield: 85.3%, MS (M+1) = 361.27.

 Step C: 4-{6-[({4-methyl-3-[(4-Pyridin-3-ylpyrimidin-2-yl)amino]phenyl}amino)carbonyl]-1,2, Tert-butyl 3,4-tetrahydronaphthalen-1-yl}piperazine-1-carboxylate

5-(4-tert-Butoxycarbonyl-piperazin-1-yl)-5,6,7,8-tetrahydronaphthalene-2-carboxylic acid (1.8 g, 5 mmol), 6-methyl-! ^-[(4-pyridine- 3-yl)pyrimidin-2-yl]benzene-1,3-diamine (1.83 g, 5.5 mmol), EDCI (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide) (1.15 g, 6 mmol), DMAP (4,4-methylaminopyridine) (0.31 g, 2.5 mmol) was added to a mixture of 30 ml of tetrahydrofuran and 10 ml of DMF, and reacted at room temperature for 6 hours. The mixture was evaporated to dryness to dryness eluted eluted eluted eluted eluted eluted eluted eluted eluted Column chromatography was carried out under a 50% ethyl acetate / dichloromethane / 1% triethylamine mobile phase to afford 2.76 g of the desired product.

MS(M+l)=620.43o 1H-NMR (DMSO-J ₆ ppm): δ 10.01 (s, H); 9.23 (s, IH); 8.94 (s, IH); 8.61 (d, J) = 4.8 Hz, IH); 8.51 (d, J = 5.2 Hz, IH); 8.45 (d, J = 8.4 Hz, IH); 7.98 (s, lH); 7.74 (s, lH); 7.72 (d, J = 8.0 Hz, IH); 7.55 (dd, J = 8.0 Hz, 4.8 Hz, IH); 7.47 (d, J = 8.4 Hz, IH); 7.44 (d, J = 5.2 Hz, IH); 7.19 (d, J = 7.6 Hz, IH); 7.03 (d, J = 9.0 Hz, IH); 4.32 (t, J = 8.0 Hz, IH); 3.12-3.31 (m, 4H); 2.51-2.83 (m, 4H); 2.45 (m, 2H); 2.35 (m, 2H); 2.3 l (m, 2H); 2.08 (s, 3H); 1.95 (m, 2H); 1.77 (m, 2H); 1.46 (s, 9H).

 [0054]

Example 3 N-{4-methyl-3-[(4-pyridin-3-yl)pyrimidin-2-amino]phenyl}-5-(piperazine small group) -5,6,7,8-tetra Preparation of hydrogen naphthalene-2-amide

Its structural formula is as follows:

4-{6-[({4-methyl-3-[(4-Pyridin-3-ylpyrimidin-2-yl)amino]phenyl}amino)carbonyl]-1,2,3,4-tetra Hydronaphthalen-1-yl}piperazine-1-carboxylic acid tert-butyl ester (1.86 g, 3 mmol) was dissolved in a mixed solution of 10 ml of trifluoroacetic acid: dichloromethane = 1:4. After stirring at room temperature for half an hour, 20 ml of an aqueous sodium carbonate solution was added to adjust the pH to be weakly basic, and the organic phase was dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the product 1.48 g, yield 95.1%.

MS(M+l)=520.22o 1H-NMR (DMSO-J ₆ ppm): δ 10.03 (s, IH); 9.22 (s, IH); 8.96 (s, lH); 8.64 (d, J) =4.8 Hz, IH); 8.52 (d, J = 5.2 Hz, IH); 8.36 (d, J = 8.0 Hz, IH); 8.02 (s, lH); 7.76 (s, lH); 7.77 (d, J = 8.0 Hz, IH); 7.55 (dd, J = 8.0 Hz, 4.8 Hz, IH); 7.46 (d, J = 8.2 Hz, IH); 7.35 (d, J = 5.2 Hz, IH); 7.33 (d, J = 8.0 Hz, IH); 7.15 (d, J = 8.8 Hz, IH); 4.26 (t, J = 6.8 Hz, IH); 2.87 (m, 2H); 2.66 (m, 4H); 2.38 (m, 2H); 2.28 (m, 2H); 2.22 (s, 3H); 1.86 (m, 2H); 1.75 (m, 2H).

[0056]

0-8'Δ'9'ς- [耷 翁 耷 (耷 ώ -t - ε-3⁄43⁄4ϋ-ΐ7)]-ε-3⁄4ώ-ΐ7}-Ν ς m^.

 [8SOO]

•(Ηε 's)orz -iuzi ^)οτ-ζτ: (HI '^H 8"9=r 'ι)ιε·ΐ: (m ' ^Ζ Η 8"8=r 'Ρ)ΔΓΔ: (HI 9 -Δ=Γ 'P) B: (m ' ^Ζ Η ζ'ς=ί ^IVL: (m ' ^Ζ -8 -8=r 'ρ)9 · B: (m ' ^Ζ Η 8 o-8=r ' ΡΡ)Ο : (HI 8"8=r

'Ρ)乙 9·8 •(Hl's)86"8 '(HI 's) ςζ'6 '(HI 's) PVOI 9 ： (uidd -Q a) ¾WN"H_T °in=(I+W)SW [乙 SOO] 'Ρ'5Δ"Δ: (Hi's) B; B: (HI's)S0'8 '(Ηΐ 'ΖΗ

'Ρ) B 9·8 • (Hl's) 86"8 '(HI 's) ςζ '6 '(HI 's) PVOI 9 : (uidd -Q a) 3⁄4WN"H _T °in=(I+W) SW [B SOO]

mm °%8 6

Mm

 ^^tp^ ' 聱 o *ώγαί 'mm mm^ ^ ^ ι 止 ^ 聱 τ\ '3⁄4ί3⁄4

'3⁄4ί o)3⁄4ίϋ3⁄40-8'Δ'9'ς-(3⁄4-ΐ-3⁄43⁄4)-ς-[3⁄4*(3⁄43⁄4-ζ-3⁄43⁄43⁄4-ε-3⁄4 ϋ-ΐ7)-ε-3⁄4ώ-ΐ7] -Ν

0-8'Δ'9'ς- 3⁄4-i-Ugata (Member 7鹜-z -{3⁄43⁄4 [耷葛耷ε-3⁄4 ϋ-ΐ7)]-ε-耷P m^. l78l.00/ClOZ OAV N-[4-Methyl-3-(4-pyridin-3-ylpyrimidin-2-yl)phenyl]-5-(piperazin-1-yl)-5,6,7,8-tetrahydro Naphthalene-2-amide (0.52 g, 1 mmol) was dissolved in acetonitrile (10 mL), chloroacetonitrile (0.9 g, 1.2 mmol), triethylamine (121 mg, 1.2 mmol) After stirring for 1 hour, the reaction mixture was concentrated under reduced pressure. Toluene (20 ml) was evaporated. The yield was 90.8%.

MS(M+l)=559.32o 1H-NMR (DMSO-J ₆ ppm): δ 10.10 (s, IH); 9.33 (s, IH); 8.96 (s, lH); 8.77 (d, J) =4.8 Hz, IH); 8.48 (d, J=5.2 Hz, IH); 8.43 (d, J=8.4 Hz, IH); 8.11(s,lH); 7.78(s,lH); 7.74(d, J = 8.8 Hz, IH); 7.52 (dd, J = 8.0 Hz, 4.8 Hz, IH); 7.47 (d, J = 8.2 Hz, IH); 7.44 (d, J = 5.2 Hz, IH); 7.28 (d, J = 7.6 Hz, IH); 7.15 (d, J = 8.8 Hz, IH); 4.33 (t, J = 6.8 Hz, IH); 3.48 (s, 2H); 2.3-3. l(m, 10H); 2.18(s, 3H); 2.14(m, 2H); 1.84 (m, 2H).

[0060]

Example 6 Preparation of Compound 2 and Its Pharmaceutically Acceptable Salt

 (i) N-{4-methyl-3-[(4-pyrimidin-5yl)pyrimidin-2-amino]phenyl}-5-(4-methylpiperazin-1-yl)-5,6 , 7,8-tetrahydronaphthalene-2-amide (Compound 2),

 Compound 2

 Step A: Synthesis of 5-Acetylpyrimidine

Dissolve 1-(pyrimidin-5-yl)ethanol (2.48 g, 20 mmol) in 50 ml of dichloromethane, add active manganese dioxide (3 g), react at room temperature for 3 hours, filter, and decompress the filtrate. Separation by column chromatography gave the product 1 g, yield 45%, MS (M+1) = 123.11.

Step B _: Synthesis of (2E)-3-(dimethylamino)succinimide (pyrimidin-5-yl)prop-2-enionone

5-Acetylpyrimidine (1 g, 8.2 mmol) and hydrazine, hydrazine-dimethylformamide dimethyl acetal (1.3 g, 11 mmol) were dissolved in 20 mL of isopropanol and stirred under reflux 24 hour. After cooling to room temperature, the solution was concentrated under reduced pressure. The residue was added to diethyl ether. After cooling in an ice water bath for several hours, the solid was collected by filtration and washed with cold diethyl ether. The product was dried, 1.05 g, yield 61.9%, MS (M+1) = 178.34.

将 (2E)-3- (二甲氨基) -1- (嘧啶 -5-基)丙 -2-烯 -1-酮 (1克， 5.6毫摩尔)和 N-(2-甲基 -5-硝基苯)胍硝 酸盐 (1.44克， 5.6毫摩尔)悬浮在 20毫升异丙醇中。 加入氢氧化钠 (0.28克， 7毫摩尔)。 混合 液在回流搅拌 8小时冷却到室温。 过滤收集固体， 并用异丙醇和乙醚洗涤。 过滤收集固体， 真空干燥后得到产物 1.26克， 产率 73.5%， MS(M+l)=309.21 o Step C _: Synthesis of N-(2-methyl-5--4,5'-dipyrimidin-2-amine

(2E)-3-(Dimethylamino)-1-(pyrimidin-5-yl)prop-2-en-1-one (1 g, 5.6 mmol) and N-(2-methyl-5- Nitrobenzene) hydrazine nitrate (1.44 g, 5.6 mmol) was suspended in 20 ml of isopropanol. Sodium hydroxide (0.28 g, 7 mmol) was added. The mixture was stirred at reflux for 8 hours and cooled to room temperature. The solid was collected by filtration and washed with isopropyl alcohol and diethyl ether. The solid was collected by filtration and dried in vacuo to yield 1.26 g,yield: 73.5%, MS (M+l) = 309.21.

Step D: Synthesis of !^-([4,5-dipyrimidin-2-yl)-6-methylbenzene-1,3-diamine

Dissolve stannous chloride dihydrate (3.6 g, 16 mmol) in 10 ml of concentrated hydrochloric acid and add this solution to N-(2-methyl-5-nitrophenyl)-4 with vigorous stirring. , 5 dipyrimidin-2-amine, after stirring for 2 hours, the mixture was poured into ice water, and the mixture was neutralized with potassium carbonate to pH = 8, and then extracted with ethyl acetate three times, and the combined extraction was carried out. The solution was washed with brine, dried over anhydrous sodium sulfate]]]]]] MS (M+1) = 279.33.

Step E: N-{4-methyl-3-[(4-pyrimidin-5yl)pyrimidin-2-amino]phenyl}-5-(4-methylpiperazin-1-yl)- Synthesis of 5,6,7,8-tetrahydronaphthalene-2-amide (Compound 2)

5-(4-Methylpiperazine small)-5,6,7,8-tetrahydronaphthalene-2-carboxylic acid (1.37 g, 5 mmol), ! ^-([4,5-Dipyrimidin]-2-yl)-6-methylbenzene-1,3-diamine (1.53 g, 5.5 mmol), EDCI (1-ethyl-3-(3-di) Methylaminopropyl)carbodiimide) (1.15 g, 6 mmol), DMAP (4,4-methylaminopyridine) (0.31 g, 2.5 mmol) was added to 30 ml of tetrahydrofuran and 10 ml of DMF mixture. After reacting for 4 hours at room temperature, the reaction solution was evaporated under reduced pressure to tetrahydrofuran, and ethyl acetate 50 was added. ML, and washed with brine 3 times (60, 60, 30 ml), the ethyl acetate phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was evaporated under reduced pressure. The target product was isolated under the mobile phase of formazan/1% triethylamine, ie, Compound 2, 1.57 g, yield 58.8%.

MS (M+1) = 535.54. 1H-NMR (DMSO-J ₆ ppm): δ 10.04 (s, IH); 9.45 (s, 2H); 9.32 (s, lH); 9.11 (s, lH); 8.57 (d, J = 6.0 Hz, IH 8.14(s, IH); 7.75(s, IH); 7.73(d, J=9.0 Hz, IH); 7.49(d, J=6.0 Hz, IH); 7.45(d, J=9.0 Hz, IH 7.34(d, J=9.0 Hz, IH); 7.17(d, J=9.0 Hz, IH); 4.31(t, J=6.0 Hz, IH); 2.3-3.0(m, 10H); 2.16(s , 3H); 2.08(s, 3H); 1.98(m, 2H); 1.88 (m, 2H).

 (b) N-{4-methyl-3-[(4-pyrimidin-5yl)pyrimidin-2-amino]phenyl}-5-(4-methylpiperazine small group)-5, Synthesis of 6,7,8-tetrahydronaphthalene-2-amide mesylate (amine phenylinibine)

Its structural formula is as follows:

The reaction flask was added to the compound 2 (5.34 g, 10 mmol), anhydrous methanol 100 ml, methanesulfonic acid (1.01 g, 10.5 mmol), and refluxed at 70 ° C for 1 hour, then added pharmaceutically decolorized charcoal 2 g After refluxing for an additional 1 hour, it was suction filtered, and the filtrate was concentrated under reduced pressure. Filtration and drying to give the product N-{4-methyl-3-[(4-pyridin-3-yl)pyrimidin-2-amino]phenyl}-5-(4-methylpiperazin-1-yl)- 5,6,7,8-tetrahydronaphthalene-2-carboxamide methanesulfonate 5.72 g, yield 90.8%. Melting point: 201~203 °C. Elemental analysis: C ₃₂ H ₃₈ N ₈ 0 ₄ calc.

[0068]

Example 7 N-{ 4-methyl-3-[(4-pyridin-3-yl)pyrimidin-2-amino]phenyl}-5-(3-methylimidazolium small group) -5,6,7 Synthesis of 8-tetrahydronaphthalene-2-amide

Its structural formula is as follows:

Step A: Synthesis of 1-methylimidazolidine

 I

N-methylethylenediamine (1 g, 13.5 mmol) was added to formaldehyde (0.4 g, 13.5 mmol), potassium carbonate (6.4 g, 47.2 mmol), and magnesium sulfate (5.6 g, 47.2 mmol). The mixture was stirred at room temperature for 18 hours, filtered, and the filtrate was concentrated under reduced pressure. The purified product was purified by methylene chloride column: methylene chloride: methanol = 9:1 to give the product 1-methylimidazolidine. , yield 70%, MS (M+l) = 87.11 o

 Step B: Synthesis of methyl 5-(3-methylimidazolidine-1-yl)-5,6,7,8-tetrahydronaphthalene-2-carboxylate

Methyl 5-chloro-5,6,7,8-tetrahydronaphthalene-2-carboxylate (2.41 g, 10 mmol) was dissolved in 20 mL of DMF solvent and potassium carbonate (2.78 g, 20 mmol) , 1-methylimidazolidine (1.72 g, 20 mmol), after reacting at 50 ° C for 5 hours, the reaction solution was added to ethyl acetate (80 ml), pH was adjusted to neutral with dilute aqueous hydrochloric acid, and acetic acid was separated. The ethyl ester phase was washed with saturated brine (3×80 ml), dried over anhydrous sodium sulfate, filtered and evaporated. 2.03 g, yield 74.1%, MS (M+1) = 275.28.

Step C: Synthesis of 5-(3-methylimidazolidine-1-yl)-5,6,7,8-tetrahydronaphthalene-2-carboxylic acid

Dissolve methyl 5-(3-methylimidazol-1-yl)-5,6,7,8-tetrahydronaphthalene-2-carboxylate (2.6 g, 10 mmol) in 30 mL of methanol and 20 mL In a solution of tetrahydrofuran, 2N NaOH (10 ml, 20 mmol) was added, and the reaction was carried out for 10 hours at room temperature. After adjusting pH = 3 to 4 with 2N aqueous hydrochloric acid solution, the organic solvent was evaporated under reduced pressure and ethyl acetate (60 ml) The mixture was washed with saturated brine (3×60 mL), EtOAc EtOAcjjjjjjjjjj : 2.21 g of the product was isolated under the conditions of 5 drops, the yield was 85.0%.

 Step D: Synthesis of 5-(3-methylimidazolidine-1-yl)-1,2,3,4-tetrahydronaphthalene-5-acyl chloride hydrochloride

SOC1 ₂ (200 ml) was added to the reaction flask, and 5-(3-methylimidazolidine-1-yl)-5,6,7,8-tetrahydronaphthalene-2-carboxylic acid (26 g) was added. After the hour, the temperature was lowered to room temperature, and S0C1 _{2 was} evaporated under reduced pressure. Toluene solution (50 ml) was then added. After cooling to 0 ° C, a large amount of white solid was precipitated, filtered, and the solid was dried to give a product (about 34.8 g, yield 99.5) %).

Step E: N-{4-Methyl-3-[(4-pyridin-3-yl)pyrimidin-2-amino]-phenyl}-5-(4-methylpiperazine small group)-5 Synthesis of 6,6,8-tetrahydronaphthalene-2-amide

Dissolve 6-methyl-!^-[(4-pyridin-3-yl)pyrimidin-2-yl]benzene-1,3-diamine (2.77 g, 10 mmol) in pyridine (30 mL) in. 5-(3-Methylimidazolidine-1-yl)-1,2,3,4-tetrahydronaphthalen-5-acyl chloride hydrochloride (2.78 g, 10) was added portionwise with stirring at 0~5 °C. Millimoles). Add in about 30 minutes. Stir to room temperature and stir for 4 hours. The reaction mixture was poured into aq. After stirring for a while, the dichloromethane phase was separated and the aqueous extracted with dichloromethane (2×50 mL). The combined organic layers were dried with EtOAc EtOAc The residue was purified by silica gel column chromatography eluting elut elut elut elut elut eluting

δ 9.99 (s, 1H); 9.11 (s, 1H); 8.85(s,lH); 8.66(d, J=4.8 Hz, 1H); 8.52(d, J=4.8 Hz, 1H); 8.47(d, J=9.0 Hz, 1H); 8.02(s,lH); 7.74(s,lH); 7.72(d, J=9.0 Hz, 1H); 7.43(dd, J=8.0 Hz, 4.8 Hz, 1H); 7.38(d, J=8.0 Hz, 1H); 7.34(d, J=4.8 Hz, 1H); 7.27(d, J=9.0 Hz, 1H); 7.16(d, J=9.0 Hz, 1H); 4.22(t, J=9.0 Hz, 1H); 3.46 (s, 2H); 2.2-2.7(m, 6H); 2.11(s, 3H); 2.03(s, 3H); 2.0(m,2H); 1.86(m, 2H)。 [0074]

δ 9.99 (s, 1H); 9.11 (s, 1H); 8.85 (s, lH); 8.66 (d, J = 4.8 Hz, 1H); 8.52 (d, J = 4.8 Hz, 1H); 8.47 (d, J=9.0 Hz, 1H); 8.02(s,lH); 7.74(s,lH); 7.72(d, J=9.0 Hz, 1H); 7.43(dd, J=8.0 Hz, 4.8 Hz, 1H); 7.38 (d, J = 8.0 Hz, 1H); 7.34 (d, J = 4.8 Hz, 1H); 7.27 (d, J = 9.0 Hz, 1H); 7.16 (d, J = 9.0 Hz, 1H); 4.22 (t , J=9.0 Hz, 1H); 3.46 (s, 2H); 2.2-2.7(m, 6H); 2.11(s, 3H); 2.03(s, 3H); 2.0(m,2H); 1.86(m, 2H).

[0075]

Example 8 Pharmacodynamic Test of Cyclobutinib and Amphetamine

The benzophenidine and ethamidine provided in Example 1 and Example 6 were test samples, and showed excellent antitumor effects as shown by the following pharmacodynamic tests.

(1) Method for measuring growth inhibitory activity (GI _5Q ) for various cancer cells:

The tumor cells were trypsinized, dispersed into individual cells, and suspended in RPMI1640 medium containing penicillin (25 U/ml) and streptomycin (25 g/ml). The cells were seeded in a 96-well culture plate (Corning Incorporated), cultured at 37 V, air containing 5% CO ₂ at a relative humidity of 100% for 24 hours, and the culture solution was discarded, and a series of test samples were added. The culture medium is set to parallel holes at each concentration. After culturing for 24 hours, the culture solution containing the test sample is discarded, and after adding the conventional culture solution for 48 hours, the culture solution is discarded, and then the thiazole blue (MTT) is replaced. , American Sigma product) culture solution, MTT final concentration is 0.5g / L, continue to incubate for 4 hours, add dimethyl sulfoxide solution, 1 hour after the purple crystal is completely dissolved, in SK601 type microplate reader (Japan Seikagaku's product) detects the optical density (OD) at 570 nm / 630 nm. The half-growth inhibition rate of the test sample on the tumor was calculated as follows:

 (T-To) I (C-To) X 100

Note: C indicates the OD value of the control cells.

 T represents the OD value of the cells in the test sample group

T ₀ represents the OD value of the control plate cells when the test sample is added

 The inhibitory effect of the test sample on various cancer cells is shown in Table 1.

[0077] Inhibition of tumor cells by compound 1 and compound 2

Type of cancer Cancer name

 Compound 1 Compound 2 Lung cancer A-549 0. 6 0. 8

 HOP- 62 0. 3 6. 2

HOP- 92 5. 8 9. 2

NCI-H460 11. 7 26. 2 Liver cancer SMMC-7721 〉30 〉30 Leukemia CCRF-CEM 0. 03 0. 08

 HL-60 (TB) 0. 2 0. 9

MOLT- 4 5. 1 6. 3

RPMI-8226 3. 7 2. 8

SR 0. 8 0. 7

K-502 5. 6 4. 4

K-562 0. 9 0. 7

P388 4. 6 5. 6 Breast cancer T-47D 〉30 〉30

 HS578T 〉30 〉30 Colon cancer HT-29 11. 6 17. 2

 HCT-15 8. 7 10. 6

KM12 28. 6 〉30 Ovarian cancer OVCAR-3 0. 5 0. 8

 OVCAR-4 0. 6 1. 4

MKN-45 〉30 〉30 Neurocarcinoma SF-295 〉30 〉30

 SF-593 〉30 〉30

SNB-75 〉30 〉30

¹ disease A498 23. 2 12. 6

ACHN 7. 8 5. 8 TK-10 11. 3 19. 6

(2) Inhibition of human cancer-bearing mouse leukemia K-562

The tumor tissue in the vigorous growth period was cut into 1.5 mm ³ and inoculated subcutaneously in the right axilla of the nude mice under aseptic conditions. The nude mice xenografts were measured for the diameter of the transplanted tumor using a vernier caliper, and the animals were randomly divided into groups after the tumors were grown to 100 to 300 mm ³ . The method of measuring the tumor diameter is used to dynamically observe the antitumor effect of the test substance. The number of tumor diameters measured twice a week, and each mouse was weighed at the same time. The experimental group was administered intravenously three times a week, and the positive control group was administered intravenously three times a week. The negative control group was given the same amount of physiological saline at the same time. The formula for calculating the tumor volume (TV) is:

TV=1 I 2 X aX b ²

Where a and b represent the length and width, respectively. Based on the measured results, the relative tumor volume (RTV) was calculated and calculated as: RTV=V _t /V _Q . Where V _Q is the tumor volume measured at the time of sub-cage administration (g卩d _Q ), and 1⁄4 is the tumor volume at each measurement. The evaluation index of antitumor activity is the relative tumor growth rate T / C (%), and the calculation formula is as follows:

 TRTV

 T / C ( ) = X 100

TRTV: experimental group RTV; C _rtv : negative control group RTV.

 [0078] The inhibitory effect of the test sample on human leukemia K-562, the results are shown in Table 2.

 Table 2 Inhibition of X-ray xenografted on human leukemia K-562 in nude mice by Compound 1 and Compound 2

 Compound 1 compound 2 imatinib

U quantity (mg · kg" ¹ ) 80 100 150

 Relative tumor growth rate T/C (%) 2. 6 10. 8 22. 3

(3) Inhibition of human cancer-bearing mouse leukemia P388

Under sterile conditions, the P388 leukemia ascites inoculated in mice for 7 days was diluted 1:15 to adjust the number of leukemia cells to 1 X 10 ⁷ /mL, 0.2 mL per mouse, and the next day was randomly divided into 6 groups. Each of the compound 1 and compound 2 groups was administered intraperitoneally once a day for 10 days, and the imatinib group was administered intraperitoneally once every other day for 5 times. The control group was given the same volume of normal saline. Animal deaths were recorded daily for 30 days over 30 days. Calculate the life extension rate according to the formula: Life extension rate (%) = (average survival days of the drug group - average survival days of the control group) / average survival days of the control group X 100%.

 [0080] The inhibitory effect of the test sample on human leukemia P388, the results are shown in Table 3.

 Table 3 Inhibition of xenonibine and imifenib on xenografted human leukemia P388 in nude mice

 Compound 1 compound 2 imatinib

U quantity (mg · kg" ¹ ) 80 100 150

 Life extension rate (%) 80. 3% 78. 4% 66. 7% The compound 1 and the compound 2 provided by the present invention are more excellent as shown by the above pharmacological test results as compared with the control compound imatinib. Inhibition of tumors.

 [0082]

Example 9 Safety Test of Cyclobutinib and Ambrotinib (Acute Toxicity)

The results of administration of circophentinib and etimicin in the tail vein of mice showed that both cimetidine and etimicin were toxic to the circulatory system, and the animals in the high-dose group were shocked 0.5 to 2 hours after administration. Jumping, then you can see the lack of energy, body tremor, less moving part of the mouse draining feces, there is light yellow thin feces pollution around the anus. The death occurred within 5 to 12 hours after administration. The dead mice were dissected, and the intestinal swell of the mice was visually observed. The contents of the water samples were cut open, the liver became white, and the other organs showed no abnormalities. Liver histopathology Examination showed hepatic cell spot necrosis. Surviving mice in each group were observed for 14 consecutive days. No abnormalities were observed in feeding, drinking water, general state and activity. After the observation period, the mice were sacrificed, and no abnormalities were observed in the liver and main organs by naked eyes.

[0083] Cyclobutinib is toxic to the respiratory system and causes respiratory failure in mice after administration. Its LD ₅₀ is 420.6 mg / kg, and the 95 % confidence limit is 340.3 mg / kg ~ 500.9 mg / kg. The toxic target organ is mainly the liver.

[0084] Amphetamine is toxic to the respiratory system and causes respiratory failure in mice after administration. Its LD ₅₀ is 463.5 mg / kg, and the 95 % confidence limit is 370.9 mg / kg ~ 556.1 mg / kg. The toxic target organs are mainly the lungs.

As a result of the above pharmacological test, the compound of the present invention exhibits an excellent antitumor effect, and as an antitumor agent, it is effective for preventing and treating diseases, particularly cancer. When the compound of the present invention is used in such a use, it can be formulated into an effective amount and a pharmaceutically acceptable carrier or excipient containing the compound of the present invention.

The form of administration of the compound of the present invention as an antitumor agent can be selected from various forms, and examples thereof include an oral preparation such as a tablet, a capsule, a powder, a granule or a liquid, or, for example, a solution or suspension. A liquid non-oral preparation, an injection, a suppository, an ointment, or the like which is sterilized by a liquid or the like.

The solid preparation may be directly prepared in the form of a tablet, a capsule, a granule or a powder, but may also be produced using a suitable additive. Examples of such an additive include sugars such as lactose and glucose, and starches such as corn, wheat, and rice, and fatty acids such as stearic acid, such as magnesium metasilicate aluminate or anhydrous calcium phosphate. Inorganic salt, case A synthetic polymer such as polyvinylpyrrolidone or polyalkylene glycol, for example, a fatty acid salt such as calcium stearate or magnesium stearate, or an alcohol such as stearyl alcohol or benzyl alcohol, such as methyl fiber. Synthetic cellulose derivatives such as carboxymethylcellulose, ethylcellulose or hydroxypropylmethylcellulose, and other additives which are generally used, such as gelatin, talc, vegetable oil, gum arabic, and the like.

 The solid preparation of these tablets, capsules, granules, powders and the like may usually contain 0.1 to 99% (w/w), preferably 0.1 to 50% (w/w) of the active ingredient.

 The liquid preparation can be produced in the form of a liquid, an alcohol or a liquid preparation such as soybean oil, peanut oil or sesame oil in a form of a suspension, a syrup, an injection or a drip, using a suitable additive which is usually used.

[0090] Particularly, as a suitable solvent for administration by parenteral injection, intravenous injection or subcutaneous injection, for example, distilled water for injection, physiological saline, aqueous glucose solution, ethanol, polyethylene glycol, or the like may be mentioned. A liquid for intravenous injection (for example, an aqueous solution of citric acid and sodium citrate) or an electrolyte solution (for intravenous and intravenous injection), or the like, or a mixed solution of these.

 In addition to these pre-dissolved injections, it is also possible to form a form which is dissolved at the time of use by adding a powder or a suitable additive. These injection solutions may usually contain 0.1 to 20% (w/w), preferably 0.5 to 5% (w/w) of the active ingredient.

 Further, a dosage form of a suspending agent, a syrup or the like for oral administration may usually contain 0.5 to 10% (w/w) of an active ingredient.

 The preferred amount of the compound of the present invention can be administered depending on the kind of the compound to be used, the type of the compound to be compounded, the frequency of application and the specific site to be treated, the severity of the condition, the age of the patient, the diagnosis of the doctor, and the tumor. The type varies, but as a general target, for example, the dose per one adult per day can be in the range of l to 300 mg at the time of oral administration, and in the case of parenteral administration, at the time of intravenous injection. Preferably, it is in the range of from l to 150 mg per day. In addition, the number of administrations varies depending on the administration method and symptoms, but it is 1 to 3 times a day. Further, an administration method such as intermittent administration such as administration every other day or administration for two days may be used.

Claims

1. The expression represented by the general formula [ I ]

In the formula: Is a saturated cyclic amino group; the saturated cyclic amino group is selected from piperidinyl, piperazinyl, imidazolidinyl, pyrrolidinyl, azacyclobutane or morpholinyl, or each 1, 2 3 or 4 substituents, which are a hydrogen atom, a halogen, an amino group, a cyano group, a C ₆ alkyl group, a d- ₆ hydroxyalkyl group, a d- ₆ halogenated alkyl group or a d- ₆ cyanoalkyl group; Pyridyl, pyrimidinyl, pyrazinyl, triazinyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, isoxazolyl, triazolyl, or pyrazolyl, or optional for each group substituted with 1, 2, 3, or R ₂ is substituted, the R ₂ is independently selected from halo, cyano, amino, C ^ ₆ alkyl, d- ₆ hydroxyalkyl, d- ₆ haloalkyl, or — ₆ cyanoalkyl; E is selected from pyridyl, pyrimidinyl, pyrazinyl, triazinyl, thiazolyl, isothiazolyl, imidazolyl, oxazolyl, isoxazolyl, triazolyl, pyrazolyl, oxazolyl, Pyrrolopyridinyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl, quinolinyl, isoquinolyl, quinazolinyl, piperazinyl or morpholinyl, or each group optionally being 1, 2 or substituted with three R _3, said R ₃ is independently selected from halo, cyano, amino, C ^ ₆ alkyl, d- ₆ hydroxyalkyl, d- ₆ haloalkyl, or - ₆ alkoxy cyano base.  2. An acceptable salt according to claim 1, characterized by having the structure of the formula [ Π ]:

In the formula: R4 and R _{5 are} independently selected from hydrogen, cyano, amino, d- ₆ alkyl, d- ₆ hydroxyalkyl, d- ₆ haloalkyl, cyanoalkyl, or d- _6; R ₆ is independently selected from hydrogen atoms, C ^ ₆ alkyl, C ₂ - ₆ hydroxyalkyl, C ₂ - ₆ haloalkyl, d- ₆ cyanoalkyl, acyl and carboxylate; p is 0, 1, or 2.  The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein D is pyrimidinyl and E is pyridyl, having the formula [III]:

 [III] The compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein R 4 and R ₅ are a hydrogen atom, R ₆ is a methyl group, and p is 1, and the knot thereof is:

The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein D is pyrimidinyl and E is pyrimidinyl, having the formula [IV]:

 [IV] 6. The compound of claim 5 and a pharmaceutically acceptable salt thereof as claimed in claim, wherein, R4 and R ₅ is a hydrogen atom, R ₆ is a methyl group, p is 1, and its knot:

The use of the compound according to claim 1, 2, 3, 4, 5 or 6 and a pharmaceutically acceptable salt thereof as an active ingredient for the preparation of an antitumor drug.  A pharmaceutical composition comprising a compound according to any one of claims 1 to 6 and a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.