CN105418615B - Heterocyclic carbamate derivatives and preparation and application - Google Patents

Abstract

The benzamide derivative and its preparation and application belong to the field of medicinal chemistry. The present invention relates to N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl) benzamide derivative represented by formula and preparation and application. R ¹ means -OCH ₃ , -F, -CF ₃ , -CH ₂ CH ₃ , -H, R ² represents -CF ₃ , -F, -Cl or -H. The preparation is to prepare 2-methyl-5-nitrophenylhydrazine, and then react to obtain 4-bromo-1-(2-methyl-5-nitrophenyl) pyrazole, then reduce, Boc anhydride protection, and Reaction of boronic acid pinacol ester, followed by coupling to generate N-Boc-3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline , and finally de-Boc and aminolysis. The compounds of the present invention have inhibitory effect on Bcr-Abl kinase.

Description

Benzamide derivatives and their preparation and application

technical field

The invention belongs to the field of medicinal chemistry, and relates to benzamide derivatives, preparation and application, in particular to N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl) )-4-methylphenyl)benzamide derivative and its preparation method and application as Bcr-Abl tyrosine kinase inhibitor.

Background technique

Chronic myelogenous leukemia (CML) is a malignant bone marrow hematopoietic stem cell clonal proliferative disease, 95% of patients have characteristic Ph chromosome and its molecular marker Bcr-Abl fusion gene. Studies have shown that the fusion gene Bcr-Abl is a molecular biological feature of chronic myelogenous leukemia, and its product Bcr-Abl kinase is a key factor in the pathogenesis.

Currently, the most commonly used small molecule inhibitors (TKIs) against Bcr-Abl tyrosine kinase include: the first-generation drug imatinib; the second-generation drugs dasatinib, nilotinib and bosutinib Ni; the third-generation drug Ponatinib. Imatinib is an oral anti-chronic myelogenous leukemia small molecule tyrosine kinase inhibitor developed by Swiss Novartis. It has created a new era of targeting kinases to treat diseases. Imatinib can bind to the ATP site of the Bcr-Abl kinase region to prevent phosphorylation of amino acid residues, thereby blocking signal transduction pathways, inhibiting cell proliferation, and can effectively relieve CML. After treatment, the 5-year survival rate of patients can reach 90%, and it can specifically act on CML cancer cells, and has almost no damage to normal cells, and the toxic and side effects are greatly reduced. With the use of Imatinib, mutations in the Bcr-Abl gene lead to the emergence of drug resistance, which greatly reduces the efficacy of Imatinib. Dasatinib is a multi-targeted oral kinase inhibitor targeting Bcr-Abl tyrosine kinase, Src kinase family (Src, Lck, Fyn), c-Kit and PDGFR-B, etc. The drug was developed by Bristol-Myers Squibb on June 28, 2006 and was approved by the FDA for marketing. Like Imatinib, it competitively binds to the ATP site of the Bcr-Abl kinase region, but the activity of inhibiting Bcr-Abl kinase is 300 times that of Imatinib. Although it can overcome the resistance of multiple Imatinib, it is ineffective against Bcr-Abl. Nilotinib (Nilotinib), a novel aniline derivative of Imatinib, was approved by the US FDA on October 29, 2007. Its affinity for Bcr-Abl kinase is 20 times stronger than that of Imatinib, and it has broad activity against patients with Imatinib resistance (except T315I mutation). Most patients using Nilotinib for CML will have common adverse reactions such as gastrointestinal reactions, bone marrow suppression, and hyperbilirubinemia. Aiming at the drug resistance caused by Imatinib, Dasatinib and Nilotinib, Wyeth Pharmaceutical Company of the United States developed 4-substituted aniline-3-quinoline plus nitrile drug Bosutinib for the treatment of CML, which was released in September 2012. It was approved by the FDA on March 4. Bosutinib can not only inhibit the autophosphorylation of Scr protein in various human tumor cells, but also inhibit the autophosphorylation of Bcr-Abl protein, and in the anti-proliferation test against KU812 and K562 cells (CML cells containing Bcr-Abl) , the activity of Bosutinib is significantly higher than that of Imatinib, and the oral bioavailability of Bosutinib is higher. However, patients receiving Bosutinib treatment often experience adverse reactions such as abdominal pain, diarrhea, thrombocytopenia, fever and fatigue. Ponatinib is an oral multi-target inhibitor that can act on Abl, PDGFRα, VEGFR2, FGFR1 and Src kinases. Due to its unique mechanism of action, taking it can inhibit the activity of Bcr-Abl kinase including T315I mutation. However, according to the latest FDA report, patients taking Ponatinib will have serious vascular problems, so the safety evaluation of the drug is expected.

Imatinib and Ponatinib have shown very good curative effects in the treatment of CML, but they are both expensive. According to statistics, patients taking Imatinib and Ponatinib need to spend 92,000 and 138,000 dollars per year, respectively. Therefore, it is imperative to seek low-cost alternatives to Imatinib and Ponatinib.

Contents of the invention

The object of the present invention is to provide N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)benzamide derivatives Its preparation method and application as Bcr-Abl tyrosine kinase inhibitor.

The present invention provides N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)benzene represented by formula (I) formamide derivatives,

R ¹ represents -OCH ₃ , -F, -CF ₃ , -CH ₂ CH ₃ , -H,

R ² represents -CF ₃ , -F, -Cl or -H.

All the compounds of the present invention have inhibitory effect on Bcr-Abl tyrosine kinase, and the inhibitory effect of some compounds is remarkable.

The present invention also provides N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)benzamide derivatives The preparation method comprises the following steps:

(a) Dissolving 2-methyl-5-nitroaniline in ethanol, adding 37% concentrated hydrochloric acid, placing the above system under ice bath conditions, then adding the aqueous solution of sodium nitrite dropwise to the above system, Preferably, the reaction temperature is 0-5°C, and the reaction time is 0.5-1 hour to obtain 2-methyl-5-nitrophenylhydrazine hydrochloride represented by formula (II);

(b) tin protochloride dihydrate is dissolved in concentrated hydrochloric acid (mass percentage concentration 37%), is placed under ice-bath condition, then 2-methyl-5-nitrophenylhydrazine hydrochloride is added dropwise to In the above-mentioned system, preferred formula (II) compound and the substance ratio of stannous chloride dihydrate are 1: (1.5～2), reaction temperature 0～10 ℃, reaction time 1～2 hour, add NaOH solution (such as Concentration 1mol/L) and, obtain the 2-methyl-5-nitrophenylhydrazine represented by formula (III);

(c) reacting the compound of formula (III) and 2-bromomalonaldehyde in ethanol, the preferred formula (III) compound and the ratio of substances of 2-bromomalonaldehyde are 1: (1.1～1.5), the temperature of reaction 55-65°C, the reaction time is 2-4 hours, and 4-bromo-1-(2-methyl-5-nitrophenyl)pyrazole represented by formula (IV) is obtained;

(d) dissolving the compound of formula (IV) in ethanol, adding an aqueous solution of ammonium chloride and iron powder, preferably at a reaction temperature of 80 to 90° C., and a reaction time of 2 to 3 hours to obtain 3-(4) represented by formula (V). -Bromo-1-pyrazolyl)-4-methylaniline;

(e) reacting the compound of formula (V) and Boc anhydride in tetrahydrofuran, preferably the molar ratio of the compound of formula (V) and Boc anhydride is 1: (1.1～1.4), the reaction temperature is 66～70°C, and the reaction time is 2 ~3 hours, N-Boc-3-(4-bromo-1-pyrazolyl)-4-methylaniline represented by formula (VI) was obtained;

(f) Dissolve the compound of formula (VI) in dioxane, add biboronic acid pinacol ester, potassium acetate and two (triphenylphosphine) palladium dichloride, preferred formula (VI) compound, biboronic acid pinacol The molar ratio of the alcohol ester, potassium acetate and bis(triphenylphosphine)palladium dichloride is 1:(1.1~1.5):3:(0.1~0.2), the reaction temperature is 100~110°C, and the reaction time is 15 After ~18 hours, N-Boc-3-(4-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl)) represented by formula (VII) was obtained -1-pyrazolyl)-4-methylaniline;

(g) reacting imidazo[1,2-b]pyridazine, NBS and a catalytic amount of AIBN in chloroform, preferably with a molar ratio of imidazo[1,2-b]pyridazine, NBS and AIBN being 1: (1.5～2): (0.1～0.2), the reaction temperature is 65°C, and the reaction time is 2～3 hours to obtain 3-bromoimidazo[1,2-b]pyridazine represented by formula (VIII);

(h) Formula (VII) compound and formula (VIII) compound are dissolved in the mixed solution of dioxane and water, add tetrakis (triphenylphosphine) palladium and sodium carbonate, preferred formula (VII) compound, formula ( VIII) The molar ratio of compound, sodium carbonate and four (triphenylphosphine) palladium is 1: 2: 3: (0.1～0.2), 110～120 ℃ of temperature of reaction, 7～10 hours of reaction times, obtain formula ( IX) N-Boc-3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline;

(i) react formula (IX) and trifluoroacetic acid in dichloromethane, preferably reaction time 5～6 hours, obtain 3-(4-(3-imidazo[1,2-b] represented by formula (X) ]pyridazinyl)-1-pyrazolyl)-4-methylaniline;

(j) Reaction of formula (X) compound, 4-chloromethylbenzoyl chloride and triethylamine in dichloromethane, preferred formula (X) compound, 4-chloromethylbenzoyl chloride and triethylamine The amount ratio is 1: (2～3): (0.2～0.5), and the reaction time is 2～3 hours, obtains the 4-chloromethyl-N-(3-(4-(3-imidazole) represented by formula (XI) [1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)benzamide;

(K) Reaction of formula (XI) compound, nucleophile and potassium iodide in DMF, preferred formula (XI) compound, nucleophile and potassium iodide are 1: (2～3): (0.1～0.2 ), reaction temperature, 80～90 ℃, reaction time 2～3 hours, obtain N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1 represented by formula (I) -pyrazolyl)-4-methylphenyl)benzamide derivatives;

R ¹ means

The nucleophile is:

Or (1) formula (X) compound, substituted benzoyl chloride and triethylamine are reacted in methylene chloride, and the ratio of substance of preferred formula (X) compound, substituted benzoyl chloride and triethylamine is 1:( 2～3): (0.2～0.5), reaction time 2～3 hours, obtain the N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)- 1-pyrazolyl)-4-methylphenyl)benzamide derivatives;

R ¹ represents -OCH ₃ , -F, -CF ₃ , -CH ₂ CH ₃ or -H, and R ² represents -CF ₃ , -F, -Cl or -H.

Substituted benzoyl chloride structural formula is:

The method of the invention uses common industrial reagents and conventional production conditions, and the reaction conditions are mild and the steps are simple.

The chemical reaction formula in the above-mentioned synthetic process is as follows:

Wherein, R ¹ represents -OCH ₃ , -F, -CF ₃ , -CH ₂ CH ₃ or -H, and further represents

R ² represents -CF ₃ , -F, -Cl or -H.

The method of the present invention is at first with 2-methyl-5-nitroaniline as raw material, generates 2-methyl-5-nitrophenylhydrazine through diazotization and reduction reaction, carries out condensation relation with 2-bromomalonaldehyde then Ring, to obtain 4-bromo-1-(2-methyl-5-nitrophenyl) pyrazole intermediate, then reduced, Boc anhydride protection, palladium catalyzed and biboronic acid pinacol ester reaction, to obtain N- Boc-3-(4-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl))-1-pyrazolyl)-4-methylaniline , followed by palladium-catalyzed coupling with 3-bromoimidazo[1,2-b]pyridazine to generate N-Boc-3-(4-(3-imidazo[1,2-b]pyridazinyl)-1 -pyrazolyl)-4-methylaniline intermediate, and finally this intermediate undergoes de-Boc and ammonolysis under alkaline conditions to generate the compound of formula (I). The compound of the present invention has inhibitory effect on Bcr-Abl kinase.

Detailed ways

The present invention will be further described below in conjunction with the examples, but the present invention is not limited to the following examples.

Example 1

N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-3-trifluoromethylbenzamide preparation

(a) Preparation of 2-methyl-5-nitrophenylhydrazine hydrochloride

The raw material 2-methyl-5-nitroaniline (6.0g, 39.4mmol) was dissolved in a three-necked round-bottomed flask containing 250mL of ethanol, and 10mL of 37% concentrated hydrochloric acid was added, and the above system was placed in an ice bath, and then Sodium nitrite (2.8g, 40.6mmol) was dissolved in 15mL of distilled water, and slowly added to the above reaction system with a constant pressure dropping funnel, and the temperature was controlled at 0-5°C during the dropping process. The system continued to react under ice-bath conditions for 0.5-1 hour, and the reaction progress was detected by TLC. After the raw materials disappeared, the reaction was stopped, and the next step was directly carried out.

(b) Preparation of 2-methyl-5-nitrophenylhydrazine

14mL of 37% concentrated hydrochloric acid was added to a three-necked flask, 20mL of distilled water was added thereto, and stannous chloride dihydrate (18.0g, 79.6mmol) was weighed and added to the flask, the above system was placed in an ice bath, and then Slowly add the 2-methyl-5-nitrophenylhydrazine hydrochloride obtained in step (a) into the above reaction system through a constant pressure dropping funnel, and control the temperature at 0-10°C during the dropping process. , continue to react the reaction system under ice-bath conditions for 1 to 2 hours, TLC to detect the reaction progress, stop the reaction after the disappearance of the raw materials. Add 1mol/L NaOH solution to neutralize, distill under reduced pressure to remove part of the solvent, add 50mL of water, extract with 50mL×3 ethyl acetate, combine organic phases, dry over anhydrous magnesium sulfate, filter, desolvate under reduced pressure, ethyl acetate Recrystallization gave a yellow solid product.

The pure product is a yellow solid, two-step combined yield: 65%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 7.84(d, J=2.4Hz, 1H), 7.37(dd, J=8.1, 2.4Hz, 1H), 7.16(d, J=8.1Hz, 1H ),6.88(s,1H),4.22(s,2H),2.13(s,3H).

(c) Preparation of 4-bromo-1-(2-methyl-5-nitrophenyl)pyrazole

Dissolve 2-methyl-5-nitrophenylhydrazine (12.0g, 71.9mmol) and 2-bromomalondialdehyde (13.0g, 86.3mmol) in 50mL of ethanol, raise the temperature of the system to 55～65℃, and react for 2～ After 4 hours, TLC was used to detect the progress of the reaction, and the reaction was stopped after the disappearance of the starting material. The system was desolvated under reduced pressure, and the residue was purified by column chromatography (eluent: petroleum ether/ethyl acetate) to obtain a light yellow solid.

The pure product is light yellow solid, yield: 40%

¹ H NMR (CDCl ₃ , 400MHz, δppm): 8.10-8.36(m, 2H), 7.74(d, J=8.2Hz, 2H), 7.53(d, J=8.7Hz, 1H), 2.43(s, 3H ).

ESI-MS m/z:281.4[M+H] ⁺ .

(d) Preparation of 3-(4-bromo-1-pyrazolyl)-4-methylaniline

Dissolve 4-bromo-1-(2-methyl-5-nitrophenyl)pyrazole (3.5 g, 12.5 mmol) in ethanol, then dissolve ammonium chloride (2.67 g, 49.9 mmol) in 6 mL of water , and then add an aqueous solution of ammonium chloride to the above system, the system is heated to 55-60°C, 2.8g of iron powder is added, and the reaction is refluxed for 2-3 hours. The reaction process is detected by TLC, and the reaction is stopped after the disappearance of the raw materials. Filter through celite and concentrate the filtrate under reduced pressure. The residue is purified by column chromatography (eluent: petroleum ether/ethyl acetate) to obtain a light yellow oily liquid.

The pure product is light yellow oily liquid, yield: 80%

ESI-MS m/z:251.3[M+H] ⁺ .

(e) Preparation of N-Boc-3-(4-bromo-1-pyrazolyl)-4-methylaniline

3-(4-Bromo-1-pyrazolyl)-4-methylaniline (3.2g, 12.7mmol) was dissolved in tetrahydrofuran solution, the above system was placed in ice bath, and 3.5mL Boc anhydride was added dropwise Add it to the above system, drop it, and reflux for 2 to 3 hours. TLC detects the reaction process. After the raw materials disappear, the reaction solution is cooled to room temperature, and the solvent is removed under reduced pressure. The residue is subjected to column chromatography (eluent: petroleum ether/ ethyl acetate) to obtain a pale yellow solid.

The pure product is light yellow solid, yield: 90%

¹ H NMR(DMSO-d ₆ ,400MHz,δppm):9.55(s,1H),8.32(s,1H),7.83(s,1H),7.55(s,1H),7.38(d,J=8.3Hz ,1H),7.26(d,J=8.4Hz,1H),2.10(s,3H),1.47(s,9H).

ESI-MS m/z:351.3[M+H] ⁺ .

(f) N-Boc-3-(4-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl))-1-pyrazolyl)- Preparation of 4-methylaniline.

Dissolve N-Boc-3-(4-bromo-1-pyrazolyl)-4-methylaniline (3.0g, 8.5mmol) in 20mL of dioxane, and then add bis(triphenyl Phosphine)palladium dichloride (0.6g, 0.85mmol), potassium acetate (2.5g, 25.5mmol) and pinacol diborate (2.6g, 10.2mmol). Under the protection of argon, heat to reflux for 15-18 hours. TLC detects the progress of the reaction. After the disappearance of the raw materials, the reaction solution is cooled to room temperature, desolvated under reduced pressure, and the residue is subjected to column chromatography (eluent: petroleum ether/ethyl acetate ester) to a pale yellow solid.

The pure product is light yellow solid, yield: 50%

ESI-MS m/z:400.2[M+H] ⁺ .

(g) Preparation of 3-bromoimidazo[1,2-b]pyridazine

Add imidazo[1,2-b]pyridazine (6.0g, 50.4mmol), NBS (6.0g, 75.6mmol) and a catalytic amount of AIBN into 50mL of chloroform, heat to reflux for 2 hours, and detect the reaction progress by TLC , after the disappearance of the raw materials, the reaction solution was cooled to room temperature and stirred in an ice bath, a precipitate was precipitated, filtered and the filtrate was concentrated under reduced pressure to obtain the pure product.

The pure product is light yellow solid, yield: 75%

¹ H NMR (CDCl ₃ , 400MHz, δppm): 8.50(d, J=3.6Hz, 1H), 8.01-7.98(m, 1H), 7.84(s, 1H), 7.15-7.12(m, 1H).

ESI-MS m/z:200.1[M+H] ⁺ .

(h) Preparation of N-Boc-3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline

N-Boc-3-(4-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl))-1-pyrazolyl)-4- Add methylaniline (2.5g, 6.3mmol) into the flask, add 30mL of dioxane, 6mL of water and stir to dissolve, add 3-bromoimidazo[1,2-b]pyridazine (2.5g, 12.6mmol), four (Triphenylphosphine) palladium (0.73g, 0.63mmol) and sodium carbonate (2.67g, 25.2mmol) were stirred evenly, then heated to reflux under the protection of argon, and the reaction process was detected by TLC. After reacting for 7-10 hours, cool to room temperature . The reaction solution was filtered with diatomaceous earth and precipitated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether/ethyl acetate).

The pure product is light yellow solid, yield: 65%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 9.53(s, 1H), 8.69(s, 1H), 8.66(d, J=4.4Hz, 1H), 8.45(s, 1H), 8.21(dd ,J=13.8,6.9Hz,2H),7.66(s,1H),7.42(d,J=8.4Hz,1H),7.31–7.24(m,2H),2.20(s,3H),1.48(s, 8H).

ESI-MS m/z:391.2[M+H] ⁺ .

(i) Preparation of 3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline

Dissolve N-Boc-3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline (1.0 g, 2.56 mmol) in 30 mL di Add 3 mL of trifluoroacetic acid dropwise to the above system in an ice bath in methyl chloride. After the drop is complete, react at room temperature for 5 to 6 hours. TLC detects the reaction progress, and stops the reaction after the disappearance of the raw materials. Part of the solvent was removed by distillation under reduced pressure, the residue was neutralized with saturated aqueous sodium bicarbonate solution, extracted with 30mL×3 dichloromethane, the organic phases were combined, dried over anhydrous magnesium sulfate, filtered, and precipitated under reduced pressure, the residue was passed through the column layer Purify by analysis (eluent: petroleum ether/ethyl acetate).

The pure product is light yellow solid, yield: 85%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 8.66(dd, J=4.4, 1.4Hz, 1H), 8.64(s, 1H), 8.41(s, 1H), 8.21(s, 1H), 8.21 –8.18(m,1H),7.25(dd,J=9.2,4.5Hz,1H),7.04(d,J=8.2Hz,1H),6.66(d,J=2.3Hz,1H),6.60(dd, J＝8.2,2.3Hz,1H),5.23(s,2H),2.09(s,3H).

ESI-MS m/z:291.1[M+H] ⁺ .

(j) N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-3-trifluoromethylbenzene Preparation of formamide

3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline (0.2 g, 0.69 mmol) was dissolved in 5 mL of dichloromethane, Add 50 μL of triethylamine, put the above system in ice bath, then dissolve 3-trifluoromethylbenzoyl chloride (0.43g, 2.07mmol) in 3mL of dichloromethane, and then slowly add it dropwise to the above ice In the bath system, after dropping, continue to react in the ice bath for 2-3 hours, TLC detects the reaction progress, and stops the reaction after the disappearance of the raw materials. The reaction solution was filtered, the insoluble matter was washed with water and dichloromethane respectively, and finally purified by column chromatography (eluent: dichloromethane/methanol) to obtain pure product.

The pure product is yellow-green fluorescent solid, yield: 80%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 8.94(d, J=4.0Hz, 1H), 8.81(s, 1H), 8.51(d, J=10.2Hz, 2H), 8.39(d, J =9.3Hz,1H),8.27–8.20(m,2H),7.95(d,J=7.0Hz,2H),7.76(dd,J=14.6,7.2Hz,2H),7.66(dd,J=9.3, 4.5Hz, 1H), 7.44(d, J=8.4Hz, 1H), 2.23(s, 3H).

HRMS(ESI)m/z:C ₂₄ H ₁₇ F ₃ N ₆ O, calculated 463.1494[M+H] ⁺ ,found 463.1513[M+H] ⁺ .

Example 2

N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4-trifluoromethylbenzamide preparation

Steps (a), (b), (c), (d), (e), (f), (g), (h), (i) are the same as in Example 1.

(j) N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4-trifluoromethylbenzene Preparation of formamide

3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline (0.2 g, 0.69 mmol) was dissolved in 5 mL of dichloromethane, Add 50 μL of triethylamine, put the above system in ice bath, then dissolve 4-trifluoromethylbenzoyl chloride (0.43g, 2.07mmol) in 3mL of dichloromethane, then slowly add it dropwise to the above ice In the bath system, after dropping, continue to react in the ice bath for 2-3 hours, TLC detects the reaction progress, and stops the reaction after the disappearance of the raw materials. The reaction solution was filtered, the insoluble matter was washed with water and dichloromethane respectively, and finally purified by column chromatography (eluent: dichloromethane/methanol) to obtain pure product.

The pure product is yellow-green fluorescent solid, yield: 80%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 8.92(d, J=3.5Hz, 1H), 8.80(s, 1H), 8.50(d, J=2.6Hz, 2H), 8.38(d, J =9.2Hz,1H),8.11(d,J=8.1Hz,2H),7.98(d,J=1.9Hz,1H),7.89(d,J=8.3Hz,2H),7.76–7.71(m,1H ),7.63(dd,J=9.2,4.5Hz,1H),7.44(d,J=8.5Hz,1H),2.23(s,3H).

HRMS(ESI)m/z:C ₂₄ H ₁₇ F ₃ N ₆ O, calculated 463.1494[M+H] ⁺ , found 463.1512[M+H] ⁺ .

Example 3

Preparation of N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4-fluorobenzamide

Steps (a), (b), (c), (d), (e), (f), (g), (h), (i) are the same as in Example 1.

(j) N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4-fluorobenzamide preparation

3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline (0.2 g, 0.69 mmol) was dissolved in 5 mL of dichloromethane, Add 50 μL of triethylamine, place the above system in an ice bath, then dissolve 4-fluorobenzoyl chloride (0.33 g, 2.07 mmol) in 3 mL of dichloromethane, and then slowly add it dropwise to the above ice bath system , After dropping, continue to react in ice bath for 2-3 hours, TLC to detect the reaction progress, stop the reaction after the disappearance of the raw materials. The reaction solution was filtered, the insoluble matter was washed with water and dichloromethane respectively, and finally purified by column chromatography (eluent: dichloromethane/methanol) to obtain pure product.

The pure product is yellow-green fluorescent solid, yield: 80%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 10.50(s, 1H), 8.78(s, 1H), 8.75(d, J=4.0Hz, 1H), 8.50(s, 1H), 8.34(s ,1H),8.27(d,J=9.5Hz,1H),8.08(dd,J=8.5,5.6Hz,2H),8.01(s,1H),7.82(d,J=8.4Hz,1H),7.45 –7.34(m,4H),2.27(s,3H).

HRMS(ESI)m/z:C ₂₃ H ₁₇ FN ₆ O, calculated 413.1526[M+H] ⁺ ,found 413.1541[M+H] ⁺ .

Example 4

Preparation of N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-3-chlorobenzamide

Steps (a), (b), (c), (d), (e), (f), (g), (h), (i) are the same as in Example 1.

(j) N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-3-chlorobenzamide preparation

3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline (0.2 g, 0.69 mmol) was dissolved in 5 mL of dichloromethane, Add 50 μL of triethylamine, place the above system in an ice bath, then dissolve 3-chlorobenzoyl chloride (0.36 g, 2.07 mmol) in 3 mL of dichloromethane, and then slowly add it dropwise to the above ice bath system , After dropping, continue to react in ice bath for 2-3 hours, TLC to detect the reaction progress, stop the reaction after the disappearance of the raw materials. The reaction solution was filtered, the insoluble matter was washed with water and dichloromethane respectively, and finally purified by column chromatography (eluent: dichloromethane/methanol) to obtain pure product.

The pure product is yellow-green fluorescent solid, yield: 80%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 10.55(s, 1H), 8.76(s, 1H), 8.67(d, J=3.4Hz, 1H), 8.49(s, 1H), 8.25(s ,1H),8.20(d,J=9.0Hz,1H),8.04(s,1H),7.98(s,1H),7.94(d,J=7.6Hz,1H),7.82(d,J=8.3Hz ,1H),7.68(d,J=7.5Hz,1H),7.58(t,J=7.9Hz,1H),7.43(d,J=8.4Hz,1H),7.26(dd,J=9.1,4.4Hz ,1H),2.28(s,3H).

HRMS(ESI)m/z:C ₂₃ H ₁₇ FN ₆ O, calculated 413.1526[M+H] ⁺ ,found 413.1541[M+H] ⁺ .

Example 5

Preparation of N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4-methoxybenzamide

Steps (a), (b), (c), (d), (e), (f), (g), (h), (i) are the same as in Example 1.

(j) N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4-methoxybenzyl Preparation of amides

3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline (0.2 g, 0.69 mmol) was dissolved in 5 mL of dichloromethane, Add 50 μL of triethylamine, place the above system in an ice bath, then dissolve 4-methoxybenzoyl chloride (0.35 g, 2.07 mmol) in 3 mL of dichloromethane, and then slowly add it dropwise to the above ice bath In the system, after dropping, continue the ice-bath reaction for 2-3 hours, TLC to detect the reaction progress, stop the reaction after the disappearance of the raw materials. The reaction solution was filtered, the insoluble matter was washed with water and dichloromethane respectively, and finally purified by column chromatography (eluent: dichloromethane/methanol) to obtain pure product.

The pure product is yellow-green fluorescent solid, yield: 80%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 10.28(s, 1H), 8.75(s, 1H), 8.68(d, J=3.2Hz, 1H), 8.49(s, 1H), 8.25(s ,1H),8.21(d,J=9.1Hz,1H),7.99(d,J=8.2Hz,3H),7.81(d,J=8.3Hz,1H),7.40(d,J=8.4Hz,1H ),7.27(dd,J=9.2,4.4Hz,1H),7.08(d,J=8.7Hz,2H),3.84(s,3H),2.27(s,3H).

HRMS(ESI)m/z:C ₂₄ H ₂₁ N ₆ O ₂ ,calculated 425.1726[M+H] ⁺ ,found 425.1742[M+H] ⁺ .

Example 6

N-(3-(4-(3-(imidazo[1,2-b]pyridazinyl))-1-pyrazolyl)-4-methylphenyl)-4-ethylbenzamide preparation

Steps (a), (b), (c), (d), (e), (f), (g), (h), (i) are the same as in Example 1.

(j) N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4-ethylbenzamide preparation of

3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline (0.2 g, 0.69 mmol) was dissolved in 5 mL of dichloromethane, Add 50 μL of triethylamine, put the above system in ice bath, then dissolve 4-ethylbenzoyl chloride (0.35 g, 2.07 mmol) in 3 mL of dichloromethane, and then slowly add it dropwise to the above ice bath system In the middle, after dropping, continue to react in ice bath for 2-3 hours, TLC detects the progress of the reaction, and stops the reaction after the disappearance of the raw materials. The reaction solution was filtered, the insoluble matter was washed with water and dichloromethane respectively, and finally purified by column chromatography (eluent: dichloromethane/methanol) to obtain pure product.

The pure product is yellow-green fluorescent solid, yield: 80%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 10.34(s, 1H), 8.75(s, 1H), 8.67(d, J=4.1Hz, 1H), 8.48(s, 1H), 8.24(s ,1H),8.21(d,J=9.0Hz,1H),8.00(s,1H),7.91(d,J=8.0Hz,2H),7.82(d,J=8.4Hz,1H),7.43–7.34 (m,3H),7.26(dd,J=9.1,4.5Hz,1H),2.69(d,J=7.4Hz,2H),2.27(s,3H),1.23(d,J=7.5Hz,3H) .

HRMS(ESI) m/z: C ₂₅ H ₂₃ N ₆ O, calculated 423.1933[M+H] ⁺ ,found 423.1949[M+H] ⁺ .

Example 7

Preparation of N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-3-fluorobenzamide

Steps (a), (b), (c), (d), (e), (f), (g), (h), (i) are the same as in Example 1.

(j) N-(3-(4-(3-(imidazo[1,2-b]pyridazinyl))-1-pyrazolyl)-4-methylphenyl)-3-fluorobenzyl Preparation of amides

3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline (0.2 g, 0.69 mmol) was dissolved in 5 mL of dichloromethane, Add 50 μL of triethylamine, place the above system in an ice bath, then dissolve 3-fluorobenzoyl chloride (0.33 g, 2.07 mmol) in 3 mL of dichloromethane, and then slowly add it dropwise to the above ice bath system , After dropping, continue to react in ice bath for 2-3 hours, TLC to detect the reaction progress, stop the reaction after the disappearance of the raw materials. The reaction solution was filtered, the insoluble matter was washed with water and dichloromethane respectively, and finally purified by column chromatography (eluent: dichloromethane/methanol) to obtain pure product.

The pure product is yellow-green fluorescent solid, yield: 80%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 8.91(d, J=4.1Hz, 1H), 8.80(s, 1H), 8.49(d, J=2.4Hz, 2H), 8.37(d, J =9.4Hz,1H),7.97(s,1H),7.79(d,J=7.9Hz,1H),7.74(d,J=8.4Hz,2H),7.64–7.54(m,2H),7.49–7.39 (m,2H),2.23(s,3H).

HRMS(ESI)m/z:C ₂₃ H ₁₈ FN ₆ O, calculated 413.1526[M+H] ⁺ , found 413.1552[M+H] ⁺ .

Example 8

N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4–(1-(4-methyl Preparation of piperazinyl)methyl)benzamide

Steps (a), (b), (c), (d), (e), (f), (g), (h), (i) are the same as in Example 1.

(j) 4-Chloromethyl-N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)benzyl Preparation of amides

3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline (0.6 g, 2.07 mmol) was dissolved in 10 mL of dichloromethane, Add 0.1mL triethylamine, place the above system in ice bath, then dissolve 4-chloromethylbenzoyl chloride (0.99g, 6.21mmol) in 5mL dichloromethane, and then slowly add it dropwise to the above ice In the bath system, after dropping, continue to react in the ice bath for 2-3 hours, TLC detects the reaction progress, and stops the reaction after the disappearance of the raw materials. The reaction solution was filtered, the insoluble matter was washed with water and dichloromethane respectively, and finally purified by column chromatography (eluent: dichloromethane/methanol) to obtain pure product.

The pure product is yellow-green fluorescent solid, yield: 80%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 10.52(s, 1H), 8.89(d, J=3.8Hz, 1H), 8.82(s, 1H), 8.52(d, J=4.0Hz, 2H ),8.40(d,J=9.0Hz,1H),8.04(s,1H),8.00(d,J=8.1Hz,2H),7.82(d,J=7.2Hz,1H),7.60(d,J =8.1Hz, 2H), 7.56(dd, J=9.3, 4.4Hz, 1H), 7.43(d, J=8.3Hz, 1H), 4.85(s, 2H), 2.27(s, 3H).

ESI-MS m/z:443.1[M+H] ⁺ .

(k) N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4–(1-(4 -Preparation of -methylpiperazinyl)methyl)benzamide

4-chloromethyl-N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)benzamide ( 0.2g, 0.45mmol) was dissolved in 5mL N,N-dimethylformamide, a catalytic amount of potassium iodide was added, stirred at room temperature for 15 minutes, and then 1-methylpiperazine (0.13g, 1.35mmol) was added to the reaction system , the temperature of the system was raised to 90° C., and the reaction was carried out for 2 to 3 hours. The progress of the reaction was detected by TLC, and the reaction was stopped after the disappearance of the raw materials. The reaction solution was cooled to room temperature, 20 mL of water was added thereto, extracted with 20×3 mL of dichloromethane, and the organic phases were combined. Then the organic phase was washed three times with saturated ammonium chloride solution, dried over anhydrous magnesium sulfate, and precipitated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol) to obtain a pure product.

The pure product is yellow-green fluorescent solid, yield: 80%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 10.42(s, 1H), 8.76(s, 1H), 8.68(d, J=3.3Hz, 1H), 8.49(s, 1H), 8.28–8.17 (m,2H),8.01(s,1H),7.95(d,J=8.0Hz,2H),7.82(d,J=8.2Hz,1H),7.44(dd,J=17.4,8.2Hz,3H) ,7.27(dd,J=9.1,4.3Hz,1H),3.56(s,2H),2.45(s,6H),2.27(s,8H).

HRMS(ESI)m/z:C ₂₉ H ₃₁ N ₈ O, calculated 507.2621[M+H] ⁺ , found 507.2619[M+H] ⁺ .

Example 9

N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4-(1-morpholinyl)methyl Preparation of phenylbenzamide

Steps (a), (b), (c), (d), (e), (f), (g), (h), (i) are the same as in Example 1, and step (j) is the same as in Example 8

(k) N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4-(1-morpholine Base) the preparation of methyl benzamide

4-chloromethyl-N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)benzamide ( 0.2g, 0.45mmol) was dissolved in 5mL N,N-dimethylformamide, a catalytic amount of potassium iodide was added, stirred at room temperature for 15 minutes, then morpholine (0.12g, 1.35mmol) was added to the reaction system, and the system was heated To 90°C, react for 2-3 hours, check the progress of the reaction by TLC, and stop the reaction after the disappearance of the raw materials. The reaction solution was cooled to room temperature, 20 mL of water was added thereto, extracted with 20×3 mL of dichloromethane, and the organic phases were combined. Then the organic phase was washed three times with saturated ammonium chloride solution, dried over anhydrous magnesium sulfate, and precipitated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol) to obtain a pure product.

The pure product is yellow-green fluorescent solid, yield: 80%

¹ H NMR (DMSO-d ₆ , 400MHz, δppm): 10.40(s, 1H), 8.76(s, 1H), 8.68(d, J=3.8Hz, 1H), 8.49(s, 1H), 8.25(s ,1H),8.21(d,J=9.3Hz,1H),8.00(s,1H),7.94(d,J=8.0Hz,2H),7.81(d,J=7.9Hz,1H),7.48(d ,J=7.9Hz,2H),7.42(d,J=8.7Hz,1H),7.27(dd,J=8.9,4.1Hz,1H),3.59(s,4H),3.55(s,2H),2.37 (s,4H),2.27(s,3H).

HRMS(ESI)m/z:C ₂₈ H ₂₈ N ₇ O ₂ ,calculated 494.2304[M+H] ⁺ ,found 494.2303[M+H] ⁺ .

Example 10

N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4-(1-piperidinyl)methyl Preparation of phenylbenzamide

Steps (a), (b), (c), (d), (e), (f), (g), (h), (i) are the same as in Example 1, and step (j) is the same as in Example 8

(k) N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)-4-(1-piperidine Base) the preparation of methyl benzamide

4-chloromethyl-N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)benzamide ( 0.2g, 0.45mmol) was dissolved in 5mL N,N-dimethylformamide, a catalytic amount of potassium iodide was added, stirred at room temperature for 15 minutes, then piperidine (0.11g, 1.35mmol) was added to the reaction system, and the system was heated To 90°C, react for 2-3 hours, check the progress of the reaction by TLC, and stop the reaction after the disappearance of the raw materials. The reaction solution was cooled to room temperature, 20 mL of water was added thereto, extracted with 20×3 mL of dichloromethane, and the organic phases were combined. Then the organic phase was washed three times with saturated ammonium chloride solution, dried over anhydrous magnesium sulfate, and precipitated under reduced pressure, and the residue was purified by column chromatography (eluent: dichloromethane/methanol) to obtain a pure product.

The pure product is yellow-green fluorescent solid, yield: 80%

¹ H NMR(DMSO-d ₆ ,400MHz,δppm):8.76(s,1H),8.68(s,1H),8.49(s,1H),8.25(s,1H),8.21(d,J＝9.1Hz ,1H),8.00(s,1H),7.92(d,J=8.0Hz,2H),7.81(d,J=8.1Hz,1H),7.47–7.41(m,3H),7.27(dd,J= 9.1,4.4Hz,1H),3.50(s,2H),2.33(s,4H),2.27(s,3H),1.50(s,6H).

HRMS(ESI)m/z:C ₂₉ H ₃₀ N ₇ O, calculated 492.2512[M+H] ⁺ , found 492.2510[M+H] ⁺ .

Example 11

The Kinase-Glo kinase luminescence method was used to measure protein kinase activity in vitro, and the specific method is as follows.

Sample assay preparation:

(1) The sample compound was dissolved in DMSO, then diluted with DMSO and quantified to a concentration of 500 μM and transferred to the quantification plate and the solution was transferred to the dose plate. Compounds were serially diluted in DMSO at 5-fold concentration. Each concentration was then diluted 10-fold with reaction buffer solution to obtain a 10× final concentration. The compounds with a concentration ranging from 0.03 to 50 μM were transferred to the Bcr-Abl activity assay plate at a dose of 1 μL/well.

(2) Prepare the positive control substance STSP into a stock solution with a concentration of 10 mM with DMSO, and dilute to 100 μM with DMSO. First serially dilute with DMSO at a 5-fold concentration. Each concentration was then diluted 10-fold with reaction buffer solution to obtain a 10× final concentration. The compounds with a concentration ranging from 0.00064 to 10 μM were transferred to the Bcr-Abl activity assay plate at a dose of 1 μL/well.

(3) For HPE and ZPE wells, dilute 2 μL DMSO 10 times with reaction buffer solution to obtain 10% DMSO dilution, and then transfer it to the activity assay plate at a dose of 1 μL/well.

_IC50 Determination of Compound Inhibition of Bcr-Abl:

(1) Preparation of samples required for determination, compound without HPE kinase, but containing ATP, substrate and 1% DMSO; ZPE (0% effect): no compound but containing kinase, ATP, substrate and 1% DMSO solution; Positive Compound Wells: Contains Kinase, ATP, Substrate, and Positive Compounds at various concentrations. Test compound: Contains kinase, ATP, substrate and different concentrations of test compound.

(2) Preparation of reagents required for determination, 4×ATP: Dilute ATP 4 times in the test buffer solution to obtain a working solution; 4× substrate: Dilute Poly (glucose:tyrosine) 4 times in the test buffer solution Obtain working solution; 2.5×Bcr-Abl kinase: Kinase was diluted 2.5 times with assay dilution buffer solution to obtain working solution.

(3) Kinase reaction Add 10× compound to 384-well assay plate, 1 μL/well. For HPE and ZPE wells, add the same volume (1 μL/well) of 10% DMSO solution; add 2.5×Bcr-Abl kinase to the assay plate, 4 μL/well. For HPE and ZPE wells, add the same volume (4 μL/well) of assay buffer solution; put the assay plate into a centrifuge and centrifuge at 1 000 rpm for 1 minute to mix the system; pre-incubate the assay plate at 30°C for 30 minutes; mix equally A volume of 4xATP and 4xsubstrate yields a 2xATP-substrate mixture. The mixture is the reaction mixture of Bcr-Abl kinase activity assay receptor; add 2×ATP-substrate mixture to the assay plate, 5 μL/well; put the assay plate in a centrifuge and centrifuge at 1 000 rpm for 1 minute to mix the system; assay plate Pre-incubate at 30°C for 1 hour; add Kinase glo plus to each well, 10 μL/well, and incubate the assay plate at 27°C for 20 minutes; read the fluorescence intensity value with Envision. Place Kinase glo plus reagent at room temperature for 30 minutes before use.

(4) Analysis and processing of raw data Prism 5.0 was used to analyze the raw data; the compound inHibitiory rate was calculated as Compound inHibitiory rate=("Compound" reading-ZEP)/(HPE-ZPE)×100%, and the measurement results are shown in Table 1.

Table 1: Using STSP as a positive control, the activities of 10 synthesized target compounds were determined

.

Claims (2)

1. The preparation method of N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)benzamide derivatives, The structural formula of N-(3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)benzamide derivatives is the following formula (IA) or (IB): R ¹ in (IA) means In (IB), R ¹ represents -OCH ₃ , -F, -CF ₃ , -CH ₂ CH ₃ or -H; R ² represents -CF ₃ , -F, -Cl or -H; it is characterized by comprising the following steps : (a) Dissolve 2-methyl-5-nitroaniline in ethanol, add concentrated hydrochloric acid, place the above system under ice bath conditions, then add the aqueous solution of sodium nitrite to the above system to obtain formula (II) Represented 2-methyl-5-nitrophenylhydrazine hydrochloride; (b) Dissolve stannous chloride dihydrate in 37% concentrated hydrochloric acid, place it in an ice bath, then add the compound of formula (II) dropwise to the above system, react for 0.5 to 2 hours, then add NaOH solution Neutralize to obtain 2-methyl-5-nitrophenylhydrazine represented by formula (III); (c) reacting the compound of formula (III) with 2-bromomalonaldehyde in ethanol to obtain 4-bromo-1-(2-methyl-5-nitrophenyl)pyrazole represented by formula (IV); (d) dissolving the compound of formula (IV) in ethanol, adding an aqueous solution of ammonium chloride and iron powder to obtain 3-(4-bromo-1-pyrazolyl)-4-methylaniline represented by formula (V) ; (e) reacting the compound of formula (V) and Boc anhydride in tetrahydrofuran to obtain N-Boc-3-(4-bromo-1-pyrazolyl)-4-methylaniline represented by formula (VI); (f) Dissolve the compound of formula (VI) in dioxane, add biboronic acid pinacol ester, potassium acetate and two (triphenylphosphine) palladium dichloride, react to obtain the N- Boc-3-(4-(2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolanyl))-1-pyrazolyl)-4-methylaniline ; (g) reacting imidazo[1,2-b]pyridazine, NBS and a catalytic amount of AIBN in chloroform to obtain 3-bromoimidazo[1,2-b]pyridazine represented by formula (VIII); (h) Formula (VII) compound and formula (VIII) compound are dissolved in the mixed solution of dioxane and water, add tetrakis (triphenylphosphine) palladium and sodium carbonate, react to obtain the N represented by formula (IX) -Boc-3-(4-(3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylaniline; (i) react formula (IX) and trifluoroacetic acid in dichloromethane to obtain 3-(4-(3-imidazo[1,2-b]pyridazinyl)-1- Pyrazolyl)-4-methylaniline; (j) react formula (X) compound, 4-chloromethylbenzoyl chloride and triethylamine in dichloromethane to obtain 4-chloromethyl-N-(3-(4- (3-imidazo[1,2-b]pyridazinyl)-1-pyrazolyl)-4-methylphenyl)benzamide; (k) react with formula (XI) compound, nucleophile and potassium iodide in DMF, obtain formula (IA) representation N-(3-(4-(3-(imidazo[1,2-b]pyridazinyl))-1-pyrazolyl)-4-methylphenyl)benzamide derivatives, R ¹ means Or (1) react formula (X) compound, substituted benzoyl chloride and triethylamine in dichloromethane, obtain N-(3-(4-(3-imidazo[1,2 -b] pyridazinyl) -1-pyrazolyl) -4-methylphenyl) benzamide derivatives; R ¹ represents -OCH ₃ , -F, -CF ₃ , -CH ₂ CH ₃ or -H, R ² represents -CF ₃ , -F, -Cl or -H; Step (k) nucleophile is: Step (1) substituted benzoyl chloride structural formula is: 2. according to the method for claim 1, it is characterized in that, step (a) reaction temperature 0～5 ℃, reaction time 0.5～1 hour; Quantitative ratio is 1: (1.5～2), temperature of reaction 0～10 ℃, reaction time 1～2 hour; Step (c) formula (III) compound and the substance quantity ratio of 2-bromomalondialdehyde are 1:( 1.1～1.5), reaction temperature 55～65 ℃, reaction time 2～4 hours; Step (d) reaction temperature 80～90 ℃, reaction time 2～3 hours; Step (e) formula (V) compound and Boc acid anhydride The amount ratio of substance is 1: (1.1～1.4), reaction temperature 66～70 ℃, reaction time 2～3 hours; Step (f) formula (VI) compound: biboronic acid pinacol ester: potassium acetate: two (three The molar ratio of phenylphosphine) palladium dichloride is 1:(1.1～1.5):3:(0.1～0.2), the reaction temperature is 100～110°C, and the reaction time is 15～18 hours; step (g) imidazo [1,2-b] Pyridazine: NBS: AIBN substance molar ratio is 1: (1.5～2): (0.1～0.2), reaction temperature 65 ℃, reaction time 2～3 hours; step (h) formula ( VII) compound: compound of formula (VIII): sodium carbonate: tetrakis(triphenylphosphine) palladium molar ratio is 1:2:3:(0.1～0.2), reaction temperature is 110～120°C, reaction time is 7～ 10 hours; step (i) normal temperature reaction 5～6 hours; step (j) formula (X) compound: 4-chloromethylbenzoyl chloride: the molar ratio of the substance of triethylamine is 1: (2～3): (0.2～0.5), reaction time 2～3 hours; Step (k) formula (XI) compound: nucleophile: the substance molar ratio of potassium iodide is 1: (2～3): (0.1～0.2), temperature of reaction 80～90 DEG C, reaction time 2～3 hours; Step (1) formula (X) compound: substituted benzoyl chloride: the molar ratio of the substance of triethylamine is 1: (2～3): (0.2～0.5), The reaction time is 2 to 3 hours.