WO2022126441A1 - Oxindole derivative and pharmaceutical use thereof - Google Patents

Abstract

Provided are an oxindole derivative and a pharmaceutical use thereof. The structure of the oxindole derivative is shown in formula (A). Experimental results show that the provided compound has significantly improved pharmacokinetic properties compared with BIBF1120, has an excellent inhibition effect on VEGFR, FGFR and PDGFR, can be used as an inhibitor of VEGFR, FGFR and/or PDGFR, as an angiogenesis inhibitor, and as a drug for preventing and/or treating various tumors including pharyngeal squamous cell carcinoma, and has wide application prospects.

Description

Indolinone derivatives and their pharmaceutical uses technical field

The invention belongs to the technical field of chemical medicine, and particularly relates to an indolinone derivative and its pharmaceutical use.

Background technique

Nintedanib (BIBF1120) is an orally active triple kinase inhibitor that simultaneously acts on 3 key receptor families involved in angiogenesis - vascular endothelial growth factor receptor (VEGFR), fibroblast growth factor receptor (FGFR) and platelet-derived growth factor receptor (PDGFR). The structural formula of BIBF1120 is as follows:

As a novel angiogenesis inhibitor, BIBF1120 can inhibit the formation of blood vessels around and inside the tumor, thereby reducing the oxygen and nutrient supply of the tumor, causing tumor cells to shrink and die, and achieve anti-tumor effect. Compared with traditional chemotherapy methods, this anti-tumor treatment method has the advantage of less side effects. Studies have found that BIBF1120 has a good effect on a variety of tumors such as liver cancer, lung cancer, rectal cancer, uterine cancer, brain cancer, metastatic colorectal cancer, and pharyngeal squamous cell carcinoma.

However, the antitumor effect and pharmacokinetic properties of BIBF1120 still need to be further improved. Due to the complex metabolic process of biological systems, the pharmacokinetic properties of drugs in vivo are affected by many factors and show corresponding complexity. Compared with the corresponding non-deuterated drugs, the changes in the pharmacokinetic properties of deuterated drugs exhibit great contingency and unpredictability. Rather than prolonging the half-life, deuterium at some sites may shorten it (Scott L.Harbeson, Roger D.Tung.Deuterium in Drug Discovery and Development, P405-406.), deteriorating its pharmacokinetic properties; On the other hand, the hydrogen at certain positions on the drug molecule is not easily deuterated due to steric hindrance and other reasons. Therefore, the deuteration of the drug is not arbitrary, and the deuterated sites are unpredictable.

Therefore, more angiokinase inhibitors with novel structure, better inhibitory activity and better pharmacokinetic properties have been developed, which are effective against liver cancer, lung cancer, rectal cancer, uterine cancer, brain cancer, metastatic intestinal cancer, pharyngeal squamous cell carcinoma, etc. The treatment of this disease is of great significance.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide an indolinone derivative and its use in the preparation of VEGFR, FGFR and/or PDGFR inhibitors.

The present invention specifically provides a compound represented by formula (A) or a pharmaceutically acceptable salt thereof:

Wherein, R ₁ and R ₂ are independently selected from C1-C6 alkyl and deuterated C1-C6 alkyl;

R ₃ is selected from C1-C6 alkyl, halogenated C1-C6 alkyl,

R ₄ to R ₁₁ are each independently selected from H or deuterium;

And when R ₂ is methyl, R ₃ is methyl, and R ₄ to R ₁₁ are hydrogen, R ₁ is not methyl.

Further, R ₁ and R ₂ are each independently selected from C1-C3 alkyl and deuterated C1-C3 alkyl;

所述卤素优选为氟。 R ₃ is selected from C1-C2 alkyl, halogenated C1-C2 alkyl,

The halogen is preferably fluorine.

Further, R ₁ and R ₂ are each independently selected from methyl or deuterated methyl, and the deuterated methyl is preferably -CD ₃ ;

R ₃ is selected from CF ₃ , CHF ₂ , CH ₂ F, CH ₂ CF ₃ , CH ₂ CHF ₂ , CH ₂ CH ₂ F,

R ₄ to R ₁₁ are all H or all D.

Further, the compound is selected from one of the following compounds:

The present invention also provides a method for preparing the above-mentioned compound or a pharmaceutically acceptable salt thereof, characterized in that: the method comprises using compound (A-1) and compound (A-2) as raw materials, and reacting to obtain a compound represented by formula (A);

Among them, R ₁ to R ₁₁ are as described above.

Further, the molar ratio of compound (A-1) and compound (A-2) is 1:1; the solvent of the reaction is methanol.

The present invention also provides an antitumor drug, which is a preparation prepared by using the above-mentioned compound or a pharmaceutically acceptable salt thereof as an active ingredient and adding auxiliary materials commonly used in the pharmaceutical field.

The present invention also provides the use of the above-mentioned compounds or their pharmaceutically acceptable salts in the preparation of VEGFR, FGFR and/or PDGFR inhibitors; the VEGFR is preferably VEGFR2, the FGFR is preferably FGFR1, and the PDGFR is preferably PDGFRα .

The present invention also provides the use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of an angiogenesis inhibitor.

The present invention also provides the use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and/or treating tumors, the tumors are preferably liver cancer, lung cancer, rectal cancer, uterine cancer, brain cancer, metastatic colorectal cancer , Pharyngeal squamous cell carcinoma.

Definitions of terms used in the present invention: Unless otherwise specified, the initial definitions of groups or terms provided herein apply to the group or term throughout the specification; for terms that are not specifically defined herein, the content and context of the disclosure should be used. , give their meanings that those skilled in the art can give them.

The minimum and maximum carbon content in the hydrocarbon group is indicated by the prefix. For example, a C1 _{- C6} alkyl group refers to a straight or branched chain alkyl group containing 1 to 6 carbon atoms.

The deuterated C1-C6 alkyl group refers to a group in which one or more hydrogens in the C1-C6 alkyl group are replaced by deuterium.

The halogenated C1-C6 alkyl group refers to a group in which one or more hydrogens in the C1-C6 alkyl group are replaced by halogen.

Halogen is fluorine, chlorine, bromine or iodine.

The experimental results show that the compounds provided by the present invention have significantly improved pharmacokinetic properties than BIBF1120, and have simultaneous effects on vascular endothelial growth factor receptor (VEGFR), fibroblast growth factor receptor (FGFR) and platelet-derived growth factor receptor. (PDGFR) all have excellent inhibitory effects, and can be used as VEGFR, FGFR and/or PDGFR inhibitors, as angiogenesis inhibitors, as drugs for the prevention and/or treatment of various tumors including pharyngeal squamous cell carcinoma, with a broad range of application prospects.

Obviously, according to the above-mentioned content of the present invention, according to the common technical knowledge and conventional means in the field, without departing from the above-mentioned basic technical idea of the present invention, other various forms of modification, replacement or change can also be made.

The above content of the present invention will be further described in detail below through the specific implementation in the form of examples. However, this should not be construed as limiting the scope of the above-mentioned subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention belong to the scope of the present invention.

Description of drawings

Figure 1 is the tumor growth curve of each group in the in vivo experiment, wherein 6004 is the compound ZLF6004, and the control group is the solvent control group.

Detailed ways

The raw materials and equipment used in the specific embodiments of the present invention are all known products, which are obtained by purchasing commercially available products.

Example 1 Methyl-(Z)-3-(((4-(2-(4-(fluoromethyl)piperazine-1-substituted)-N-methylacetamido)anilino)(phenyl) Preparation of Methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6001)

The synthetic route is as follows:

Step 1: the preparation of methyl m-nitrobenzoate (2)

1.0 g of m-nitrobenzoic acid was added to 8 mL of methanol, 4% concentrated sulfuric acid was added, the reaction was microwaved at 100 °C for 10 minutes, methylene chloride was added, washed with saturated NaHCO ₃ and saturated brine successively, dried over anhydrous sodium sulfate, and extracted. Filter and spin dry to obtain 0.95 g of product. Yield 85.8%.

Step 2: Preparation of methyl 4-(2-methoxy-2-acetyl)-3 nitrobenzoate (3)

1.97g potassium tert-butoxide (18mmol) was dissolved in 20mL DMF, cooled to -10°C, and m-nitrobenzoic acid (1.36g, 7.5mmol) methyl ester and methyl chloroacetate (0.73mL, 4mL DMF) were dissolved in 4mL DMF. 8.3mmol) mixed solution was added dropwise to the above solution, after the dropwise addition, the reaction was performed at -10°C until no raw materials were detected, and the reaction solution was poured into a mixed solution of ice water (25mL) and concentrated hydrochloric acid (8.5mL). , extracted with ethyl acetate, washed with saturated brine, dried, and spin-dried. Column chromatography gave crude product 1.20 g (63.3%).

Step 3: Preparation of methyl 2-indolone-6-carboxylate (4)

4-(2-Methoxy-2-acetyl)-3 nitrobenzoic acid (3) (0.97 g, 3.83 mmol) was dissolved in 20 mL of acetic acid, 130 mg of 10% Pd/C was added, and the hydrogen pressure was The reaction was carried out at room temperature for 2.5h under the condition of 50psi. The catalyst was removed by filtration, the solvent was evaporated, triturated with ethyl acetate, washed, filtered, and dried under vacuum at 100°C to obtain 2.8 g of product 4 (38.2%).

Step 4: Preparation of methyl 1-chloroacetyl-2-indolone-6-carboxylate (5)

At room temperature, methyl 2-indolinone-6-carboxylate 4 (2.0 g, 10.35 mmol) was suspended in 12 ml of toluene, and chloroacetic anhydride (2.7 g, 15.5 mmol) was added to the above suspension, heated to reflux for 3 h, Cooled to 80°C, methylcyclohexane (6ml) was added in 30min, the suspension was cooled to room temperature with stirring, the crude product was obtained by suction filtration, washed with cold methanol (4ml), and dried to obtain 2.55g (92.6%) of white solid.

Step 5: Preparation of (E)-1-chloroacetyl-3(methoxy(phenyl)methylene)-2-indolone-6carboxylic acid methyl ester (6)

At ambient temperature, methyl 1-chloroacetyl-2-indolinone-6-carboxylate 5 (0.6 g, 2.3 mmol) was suspended in toluene (5 ml), to which was added acetic anhydride (0.81 g, 7.85 mmol), The system was heated to 110 °C, trimethyl orthobenzoate (1.0 g, 5.4 mmol) was added within 1 h, the reaction was continued for 3 h, the volatile products were volatilized, toluene (4 ml) was added to keep the system concentration unchanged, and the system was cooled to 5 ° C, After stirring for 1 h, the crude product was obtained by suction filtration, washed with a mixture of toluene, toluene and ethyl acetate (1:1) successively, and dried to obtain 0.8 g (93.0%) of a light yellow solid.

Step 6: Preparation of (E)-methyl 3-(methoxy(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (7)

A solution of potassium hydroxide (82mg, 1.2mmol) in methanol (1ml) was added to (E)-1-chloroacetyl-3(methoxy(phenyl)methylene)-2-indole at 63°C Keto-6 methyl formate 6 (1.6g, 2mmol) in methanol (3ml) suspension, the reaction system was stirred for 30min, cooled to 0°C, stirred at 0°C for 2h, suction filtered, washed with methanol, and dried to obtain 0.95g (74.9%) yellow solid.

Step 7: 2-Chloro-N-methyl-N-(4-nitrophenyl)acetamide (9)

N-methyl-4-nitroaniline (1.25 g, 8.22 mmol) was added to ethyl acetate (3 mL), heated to 60°C, and chloroacetic anhydride (1.63 g, 9.5 mmol) ethyl acetate ( 5mL) solution, refluxed for 1h, cooled to 75°C, then added 8mL of cyclohexane, crystals formed at 60°C, cooled to 0°C, stirred for 1h, filtered, washed with cyclohexane, and dried to obtain 1.48g (78.8g) of white crystals. %).

Step 8: Preparation of N-(4-nitrophenyl)-Nmethyl-2-(4-Boc-piperazine-1-substituted)acetamide (10)

1 g of 2-chloro-N-methyl-N-(4-nitrophenyl)acetamide 8 was dissolved in 8 mL of toluene, heated to 40°C, and N-Boc-piperazine (2.1 g, 2.5 g) was added dropwise within 30 min. eq), heated and stirred for 2h (55°C), lowered to room temperature, washed with water (5mL), diluted the toluene solution system with 10mL of isopropanol, added 100mg Pd/C (10%), passed hydrogen (4bar), and stirred at room temperature After 3 h, the catalyst was removed by filtration, the solvent was spun out, and the residue was recrystallized with ethyl acetate/petroleum ether, washed with petroleum ether, and dried to obtain 1.35 g (91.1%) of white crystals.

Step 9: Preparation of N-(4-nitrophenyl)-Nmethyl-2-(piperazine-1-substituted)acetamide (11) 1 g of N-(4-aminophenyl)-Nmethyl- 2-(4-Boc-piperazine-1-substituted)acetamide was dissolved in 10 mL of DCM and stirred in an ice bath. Trifluoroacetic acid (1.1 ml, 6.0 eq) was added dropwise within 2 min. After dropping, the mixture was stirred at room temperature for 2 h, spin-dried under reduced pressure, added with saturated aqueous sodium bicarbonate solution, extracted with DCM, dried over anhydrous magnesium sulfate, and the organic solvent was spin-dried to obtain oil. Liquid 1.40 g (94.4%).

Step 10: Preparation of N-(4-nitrophenyl)-Nmethyl-2-(4-(1-fluoro-ethyl-1-substituted)acetamide (12)

1 g of N-(4-nitrophenyl)-Nmethyl-2-(4-piperazine-1-substituted)acetamide was dissolved in 10 mL of DCM and stirred in an ice bath. 1-Fluoro-2-iodoethane (1.11 ml, 6.0 eq) was added dropwise within 2 min, stirred in an ice bath for 2 h, filtered, spin-dried under reduced pressure, added with saturated aqueous sodium bicarbonate solution, extracted with DCM, and dried over anhydrous magnesium sulfate The organic solvent was spin-dried to obtain 0.88 g (79.3%) of a yellow solid.

Step Eleven: Preparation of N-(4-aminophenyl)-Nmethyl-2-(4-(1-fluoroethyl-1-substituted)acetamide (13)

1 g of N-(4-aminophenyl)-Nmethyl-2-(4-piperazine-1-substituted)acetamide was dissolved in 10 mL of ethanol and 4 aqueous solution, and stirred in an ice bath. 130 mg of 10% Pd/C was added, and the reaction was carried out at room temperature for 2.5 h under normal pressure hydrogenation. The catalyst was removed by filtration, the solvent was evaporated, triturated with ethyl acetate, washed, filtered, and dried under vacuum at 100°C to obtain 83 g of product 13 (92.2%).

Step 12: Methyl-(Z)-3-(((4-(2-(4-(fluoromethyl)piperazine-1-substituted)-N-acetylamino)anilino)(phenyl)idene Preparation of methyl)-2-indolone-6-carboxylate (ZLF6001)

(E)-Methyl 3-(methoxy(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester 7 (0.5 mg, 1.62 mmol) and N-(4-aminophenyl) A suspension of -N methyl-2-(4-(1-fluoroethyl-1-substituted)acetamide 13 (0.48 mg, 1.70 mmol) in methanol (1.8 ml) was heated to reflux for 8 h, slowly cooled to 10 °C, 10 Continue to stir at ℃ for 1 h, filter with suction, wash with cold methanol, and dry to obtain 0.75 g (83.3%) of a yellow solid.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.63-7.54(m, 3H), 7.50(d, J=7.1Hz, 2H), 7.42( d,J＝1.3Hz,1H),7.20(d,J＝8.2Hz,1H),7.14(d,J＝8.4Hz,2H),6.89(d,J＝8.4Hz,2H),5.83(d, J=8.2Hz, 1H), 4.55(t, J=4.9Hz, 1H), 4.43(t, J=4.9Hz, 1H), 3.77(s, 3H), 3.07(s, 3H), 2.71(s, 2H), 2.59(t, J=4.8Hz, 1H), 2.53(s, 1H), 2.31(s, 4H), 2.22(s, 4H).

ESI-MS m/z: 570.6[MH] ^- .

Example 2 Methyl-(Z)-3-(((4-(2-(4-(difluoromethyl)piperazine-1-substituted)-N-methylacetamido)anilino)(phenyl ) Methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6002) preparation

The synthesis steps were performed according to the method described in Example 1, except that the raw material 1-fluoro-2-iodoethane was replaced with 1,1-difluoro-2-iodoethane to synthesize compound ZLF6002. The yield of the last reaction was 75%.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.64-7.53(m, 3H), 7.52-7.47(m, 2H), 7.42(d, J ＝1.4Hz,1H),7.20(dd,J＝8.2,1.6Hz,1H),7.14(d,J＝8.5Hz,2H),6.89(d,J＝8.6Hz,2H),6.09(tt,J =55.8,4.3Hz,1H),5.83(d,J=8.2Hz,1H),3.77(s,3H),3.06(s,3H),2.66(td,J=15.7,4.2Hz,4H),2.39 (s,4H),2.22(s,4H).

ESI-MS m/z: 588.6[MH] ^- .

Example 3 Methyl-(Z)-3-(((4-(2-(4-(trifluoromethyl)piperazine-1-substituted)-N-methylacetamido)anilino)(phenyl ) Methylene)-2-indolone-6-carboxylate methyl ester (ZLF6003) preparation

The synthesis steps were performed according to the method described in Example 1, except that the raw material 1-fluoro-2-iodoethane was replaced with trifluoro-2-iodoethane to synthesize compound ZLF6003. The yield of the last reaction was 72%.

¹ H NMR (400MHz, DMSO-d6)δ12.23(s,1H),10.96(s,1H),7.63-7.54(m,3H),7.53-7.47(m,2H),7.43(d,J= 1.2Hz, 1H), 7.20 (dd, J=8.2, 1.5Hz, 1H), 7.13 (d, J=8.5Hz, 2H), 6.89 (d, J=8.5Hz, 2H), 5.83 (d, J= 8.2Hz, 1H), 3.77(s, 3H), 3.20–3.01(m, 5H), 2.72(s, 2H), 2.48–2.39(s, 4H), 2.23(s, 4H).

ESI-MS m/z: 606.6[MH] ^- .

Example 4 Methyl-(Z)-3-(((4-(2-(4-(2-(2-methoxyethyletheryl)ethyl)piperazine-1-substituted)-N-methyl Preparation of acetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6004)

The synthesis steps were carried out according to the method described in Example 1, except that the raw material 1-fluoro-2-iodoethane was replaced with 1-iodo-2(2-methoxyethyl ether)ethane to synthesize compound ZLF6004. The yield of the last reaction was 69%.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.63-7.54(m, 3H), 7.50(d, J=7.1Hz, 2H), 7.42( d,J＝1.3Hz,1H),7.20(d,J＝8.2Hz,1H),7.14(d,J＝8.4Hz,2H),6.89(d,J＝8.4Hz,2H),5.83(d, J=8.2Hz, 1H), 4.55(t, J=4.9Hz, 1H), 4.43(t, J=4.9Hz, 1H), 3.07(s, 3H), 2.71(s, 2H), 2.59(t, J=4.8Hz, 1H), 2.53(s, 1H), 2.31(s, 4H), 2.22(s, 4H).

ESI-MS m/z: 573.7[MH] ^- .

Example 5 Methyl-d3-(Z)-3-(((4-(2-(4-(fluoromethyl)piperazine-1-substituted)-N-methylacetamido)anilino)(benzene Preparation of methyl) methylene)-2-indolone-6-carboxylate (ZLF6005)

According to the method described in Example 1, the difference is that the raw material methanol in step 1 is replaced with deuterated methanol to synthesize compound ZLF6005. The yield of the last reaction was 69%.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.64-7.53(m, 3H), 7.52-7.47(m, 2H), 7.42(d, J ＝1.4Hz,1H),7.20(dd,J＝8.2,1.6Hz,1H),7.14(d,J＝8.5Hz,2H),6.89(d,J＝8.6Hz,2H),6.09(tt,J ＝55.8, 4.3Hz, 1H), 5.83(d, J＝8.2Hz, 1H), 3.06(s, 3H), 2.66(td, J＝15.7, 4.2Hz, 4H), 2.39(s, 4H), 2.22 (s,4H).

ESI-MS m/z: 591.7[MH] ^- .

Example 6 Methyl-d3-(Z)-3-(((4-(2-(4-(difluoromethyl)piperazine-1-substituted)-N-methylacetamido)anilino)( Preparation of phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6006)

According to the method described in Example 1, the difference is that the raw material methanol in step 1 is replaced with deuterated methanol, and the raw material 1-fluoro-2-iodoethane is replaced with 1,1-difluoro-2-iodoethane alkane, synthesized compound ZLF6006. The yield of the last reaction was 71%.

¹ H NMR (400MHz, DMSO-d6) δ 12.23 (s, 1H), 10.96 (s, 1H), 7.63–7.54 (m, 3H), 7.53–7.47 (m, 2H), 7.43 (d, J= 1.2Hz, 1H), 7.20 (dd, J=8.2, 1.5Hz, 1H), 7.13 (d, J=8.5Hz, 2H), 6.89 (d, J=8.5Hz, 2H), 5.83 (d, J= 8.2Hz, 1H), 3.20–3.01 (m, 5H), 2.72 (s, 2H), 2.48–2.39 (s, 4H), 2.23 (s, 4H).

ESI-MS m/z: 609.7[MH] ^- .

Example 7 Methyl-d3-(Z)-3-(((4-(2-(4-(trifluoromethyl)piperazine-1-substituted)-N-methylacetamido)anilino)( Preparation of phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6007)

The synthesis steps were in accordance with the method described in Example 1, except that N-trideuteromethyl-p-nitroaniline was used instead of N-methyl-p-nitroaniline, and the yield in the final step was 69%.

Wherein, the synthetic route of N-trideuteromethyl-p-nitroaniline is as follows:

The synthetic route of N-trideuteromethyl-p-nitroaniline refers to Ullmann amination reference J.Org.Chem.2011,76,1180-1183.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.63-7.54(m, 3H), 7.50(d, J=7.1Hz, 2H), 7.42( d,J＝1.3Hz,1H),7.20(d,J＝8.2Hz,1H),7.14(d,J＝8.4Hz,2H),6.89(d,J＝8.4Hz,2H),5.83(d, J=8.2Hz, 1H), 4.55(t, J=4.9Hz, 1H), 4.43(t, J=4.9Hz, 1H), 3.77(s, 3H), 2.71(s, 2H), 2.59(t, J=4.8Hz, 1H), 2.53(s, 1H), 2.31(s, 4H), 2.22(s, 4H).

ESI-MS m/z: 573.7[MH] ^- .

Example 8 Methyl-d3-(Z)-3-(((4-(2-(4-(fluoromethyl)piperazine-1-substituted)-N-methylacetamido)anilino)(benzene Preparation of methyl) methylene)-2-indolone-6-carboxylate (ZLF6008)

The synthesis steps were as described in Example 1, except that N-trideuteromethyl-p-nitroaniline was used instead of N-methyl-p-nitroaniline, and 1,1-difluoro-2-iodoethane was used. The raw material 1-fluoro-2-iodoethane was replaced, and the yield of the last step was 69%.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.64-7.53(m, 3H), 7.52-7.47(m, 2H), 7.42(d, J ＝1.4Hz,1H),7.20(dd,J＝8.2,1.6Hz,1H),7.14(d,J＝8.5Hz,2H),6.89(d,J＝8.6Hz,2H),6.09(tt,J ＝55.8, 4.3Hz, 1H), 5.83(d, J＝8.2Hz, 1H), 3.77(s, 3H), 2.66(td, J＝15.7, 4.2Hz, 4H), 2.39(s, 4H), 2.22 (s,4H).

ESI-MS m/z: 591.7[MH] ^- .

Example 9 Methyl-(Z)-3-(((4-(2-(4-(fluoromethyl)piperazine-1-substituted)-N-(methyl-d3)acetamido)anilino) Preparation of (Phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6009)

The synthesis steps were performed according to the method described in Example 1, except that N-trideuteromethyl-p-nitroaniline was used instead of N-methyl-p-nitroaniline to synthesize compound ZLF6009. The yield of the last reaction was 69%.

Wherein, the synthetic route of N-trideuteromethyl-p-nitroaniline is as follows:

The synthetic route of N-trideuteromethyl-p-nitroaniline refers to Ullmann amination reference J.Org.Chem.2011,76,1180-1183.

¹ H NMR (400MHz, DMSO-d6) δ 12.23 (s, 1H), 10.96 (s, 1H), 7.63–7.54 (m, 3H), 7.53–7.47 (m, 2H), 7.43 (d, J= 1.2Hz, 1H), 7.20 (dd, J=8.2, 1.5Hz, 1H), 7.13 (d, J=8.5Hz, 2H), 6.89 (d, J=8.5Hz, 2H), 5.83 (d, J= 8.2Hz, 1H), 3.77(s, 3H), 3.18–3.01(m, 2H), 2.72(s, 2H), 2.48–2.39(s, 4H), 2.23(s, 4H).

ESI-MS m/z: 609.7[MH] ^- .

Example 10 Methyl-(Z)-3-(((4-(2-(4-(difluoromethyl)piperazine-1-substituted)-N-(methyl-d3)acetamido)anilino ) (phenyl) methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6010) preparation

The synthesis steps were as described in Example 1, except that N-trideuteromethyl-p-nitroaniline was used instead of N-methyl-p-nitroaniline, and 1,1-difluoro-2-iodoethane was used. Substitute the raw material 1-fluoro-2-iodoethane to synthesize compound ZLF6010. The yield of the last step was 69%.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.63-7.54(m, 3H), 7.50(d, J=7.1Hz, 2H), 7.42( d,J＝1.3Hz,1H),7.20(d,J＝8.2Hz,1H),7.14(d,J＝8.4Hz,2H),6.89(d,J＝8.4Hz,2H),5.83(d, J=8.2Hz, 1H), 4.55(t, J=4.9Hz, 1H), 4.43(t, J=4.9Hz, 1H), 3.77(s, 3H), 3.07(s, 3H), 2.71(s, 2H), 2.59(t, J=4.8Hz, 1H), 2.53(s, 1H).

ESI-MS m/z: 578.7[MH] ^- .

Example 11 Methyl-(Z)-3-(((4-(2-(4-(trifluoromethyl)piperazine-1-substituted)-N-(methyl-d3)acetamido)anilino ) (phenyl) methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6011) preparation

The synthesis steps were in accordance with the method described in Example 1, except that N-trideuteromethyl-p-nitroaniline was used to replace N-methyl-p-nitroaniline, and trifluoro-2-iodoethane was used to replace the raw material 1- Fluoro-2-iodoethane to synthesize compound ZLF6011. The yield of the last step was 69%.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.64-7.53(m, 3H), 7.52-7.47(m, 2H), 7.42(d, J ＝1.4Hz,1H),7.20(dd,J＝8.2,1.6Hz,1H),7.14(d,J＝8.5Hz,2H),6.89(d,J＝8.6Hz,2H),6.09(tt,J =55.8,4.3Hz,1H),5.83(d,J=8.2Hz,1H),3.77(s,3H),3.06(s,3H).

ESI-MS m/z: 596.7[MH] ^- .

Example 12 Methyl-(Z)-3-(((4-(2-(4-(2-(2-methoxyethyletheryl)ethyl)piperazine-1-substituted)-N-(methyl) Preparation of methyl-d3)acetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylate (ZLF6012)

The synthetic steps were in accordance with the method described in Example 1, except that N-trideuteromethyl-p-nitroaniline was used instead of N-methyl-p-nitroaniline, and 1-iodo-2(2-methoxyethyl ether was used instead of ) ethane replaces the raw material 1-fluoro-2-iodoethane to synthesize compound ZLF6012. The yield of the last step was 69%.

¹ H NMR (400MHz, DMSO-d6) δ 12.23 (s, 1H), 10.96 (s, 1H), 7.63–7.54 (m, 3H), 7.53–7.47 (m, 2H), 7.43 (d, J= 1.2Hz, 1H), 7.20 (dd, J=8.2, 1.5Hz, 1H), 7.13 (d, J=8.5Hz, 2H), 6.89 (d, J=8.5Hz, 2H), 5.83 (d, J= 8.2Hz, 1H), 3.77(s, 3H), 3.20–3.01(m, 5H), 2.72(s, 2H).

ESI-MS m/z: 614.7[MH] ^- .

Example 13 Methyl-(Z)-3-(((4-(2-(4-(fluoromethyl)piperazine-1-substituted-2,2,3,3,5,5,6,6 Preparation of -d8)-N-methylacetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6013)

The synthesis steps were performed according to the method described in Example 1, except that N-Boc-2,3,5,6-octadeuteropiperazine was used to replace the raw material N-Boc-piperazine to synthesize compound ZLF6013. The yield of the last step was 69%. Among them, N-Boc-2,3,5,6-octadeuteropiperazine is commercially available.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.63-7.54(m, 3H), 7.50(d, J=7.1Hz, 2H), 7.42( d,J＝1.3Hz,1H),7.20(d,J＝8.2Hz,1H),7.14(d,J＝8.4Hz,2H),6.89(d,J＝8.4Hz,2H),5.83(d, J=8.2Hz, 1H), 4.55(t, J=4.9Hz, 1H), 4.43(t, J=4.9Hz, 1H), 3.77(s, 3H), 2.71(s, 2H), 2.59(t, J=4.8Hz, 1H), 2.53(s, 1H).

ESI-MS m/z: 581.7[MH] ^- .

Example 14 Methyl-(Z)-3-(((4-(2-(4-(difluoromethyl)piperazine-1-substituted-2,2,3,3,5,5,6, Preparation of 6-d8)-N-methylacetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6014)

The synthesis procedure was as described in Example 1, except that the raw material N-Boc-piperazine was replaced with N-Boc-2,3,5,6-octadeuteropiperazine, and The compound ZLF6014 was synthesized by replacing the raw material 1-fluoro-2-iodoethane with 2-iodoethane. The yield of the last step was 62%. Among them, N-Boc-2,3,5,6-octadeuteropiperazine is commercially available.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.64-7.53(m, 3H), 7.52-7.47(m, 2H), 7.42(d, J ＝1.4Hz,1H),7.20(dd,J＝8.2,1.6Hz,1H),7.14(d,J＝8.5Hz,2H),6.89(d,J＝8.6Hz,2H),6.09(tt,J =55.8,4.3Hz,1H),5.83(d,J=8.2Hz,1H),3.77(s,3H),2.66(td,J=15.7,4.2Hz,4H).

ESI-MS m/z: 599.7[MH] ^- .

Example 15 Methyl-(Z)-3-(((4-(2-(4-(trifluoromethyl)piperazine-1-substituted-2,2,3,3,5,5,6, Preparation of 6-d8)-N-methylacetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6015)

The synthetic steps were as described in Example 1, except that the raw material N-Boc-piperazine was replaced with N-Boc-2,3,5,6-octadeuteriopiperazine, and the The raw material 1-fluoro-2-iodoethane was replaced with alkane to synthesize compound ZLF6015. The yield of the last step was 62%. Among them, N-Boc-2,3,5,6-octadeuteropiperazine is commercially available.

¹ H NMR (400MHz, DMSO-d6) δ 12.23 (s, 1H), 10.96 (s, 1H), 7.63–7.54 (m, 3H), 7.53–7.47 (m, 2H), 7.43 (d, J= 1.2Hz, 1H), 7.20 (dd, J=8.2, 1.5Hz, 1H), 7.13 (d, J=8.5Hz, 2H), 6.89 (d, J=8.5Hz, 2H), 5.83 (d, J= 8.2Hz, 1H), 3.77(s, 3H), 3.17–3.01(m, 2H), 2.72(s, 2H).

ESI-MS m/z: 617.7[MH] ^- .

Example 16 Methyl-(Z)-3-(((4-(2-(4-(2-(2-methoxyethyletheryl)ethyl)piperazine-1-substituted-2,2,3 Preparation of ,3,5,5,6,6-d8)-N-methylacetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6016)

The synthesis steps were in accordance with the method described in Example 1, except that the raw material N-Boc-piperazine was replaced with N-Boc-2,3,5,6-octadeuteropiperazine, and the raw material N-Boc-piperazine was replaced with 1-iodo-2(2 -Methoxyethyl ether)ethane replaces the raw material 1-fluoro-2-iodoethane to synthesize compound ZLF6016. The yield of the last step was 62%. Among them, N-Boc-2,3,5,6-octadeuteropiperazine is commercially available.

¹ H NMR (400MHz, DMSO-d6) δ 12.23 (s, 1H), 10.96 (s, 1H), 7.65–7.54 (m, 3H), 7.53–7.47 (m, 2H), 7.42 (d, J= 1.3Hz, 1H), 7.20 (dd, J=8.2, 1.5Hz, 1H), 7.13 (d, J=8.5Hz, 2H), 6.89 (d, J=8.6Hz, 2H), 5.83 (d, J= 8.2Hz, 1H), 3.77(s, 3H), 3.53-3.44(m, 4H), 3.44-3.37(m, 2H), 3.23(s, 3H), 2.71(s, 2H), 2.42(s, 2H) ).

ESI-MS m/z: 637.8[MH] ^-

Example 17 Methyl-(Z)-3-(((4-(fluoromethyl)piperazine-1-substituted-2,2,3,3,5,5,6,6-d8)-N- Preparation of (methyl-d3)acetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6017)

The synthesis steps were in accordance with the method described in Example 1, except that the raw material N-Boc-piperazine was replaced with N-Boc-2,3,5,6-octadeuteropiperazine, and the raw material N-Boc-piperazine was replaced with N-trideuteriomethyl Nitroaniline was used to replace N-methyl-p-nitroaniline to synthesize compound ZLF6017. The yield of the last step was 62%. Among them, N-Boc-2,3,5,6-octadeuteropiperazine is commercially available.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.63-7.54(m, 3H), 7.50(d, J=7.1Hz, 2H), 7.42( d,J＝1.3Hz,1H),7.20(d,J＝8.2Hz,1H),7.14(d,J＝8.4Hz,2H),6.89(d,J＝8.4Hz,2H),5.83(d, J=8.2Hz, 1H), 4.55(t, J=4.9Hz, 1H), 4.43(t, J=4.9Hz, 1H), 3.07(s, 3H), 2.71(s, 2H), 2.59(t, J=4.8Hz, 1H), 2.53(s, 1H).

ESI-MS m/z: 581.7[MH] ^- .

Example 18 Methyl-(Z)-3-(((4-(difluoromethyl)piperazine-1-substituted-2,2,3,3,5,5,6,6-d8)-N Preparation of -(methyl-d3)acetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6018)

The synthesis steps were as described in Example 1, except that the raw material N-Boc-piperazine was replaced with N-Boc-2,3,5,6-octadeuteropiperazine, and the The raw material 1-fluoro-2-iodoethane was replaced with alkane, and the compound ZLF6018 was synthesized with N-trideuteromethyl-p-nitroaniline instead of N-methyl-p-nitroaniline. The yield of the last step was 62%. Among them, N-Boc-2,3,5,6-octadeuteropiperazine is commercially available.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.64-7.53(m, 3H), 7.52-7.47(m, 2H), 7.42(d, J ＝1.4Hz,1H),7.20(dd,J＝8.2,1.6Hz,1H),7.14(d,J＝8.5Hz,2H),6.89(d,J＝8.6Hz,2H),6.09(tt,J =55.8,4.3Hz,1H),5.83(d,J=8.2Hz,1H),3.06(s,3H),2.66(td,J=15.7,4.2Hz,4H).

ESI-MS m/z: 599.7[MH] ^- .

Example 19 Methyl-(Z)-3-(((4-(trifluoromethyl)piperazine-1-substituted-2,2,3,3,5,5,6,6-d8)-N Preparation of -(methyl-d3)acetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6019)

The synthetic steps were as described in Example 1, except that the raw material N-Boc-piperazine was replaced with N-Boc-2,3,5,6-octadeuteriopiperazine, and the The raw material 1-fluoro-2-iodoethane was replaced by alkane, and the compound ZLF6019 was synthesized by N-trideuteromethyl-p-nitroaniline instead of N-methyl-p-nitroaniline. The yield of the last step was 62%. Among them, N-Boc-2,3,5,6-octadeuteropiperazine is commercially available.

¹ H NMR (400MHz, DMSO-d6) δ 12.23 (s, 1H), 10.96 (s, 1H), 7.63–7.54 (m, 3H), 7.53–7.47 (m, 2H), 7.43 (d, J= 1.2Hz, 1H), 7.20 (dd, J=8.2, 1.5Hz, 1H), 7.13 (d, J=8.5Hz, 2H), 6.89 (d, J=8.5Hz, 2H), 5.83 (d, J= 8.2Hz, 1H), 3.20–3.01(m, 5H), 2.72(s, 2H).

ESI-MS m/z: 617.7[MH] ^- .

Example 20 Methyl-(Z)-3-(((4-(2-(4-(2-(2-methoxyethyl etheryl)ethyl)piperazine-1-substituted-2,2,3 ,3,5,5,6,6-d8)-N-(methyl-d3)acetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6020 ) preparation

The synthesis steps were in accordance with the method described in Example 1, except that the raw material N-Boc-piperazine was replaced with N-Boc-2,3,5,6-octadeuteropiperazine, and the raw material N-Boc-piperazine was replaced with 1-iodo-2( The raw material 1-fluoro-2-iodoethane was replaced by 2-methoxyethyl ether)ethane, and the compound ZLF6020 was synthesized by N-trideuteromethyl-p-nitroaniline instead of N-methyl-p-nitroaniline. The yield of the last step was 62%. Among them, N-Boc-2,3,5,6-octadeuteropiperazine is commercially available.

¹ H NMR (400MHz, DMSO-d6) δ 12.23 (s, 1H), 10.96 (s, 1H), 7.65–7.54 (m, 3H), 7.53–7.47 (m, 2H), 7.42 (d, J= 1.3Hz, 1H), 7.20 (dd, J=8.2, 1.5Hz, 1H), 7.13 (d, J=8.5Hz, 2H), 6.89 (d, J=8.6Hz, 2H), 5.83 (d, J= 8.2Hz, 1H), 3.77(s, 3H), 3.53-3.44(m, 4H), 3.44-3.37(m, 2H), 3.06(s, 3H), 2.71(s, 2H), 2.42(s, 2H) ).

ESI-MS m/z: 637.8[MH] ^- .

Example 21 Methyl-d3-(Z)-3-(((4-(fluoromethyl)piperazine-1-substituted-2,2,3,3,5,5,6,6-d8)- Preparation of N-methylacetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6021)

The synthesis steps are in accordance with the method described in Example 1, except that the raw material methanol in step 1 is replaced with deuterated methanol, and the raw material N-Boc-2,3,5,6-octadeuteriopiperazine is used to replace the raw material N- Boc-piperazine, synthesized compound ZLF6021. The yield of the last step was 62%. Among them, N-Boc-2,3,5,6-octadeuteropiperazine is commercially available.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.63-7.54(m, 3H), 7.50(d, J=7.1Hz, 2H), 7.42( d,J＝1.3Hz,1H),7.20(d,J＝8.2Hz,1H),7.14(d,J＝8.4Hz,2H),6.89(d,J＝8.4Hz,2H),5.83(d, J=8.2Hz, 1H), 4.55(t, J=4.9Hz, 1H), 4.43(t, J=4.9Hz, 1H), 2.71(s, 2H), 2.59(t, J=4.8Hz, 1H) ,2.53(s,1H),2.31(s,4H),2.22(s,4H).

ESI-MS m/z: 576.7[MH] ^- .

Example 22 Methyl-d3-(Z)-3-(((4-(difluoromethyl)piperazine-1-substituted-2,2,3,3,5,5,6,6-d8) Preparation of -N-methylacetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6022)

The synthesis steps are in accordance with the method described in Example 1, except that the raw material methanol in step 1 is replaced with deuterated methanol, and the raw material N-Boc-2,3,5,6-octadeuteriopiperazine is used to replace the raw material N- Boc-piperazine, the raw material 1-fluoro-2-iodoethane was replaced with difluoro-2-iodoethane to synthesize compound ZLF6022. The yield of the last step was 62%. Among them, N-Boc-2,3,5,6-octadeuteropiperazine is commercially available.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.64-7.53(m, 3H), 7.52-7.47(m, 2H), 7.42(d, J ＝1.4Hz,1H),7.20(dd,J＝8.2,1.6Hz,1H),7.14(d,J＝8.5Hz,2H),6.89(d,J＝8.6Hz,2H),6.09(tt,J =55.8,4.3Hz,1H),5.83(d,J=8.2Hz,1H),3.06(s,3H),2.66(td,J=15.7,4.2Hz,4H).

ESI-MS m/z: 599.7[MH] ^- .

Example 23 Methyl-d3-(Z)-3-(((4-(trifluoromethyl)piperazine-1-substituted-2,2,3,3,5,5,6,6-d8) Preparation of -N-methylacetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6023)

The synthesis steps are in accordance with the method described in Example 1, except that the raw material methanol in step 1 is replaced with deuterated methanol, and the raw material N-Boc-2,3,5,6-octadeuteriopiperazine is used to replace the raw material N- Boc-piperazine, the starting material 1-fluoro-2-iodoethane was replaced with trifluoro-2-iodoethane to synthesize compound ZLF6023. The yield of the last step was 62%. . Among them, N-Boc-2,3,5,6-octadeuteropiperazine is commercially available.

¹ H NMR (400MHz, DMSO-d6) δ 12.23 (s, 1H), 10.96 (s, 1H), 7.63–7.54 (m, 3H), 7.53–7.47 (m, 2H), 7.43 (d, J= 1.2Hz, 1H), 7.20 (dd, J=8.2, 1.5Hz, 1H), 7.13 (d, J=8.5Hz, 2H), 6.89 (d, J=8.5Hz, 2H), 5.83 (d, J= 8.2Hz, 1H), 3.20–3.01(m, 5H), 2.72(s, 2H).

ESI-MS m/z: 617.7[MH] ^- .

Example 24 Methyl-d3-(Z)-3-(((4-(2-(4-(2-(2-methoxyethyletheryl)ethyl)piperazine-1-substituted-2,2 ,3,3,5,5,6,6-d8)-N-methylacetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6024) preparation

The synthesis steps are in accordance with the method described in Example 1, except that the raw material methanol in step 1 is replaced with deuterated methanol, and the raw material N-Boc-2,3,5,6-octadeuteriopiperazine is used to replace the raw material N- Boc-piperazine, synthesized compound ZLF6024. The yield of the last step was 62%, and the starting material 1-fluoro-2-iodoethane was replaced with 1-iodo-2(2-methoxyethyl ether)ethane. Among them, N-Boc-2,3,5,6-octadeuteropiperazine is commercially available.

¹ H NMR (400MHz, DMSO-d6) δ 12.23 (s, 1H), 10.96 (s, 1H), 7.65–7.54 (m, 3H), 7.53–7.47 (m, 2H), 7.42 (d, J= 1.3Hz, 1H), 7.20 (dd, J=8.2, 1.5Hz, 1H), 7.13 (d, J=8.5Hz, 2H), 6.89 (d, J=8.6Hz, 2H), 5.83 (d, J= 8.2Hz, 1H), 3.77(s, 3H), 3.53-3.44(m, 4H), 3.44-3.37(m, 2H), 3.06(s, 3H), 2.71(s, 2H), 2.42(s, 2H) ).

ESI-MS m/z: 637.8[MH] ^- .

Example 25 Methyl-d3-(Z)-3-(((4-(2-(4-(fluoromethyl)piperazine-1-substituted)-N-(methyl-d3)acetamido)aniline Preparation of methyl)(phenyl)methylene)-2-indolinone-6-carboxylate (ZLF6025)

The synthesis steps are in accordance with the method described in Example 1, except that the raw material methanol in step 1 is replaced with deuterated methanol, and N-trideuteromethyl-p-nitroaniline is used instead of N-methyl-p-nitroaniline, Compound ZLF6025 was synthesized. The yield of the last step was 62%.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.63-7.54(m, 3H), 7.50(d, J=7.1Hz, 2H), 7.42( d,J＝1.3Hz,1H),7.20(d,J＝8.2Hz,1H),7.14(d,J＝8.4Hz,2H),6.89(d,J＝8.4Hz,2H),5.83(d, J=8.2Hz, 1H), 4.55(t, J=4.9Hz, 1H), 4.43(t, J=4.9Hz, 1H), 2.71(s, 2H), 2.59(t, J=4.8Hz, 1H) ,2.53(s,1H),2.31(s,4H),2.22(s,4H).

ESI-MS m/z: 576.7[MH] ^- .

Example 26 Methyl-d3-(Z)-3-(((4-(2-(4-(difluoromethyl)piperazine-1-substituted)-N-(methyl-d3)acetamido) Preparation of anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6026)

The synthesis steps are in accordance with the method described in Example 1, except that the raw material methanol in step 1 is replaced with deuterated methanol, and N-trideuteromethyl-p-nitroaniline is used instead of N-methyl-p-nitroaniline, The raw material 1-fluoro-2-iodoethane was replaced with difluoro-2-iodoethane to synthesize compound ZLF6026. The yield of the last step was 62%.

¹ H NMR (400MHz, DMSO-d ₆ ) δ 12.23(s, 1H), 10.96(s, 1H), 7.64-7.53(m, 3H), 7.52-7.47(m, 2H), 7.42(d, J ＝1.4Hz,1H),7.20(dd,J＝8.2,1.6Hz,1H),7.14(d,J＝8.5Hz,2H),6.89(d,J＝8.6Hz,2H),6.09(tt,J =55.8,4.3Hz,1H),5.83(d,J=8.2Hz,1H),2.66(td,J=15.7,4.2Hz,4H),2.39(s,4H),2.22(s,4H).

ESI-MS m/z: 594.7[MH] ^- .

Example 27 Methyl-d3-(Z)-3-(((4-(2-(4-(trifluoromethyl)piperazine-1-substituted)-N-(methyl-d3)acetamido) Preparation of anilino)(phenyl)methylene)-2-indolinone-6-carboxylic acid methyl ester (ZLF6027)

The synthesis steps are in accordance with the method described in Example 1, except that the raw material methanol in step 1 is replaced with deuterated methanol, and N-trideuteromethyl-p-nitroaniline is used instead of N-methyl-p-nitroaniline, The starting material 1-fluoro-2-iodoethane was replaced with trifluoro-2-iodoethane to synthesize compound ZLF6027. The yield of the last step was 62%.

¹ H NMR (400MHz, DMSO-d6) δ 12.23 (s, 1H), 10.96 (s, 1H), 7.63–7.54 (m, 3H), 7.53–7.47 (m, 2H), 7.43 (d, J= 1.2Hz, 1H), 7.20 (dd, J=8.2, 1.5Hz, 1H), 7.13 (d, J=8.5Hz, 2H), 6.89 (d, J=8.5Hz, 2H), 5.83 (d, J= 8.2Hz, 1H), 3.18–3.01 (m, 2H), 2.72 (s, 2H), 2.48–2.39 (s, 4H), 2.23 (s, 4H).

ESI-MS m/z: 612.7[MH] ^- .

Example 28 Methyl-d3-(Z)-3-(((4-(2-(4-(2-(2-methoxyethyletheryl)ethyl)piperazine-1-substituted)-N- Preparation of (methyl-d3)acetamido)anilino)(phenyl)methylene)-2-indolone-6-carboxylic acid methyl ester (ZLF6028)

The synthesis steps are in accordance with the method described in Example 1, except that the raw material methanol in step 1 is replaced with deuterated methanol, and N-trideuteromethyl-p-nitroaniline is used instead of N-methyl-p-nitroaniline, The raw material 1-fluoro-2-iodoethane was replaced with 1-iodo-2(2-methoxyethyl ether)ethane to synthesize compound ZLF6028. The yield of the last step was 62%.

¹ H NMR (400MHz, DMSO-d6) δ 12.23 (s, 1H), 10.96 (s, 1H), 7.65–7.54 (m, 3H), 7.53–7.47 (m, 2H), 7.42 (d, J= 1.3Hz, 1H), 7.20 (dd, J=8.2, 1.5Hz, 1H), 7.13 (d, J=8.5Hz, 2H), 6.89 (d, J=8.6Hz, 2H), 5.83 (d, J= 8.2Hz, 1H), 3.77(s, 3H), 3.53-3.44(m, 4H), 3.44-3.37(m, 2H), 2.71(s, 2H), 2.42(s, 2H), 2.29(s, 4H) ),2.21(s,4H).

ESI-MS m/z: 632.8[MH] ^- .

The beneficial effects of the present invention are demonstrated below through specific test examples.

Test Example 1 Inhibitory activity test of the compounds of the present invention on VEGFR2, FGFR1 and PDGFRα kinases

The purpose of this experiment is to detect the inhibitory activity of the compounds of the invention on kinases in vitro,

1 Experimental material

20 mM 3-(N-morpholino)propanesulfonic acid (MOPS); 1 mM ethylenediaminetetraacetic acid (EDTA); 0.01% Brij-35; 5% Glycerol; 0.1% mercaptoethanol (mercptoethanol); 1 mg/ml bovine serum albumin (BSA); 10 mM manganese dichloride solution (MnC12); 10 mM magnesium acetate and y-33p-ATP solution; stop buffer (3% phosphate buffer) ; washing buffer (75 mM phosphate solution); methanol; Filtermat A membrane; VEGFR2, FGFR1 and PDGFRα kinases, compounds prepared in the Examples.

2 Experimental methods

Using isotope labeling method, VEGFR2, FGFR1 and PDGFRα kinase crystalline forms were tested for in vitro activity inhibition respectively. Have the following steps:

In a reaction tube, add buffer (8mM MOPS, pH 7.0, 0.2mM EDTA, 10mM MnC12), test kinase (5-10mU), test kinase substrate, and 10mM magnesium acetate and γ33P-ATP in sequence solutions, test compounds at various concentrations. The reaction was initiated by the addition of MgATP (the final concentration of ATP is the Km value of the corresponding kinase) and incubated at room temperature for 40 minutes. The reaction was finally terminated with 5 μL of 3% phosphate buffer and 10 μL of the reaction was titrated to The filtermat A membrane was washed three times with 75mM phosphate solution for 5 minutes each, and then washed once with methanol. Finally, the filtermat A membrane was dried and counted. The magnitude of the scintillation count reflects the phosphorylated substrate. The degree of inhibition of kinase activity can be characterized. BIBF1120 is used as a positive control.

The kinase inhibitory activity of the test compound was expressed by the half inhibitory concentration (IC ₅₀ ), and the IC ₅₀ value was obtained by calculating the inhibitory rate of the test compound on the kinase activity at different concentrations.

3 Experimental results

Through the above experimental methods, the IC ₅₀ values of the compounds obtained in the examples of the present invention against VEGFR2, FGFR1 and PDGFRα kinase inhibitory activities were tested, and the results are shown in Table 1.

Table 1 _IC50 values of compounds for VEGFR2, FGFR1 and PDGFRα kinase inhibitory activities

In Table 1, A: 1-100 nM, B: 100-500 nM, C: >500 nM.

The experimental results show that the compounds of the present invention have strong inhibitory activities on VEGFR2, FGFR1 and PDGFRα.

Test Example 2 In vivo antitumor experiment of the compound ZLF 6004 of the present invention

1 Experimental material

Fetal bovine serum, culture medium, trypsin, etc. were purchased from Gibco BRL company (Invitrogen Corporation, USA), culture medium was purchased from ATCC (American Type Culture Collection), human pharyngeal squamous cell carcinoma cell line FaDu was purchased from American ATCC company, NOD- Balb/c mice were purchased from Beijing Huafukang Animal Experiment Center.

2 Experimental methods

Using 6-8 weeks old NOD-Balb/c mice, human pharyngeal squamous cell carcinoma FaDu cells were inoculated into the subcutaneous posterior flanks of mice at a concentration of about 1×10 ⁷ cells/0.1ml/mice, and the tumors grew to After a certain volume, mice were randomized and administered orally by gavage.

FaDu experimental groups: (1) low-dose compound ZLF6004, 50 mg/kg q.d; (2) high-dose compound ZLF6004, 1000 mg/kg q.d; (3) solvent control group. The solvent is a mixed solvent of 12.5% EL (castor oil polyoxyethylene ether) + 12.5% EtOH + 75% water. 8 mice per group.

Observation indicators: The long diameter and short neck of the tumor were measured every 3 days, and the tumor volume (long diameter × short diameter ² × 0.52) was calculated, and the average value was calculated; and the mice were observed for diarrhea, cramps, rash, and body weight. significantly reduced the response.

3 Experimental results

During the administration, no adverse reactions such as weight loss, rash and diarrhea were found in the mice, indicating that the compound ZLF6004 of the present invention has very low toxicity within the administration dose range under the test dose.

The tumor growth curves of each group measured in the experiment are shown in Figure 1. The experimental results show that the compound ZLF6004 of the present invention has an obvious growth inhibitory effect on pharyngeal squamous cell carcinoma tumors in vivo.

Test Example 3 Pharmacokinetic Experiment of Compound ZLF 6004

1 Experimental method

Male SD rats were randomly divided into BIBF1120 oral administration group and 6004 oral administration group, with 8 rats in each group, and the administration dose was 50 mg/kg; 0.25h, 0.5h, 1h, 2h, 4h after administration , 6h, 8h, 10h, 24h sequentially collect blood samples for processing, using LC/MS detection. The blood drug concentration-time data obtained by detection is used to calculate the pharmacokinetic parameters as shown in Table 2 below using Phoenix WinNonlin7.0.

2 Experimental results

Table 2 Pharmacokinetic properties test results

It can be seen from the results in Table 2 that, compared with BIBF1120, the half-life (T _1/2 ), time to peak (T _max ), maximum blood concentration (C _max ) and area under the curve of the compound ZLF6004 of the present invention are all obvious increase, indicating that the compound ZLF6004 of the present invention has significantly improved pharmacokinetic properties than BIBF1120.

In conclusion, the present invention provides a compound represented by formula (A) and a pharmaceutically acceptable salt thereof. The experimental results show that the compounds provided by the present invention have significantly improved pharmacokinetic properties than BIBF1120, and have simultaneous effects on vascular endothelial growth factor receptor (VEGFR), fibroblast growth factor receptor (FGFR) and platelet-derived growth factor receptor. (PDGFR) all have excellent inhibitory effects, and can be used as VEGFR, FGFR and/or PDGFR inhibitors, as angiogenesis inhibitors, as drugs for the prevention and/or treatment of various tumors including pharyngeal squamous cell carcinoma, with a broad range of application prospects.

Claims (10)

A compound represented by formula (A) or a pharmaceutically acceptable salt thereof:

Wherein, R ₁ and R ₂ are independently selected from C1-C6 alkyl and deuterated C1-C6 alkyl; R ₃ is selected from C1-C6 alkyl, halogenated C1-C6 alkyl,

R ₄ to R ₁₁ are each independently selected from H or deuterium; And when R ₂ is methyl, R ₃ is methyl, and R ₄ to R ₁₁ are hydrogen, R ₁ is not methyl. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ are independently selected from C1-C3 alkyl and deuterated C1-C3 alkyl; R ₃ is selected from C1-C2 alkyl, halogenated C1-C2 alkyl,

The halogen is preferably fluorine. The compound of claim 2 or a pharmaceutically acceptable salt thereof, wherein R ₁ and R ₂ are each independently selected from methyl or deuterated methyl, and the deuterated methyl is preferably - _CD3 ; R ₃ is selected from CF ₃ , CHF ₂ , CH ₂ F, CH ₂ CF ₃ , CH ₂ CHF ₂ , CH ₂ CH ₂ F,

R ₄ to R ₁₁ are all H or all D. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, wherein the compound is selected from one of the following compounds:

The method for preparing a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, characterized in that: the method comprises using compound (A-1) and compound (A-2) as raw materials to carry out Reaction to obtain the compound represented by formula (A);

wherein R ₁ to R ₁₁ are as described in any one of claims 1 to 4 . The preparation method according to any one of claims 5, characterized in that: the molar ratio of compound (A-1) and compound (A-2) is 1:1; the solvent of the reaction is methanol. An antitumor drug is characterized in that: it is a preparation prepared by using the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof as an active ingredient, and adding auxiliary materials commonly used in the pharmaceutical field. The use of the compound described in any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof in the preparation of a VEGFR, FGFR and/or PDGFR inhibitor; the VEGFR is preferably VEGFR2, the FGFR is preferably FGFR1, and the VEGFR is preferably FGFR1. PDGFR is preferably PDGFRα. Use of the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof in the preparation of an angiogenesis inhibitor. Use of the compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing and/or treating tumors, the tumors are preferably liver cancer, lung cancer, rectal cancer, uterine cancer, and brain cancer Metastatic bowel cancer, pharyngeal squamous cell carcinoma.

Images