CN103922992B - A kind of antitumour activity indolone derivatives, preparation method and use - Google Patents

Abstract

The present invention relates to a kind of indolone derivatives, described derivative under catalyzer, part and additive exist, can be obtained by cyanoindole ketone compound and boronic acid compounds one-step synthesis.Simultaneously the described derivative of Late Cambrian has excellent antitumour activity, through active testing, find that it has good cell tumour growth-inhibiting effect to human stomach cancer cell line (SGC-7901), human lung carcinoma cell line (H446) and human stomach cancer cell line (HGC-27), thus antitumor drug can be used as in field of medicaments, there is good medical research prospect and industrial application value.

Description

A kind of anticancer active indolinone derivative, its synthesis method and its application

technical field

The invention relates to a pharmaceutically active compound, in particular to an indolinone derivative with excellent anticancer activity, a synthesis method and its use, and belongs to the field of medicinal chemistry.

Background technique

Indolinone derivatives are an important class of nitrogen-containing and oxygen-containing organic compounds, which play an important role in organic and pharmaceutical synthesis. Therefore, it has attracted much attention in the fields of pharmaceutical chemical industry, organic synthesis and material chemistry.

So far, a variety of pharmaceutical compounds having an indolinone structure have been developed. For example, 7-benzoylindol-2-one is usually used to synthesize non-steroidal anti-inflammatory analgesic drug Amfenac sodium (amfenac sodium); and 7-(p-bromobenzoyl)indole-2- Ketones are commonly used to synthesize the non-steroidal anti-inflammatory analgesic drug Bromfenac sodium (bromfenac sodium):

It is precisely because of the important role of indolinone derivatives that people have carried out a lot of research on their synthesis and developed a variety of synthetic methods, mainly including the following chemical synthesis methods:

In 1979, William J.Welstead, Jr. et al. reported the synthetic method shown in the following formula. This method first uses N-aminoindol-2-one as starting material, and in the mixed solvent of acetic acid and ethanol with benzene The dehydration condensation reaction of ethyl methyl ketone generates imine intermediate I; the rearrangement and ring closure reaction in the molecule of imine intermediate I under acidic conditions construct 2-(2-methyl-3 -Phenyl-7 indolyl ethyl acetate intermediate II; then use ozone as an oxidizing agent to oxidize intermediate II to generate intermediate III after indole ring opening; finally, intermediate III carries out intramolecular decomposition under acidic conditions Cyclization reaction obtains target product 7-benzoyl indol-2-ketone (3a).This method needs to go through four-step reaction and just can synthesize 7-benzoyl and indol-2-ketone, and technique is comparatively complicated, and its total income The rate is low; and ozone has been used in the reaction process, which requires special equipment and complicated operations. Its reaction formula is as follows:

In 1984, David A. Walsh and others reported the synthesis method of 7-benzoyl and indol-2-one: first, at ^-70 °C, 2-methylthioglycolate ethyl ester and 2-amino The reaction of diphenylindolinone generates sulfur salt (IV); followed by a rearrangement reaction in the presence of alkali (such as triethylamine) to generate 3-substituted 2-aminodiphenylindolinone intermediate V; in acidic conditions The following intermediate V undergoes an intramolecular cyclization reaction to generate 7-benzoyl-3-(methylmercapto)indol-2-one (VI); finally 7-benzoyl-3-(methylmercapto)indole -2-one (VI) was reduced in the presence of Raney nickel to give the target product 7-benzoylindol-2-one (3a). This method also needs to undergo four-step reactions to synthesize 7-benzoyl and indol-2-ones. The process is relatively complicated, and its total yield is low, and it needs to be at low temperature (at -70°C) during the first step reaction. The reaction conditions are relatively harsh. Its reaction formula is as follows:

In 1992, R.G.Poser reported a new method for the synthesis of 7-aryl indol-2-ketone derivatives: using p-bromobenzonitrile and indoline as raw materials, using boron trichloride and aluminum trichloride as Catalyst, carry out Friede-Crafts acylation reaction, generate 7-(p-bromobenzoyl) indoline (VII); Then generate 7-(p-bromobenzoyl) indole (VIII) through manganese dioxide oxidation reaction; Then use N-chlorosuccinimide (NCS) as a chlorination reagent to carry out chlorination reaction to generate chloro 7-(p-bromobenzoyl)indole (IV); finally carry out acidolysis reaction in the presence of phosphoric acid , to generate the target product 7-(p-bromobenzoyl)indol-2-one (3b). But this method still needs four-step reactions to realize the synthesis of 7-(p-bromobenzoyl)indol-2-one (3b), so the overall yield is low, about 28%. Its reaction formula is as follows:

Recently, Xia Zekuan, Shandong Institute of Pharmaceutical Industry and others have improved the above reaction process, using self-made active manganese dioxide as an oxidant, and the yield of the oxidation product 7-(p-bromobenzoyl)indole (VIII) can be increased to 92.4% (document 85%); In addition, also changed the aftertreatment method of chlorination reaction, got rid of diethyl ether, increased dichloromethane extraction, the yield of chlorinated 7-(p-bromobenzoyl) indole (IV) There is a significant increase to 97.9% (64% in literature). Although this method improves the overall yield of 7-(p-bromobenzoyl)indol-2-one (3b), it is still very low, about 33%.

But so far, people have not found that indolinone derivatives have anticancer activity, and there is still no clear understanding of their active targets and substituents. But on the other hand, due to the excellent biological activity of indolinone compounds, it is still the current research method and the Emphasis, this is also the basis and motivation for the present invention to be accomplished.

Contents of the invention

In view of this, in order to find novel indolinone derivatives with excellent pharmaceutical activity and biological activity, the present inventors conducted in-depth research and completed the present invention after devoting a lot of creative work.

The present invention relates to a drug with excellent anti-cancer activity, especially good cell tumor growth inhibitory effect on human gastric cancer cell line (SGC-7901), human lung cancer cell line (H446) and human gastric cancer cell line (HGC-27). The indolinone derivative also relates to a synthesis method and application of the derivative.

Specifically, in the first aspect, the present invention relates to an indolinone derivative shown in formula (I) having excellent anticancer activity:

Each R is independently selected from H, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, halogen, halogenated C ₁ -C ₆ alkyl or halogenated C ₁ -C ₆ alkoxy _;

n is independently an integer of 0-5.

Preferably, the indolinone derivative is preferably a compound of the following formula (II):

Wherein, R ₁ and n have the same definition as above.

In the indolinone derivatives of the present invention, unless otherwise specified, throughout, the meaning of C ₁ -C ₆ alkyl refers to a straight chain or branched chain alkyl group with 1-6 carbon atoms, which includes C ₁ alkyl, C ₂ alkyl, C ₃ alkyl, C ₄ alkyl, C ₅ alkyl or C ₆ alkyl, non-limiting examples may be methyl, ethyl, n-propyl, isopropyl , n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl or n-hexyl, etc.

In the indolinone derivatives of the present invention, unless otherwise specified, throughout, C ₁ -C ₆ alkoxy refers to the group after the above-mentioned "C ₁ -C ₆ alkyl" is connected to an O atom .

In the indolinone derivatives of the present invention, unless otherwise specified, halogen or halogen in halogeno can be, for example, F, Cl, Br or I throughout.

In the indolinone derivatives of the present invention, unless otherwise specified, throughout, the meaning of halogenated C ₁ -C ₆ alkyl refers to the above-defined "C ₁ -C ₆ alkyl" substituted by halogen, Non-limiting examples are trifluoromethyl, pentafluoroethyl, difluoromethyl, chloromethyl and the like.

In the indolinone derivatives of the present invention, unless otherwise specified, throughout, the meaning of halogenated C ₁ -C ₆ alkoxy refers to the above-mentioned "C ₁ -C ₆ alkoxy" substituted by halogen. ", non-limiting examples are trifluoromethoxy, pentafluoroethoxy, difluoromethoxy, chloromethoxy and the like.

In the indolinone derivatives of the present invention, n is an integer of 0-5, for example, it can be 0, 1, 2, 3, 4 or 5, that is, the benzene ring can be replaced by 0, 1, 2, 3, 4 or 5 R ₁ that are each the same or different are substituted.

Among the indolinone derivatives of the present invention, R ₁ is preferably C ₁ -C ₆ alkoxy, most preferably methoxy.

In the indolinone derivatives of the present invention, n is preferably an integer of 1-3, most preferably 3.

Among the indolinone derivatives of the present invention, the compound of formula (I) is preferably the compound of the following formula (III), (VI) or (V):

Among the indolinone derivatives of the present invention, the compound of formula (I) is most preferably the compound of the following formula (V):

In a second aspect, the present invention relates to a synthetic method of the above-mentioned indolinone derivatives, the method comprising: in the presence of a palladium catalyst, a ligand and an additive, in a solvent, formula (VI) compound and formula (VII) Reaction, thereby obtain the indolinone derivative of formula (I);

Wherein, R ₁ and n are as defined above.

In the synthetic method of indolinone derivative of the present invention, when formula (VI) is following formula (VIII), just can obtain formula (II) compound:

In the synthesis method of indolinone derivatives of the present invention, the palladium catalyst is palladium trifluoroacetate (Pd(O ₂ CCF ₃ ) ₂ ), palladium acetate (Pd(OAc) ₂ ), PdCl ₂ , PdCl ₂ ( PPh ₃ ) ₂ , PdCl ₂ (dppf) ₂ , PdCl ₂ (dppe), PdCl ₂ (cod), PdCl ₂ (py) ₂ , PdCl ₂ (MeCN) ₂ , Pd(acac) ₂ , Pd(PPh ₃ ) ₄ , Pd ₂ (dba) ₃ , a mixture of any one or more of Pd ₂ (dba) ₂ ; preferably palladium trifluoroacetate, palladium acetate or PdCl ₂ ; most preferably palladium trifluoroacetate.

In the synthesis method of the indolinone derivatives of the present invention, the ligand is a bidentate nitrogen-containing ligand or a phosphine ligand, for example, any one or a mixture of any of the following L1-L12:

Among them, L1-L9 is preferred, L1, L2, L3, and L8 are more preferred, and L2 is most preferred.

In the synthesis method of indolinone derivatives of the present invention, the additive is an acid, such as trifluoroacetic acid, benzoic acid, trifluoromethanesulfonic acid, p-toluenesulfonic acid, p-nitrobenzenesulfonic acid, methanesulfonic acid acid, sulfuric acid, hydrochloric acid or potassium bifluoride; preferably trifluoroacetic acid, p-toluenesulfonic acid, p-nitrobenzenesulfonic acid or methanesulfonic acid; most preferably methanesulfonic acid.

In the synthesis method of the indolinone derivatives of the present invention, the molar ratio of the compound of formula (VI) to the compound of (VII) can be properly selected, for example, the molar ratio can be 1:1-3, for example Can be 1:1, 1:1.5, 1:2, 1:2.5 or 1:3.

In the synthesis method of indolinone derivatives of the present invention, the molar ratio of the compound of formula (VI) to the catalyst is 1:0.02-0.1, such as 1:0.02, 1:0.04, 1:0.06, 1:0.02 0.08 or 1:0.1.

In the synthesis method of indolinone derivatives of the present invention, the molar ratio of the compound of formula (VI) to the ligand is 1:0.05-0.1, for example, 1:0.05, 1:0.07 or 1:0.1.

In the synthesis method of indolinone derivatives of the present invention, the molar ratio of the compound of formula (VI) to the additive is 1:5-15, for example, 1:5, 1:10 or 1:15.

In the synthesis method of the indolinone derivatives of the present invention, the reaction temperature is 60-120°C, for example, non-limitingly 60°C, 70°C, 80°C, 90°C, 100°C, 110°C or 120°C.

In the synthesis method of the indolinone derivatives of the present invention, the solvent is a mixture of an organic solvent and water, wherein the organic solvent can be, for example, tetrahydrofuran (THF), 2-methyltetrahydrofuran (2-MeTHF), toluene , xylene, dioxane, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), isopropanol, ethanol, n-butanol, chlorobenzene, etc., organic solvents and water The volume ratio is 1-3:1, for example 1:1, 2:1 or 3:1, preferably 2:1.

In the synthesis method of the indolinone derivatives of the present invention, the reaction time is not particularly limited, for example, the appropriate reaction time can be determined by tracking the residual percentage of the target product or raw material through TLC or GC-MS, which is usually 15 - 40 hours, such as non-limitatively 15 hours, 17 hours, 19 hours, 21 hours, 23 hours, 25 hours, 27 hours, 29 hours, 30 hours, 32 hours, 34 hours, 36 hours, 38 hours or 40 hours Hour.

In the synthesis method of the present invention, the post-treatment after the reaction can be any one or a combination of treatment means in crystallization, recrystallization, column chromatography purification, extraction and the like. As an exemplary post-treatment method, for example, after the reaction, add water to the reaction mixture, extract 2-4 times with ether, combine the ether layers, and dry with anhydrous sodium sulfate, filter and evaporate to remove ether Concentrate, pass the concentrate through a 300-400 mesh silica gel column, and use a mixture of ethyl acetate/petroleum ether as an eluent, wherein the volume ratio of ethyl acetate to petroleum ether is 1:5-15, thereby obtaining the target product formula (I) Compounds.

In the third aspect, the present invention relates to the novel use of indolinone derivatives represented by the above formula (I) as anticancer drugs.

The present inventors have found that the indolinone derivatives shown in the above formula (I) have excellent anticancer activity, especially for human gastric cancer cell line (SGC-7901), human lung cancer cell line (H446) and human gastric cancer cell line ( HGC-27) has a good inhibitory effect on cell tumor growth, so it can be used as an antitumor drug in the medical field.

Wherein, the following formula (V) compound has the most preferred anticancer activity:

In the fourth aspect, the present invention relates to the use of indolinone derivatives represented by the above formula (I), especially the compounds of formulas (III)-(V) in the preparation of anticancer drugs.

The (I) indolinone derivatives, especially the compounds of formula (III)-(V), have good anticancer activity, especially for human gastric cancer cell line (SGC-7901), human lung cancer cell line (H446) and human Gastric cancer cell line (HGC-27) has excellent inhibitory effect.

In the fifth aspect, the present invention also relates to a pharmaceutical composition for treating cancer, especially lung cancer or gastric cancer, which comprises the indolinone derivative of formula (I) as an active ingredient, especially comprising formula (III)-(V) compound.

In addition to the above active compounds, the pharmaceutical composition may also contain pharmaceutically acceptable carriers, excipients and/or adjuvants.

In the present invention, "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not cause significant irritation to the organism and does not eliminate the biological activity and properties of the administered compound. Typically, these include properties deemed acceptable by the patient for pharmacological/toxicological considerations and by the pharmaceutical chemist for physical/chemical considerations of composition, formulation, stability, patient acceptance, and bioavailability and/or substances.

"Excipient" or "adjuvant" refers to an inert substance added to a pharmaceutical composition to further facilitate the administration of a compound, examples of which include, but are not limited to: calcium carbonate, calcium phosphate, various sugars, and various Starches, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders, disintegrants, and the like.

The indolinone derivatives of the formula (I) of the present invention, especially the compounds of the formulas (III)-(V) exhibit excellent biological activity in in vitro anticancer tests, so they have great development in the clinical treatment of tumor diseases potential and application prospects.

Detailed ways

The present invention will be described in detail below through specific examples, but the use and purpose of these exemplary embodiments are only used to exemplify the present invention, and do not constitute any form of any limitation to the actual protection scope of the present invention, nor will the present invention The scope of protection is limited to this.

Example 1: 7-(4-bromobenzoyl)-indol-2-one

At room temperature, add 100ml of solvent (a mixture of 2-MeTHF and water, the volume ratio of which is 2:1), 20mmol of 7-cyano-indol-2-one, 40mmol of p-bromophenylboronic acid, 1mmol of Palladium trifluoroacetate, 1.5 mmol of the above-mentioned ligand L2 and 200 mmol of methanesulfonic acid; the temperature was raised to 80° C. under stirring, and the reaction was kept for 35 hours.

After the reaction is finished, add 100ml of water to the reaction mixture, extract 2-4 times with ether, combine the ether layers, and dry with anhydrous sodium sulfate, filter, evaporate and remove the ether for concentration, pass the concentrate through a 300-400 mesh silica gel column , using a mixture of ethyl acetate/petroleum ether as the eluent, wherein the volume ratio of ethyl acetate to petroleum ether was 1:10, and the eluate was concentrated to obtain the title compound as a yellow solid, named A. Its yield was 74.8% with a purity of 98.9% (HPLC).

Melting point: 195-196°C.

NMR: ¹ H NMR (CDCl ₃ , 500MHz) δ9.49(s, 1H), 7.65(d, J=8.6Hz, 2H), 7.59(d, J=8.5Hz, 2H), 7.49(d, J =8.1Hz, 1H), 7.43(d, J=7.3Hz, 1H), 7.04(t, J=7.7Hz, 1H), 3.57(s, 2H);

¹³ C NMR (CDCl ₃ , 125 MHz) δ195.6, 176.4, 145.7, 136.7, 131.7 (2C), 130.9 (2C), 130.7, 129.2, 127.2, 121.1, 117.6, 112.0, 35.1.

Example 2: 7-(3-methoxybenzoyl)-indol-2-one

At room temperature, add 100ml of solvent (a mixture of 2-MeTHF and water, the volume ratio of which is 2:1), 20mmol of 7-cyano-indol-2-one, and 20mmol of m-methoxyphenylboronic acid to the reaction vessel , 0.4 mmol of palladium trifluoroacetate, 1 mmol of the above-mentioned ligand L2 and 100 mmol of methanesulfonic acid; the temperature was raised to 60° C. under stirring, and the reaction was kept for 40 hours.

After the reaction is finished, add 100ml of water to the reaction mixture, extract 2-4 times with ether, combine the ether layers, and dry with anhydrous sodium sulfate, filter, evaporate and remove the ether for concentration, pass the concentrate through a 300-400 mesh silica gel column , using a mixture of ethyl acetate/petroleum ether as the eluent, wherein the volume ratio of ethyl acetate to petroleum ether was 1:5, and the eluate was concentrated to obtain the title compound as a yellow solid, named B. Its yield was 81.9% with a purity of 99.2% (HPLC).

Melting point: 131-133°C.

NMR: ¹ HNMR(CDCl ₃ ,500MHz)δ9.69(s,1H),7.56(d,J＝8.1Hz,1H),7.43-7.38(m,2H),7.29-7.26(m,2H), 7.14-7.12(m,1H),7.04(t,J=7.7Hz,1H),3.85(s,3H),3.57(s,2H);

¹³ C NMR (CDCl ₃ , 125 MHz) δ196.4, 176.8, 159.6, 145.5, 139.2, 131.0, 129.3, 129.0, 127.0, 121.9, 121.0, 118.3, 118.0, 114.0, 55.5, 35.1.

Example 3: 7-(3,4-dimethoxybenzoyl)-indol-2-one

At room temperature, add 100ml of solvent (a mixture of 2-MeTHF and water, the volume ratio of which is 2:1), 20mmol of 7-cyano-indol-2-one, 60mmol of 3,4-dimethoxy Phenylboronic acid, 1.5 mmol of palladium trifluoroacetate, 2 mmol of the above-mentioned ligand L2 and 300 mmol of methanesulfonic acid; the temperature was raised to 100° C. under stirring, and the reaction was kept for 30 hours.

After the reaction is finished, add 100ml of water to the reaction mixture, extract 2-4 times with ether, combine the ether layers, and dry with anhydrous sodium sulfate, filter, evaporate and remove the ether for concentration, pass the concentrate through a 300-400 mesh silica gel column , using a mixture of ethyl acetate/petroleum ether as the eluent, wherein the volume ratio of ethyl acetate to petroleum ether was 1:15, and the eluate was concentrated to obtain the title compound as a yellow solid, named as C. Its yield was 88.7% with a purity of 98.6% (HPLC).

Melting point: 154-156°C.

NMR: ¹ H NMR (CDCl ₃ , 500MHz): δ9.36(s, 1H), 7.57(d, J=8.8Hz, 1H), 7.40(d, J=7.3Hz, 1H), 7.36-7.34( m, 2H), 7.04(t, J=7.7Hz, 1H), 6.92(d, J=8.0Hz, 1H), 3.96(s, 3H), 3.93(s, 3H), 3.56(s, 2H);

¹³ C NMR (CDCl ₃ , 125 MHz) δ195.1, 176.5, 152.9, 149.1, 145.3, 130.6, 130.5, 128.5, 127.0, 124.5, 120.8, 118.4, 112.0, 109.9, 56.1, 56.1, 35.3.

Example 4: 7-(3,4,5-trimethoxybenzoyl)-indol-2-one

At room temperature, add 100ml of solvent (a mixture of 2-MeTHF and water, the volume ratio of which is 2:1), 20mmol of 7-cyano-indol-2-one, 30mmol of 3,4,5-trimethyl Oxyphenylboronic acid, 2 mmol of palladium trifluoroacetate, 1 mmol of the above-mentioned ligand L2 and 150 mmol of methanesulfonic acid; the temperature was raised to 120° C. under stirring, and the reaction was kept for 25 hours.

After the reaction is finished, add 100ml of water to the reaction mixture, extract 2-4 times with ether, combine the ether layers, and dry with anhydrous sodium sulfate, filter, evaporate and remove the ether for concentration, pass the concentrate through a 300-400 mesh silica gel column , a mixture of ethyl acetate/petroleum ether was used as the eluent, wherein the volume ratio of ethyl acetate to petroleum ether was 1:7, and the eluate was concentrated to obtain the title compound as a yellow solid, named D. Its yield was 62.8% with a purity of 98.5% (HPLC).

Melting point: 156-158°C.

NMR: ¹ H NMR (CDCl ₃ , 500MHz): δ9.45(s, 1H), 7.59(d, J=8.1Hz, 1H), 7.43(d, J=7.3Hz, 1H), 7.05(t, J=7.4Hz, 1H), 6.97(s, 2H), 3.93(s, 3H), 3.88(s, 6H), 3.58(s, 2H);

¹³ C NMR (CDCl ₃ , 125 MHz) δ195.7, 176.5, 153.0 (2C), 145.5, 141.8, 133.0, 130.7, 128.9, 127.1, 120.9, 118.0, 107.1 (2C), 61.0, 56.3 (2C), 35.2.

Comparative example 1-36

In addition to replacing the catalyst palladium trifluoroacetate with Pd(OAc) ₂ , PdCl ₂ , Pd ₂ (dba) ₃ , Pd(dba) ₂ , Pd(pyridyl) ₂ , Pd(NH ₃ ) ₄ Cl ₂ , In addition to bis(tri-tert-butylphosphine)palladium(0), Pd(acac) ₂ , and Pd(PPh ₃ ) ₄ , Comparative Examples 1-9 were implemented in the same manner as in Example 1;

In addition to replacing the catalyst palladium trifluoroacetate with Pd(OAc) ₂ , PdCl ₂ , Pd ₂ (dba) ₃ , Pd(dba) ₂ , Pd(pyridyl) ₂ , Pd(NH ₃ ) ₄ Cl ₂ , In addition to bis(tri-tert-butylphosphine)palladium(0), Pd(acac) ₂ , and Pd(PPh ₃ ) ₄ , Comparative Examples 10-18 were implemented in the same manner as in Example 2;

In addition to replacing the catalyst palladium trifluoroacetate with Pd(OAc) ₂ , PdCl ₂ , Pd ₂ (dba) ₃ , Pd(dba) ₂ , Pd(pyridyl) ₂ , Pd(NH ₃ ) ₄ Cl ₂ , In addition to bis(tri-tert-butylphosphine)palladium(0), Pd(acac) ₂ , and Pd(PPh ₃ ) ₄ , Comparative Examples 19-27 were implemented in the same manner as in Example 3;

In addition to replacing the catalyst palladium trifluoroacetate with Pd(OAc) ₂ , PdCl ₂ , Pd ₂ (dba) ₃ , Pd(dba) ₂ , Pd(pyridyl) ₂ , Pd(NH ₃ ) ₄ Cl ₂ , Except for bis(tri-tert-butylphosphine)palladium(0), Pd(acac) ₂ , and Pd(PPh ₃ ) ₄ , Comparative Examples 28-36 were carried out in the same manner as in Example 4, respectively.

The yields of the corresponding indolinone compounds are shown in the table below.

Comparative example 37-80

Using L1, L3-L12 respectively to implement Example 1 to obtain comparative examples 37-47;

Using L1, L3-L12 respectively to implement Example 2 to obtain comparative examples 48-58;

Using L1, L3-L12 respectively to implement Example 3 to obtain comparative examples 59-69;

Example 4 was implemented using L1, L3-L12 respectively to obtain comparative examples 70-80.

The yields of the corresponding indolinone compounds are shown in the table below.

Comparative example 81-88

Except that the solvent therein is replaced by the following organic solvent/water mixture (wherein the volume ratio of organic solvent to water is 2:1), comparative examples 81-88 are respectively implemented in the same manner as in Examples 1-4, which Correspondence and the productive rate of corresponding ketone compound are shown in the following table:

Inspection of Additives

Except that methanesulfonic acid wherein is replaced by trifluoroacetic acid, acetic acid, benzoic acid, trifluoromethanesulfonic acid, p-toluenesulfonic acid, p-nitrobenzenesulfonic acid, sulfuric acid, hydrochloric acid and potassium bifluoride, according to embodiment 1 respectively -4 investigated the effect of these additives, found that when being hydrochloric acid and potassium bifluoride, the reaction of embodiment 1-4 all can not carry out, and when being other above-mentioned additives, the reaction of embodiment 1-4 all can carry out, However, there is a substantial drop in yield, typically by 25-35%.

Biological activity test

MTT method was used to measure the inhibitory effect of indolinone derivatives of the present invention on tumor cell growth, and the tested cells included human gastric cancer cell line (SGC-7901), human lung cancer cell line (H446) and human gastric cancer cell line (HGC-7901). -27), the anticancer drug cisplatin was used as a positive control drug.

active test material

Cells: SGC-7901: human gastric cancer cell line; H446: human lung cancer cell line; HGC-27: human gastric cancer cell line; the above tumor cell lines were purchased from the Cell Bank of Shanghai Institute of Biological Sciences, Chinese Academy of Sciences.

Reagent: DMEM medium: product of Gibco Company; MTT: product of Sigma Company, subpackaged by Shanghai Shisheng Cell Biotechnology Co., Ltd.; fetal bovine serum: product of Gibco Company; penicillin-streptomycin: product of Gibco Company; EDTA-trypsin digestion solution : product of Gibco Company; positive control (cisplatin): product of Qilu Pharmaceutical Co., Ltd., product batch number: 1010011DC.

Experimental procedure

(1) Sample preparation

Dissolve every 1 mg of the test compound with 20 μL DMSO, take 2 μL of the dissolved solution and dilute it with 1000 μL culture medium (the culture medium is a DMEM medium containing 10% fetal bovine serum mass concentration), so that its concentration is 100 μg/mL, and then use The same culture solution was serially diluted to a concentration of 1-10 μg/mL.

(2) Preparation of culture medium

Prepare DMEM medium so that every 1000 mL of DMEM medium contains 800,000 units of penicillin, 1.0 g of streptomycin and 10% mass inactivated fetal bovine serum.

(3) Cell culture: the above-mentioned tumor cells were inoculated in the culture medium prepared in step (2), cultured in a 37° C., 5% CO ₂ incubator, and passaged for 3-5 days.

(4) Determination of the inhibitory effect of the sample on tumor cell growth

Human gastric cancer cell line (SGC-7901), human lung cancer cell line (H446), and human gastric cancer cell line (HGC-27) were digested with EDTA-trypsin digestion solution, and diluted with medium to 1×10 ⁵ /mL , added to a 96-well cell culture plate, 100 μL per well, and cultured in a 37°C, 5% CO ₂ incubator. Discard the original medium after 24 hours, add the medium containing the test sample, 200 μL per well, add 3 wells for each concentration, culture in a 37°C, 5% _CO2 incubator, and add to the cell culture well after 72 hours 5 mg/mL MTT, 10 μL per well, incubate at 37°C for 4 hours, add DMSO, 150 μL per well, shake with a shaker to completely dissolve formazan, and use a microplate reader to perform colorimetry at a wavelength of 570 nm. The above-mentioned cancer cells cultured in the medium containing no samples and the same concentration of DMSO under the same conditions were respectively used as controls, and the median lethal concentration (IC ₅₀ ) of the samples on the growth of tumor cells was calculated.

After the above-mentioned steps are measured, the _IC50 of the compound of the present invention is shown in the following table:

It can be seen from the above table that the compound D of the present invention has particularly excellent anti-gastric cancer and anti-lung cancer activities, and its _IC50 is much lower than that of cisplatin, especially for the inhibitory activity of H446, which is three orders of magnitude lower than that of cisplatin. The inhibitory activity of SGC-7901 and HGC-27 is also two orders of magnitude lower than that of cisplatin, so it has great application potential and clinical value in anticancer drugs.

In summary, the present invention provides a kind of indolinone derivative with anticancer activity, and at the same time, it is found that this type of compound has excellent anti-gastric cancer and anti-lung cancer activities. This is the first discovery of the use of this type of compound, so as to provide The research and development of anticancer drugs has expanded the substrates and possibilities, and has great clinical value and research and development potential.

It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the protection scope of the present invention. In addition, it should also be understood that after reading the technical content of the present invention, those skilled in the art can make various changes, modifications and/or variations to the present invention, and all these equivalent forms also fall within the appended claims of the present application. within the defined scope of protection.

Claims (6)

1. an indolinone derivative with anticancer activity, characterized in that: the indolinone derivative is the following formula (III), (IV) or (V) compound: 2. indolinone derivatives as claimed in claim 1, is characterized in that: described indolinone derivatives are following formula (V) compound: 3. the synthetic method of indolinone derivative as claimed in claim 1 or 2, described method comprises: in the presence of palladium catalyst, part and additive, in solvent, formula (VI) compound and formula (VII) Carry out reaction, thereby obtain described indolinone derivative; Wherein, R ₁ and n are as defined in claim 1 or 2 respectively; The palladium catalyst is palladium trifluoroacetate; The ligand is the following formula L2: The additive is trifluoroacetic acid, p-toluenesulfonic acid, p-nitrobenzenesulfonic acid or methanesulfonic acid; The solvent is a mixture of an organic solvent and water, wherein the organic solvent is tetrahydrofuran, 2-methyltetrahydrofuran, toluene, xylene, dioxane, isopropanol, ethanol, and the volume ratio of the organic solvent to water is 1-3 :1. 4. Use of the derivative according to any one of claims 1-2 in the preparation of anticancer drugs, the cancer being lung cancer or gastric cancer. 5. A pharmaceutical composition comprising the indolinone derivative according to any one of claims 1-2 as an active ingredient. 6. The pharmaceutical composition according to claim 5, characterized in that it comprises a compound of formula (IV) or (V).