An isoindolinone derivative, preparation method, pharmaceutical composition and application

Abstract

The present invention relates to an isoindolinone derivative, a preparation method, a pharmaceutical composition and an application. The derivative is a compound of formula I or a pharmaceutically acceptable salt of the compound of formula I, wherein Ra is one of a substituted or unsubstituted aryl, a substituted or unsubstituted C1-C6 alkyl; Rb and Rc are each one of a substituted or unsubstituted C1-C6 alkyl. Compared with the prior art, the isoindolinone derivative in the present invention has a novel structure, has significant anti-cancer cell proliferation activity, and the activities of multiple compounds are superior to those of doxorubicin.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and particularly relates to an isoindolinone derivative, a preparation method, a pharmaceutical composition and an application thereof. Background Art

[0002] With the continuous improvement of diagnosis and treatment means, diseases such as tumors are more and more easily detected and diagnosed. Malignant tumors are one of the most dangerous diseases affecting human health, causing great harm to patients both physically and psychologically. Surgical resection is the most commonly used cancer treatment method at present, but it has great trauma and is not suitable for the elderly or weak patients. With the continuous increase in the number of cancer diseases detected by diagnosis and the continuous deepening of scientific understanding of cancer, the cancer pharmaceutical field is developing rapidly. Although there are already various drugs for treating cancer on the market, they all have more or less defects, such as low bioavailability, large toxic and side effects, etc. Therefore, discovering novel compound lead structures with anti-cancer activity, exploring new action targets, and then rapidly discovering highly active compounds through structural modification and optimization have important significance for the discovery of new anti-cancer drugs.

[0003] Isoindolinone compounds are a class of important alkaloids and also important structural motifs. Their structures also contain a very important γ-butyrolactam skeleton, which is commonly found in many synthetic drugs and natural products with a wide range of biological activities and is also a key intermediate for preparing various functionalized derivatives. Isoindolinone derivatives have a wide range of biological activities, such as antifungal, anti-anxiety, anti-inflammatory, antiviral effects, etc.

[0004] In previous studies, the inventors found that a novel class of γ-butyrolactam derivatives has good anti-tumor activity (Patent Authorization Number: ZL201911018651.2). Based on this structure, a reasonable design was carried out: fusing the benzene ring and butyrolactam to obtain a class of isoindolinone structural skeletons.

[0005] Screening and discovering compounds with significant inhibitory activity against cancer cell proliferation from a large number of isoindolinone compounds will have good practical significance and value, and is also an urgent problem to be solved by current researchers. Summary of the Invention

[0006] The purpose of the present invention is to provide an isoindolinone derivative, a preparation method, a pharmaceutical composition and an application thereof to solve the above problems. The present invention is designed based on the structure to obtain a novel isoindolinone derivative with anti-cancer activity.

[0007] The purpose of the present invention is achieved by the following technical solutions:

[0008] The first aspect of the present invention provides an isoindolinone derivative, which is a compound of formula I or a pharmaceutically acceptable salt of the compound of formula I:

[0009]

[0010] Wherein:

[0011] Ra is one of a substituted or unsubstituted aryl group, a substituted or unsubstituted C1-C6 alkyl group;

[0012] Rb and Rc are each one of a substituted or unsubstituted C1-C6 alkyl group.

[0013] Further, in Ra, the substituted aryl group is an aryl group substituted with one or more substituents, and the substituted C1-C6 alkyl group is a C1-C6 alkyl group substituted with one or more substituents;

[0014] In Rb and Rc, the substituted C1-C6 alkyl group is a C1-C6 alkyl group substituted with one or more substituents, and the substituents are selected from the group consisting of a halogen, a cyano group, a nitro group, a trifluoromethyl group, an aryl group, a substituted aryl group, a heteroaryl group, a substituted heteroaryl group, a C1-C6 alkyl group, and a C1-C6 alkoxy group.

[0015] Further, in Rb and Rc, the substituted aryl group is an aryl group substituted with one or more substituents, and the substituents in the substituted aryl group are selected from the group consisting of a halogen, a cyano group, a nitro group, a trifluoromethyl group, a C1-C6 alkyl group, or a C1-C6 alkoxy group;

[0016] In Rb and Rc, the substituted heteroaryl group is a heteroaryl group substituted with one or more substituents, and the substituents are selected from the group consisting of C1-C6 alkyl groups;

[0017] In Rb and Rc, the aryl group is selected from the group consisting of a phenyl group and a naphthyl group;

[0018] In Rb and Rc, the heteroaryl group is selected from the group consisting of a furyl group, a thienyl group, a pyridyl group, and a quinolinyl group.

[0019] Further, the isoindolinone derivative is selected from one of B1 to B22:

[0020]

[0021] The second aspect of the present invention provides a method for preparing an isoindolinone derivative as described above, comprising the following steps:

[0022] In an inert solvent, phthalimide and Lawesson's reagent are reacted to obtain Compound 2. Compound 2 and diethyl halomalonate undergo an Eschenmoser coupling reaction to obtain Compound 3, and Compound 3 undergoes a nucleophilic substitution reaction to obtain an isatinone derivative:

[0023]

[0024] The third aspect of the present invention provides an application of the isatinone derivative as described above, and the isatinone derivative is used for preparing an anti-tumor drug.

[0025] Further, the tumors include: nasopharyngeal carcinoma; esophageal cancer; gastric cancer; liver cancer; breast cancer; colon cancer; prostate cancer; lung cancer; cervical cancer; leukemia; oral cancer; salivary gland tumors; malignant tumors of the nasal cavity and paranasal sinuses; laryngeal cancer; ear tumors; eye tumors; thyroid tumors; mediastinal tumors; chest wall; pleural tumors; small intestine tumors; biliary tract tumors; pancreatic and periampullary tumors; mesenteric and retroperitoneal tumors; kidney tumors; adrenal tumors; bladder tumors; prostate cancer; testicular tumors; penile cancer; endometrial cancer; ovarian malignant tumors; malignant trophoblastic tumors; vulvar and vaginal cancers; malignant lymphoma; multiple myeloma; soft tissue tumors; bone tumors; skin and appendage tumors; malignant melanoma or nervous system tumors.

[0026] The fourth aspect of the present invention provides an anti-tumor drug composition, and the drug composition contains the above isatinone derivative.

[0027] Further, the drug composition includes one or more of the isatinone derivatives, and a pharmaceutically acceptable carrier medium and / or excipient.

[0028] Further, the mass percentage content of the isatinone derivative in the composition is 0.001 - 99.99%.

[0029] The fifth aspect of the present invention provides an application of the anti-tumor drug composition as described above, and the composition is used for preparing an anti-tumor drug.

[0030] Compared with the prior art, the present invention has the following technical advantages:

[0031] 1) The present invention provides a class of isatinone derivatives with novel structures and significant anti-tumor medicinal values.

[0032] 2) The compounds and compositions in the present invention have significant anti-cancer cell proliferation activities (including but not limited to: liver cancer cell line HepG2 and lung cancer cell line A549), and the activities of multiple compounds are superior to those of doxorubicin. Detailed Embodiments

[0033] A class of structurally novel isoindolinone derivatives synthesized in the present invention, which have significant anti-cancer activity. After being tested by the MTT method, these compounds showed significant inhibitory activity against the proliferation of the liver cancer cell line HepG2 and the lung cancer cell line A549.

[0034] Group definitions:

[0035] The term "alkyl" refers to a group formed by removing one hydrogen atom from an alkane molecule; the term "alkylene" refers to a group formed by removing two hydrogen atoms from an alkane molecule.

[0036] The term "halogen" refers to fluorine, chlorine, bromine, or iodine. The term "halogenated" refers to a group substituted by one or more of the above-mentioned halogen atoms, which may be the same or different, such as trifluoromethyl, pentafluoroethyl, heptafluoroisopropyl, or similar groups.

[0037] The term "inert acid solvent" refers to various acidic solvents that do not react with the raw materials, including various straight-chain, branched-chain, or cyclic alkyl or aryl carboxylic acids, such as formic acid, acetic acid, propionic acid, isopropyl acid, butyric acid, benzoic acid, phenylacetic acid, etc.

[0038] The term "pharmaceutically acceptable salt" refers to the salts formed by the compounds of formula I in the present invention and pharmaceutically acceptable inorganic acids and organic acids. Among them, the preferred inorganic acids include (but are not limited to): hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, sulfuric acid; the preferred organic acids include (but are not limited to): formic acid, acetic acid, propionic acid, succinic acid, 1,5-naphthalenedisulfonic acid, asiatatic acid, oxalic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, valeric acid, diethylacetic acid, malonic acid, succinic acid, fumaric acid, pimelic acid, adipic acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, nicotinic acid, isonicotinic acid, methanesulfonic acid, p-toluenesulfonic acid, citric acid, and amino acids.

[0039] Pharmaceutical compositions and administration methods:

[0040] The "composition" used in the present invention refers to any mixture. It can be a solution, mixture, liquid, powder, ointment, aqueous, non-aqueous, or any combination thereof.

[0041] The compounds of the present invention, their pharmaceutically acceptable salts, or compositions containing them can be administered in unit dosage forms, and the administration routes can be divided into enteral or parenteral, such as oral, intravenous injection, intramuscular injection, subcutaneous injection, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.

[0042] The pharmaceutical dosage forms can be liquid dosage forms, solid dosage forms or semi-solid dosage forms. The liquid dosage forms can be solutions (including true solutions and colloidal solutions), emulsions (including O / W, W / O type and multiple emulsions), suspensions, injections (including aqueous injections, powder injections and infusions), eye drops, nasal drops, lotions, etc.; the solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules (including hard capsules, soft capsules, enteric-coated capsules), granules, powders, pellets, dripping pills, suppositories, films, patches, aerosols (powder aerosols), sprays, etc.; the semi-solid dosage forms can be ointments, gels, pastes, etc.

[0043] The compounds of the present invention and their pharmaceutically acceptable salts can be made into ordinary preparations, and can also be made into sustained-release preparations, controlled-release preparations, targeted preparations and various particulate drug delivery systems.

[0044] In order to make the compounds of the present invention and their pharmaceutically acceptable salts into tablets, various excipients well-known in the art can be widely used, including diluents, binders, wetting agents, disintegrants, lubricants, retention agents. The diluents can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the wetting agents can be water, ethanol, isopropanol, etc.; the binders can be starch, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, acacia mucilage, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; the disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitan fatty acid ester, sodium dodecyl sulfate, etc.; the lubricants and glidants can be talc, silica, stearates, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0045] The tablets can also be further made into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer tablets and multi-layer tablets. In order to make the dosage units into capsules, the active ingredients, the compounds of the present invention and their pharmaceutically acceptable salts, can be mixed with diluents and glidants, and the mixture can be directly placed into hard capsules or soft capsules. The active ingredients, the compounds of the present invention and their pharmaceutically acceptable salts can also be first made into granules or pellets with diluents, binders and disintegrants, and then placed into hard capsules or soft capsules. The varieties of diluents, binders, wetting agents, disintegrants and glidants used for preparing the tablets of the compounds of the present invention and their pharmaceutically acceptable salts can also be used for preparing the capsules of the compounds of the present invention and their pharmaceutically acceptable salts.

[0046] To prepare the compounds of the present invention and their pharmaceutically acceptable salts into injections, water, ethanol, isopropanol, propylene glycol or their mixtures can be used as solvents, and solubilizers, cosolvents, pH regulators, and osmotic pressure regulators commonly used in the art can be added. The solubilizer or cosolvent can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc., the pH regulator can be phosphate, acetate, hydrochloric acid, sodium hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, acetate, etc. If a freeze-dried powder injection is prepared, mannitol, glucose, etc. can also be added as a support agent.

[0047] In addition, if necessary, colorants, preservatives, fragrances, flavoring agents or other additives can also be added to the pharmaceutical preparation.

[0048] To achieve the purpose of medication and enhance the therapeutic effect, the drugs or pharmaceutical compositions of the present invention can be administered by any well-known administration method.

[0049] The present invention provides a preparation method of an isoindolinone derivative, comprising the steps of:

[0050] In an inert solvent (such as toluene, xylene, benzene, DMF, acetonitrile, tetrahydrofuran, etc.), phthalimide and Lawesson's reagent are reacted to obtain compound 2, compound 2 and a halogenated malonic ester are subjected to an Eschenmoser coupling reaction to obtain compound 3, and compound 3 is subjected to a nucleophilic substitution reaction to obtain the isoindolinone derivative.

[0051]

[0052] Wherein, Ra, Rb and Rc are defined as above.

[0053] The present invention will be described in detail below with reference to specific embodiments, but it is by no means a limitation of the present invention. Features such as preparation means, materials, structures or composition ratios that are not clearly described in the present technical solution are regarded as common technical features disclosed in the prior art.

[0054] Example 1

[0055] Preparation of Intermediate 2

[0056]

[0057] In a 50 mL round-bottom flask, phthalimide (5 mmol, 736 mg), Lawesson's reagent (2 mmol, 809 mg) and 20 mL of toluene were added. The mixture was stirred at 80 °C in an oil bath for 24 hours, and the reaction process was monitored by TLC. After the reaction was completed, the solvent was removed by rotary evaporation. 15 mL of saturated brine was added, and the mixture was extracted with ethyl acetate (3×15 mL). The combined extracts were dried over anhydrous magnesium sulfate, and the solvent was removed by rotary evaporation. The crude product was purified by column chromatography (petroleum ether / ethyl acetate) to obtain 3204 mg of a pink solid with a yield of 25%. 1 H NMR (400 MHz, CDCl3) δ 10.05 (s, 1H), 7.79 (J = 7.4, 1.5 Hz, 1H), 7.66 (dd, J = 7.4, 1.5 Hz, 1H), 7.54 (m, J = 7.4, 1.6 Hz, 1H), 7.45 (m, J = 7.4, 1.6 Hz, 1H); MS (GC-MS): m / z 163.1 (M + ). 4358 mg of a green solid with a yield of 40%. 1 H NMR (500 MHz, CDCl3) δ 10.03 (s, 1H), 7.87 (m, J = 5.6, 3.4 Hz, 2H), 7.54 (m, J = 5.6, 3.5 Hz, 2H); MS (GC-MS): m / z 179.0 (M + ).

[0058] Preparation of Intermediate 3

[0059]

[0060] In a 25 mL round-bottom flask, 2,3-dihydro-3-thioxo-1H-isoindoline-1-one (2 mmol, 326 mg), dimethyl bromomalonate (2 mmol, 264 mg), anhydrous potassium carbonate (20 mmol, 2.469 g) and 10 mL of anhydrous tetrahydrofuran were added. The mixture was stirred at 60 °C in an oil bath for 6 hours, and the reaction process was monitored by TLC. After the reaction was completed, the solvent was removed by rotary evaporation. 10 mL of saturated brine was added, and the mixture was extracted with ethyl acetate (3×10 mL). The combined extracts were dried over anhydrous magnesium sulfate, and the solvent was removed by rotary evaporation. The crude product was purified by column chromatography (petroleum ether / ethyl acetate) to obtain 496 mg of a yellow solid with a yield of 95%. 1 HNMR (400 MHz, CDCl3) δ 10.05 (s, 1H), 7.90 (dd, J = 5.0, 3.3 Hz, 1H), 7.69–7.59 (m, 2H), 7.55 (dt, J = 5.0, 3.1 Hz, 1H), 3.98 (s, 3H), 3.86 (s, 3H); MS (GC-MS): m / z261.1 (M+ ).

[0061] Preparation of Target Product B1

[0062]

[0063] Add dimethyl 2-(2,3-dihydro-3-oxo-1H-isoindole-1-ylidene)malonate (0.3 mmol, 78 mg), benzyl bromide (0.225 mmol, 39 mg), anhydrous potassium carbonate (0.225 mmol, 31 mg) and 3 mL of anhydrous acetonitrile into a 10 mL round-bottom flask. Start stirring, heat the reaction solution to reflux, and continue to reflux for 3 hours. Monitor the reaction process by TLC plate. After the reaction is completed, remove the solvent by rotary evaporation, add 5 mL of saturated brine, extract with ethyl acetate (3×5 mL), combine the extracts, dry with anhydrous magnesium sulfate, remove the solvent by rotary evaporation, and purify and separate the crude product by column chromatography (petroleum ether / ethyl acetate) to obtain 53 mg of a yellow solid with a yield of 67% and a melting point of 133.2. 1 1H NMR (400 MHz, CDCl3) δ 7.94 (dd, J = 5.8, 2.8 Hz, 1H), 7.71–7.65 (m, 1H), 7.64–7.57 (m, 2H), 7.31–7.21 (m, 3H), 7.04 (d, J = 7.1 Hz, 2H), 5.26 (s, 2H), 3.85 (s, 3H), 3.51 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.24, 165.90, 164.24, 144.41, 135.38, 135.36, 133.24, 131.51, 128.49 (2C), 128.43, 127.38, 126.47 (2C), 124.45, 124.14, 107.02, 53.14, 52.63, 45.09; HRMS (EI) calcd for C 20 H 17 NO5[M] + 351.1107, found 351.1111.

[0064] Example 2

[0065] Preparation of Target Product B2

[0066]

[0067] In a 10 mL round-bottom flask, add dimethyl 2-(2,3-dihydro-3-oxo-1H-isoindole-1-ylidene)malonate (0.3 mmol, 78 mg), 4-methylbenzyl bromide (0.225 mmol, 42 mg), anhydrous potassium carbonate (0.225 mmol, 31 mg) and 3 mL of anhydrous acetonitrile. Start stirring, heat the reaction solution to reflux, and continue refluxing for 3 hours while monitoring the reaction process by TLC plate spotting. After the reaction is completed, remove the solvent by rotary evaporation, add 5 mL of saturated brine, extract with ethyl acetate (3×5 mL), combine the extracts, dry with anhydrous magnesium sulfate, remove the solvent by rotary evaporation, and purify and separate the crude product by column chromatography (petroleum ether / ethyl acetate) to obtain 50 mg of a white solid with a yield of 61% and a melting point of 144.2 °C. 1 1H NMR (400 MHz, CDCl3) δ 7.99–7.92 (m, 1H), 7.75–7.68 (m, 1H), 7.67–7.60 (m, 2H), 7.54 (d, J = 8.1 Hz, 2H), 7.19 (d, J = 8.0 Hz, 2H), 5.30 (s, 2H), 3.87 (s, 3H), 3.49 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.28, 165.96, 164.25, 144.54, 136.98, 135.42, 133.19, 132.26, 131.47, 129.18 (2C), 128.49, 126.46 (2C), 124.41, 124.10, 106.90, 53.13, 52.65, 44.84, 21.11; HRMS (EI) calcd for C 21 18 19 11 + N5O5 [M]+ 365.1263, found 365.1258.

[0068] Example 3

[0069] Preparation of the target product B3

[0070]

[0071] In a 10 mL round-bottom flask, add dimethyl 2-(2,3-dihydro-3-oxo-1H-isoindole-1-ylidene)malonate (0.3 mmol, 78 mg), 3-methylbenzyl bromide (0.225 mmol, 42 mg), anhydrous potassium carbonate (0.225 mmol, 31 mg) and 3 mL of anhydrous acetonitrile. Start stirring, heat the reaction mixture to reflux, and continue refluxing for 3 hours. Monitor the reaction process by TLC plate. After the reaction is completed, remove the solvent by rotary evaporation, add 5 mL of saturated brine, extract with ethyl acetate (3×5 mL), combine the extracts, dry with anhydrous magnesium sulfate, remove the solvent by rotary evaporation, and purify the crude product by column chromatography (petroleum ether / ethyl acetate) to obtain 74 mg of a yellow solid with a yield of 90% and a melting point of 125.5 °C. 1 1H NMR (400 MHz, CDCl3) δ 7.97–7.90 (m, 1H), 7.67 (ddt, J = 8.1, 5.5, 2.4 Hz, 1H), 7.64–7.57 (m, 2H), 7.15 (t, J = 7.6 Hz, 1H), 7.02 (d, J = 7.5 Hz, 1H), 6.85 (s, 1H), 6.82 (d, J = 7.7 Hz, 1H), 5.21 (s, 2H), 3.85 (s, 3H), 3.52 (s, 3H), 2.28 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.27, 165.94, 164.23, 144.49, 138.07, 135.42, 135.26, 133.21, 131.47, 128.47, 128.39, 128.15, 127.26, 124.42, 124.14, 123.57, 106.94, 53.13, 52.61, 45.05, 21.41; HRMS (EI) calcd for C 21 H 19 NO5 [M] + 365.1263, found 365.1261.

[0072] Example 4

[0073] Preparation of the target product B4

[0074]

[0075] Add dimethyl 2-(2,3-dihydro-3-oxo-1H-isoindole-1-ylidene)malonate (0.3 mmol, 78 mg), 4-(trifluoromethyl)benzyl bromide (0.225 mmol, 54 mg), anhydrous potassium carbonate (0.225 mmol, 31 mg) and 3 mL of anhydrous acetonitrile into a 10 mL round-bottom flask. Start stirring, heat the reaction solution to reflux, and continue refluxing for 3 hours. Monitor the reaction process by TLC plate. After the reaction is completed, remove the solvent by rotary evaporation, add 5 mL of saturated brine, extract with ethyl acetate (3×5 mL), combine the extracts, dry with anhydrous magnesium sulfate, remove the solvent by rotary evaporation, and purify and separate the crude product by column chromatography (petroleum ether / ethyl acetate) to obtain 89 mg of yellow solid, with a yield of 91%, m.p. 120.8 °C. 1 H NMR (400 MHz, CDCl3) δ 7.99–7.92 (m, 1H), 7.75–7.68 (m, 1H), 7.67–7.60 (m, 2H), 7.54 (d, J = 8.1 Hz, 2H), 7.19 (d, J = 8.0 Hz, 2H), 5.30 (s, 2H), 3.87 (s, 3H), 3.49 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 168.09, 165.71, 164.20, 144.29, 139.79, 135.23, 133.46 (2C), 131.72 (2C), 129.69 (q, J = 32.5 Hz), 128.21, 126.76, 125.48 (q, J = 3.7 Hz), 124.59, 124.26, 123.99 (q, J = 272.0 Hz), 107.04, 53.24, 52.68, 44.89; HRMS (EI) calcd for C 21 H 16 F3NO5 [M] + 419.0981, found 419.0977.

[0076] Examples 5 - 22

[0077] Referring to the methods described in Examples 1 - 4, use different starting materials to prepare the isoindolinone derivatives shown below.

[0078] 11H NMR (400 MHz, CDCl3) δ 8.03–7.92 (m, 1H), 7.70–7.62 (m, 3H), 7.55 (d, J = 7.7 Hz, 1H), 7.40 (dd, J = 14.2, 6.4 Hz, 2H), 7.30 (d, J = 7.9 Hz, 1H), 5.27 (s, 2H), 3.87 (s, 3H), 3.57 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.11, 165.66, 164.09, 144.30, 137.42, 135.23, 133.59, 131.84, 131.22, 131.02, 130.30, 129.43, 128.05, 124.53, 124.35, 118.46, 112.65, 106.92, 53.29, 52.81, 44.62; HRMS (EI) calcd for C 21 H 16 N2O5 [M] + 376.1059, found 376.1061.

[0079] 1 1H NMR (400 MHz, CDCl3) δ 8.12 (d, J = 8.1 Hz, 1H), 8.02–7.93 (m, 2H), 7.70–7.61 (m, 3H), 7.48 (t, J = 7.9 Hz, 1H), 7.39 (d, J = 7.8 Hz, 1H), 5.34 (s, 2H), 3.86 (s, 3H), 3.59 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.17, 165.69, 164.12, 148.26, 144.43, 138.05, 135.28, 133.62, 132.75, 131.85, 129.68, 128.05, 124.50, 124.39, 122.68, 121.94, 106.91, 53.29, 52.91, 44.70; HRMS (EI) calcd for C 20 H 16 N2O7 [M] + 396.0958, found 396.0955.

[0080] 11H NMR (400 MHz, CDCl3) δ 8.16 (d, J = 8.8 Hz, 2H), 8.00–7.93 (m, 1H), 7.74–7.62 (m, 3H), 7.24 (d, J = 8.8 Hz, 2H), 5.34 (s, 2H), 3.88 (s, 3H), 3.53 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.04, 165.61, 164.06, 147.24, 144.37, 143.36, 135.18, 133.64, 131.88, 128.02, 127.29 (2C), 124.58, 124.34, 123.76 (2C), 106.97, 53.31, 52.83, 44.97; HRMS (EI) calcd for C 20 H 16 N2O7 [M] + 396.0958, found 396.0954.

[0081] 1 1H NMR (400 MHz, CDCl3) δ 8.00–7.90 (m, 1H), 7.70–7.60 (m, 3H), 7.50 (d, J = 7.8 Hz, 1H), 7.39 (dd, J = 14.5, 6.7 Hz, 2H), 7.19 (d, J = 7.7 Hz, 1H), 5.31 (s, 2H), 3.85 (s, 3H), 3.51 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.18, 165.76, 164.13, 144.29, 136.68, 135.31, 133.47, 131.71, 130.80 (q, J = 32.2Hz), 129.74, 129.17, 128.16, 124.46, 124.36 (q, J = 4.0 Hz), 124.31, 123.94 (q, J = 272.3 Hz), 123.69 (q, J = 3.6 Hz), 107.06, 53.22, 52.67, 44.84; HRMS (EI) calcd for C 21 H 16 F3NO5 [M] + 419.0981, found 419.0983.

[0082] 11H NMR (400 MHz, CDCl3) δ 7.96–7.89 (m, 1H), 7.69–7.56 (m, 3H), 6.96 (d, J = 8.7 Hz, 2H), 6.79 (d, J = 8.7 Hz, 2H), 5.18 (s, 2H), 3.85 (s, 3H), 3.75 (s, 3H), 3.59 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.30, 165.99, 164.23, 158.84, 144.51, 135.43, 133.20, 131.47, 128.47, 127.86 (2C), 127.29, 124.35, 124.08, 113.91 (2C), 106.89, 55.20, 53.14, 52.68, 44.50; HRMS (EI) calcd for C 21 H 19 NO6[M] + 381.1212, found 381.1210.

[0083] 1 1H NMR (400 MHz, CDCl3) δ 7.95 (p, J = 3.6 Hz, 1H), 7.73–7.63 (m, 3H), 7.59 (d, J = 8.1 Hz, 2H), 7.18 (d, J = 8.1 Hz, 2H), 5.30 (s, 2H), 3.88 (s, 3H), 3.51 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.06, 165.64, 164.09, 144.32, 141.30, 135.19, 133.59, 132.33 (2C), 131.84, 128.07, 127.18 (2C), 124.59, 124.34, 118.57, 111.37, 107.01, 53.31, 52.78, 45.09; HRMS (EI) calcd for C 21 H 16 NO5[M] + 376.1059, found 376.1062.

[0084] 11H NMR (400 MHz, CDCl3) δ 7.98 (dd, J = 5.3, 2.4 Hz, 1H), 7.82–7.71 (m, 3H), 7.70–7.59 (m, 3H), 7.50 (s, 1H), 7.44 (dd, J = 6.2, 3.2 Hz, 2H), 7.15 (d, J = 8.5 Hz, 1H), 5.42 (s, 2H), 3.81 (s, 3H), 3.43 (s, 3H). 13 13C NMR (101 MHz, DMSO) δ 163.07, 160.63, 159.02, 139.22, 130.21, 128.04, 127.94, 127.69, 127.47, 126.30, 123.23, 123.19, 122.49, 122.39, 121.02, 120.64, 120.38, 119.20, 119.11, 118.95, 101.80, 47.86, 47.30, 40.06.; HRMS (EI) calcd for C 24 H 19 NO5 [M] + 401.1263, found 401.1266.

[0085] 1 1H NMR (400 MHz, CDCl3) δ 7.88 (dt, J = 4.5, 3.3 Hz, 1H), 7.68 (dt, J = 5.6, 3.5 Hz, 1H), 7.62–7.55 (m, 2H), 5.69 (ddt, J = 17.0, 10.6, 5.4 Hz, 1H), 5.20–5.09 (m, 2H), 4.63 (d, J = 5.4 Hz, 2H), 3.93 (s, 3H), 3.82 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 167.87, 166.12, 164.50, 145.05, 135.33, 133.13, 131.96, 131.46, 128.57, 124.32, 123.95, 117.66, 106.26, 53.20, 52.90, 44.00; HRMS (EI) calcd for C 16 H 15 NO5 [M] + 301.0950, found 301.0953.

[0086] 11H NMR (400 MHz, CDCl3) δ 7.99–7.90 (m, 1H), 7.67 (dt, J = 7.9, 3.3 Hz, 1H), 7.63–7.55 (m, 2H), 7.18 (t, J = 7.9 Hz, 1H), 6.75 (dd, J = 8.2, 2.0 Hz, 1H), 6.62 (d, J = 7.6 Hz, 1H), 6.58 (s, 1H), 5.23 (s, 2H), 3.85 (s, 3H), 3.73 (s, 3H), 3.54 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.18, 165.90, 164.22, 159.74, 144.42, 137.01, 135.37, 133.23, 131.49, 129.55, 128.39, 124.42, 124.13, 118.81, 112.61, 112.48, 106.99, 55.18, 53.12, 52.65, 45.01; HRMS (EI) calcd for C 21 H 19 NO6 [M] + 381.1212, found 381.1210.

[0087] 1 1H NMR (400 MHz, CDCl3) δ 8.16–8.09 (m, 1H), 8.00–7.94 (m, 1H), 7.82 (dt, J = 5.8, 3.6 Hz, 1H), 7.69–7.63 (m, 2H), 7.59–7.52 (m, 1H), 7.44 (t, J = 7.7 Hz, 1H), 7.11 (d, J = 7.7 Hz, 1H), 5.49 (s, 2H), 3.88 (s, 3H), 3.46 (s, 3H). 13 13C NMR (151 MHz, CDCl3) δ 163.46, 160.76, 159.43, 142.95, 139.84, 130.36, 129.03, 128.84, 127.50, 127.12, 123.50, 123.32, 122.80, 120.37, 120.31, 119.52, 102.28, 48.50, 48.06, 39.31; HRMS (EI) calcd for C 20 H 16 N2O7 [M] + 396.0958, found 396.0961.

[0088] 1 1H NMR (400 MHz, CDCl3) δ 8.97 (dd, J = 4.2, 1.7 Hz, 1H), 8.65 (d, J = 7.7 Hz, 1H), 8.14 (dd, J = 8.3, 1.6 Hz, 1H), 8.04 (d, J = 6.3 Hz, 1H), 7.77–7.71 (m, 1H), 7.52–7.46 (m, 1H), 7.46–7.40 (m, 2H), 7.40–7.35 (m, 2H), 5.24 (s, 2H), 4.05 (s, 3H), 3.92 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 178.47, 166.48, 164.33, 158.49, 149.69, 146.26, 139.74, 136.42, 136.39, 135.59, 130.88, 130.75, 130.60, 128.42, 127.90, 127.51, 126.41, 121.27, 120.14, 119.27, 52.78, 52.59, 30.63; HRMS (EI) calcd for C 23 H 18 N2O5 [M] + 402.1216, found 402.1210.

[0089] 1 1H NMR (400 MHz, CDCl3) δ 7.97–7.91 (m, 1H), 7.80–7.74 (m, 1H), 7.64–7.59 (m, 2H), 7.47 (t, J = 7.7 Hz, 1H), 7.00 (d, J = 7.6 Hz, 1H), 6.79 (d, J = 7.7 Hz, 1H), 5.26 (s, 2H), 3.86 (s, 3H), 3.58 (s, 3H), 2.49 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.29, 165.79, 164.34, 157.93, 154.83, 145.01, 136.81, 135.41, 133.22, 131.47, 128.56, 124.71, 124.03, 121.70, 117.42, 106.92, 53.07, 52.82, 47.11, 24.29; HRMS (EI) calcd for C 20 H 18 N2O5 [M] + 366.1216, found 366.1219.

[0090] 1 1H NMR (400 MHz, CDCl3) δ 7.91 (dd, J = 5.8, 2.4 Hz, 1H), 7.65 (dt, J = 7.5, 3.2 Hz, 1H), 7.59 (dd, J = 5.6, 3.2 Hz, 2H), 7.23 (dd, J = 5.0, 3.0 Hz, 1H), 6.96 (d, J = 2.7 Hz, 1H), 6.87–6.79 (m, 1H), 5.25 (s, 2H), 3.87 (s, 3H), 3.64 (s, 3H). 13 13C NMR (151 MHz, CDCl3) δ 163.29, 161.25, 159.58, 139.67, 131.55, 130.65, 128.50, 126.77, 123.70, 121.71, 121.70, 121.46, 119.62, 119.35, 117.32, 102.01, 60.28, 48.45, 48.14, 48.03, 36.36, 28.77; HRMS (EI) calcd for C 18 H 15 NO5S [M] + 357.0671, found 357.0674.

[0091] 1 1H NMR (400 MHz, CDCl3) δ 7.99–7.91 (m, 1H), 7.88–7.80 (m, 1H), 7.70–7.60 (m, 3H), 7.45 (t, J = 7.5 Hz, 1H), 7.36 (t, J = 7.5 Hz, 1H), 7.02 (d, J = 7.8 Hz, 1H), 5.32 (s, 2H), 3.90 (s, 3H), 3.41 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.08, 165.63, 164.12, 145.08, 135.19, 134.23, 133.46, 132.06, 131.75, 128.34, 127.07, 126.79 (q, J = 31.1 Hz), 126.09, 126.03 (q, J = 3.7 Hz), 125.00, 124.16, 121.63 (q, J = 274.2 Hz), 106.97, 53.21, 52.56, 43.21 (q, J = 3.8 Hz); HRMS (EI) calcd for C 21 H 16 F3NO5 [M]+ 419.0981, found 419.0986.

[0092] 1 1H NMR (400 MHz, CDCl3) δ 7.98–7.90 (m, 1H), 7.81–7.72 (m, 1H), 7.67–7.58 (m, 2H), 7.11 (dt, J = 12.9, 4.7 Hz, 3H), 6.77 (d, J = 7.0 Hz, 1H), 5.17 (s, 2H), 3.86 (s, 3H), 3.27 (s, 3H), 2.32 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.09, 165.74, 164.20, 144.29, 135.20, 135.02, 133.19, 132.90, 131.48, 130.20, 128.53, 127.05, 126.08, 124.97, 124.71, 124.07, 107.15, 53.12, 52.27, 43.51, 19.05; HRMS (EI) calcd for C 21 H 19 NO5[M] + 365.1263, found 365.1260.

[0093] 1 1H NMR (400 MHz, CDCl3) δ 7.93 (dd, J = 5.6, 2.5 Hz, 1H), 7.68 (dd, J = 5.8, 2.6 Hz, 1H), 7.65–7.56 (m, 2H), 7.25 (d, J = 8.5 Hz, 2H), 6.99 (d, J = 8.5 Hz, 2H), 5.22 (s, 2H), 3.86 (s, 3H), 3.55 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 168.14, 165.80, 164.17, 144.33, 135.31, 134.04, 133.36, 133.18, 131.62, 128.68 (2C), 128.28, 127.91 (2C), 124.47, 124.19, 106.99, 53.20, 52.71, 44.56; HRMS (EI) calcd for C 20 H 16 35 ClNO5[M] + 385.0701, found 385.0717.

[0094] 1 1H NMR (400 MHz, CDCl3) δ 8.10 (dd, J = 6.5, 1.8 Hz, 1H), 7.97 (dd, J = 6.1, 2.3 Hz, 1H), 7.72–7.64 (m, 2H), 7.49 (t, J = 7.6 Hz, 2H), 7.40 (t, J = 7.4 Hz, 1H), 7.33 (s, 2H), 3.92 (s, 3H), 3.13 (s, 3H). 13 13C NMR (151 MHz, CDCl3) δ 167.20, 165.56, 164.30, 146.97, 134.99, 133.55, 131.92, 128.97 (2C), 128.57, 128.30 (2C), 128.06, 125.53, 107.36, 53.07, 52.16; HRMS (EI) calcd for C 19 H 15 NO5[M] + 337.0950, found 337.0947.

[0095] 1 1H NMR (400 MHz, CDCl3) δ 8.08 (dd, J = 6.3, 1.9 Hz, 1H), 7.96–7.91 (m, 1H), 7.69–7.61 (m, 2H), 7.35 (t, J = 7.7 Hz, 1H), 7.18 (d, J = 7.6 Hz, 1H), 7.12 (d, J = 11.1 Hz, 2H), 3.90 (s, 3H), 3.12 (s, 3H), 2.39 (s, 3H). 13 13C NMR (151 MHz, CDCl3) δ 167.21, 165.58, 164.33, 146.99, 138.91, 134.87, 134.73, 133.48, 131.86, 128.78, 128.63, 128.57, 128.49, 125.55, 125.19, 124.14, 107.34, 53.04, 52.05, 21.27; HRMS (EI) calcd forC 20 H 17 NO5[M] + 351.1107, found 351.1104.

[0096] Verification Example 1

[0097] Antitumor activity test of the compounds of the present invention

[0098] 1. Experimental principle

[0099] Succinate dehydrogenase in the mitochondria of living cells can reduce exogenous MTT to insoluble blue-violet crystalline formazan, which deposits in the cells, while dead cells do not have this function. Dimethyl sulfoxide (DMSO) can dissolve formazan in the cells, and the light absorption value measured by an enzyme-linked immunosorbent assay (ELISA) reader can reflect the number of living cells.

[0100] 2. Antitumor activity experiment

[0101] Samples: Compounds of the examples

[0102] Cell lines: Hepatocellular carcinoma cell line HepG2 and lung cancer cell line A549

[0103] Reagents: 0.5% MTT solution, RPMI 1640 culture medium, newborn bovine serum; trypsin; 96-well culture plate; dimethyl sulfoxide;

[0104] Instruments: Laminar flow hood, incubator, Perkin Elmer fully automatic multifunctional ELISA reader

[0105] Experimental procedure:

[0106] 1) Inoculate HepG2 and A549 cells in the exponential growth phase (2×10 5 cells / mL) onto a 96-well plate and culture overnight, and then treat the cells with the drug at the set concentration.

[0107] 2) After 24 hours of drug treatment, add 20 μL of MTT reagent (5 mg / mL) to each well. Incubate at 5% CO2 and 37 °C for 4 hours, then aspirate the supernatant of the culture medium, and add 150 μL of DMSO to each well to dissolve the formazan crystals.

[0108] 3) Measure by the double-wavelength method, and measure the absorbance at 492 and 630 nm by an ELISA reader.

[0109] 3. Antitumor activity evaluation

[0110] 1) Calculation of cell viability inhibition rate

[0111] Cell viability inhibition rate = [1 - (OD 492 - OD 630 ) / (OD 492 - OD 630 )] × 100 (%)

[0112] 2) Calculation of IC 50 value

[0113] Perform linear regression on the sample concentration and cell inhibition rate, and use software to calculate the half-maximal inhibitory concentration IC of the sample on the cells50 Value

[0114] Table 1 Inhibitory IC of cell viability 50 Value

[0115]

[0116]

[0117] As can be seen from Table 1, for HepG2 cells and A549 cells, the IC50 values of 22 compounds are all lower than 10 μM; moreover, for HepG2 cells, 16 compounds exhibit activity superior to that of the positive control doxorubicin; for A549 cells, 20 compounds exhibit activity superior to that of the positive control doxorubicin. It can be seen that this type of novel-structured isoindolinone derivative has excellent anti-cancer activity.

[0118] The above description of the embodiments is to enable those of ordinary skill in the art to understand and use the invention. It is obvious that those skilled in the art can easily make various modifications to these embodiments and apply the general principles described herein to other embodiments without creative efforts. Therefore, the present invention is not limited to the above embodiments, and the improvements and modifications made by those skilled in the art without departing from the scope of the present invention should be within the protection scope of the present invention.

Claims

1. An isatinone derivative, characterized in that, The derivative is one of the compounds from B1 to B22 or a pharmaceutically acceptable salt thereof:

2. A method for preparing the isatinone derivative as described in claim 1, characterized in that, Comprising the following steps: In an inert solvent, phthalimide and Lawesson's reagent are reacted to obtain compound 2, compound 2 and dihalomalonate are subjected to an Eschenmoser coupling reaction to obtain compound 3, and compound 3 is subjected to a nucleophilic substitution reaction to obtain an isatin derivative: Wherein Ra, Rb, and Rc are the corresponding groups in the compounds from B1 to B22, and X is Br.

3. An application of the isatinone derivative as described in claim 1, characterized in that, The isatin derivative is used for preparing an anti-tumor drug.

4. An anti-tumor pharmaceutical composition, characterized in that, The pharmaceutical composition contains the isatin derivative as described in claim 1.

5. The anti-tumor pharmaceutical composition according to claim 4, characterized in that, The pharmaceutical composition comprises one or more of the isatin derivatives, and a pharmaceutically acceptable carrier medium and / or excipient.

6. The anti-tumor pharmaceutical composition according to claim 4, characterized in that, The mass percentage content of the isatin derivative in the composition is 0.001 to 99.99%.

7. An application of the anti-tumor pharmaceutical composition as described in claim 4, characterized in that, The composition is used for preparing an anti-tumor drug.

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