Oleanan cinnamide derivatives, production methods thereof, and uses thereof

Abstract

The present invention relates to the fields of medicinal chemistry and therapeutics, specifically to 2-cyano-3,12-dioxolane-1,9(11)-diene-17-phenylacrylamide derivatives and methods for preparing the same, and further to the application of the novel compounds in the production of anti-cancer drugs.

Description

Technical Field

[0001] The present invention relates to the fields of medicinal chemistry and pharmacotherapeutics, and its content includes 2-cyano-3,12-dioxolane-1,9(11)-diene-17-cinnamamide compounds and methods for their preparation. The present invention further relates to the application of such novel compounds in the manufacture of anti-cancer drugs.

Background Art

[0002] Oleanolic acid (OA) belongs to pentacyclic triterpenoid natural products, is very widely distributed in the plant kingdom, is an active ingredient in many traditional Chinese medicines, and has a wide range of biological activities. Structure optimization based on OA is one of the hotspots in the chemical research of natural drugs.

Chemical

[0003] CDDO and its derivatives (CDDOs) are semi-synthetic OA derivatives with the strongest anti-tumor and anti-inflammatory activities discovered so far. Different from single-target drugs, CDDOs are characterized by multi-function, multi-target, and acting on the entire cell signaling pathway, and can exert anti-tumor effects from multiple stages. Therefore, high activity and low drug resistance are the advantages of this type of derivative. Here, the methyl ester derivative of CDDO (methyl 2-cyano-3,12-dioxoolean-1,9(11)-dien-28-oate, CDDO-Me) has the trade name Bardoxolone Methyl, and phase I clinical trials (NCT00529438, NCT00508807) for the treatment of advanced solid tumors and malignant lymphoma were conducted in 2006. In addition, CDDO-Me was approved by the FDA in 2008 to conduct a treatment evaluation (NCT00664027) for chronic kidney disease (CKD) caused by type II diabetes. Unfortunately, due to the high fatality rate caused by heart failure, it ended in a phase III clinical study in November 2013. According to reports, this drug was approved by the FDA in 2014 and used in a clinical study of pulmonary arterial hypertension (a rare drug), and is currently in the phase III clinical study stage (phase II NCT02036970, phase III NCT02657356).

Chemical Structure

[0004] CDDO-Me has excellent anti-tumor activity, but its cardiotoxicity still deserves attention. If the activity of this type of compound can be maintained or enhanced and its toxicity can be reduced through necessary modification and transformation, it will have important practical significance.

Summary of the Invention

Means for Solving the Problems

[0005] The present invention discloses for the first time a 2-cyano-3,12-dioxoolean-1,9(11)-diene-17-cinnamamide compound, a method for producing the same, and a pharmaceutical use thereof. The method for producing the derivatives of this type in the invention has the advantages that the raw materials are inexpensive and easily available, the reagents are environmentally friendly and low in toxicity, the conditions are mild and easy to control, and the post-treatment is convenient and simple. At the same time, it has practicality and universality. According to the results of pharmacological experiments, the compounds of the present invention have excellent antitumor activities higher than or equivalent to CDDO-Me, and the myocardial toxicity of some compounds is significantly reduced. Therefore, this type of compound can be regarded as a candidate compound for antitumor drugs.

[0006] One of the objects of the present invention is to provide a 2-cyano-3,12-dioxolan-1,9(11)-diene-17-cinnamamide compound represented by Formula I,

Chemical formula

[0007] The alkoxy is an alkoxy containing 1 to 3 carbon atoms.

[0008] In some embodiments, R1 and R2 in the above Formula I are each independently selected from H or cyano, and R3 is selected from H, C 1-3 alkyl, alkoxy, or halogen.

[0009] In some embodiments, R1 and R2 in the above Formula I are each independently selected from H or cyano, and R3 is selected from F, Cl, H, and methoxy.

[0010] In some embodiments, R1 and R2 in the above formula I are each independently selected from H or cyano, and R3 is selected from Cl or H.

[0011] In some embodiments, R1 in the above formula I is cyano, R2 is H, and R3 is Cl or H.

[0012] On the other hand, the present invention provides a compound of the following formula I or a pharmaceutically acceptable salt thereof,

Chemical formula

[0013] In some embodiments, R1 and R2 in the above formula I are each independently selected from H or cyano, and R3 is H, C 1-3 alkyl, alkoxy, halogen, C substituted with halogen 1-3 alkyl, the alkoxy contains 1 to 3 carbon atoms, and the halogen is selected from F, Cl, Br, and I.

[0014] In some embodiments, R1 in the above formula I is H or cyano, R2 is H or cyano, and R3 is selected from H, methyl, methoxy, trifluoromethyl, Cl, or F.

[0015] In some embodiments, R1 in the above formula I is H or cyano, R2 is H or cyano, and R3 is Cl, H, or trifluoromethyl.

[0016] In some embodiments, the compound of Formula I is selected from the following compounds. [Table 1]

[0017] The present invention simultaneously provides a method for producing the compound represented by Formula I. 1. Perform a Curtius rearrangement on CDDO to obtain a C-17 amino derivative (CDDO-NH2) whose structure is represented by Formula II. [Chemical formula] 2. Condense Formula II and Formula III (phenylacrylic acid-based compound) to produce the compound of Formula I. [Chemical formula] The condensing agent is selected from one or more of dicyclohexylcarbodiimide, N,N-diisopropylcarbodiimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide, benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate, or benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate, and the solvent can be selected from one or more of N,N-dimethylformamide, acetone, acetonitrile, toluene, benzene, xylene, 1,4-dioxane, ethyl acetate, dichloromethane, chloroform, tetrahydrofuran, or ether, but is not limited thereto. Preferably, the reaction temperature is 0°C to 60°C.

[0018] ​A further object of the present invention is to provide a pharmaceutical composition prepared from an effective dose of the compound of the present invention or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable pharmaceutical adjuvants. The compound of the present invention can be prepared alone or with one or more pharmaceutical carriers into different dosage forms such as tablets, capsules, granules, liquid preparations, etc., and is used for clinical oral, injection or topical administration. In these different preparations, the content of the compound of the present invention may be 0.1% - 99.9%. The dosage of the compound of the present invention may be 0.001 - 10000 mg / kg / 0.3 days, and can be appropriately adjusted according to clinical needs. The "pharmaceutically acceptable salt" of the present invention refers to a pharmaceutically acceptable acid or base addition salt, or a solvate thereof.

[0019] Another object of the present invention is to provide the application of the compound of this type or a pharmaceutically acceptable salt thereof and its pharmaceutical composition in the manufacture of anti-tumor drugs. The tumor is selected from lung cancer, liver cancer, colon cancer, pancreatic cancer, breast cancer, prostate cancer, brain cancer, ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head and neck cancer, lymphoma, melanoma or leukemia. Further, the tumor is selected from lung cancer, liver cancer or breast cancer. Preferably it is lung cancer. According to a series of tumor cell test results, the compound of the present invention has broad-spectrum anti-tumor activity, and its IC 50 values are all in the nanomolar to micromolar level, corresponding to the activity of the positive control drug CDDO-Me. Here, the toxicity of compounds I-7 and I-12 to rat embryonic cardiomyocytes H9C2 (IC 50 values are 3.176 ± 1.74 μM and 3.143 ± 1.53 μM respectively) is significantly lower than that of CDDO-Me (IC50 is 0.308 ± 0.01 μM).

[0020] Another object of the present invention is to provide the application of the compounds of this type or their pharmaceutically acceptable salts and their pharmaceutical compositions in the manufacture of anti-tumor drugs. The tumors are selected from lung cancer, liver cancer, ascites tumor, brain metastatic tumor, colon cancer, pancreatic cancer, breast cancer, prostate cancer, brain cancer, ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head and neck cancer, lymphoma, melanoma or leukemia, preferably selected from lung cancer, liver cancer, breast cancer, ascites tumor, pancreatic cancer, brain metastatic tumor, and more preferably selected from non-small cell lung cancer, liver cancer, breast cancer, ascites tumor. According to a series of tumor cell test results, the compounds of the present invention have broad-spectrum anti-tumor activity, and their IC 50 values are all in the nanomolar to micromolar level, corresponding to the activity of the positive control drug CDDO-Me. Here, the toxicity of compounds I-7 and I-12 to rat embryonic cardiomyocytes H9C2 (IC 50 values are 3.176±1.74 μM and 3.143±1.53 μM respectively) is significantly lower than that of CDDO-Me (IC50 is 0.308±0.01 μM), and compounds I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, I-12, I-13, I-14, I-16, I-18 all show lower toxicity to human renal tubular epithelial cells HK-2. Still another object of the present invention is to provide the compounds of this type or their pharmaceutically acceptable salts and their pharmaceutical compositions, which are used for anti-tumor. The tumors are selected from lung cancer, liver cancer, ascites tumor, brain metastatic tumor, colon cancer, pancreatic cancer, breast cancer, prostate cancer, brain cancer, ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head and neck cancer, lymphoma, melanoma or leukemia, preferably selected from lung cancer, liver cancer, breast cancer, ascites tumor, pancreatic cancer, brain metastatic tumor, and more preferably selected from non-small cell lung cancer, liver cancer, breast cancer, ascites tumor. According to a series of tumor cell test results, the compounds of the present invention have broad-spectrum anti-tumor activity, and their IC 50 values are all in the nanomolar to micromolar level, corresponding to the activity of the positive control drug CDDO-Me. Here, the toxicity of compounds I-7 and I-12 to rat embryonic cardiomyocytes H9C2 (IC 50The values are 3.176±1.74 μM and 3.143±1.53 μM respectively, which are significantly lower than those of CDDO-Me (IC50 is 0.308±0.01 μM). Additionally, all of compounds I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, I-12, I-13, I-14, I-16, and I-18 exhibit lower toxicity against human renal tubular epithelial cells HK-2.

[0021] Another further object of the present invention is to provide a method for treating tumors, which includes administering to a subject or patient a therapeutically effective amount of the above-mentioned types of compounds or their pharmaceutically acceptable salts and their pharmaceutical compositions. The tumors are selected from lung cancer, liver cancer, ascites tumor, brain metastatic tumor, colon cancer, pancreatic cancer, breast cancer, prostate cancer, brain cancer, ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head and neck cancer, lymphoma, melanoma or leukemia, preferably selected from lung cancer, liver cancer, breast cancer, ascites tumor, pancreatic cancer, brain metastatic tumor, more preferably selected from non-small cell lung cancer, liver cancer, breast cancer, ascites tumor. According to a series of tumor cell test results, the compounds of the present invention have broad-spectrum antitumor activity, and their IC 50 values are all in the nanomolar to micromolar level and are comparable to the activity of the positive control drug CDDO-Me. Here, the toxicity of compounds I-7 and I-12 against rat embryonic cardiomyocytes H9C2 (IC 50 The values are 3.176±1.74 μM and 3.143±1.53 μM respectively, which are significantly lower than those of CDDO-Me (IC50 is 0.308±0.01 μM). Additionally, all of compounds I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, I-12, I-13, I-14, I-16, and I-18 exhibit lower toxicity against human renal tubular epithelial cells HK-2.

[0022] This specification has described and illustrated the preferred embodiments of the present invention. However, it is obvious to those skilled in the art that such embodiments are provided only in an exemplary manner. Currently, those skilled in the art can conceive of many changes, modifications, and alternatives without departing from the present invention. It should be understood that various alternatives of the above-described embodiments of the present invention can be used to implement the present invention. The appended claims are intended to limit the scope of the present invention and cover methods, structures, and their equivalent forms within the scope of these claims.

[0023] Some chemical terms Unless otherwise defined, all scientific and technical terms used in this specification have the same meaning as commonly understood by those skilled in the art to which the subject matter of the claims belongs. Unless otherwise specifically explained, all patents, patent applications, and disclosure materials cited throughout this specification are incorporated herein by reference in their entirety.

[0024] It should be understood that the above summary and the following detailed description are exemplary and for illustrative purposes only and do not limit the subject matter of the present application in any way. In the present application, unless specifically stated otherwise, the use of the singular includes the plural. Further, it should be noted that unless otherwise explained, the terms "or" and "alternatively" used herein represent "and / or". Also, the terms "comprising" and other forms, such as "including", "containing", and "having" all belong to non-limiting descriptions.

[0025] References (Carey and Sundberg “ADVANCED ORGANIC CHEMISTRY 4 THFind definitions for standard chemical terms in, e.g., “ED.” Vols. A (2000) and B (2001), Plenum Press, New York. Unless otherwise specified, use common methods within the skill of the art, such as mass spectrometry, NMR, IR, and UV / Vis spectroscopy, and pharmacological methods. Unless otherwise provided with specific definitions, the terms employed in the descriptions of analytical chemistry, organic synthetic chemistry, and drugs and medicinal chemistry in this specification are those known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, drug manufacture, formulation and delivery, and treatment of patients. For example, use the instructions for the manufacturer's kit, or carry out reactions and purification according to known methods in the art or as described in this application. Generally, based on the general methods well-known in the art according to the descriptions in the several schematic descriptions and more specific documents generally cited and considered in this specification, the above techniques and methods can be carried out. In this specification, one of ordinary skill in the art can select groups and their substituents to provide stable structural moieties and compounds.

[0026] When describing substituents by general chemical formulas written from left to right, the substituents include chemically equivalent substituents obtained when the structural formula is written from right to left. For example, CH2O is equal to OCH2.

[0027] The "compounds" in this application refer to those including all stereoisomers, geometric isomers, tautomers and isotopes. The compounds of this application may be asymmetric and, for example, have one or more stereoisomers. Unless otherwise specified, all stereoisomers include, for example, enantiomers and diastereomers. Compounds containing asymmetric substituted carbon atoms in this application may be separated in the form of optically active alcohols or in racemic form. The form of optically active alcohols can be resolved from racemic mixtures or synthesized by using chiral starting materials or chiral reagents. The compounds of this application further include tautomeric forms. The tautomeric forms result from the exchange of one single bond with an adjacent double bond together with the movement of one proton. The compounds of this application further include all isotope atoms whether they are intermediates or final compounds. Isotope atoms have the same number of atoms but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. That is, the compounds of this application include compounds in which some or all of the hydrogen (H) is substituted by tritium (T) and / or deuterium (D), and some or all of the 12 C is 13 C and / or 14 substituted by C, and compounds substituted between isotopes of other elements (such as N, O, P, S), for example 14 N and 15 N, 18 O and 17 O, 31 P and 32 P, 35 S and 36Further includes S etc. The compounds herein can have one or more stereoisomeric centers, and each isomeric center can exist in the form of R or S structure or a combination thereof. Similarly, the compounds herein can have one or more double bonds, and each double bond can exist in the form of E (trans) or Z (cis) structure or a combination thereof. Specific stereoisomers, structural isomers, diastereomers, enantiomers or epimers should be understood to include all possible isomers, such as stereoisomers, structural isomers, diastereomers, enantiomers or epimers and mixtures thereof. Therefore, the compounds herein include all structurally different stereoisomeric, structural isomeric, diastereomeric, enantiomeric or epimeric forms and their corresponding mixtures. Techniques for converting specific stereoisomers or retaining specific stereoisomers as they are, and techniques for separating mixtures of stereoisomers are well known in the art, and those skilled in the art can select appropriate methods according to specific situations.

[0028] The terms "may / optional" or "may be / optionally" refer to the possibility that the events or situations described hereinafter may or may not occur, and the description includes the occurrence of the said events or situations and the non-occurrence of the said events or situations.

[0029] C as used herein 1-3 Refers to having 1 - 3 carbon atoms in the moiety, that is, the group includes 1 carbon atom, 2 carbon atoms, 3 carbon atoms. Therefore, for example, "C1 - C4 alkyl" refers to an alkyl having 1 - 4 carbon atoms, that is, the alkyl is selected from methyl, ethyl, propyl, isopropyl, n - butyl, isobutyl, sec - butyl and tert - butyl.

[0030] As used herein, the term "alkyl", alone or in combination, refers to a straight-chain or branched-chain aliphatic hydrocarbon which may be substituted. The "alkyl" herein may preferably have from 1 to about 20 carbon atoms, for example having from 1 to about 10 carbon atoms, having from 1 to about 8 carbon atoms, or having from 1 to about 6 carbon atoms, or having from 1 to about 4 carbon atoms or from 1 to about 3 carbon atoms. Examples of alkyl herein include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, etc. The groups defined herein, for example when a numerical range appears in "alkyl", such as "C1-C6 alkyl" or "C 1-6 alkyl", refers to an alkyl composed of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms or 6 carbon atoms, and the alkyl herein also includes the case where no numerical range is specified.

[0031] "Alkyl" used in combination herein includes an alkyl bonded to another group, such as the alkyl in alkoxy.

[0032] As used herein, the term "halogen", alone or in combination, is selected from F, Cl, Br, I.

[0033] As used herein, the term "halogeno" or "halogen substitution", used alone or in combination, refers to the substitution of one or more hydrogen atoms of an optionally substituted group (e.g., an alkyl group, an alkenyl group, and an alkynyl group) with a fluorine, chlorine, bromine, iodine atom, or a combination thereof. In some embodiments, two or more hydrogen atoms (e.g., difluoromethyl, trifluoromethyl) are substituted using the same halogen atom with each other, and in other embodiments, two or more hydrogen atoms (e.g., 1-chloro-1-fluoro-1-iodoethyl) are substituted using halogen atoms that are not at all the same with each other. Non-limiting examples of a halogenated alkyl group are a fluoromethyl group and a bromoethyl group. A non-limiting example of a halogenated alkenyl group is a bromovinyl group. A non-limiting example of a haloalkynyl group is a chloroethynyl group.

[0034] As used herein, the term "treatment" and other similar synonyms include alleviating, reducing, or improving the symptoms of a disease or medical condition, suppressing a disease or medical condition, e.g., preventing the development of a disease or medical condition, alleviating a disease or medical condition, ameliorating a disease or medical condition, alleviating the symptoms caused by a disease or medical condition, or arresting the symptoms of a disease or medical condition, preventing other symptoms, and improving or preventing the potential metabolic causes that result in the symptoms. Also, the term includes the purpose of prevention. The term further includes obtaining a therapeutic effect and / or a preventive effect. The therapeutic effect refers to curing or improving the underlying disease being treated. Also, the cure or improvement of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect. For example, although a patient may still be affected by the underlying disease, an improvement in the patient's condition is observed. For the preventive effect, the composition can be administered to a patient at risk of developing a particular disease, or to a patient in whom one or more physiological symptoms of the disease appear even without a disease diagnosis.

[0035] As used herein, the terms "effective amount", "therapeutically effective amount" or "pharmaceutically effective amount" refer to an amount of at least one active substance (the compounds of the present application) sufficient to alleviate to some extent one or more symptoms of a disease or disorder being treated after administration. The result may be a reduction and / or alleviation of signs, symptoms or causes, or any other necessary change in a biological system. For example, a "effective amount" for treatment is the amount of a composition containing a compound disclosed herein necessary to provide a clinically significant alleviation of the disorder. Techniques such as dose escalation studies can be used to determine the effective amount suitable for any individual case.

[0036] As used herein, the term "acceptable" when used with respect to a formulation, composition or ingredient refers to not having a long-term harmful effect on the general health of the subject being treated.

[0037] As used herein, the term "pharmaceutically acceptable" refers to a substance (e.g., a carrier or diluent) that does not affect the biological activity or properties of the compounds of the present application and is relatively non-toxic, i.e., the substance is applied to an individual without causing an adverse biological reaction or interacting with any component contained in the composition in an adverse manner.

[0038] As used herein, the term "pharmaceutical composition" refers to a mixture of a compound of the present application and at least one pharmaceutically acceptable substance. The pharmaceutically acceptable substances include, but are not limited to, carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents and / or excipients. As used herein, the term "carrier" refers to a relatively non-toxic substance that aids in introducing the compounds of the present application into cells or tissues.

[0039] As used herein, the term "pharmaceutically acceptable salt" refers to salts that retain the biological efficacy of the free acid and free base of the specified compound and have no adverse effects biologically or otherwise. The compounds of the present application further include pharmaceutically acceptable salts. Pharmaceutically acceptable salts refer to converting the base in the parent compound into the form of a salt. Pharmaceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of bases such as amine (ammonia) groups. The pharmaceutically acceptable salts of the present application can be synthesized from the parent compound, that is, reacting the base in the parent compound with 1-4 equivalents of an acid in a single solvent system. Suitable salts are listed in Remingtong’s Pharmaceutical Scicences, 17 th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977). Unless otherwise indicated, the salts in the present application refer to acidic salts formed by organic acids / inorganic acids and basic salts formed by organic bases / inorganic bases. Also, when the basic functional group of the compound of the general formula is pyridine or imidazole (not limited to pyridine or imidazole) and the acidic functional group is carboxylic acid (not limited to carboxylic acid), an amphoteric ion (inner salt) is formed, and the inner salt is also included in the salts in the present application.

Mode for Carrying Out the Invention

[0040] To more clearly illustrate the present invention, a series of examples are given here. These examples are illustrative and should not be construed as limiting the present invention.

[0041] Example 1, Synthesis of I-1 Dissolve 0.080 g (0.170 mmol) of CDDO-NH2 in 5 mL of dichloromethane, add 0.046 g (0.260 mmol) of 3-phenylacrylic acid, stir, add 0.177 g (PyBop, 0.340 mmol) of benzotriazol-1-yloxytris(pyrrolidino)phosphonium hexafluorophosphate, 0.074 mL (0.425 mmol) of N,N-diisopropylethylamine, and react at room temperature for 12 h. After detecting the end of the reaction by TLC, add 5 mL of water and 5 mL of dichloromethane, extract and separate, extract the aqueous layer with another 5 mL of dichloromethane, and combine the organic layers. Wash the organic layer with 5 mL of saturated sodium chloride and dry over anhydrous sodium sulfate. After suction filtration, rotary evaporate the organic layer, perform column chromatography, and then rotary evaporate to obtain 0.036 g of a yellowish-white solid with a yield of 44.9%.

[0042] ESI-MS: 593.4 [M+H] + 。 1 1H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.89 (3H, s), 1.04 (3H, s), 1.05 (3H, s), 1.17 (3H, s), 1.26 (3H, s), 1.42 (3H, s), 1.47 (3H, s), 6.01 (1H, s), 6.53 (1H, d, J = 15.60 Hz), 7.35 (3H, m), 7.48 (2H, m), 7.60 (1H, d, J = 15.54 Hz), 8.08 (1H, s).

Chemical Structure

[0043] Example 2, Synthesis of I-2 Referring to the manufacturing method of Compound I-1, using 3-(4-methylphenyl)acrylic acid as the reaction reagent instead of 3-phenylacrylic acid, with other conditions unchanged, 0.024 g of the target compound I-2, a pale yellow solid, was produced with a yield of 28.8%.

[0044] ESI-MS: 607.4 [M+H] + 。 1 1H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.90 (3H, s), 1.05 (6H, s), 1.17 (3H, s), 1.27 (3H, s), 1.41 (3H, s), 1.47 (3H, s), 2.36 (3H, s), 6.01 (1H, s), 6.43 (1H, d, J = 12.60 Hz), 7.19 (2H, s), 7.38 (2H, s), 7.57 (1H, d, J = 15.27 Hz), 8.06 (1H, s).

Chem.

[0045] Example 3, Synthesis of I-3 Referring to the manufacturing method of compound I-1, using 3-(4-methoxyphenyl)acrylic acid as the reaction reagent instead of 3-phenylacrylic acid, with other conditions unchanged, 0.035 g of the target compound I-3, a yellowish-white solid, was produced, and the yield was 41.2%.

[0046] ESI-MS: 623.4 [M+H] + , 645.4 [M+Na] + 。 1 1H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.92 (3H, s), 1.06 (6H, s), 1.18 (3H, s), 1.27 (3H, s), 1.42 (3H, s), 1.48 (3H, s), 3.84 (3H, s), 6.01 (1H, s), 6.90 (1H, d, J = 8.49 Hz), 7.45 (1H, d, J = 8.43 Hz), 7.58 (4H, m), 8.05 (1H, s).

Chem.

[0047] Example 4, Synthesis of I-4 Referring to the manufacturing method of Compound I-1, using 3-(4-trifluoromethylmethylphenyl)acrylic acid as the reaction reagent instead of 3-phenylacrylic acid, with other conditions remaining unchanged, 0.018 g of the target compound I-4, a pale yellow solid, was produced, and the yield was 20.4%.

[0048] ESI-MS: 661.4 [M+H] + , 683.4 [M+Na] + . 1 H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.91 (3H, s), 1.06 (3H, s), 1.07 (3H, s), 1.18 (3H, s), 1.27 (3H, s), 1.44 (3H, s), 1.49 (3H, s), 6.03 (1H, s), 6.54 (1H, d, J = 15.30 Hz), 7.61 (4H, m), 7.63 (1H, d, J = 12.96 Hz), 8.09 (1H, s).

Chemical Structure

[0049] Example 5, Synthesis of I-5 Referring to the manufacturing method of Compound I-1, using 3-(4-fluorophenyl)acrylic acid as the reaction reagent instead of 3-phenylacrylic acid, with other conditions remaining unchanged, 0.019 g of the target compound I-5, a pale yellow solid, was produced, and the yield was 23.4%.

[0050] ESI-MS: 611.4 [M+H] + . 1H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.89 (3H, s), 1.05 (3H, s), 1.06 (3H, s), 1.17 (3H, s), 1.26 (3H, s), 1.45 (3H, s), 1.49 (3H, s), 6.01 (1H, s), 6.55 (1H, d, J = 8.58 Hz), 7.59 (2H, s), 7.64 (2H, s), 7.63 (1H, d, J = 8.61 Hz), 8.09 (1H, s).

Chem.

[0051] Example 6, Synthesis of I-6 Referring to the manufacturing method of compound I-1, using 3-(4-chlorophenyl)acrylic acid as the reaction reagent instead of 3-phenylacrylic acid, with other conditions unchanged, 0.025 g of the target compound I-6, a yellowish-white solid, was produced, and the yield was 29.1%.

[0052] ESI-MS: 627.3 [M+H] + . 1 H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.90 (3H, s), 1.05 (3H, s), 1.06 (3H, s), 1.18 (3H, s), 1.27 (3H, s), 1.44 (3H, s), 1.49 (3H, s), 6.03 (1H, s), 6.53 (1H, d, J = 8.66 Hz), 7.62 (2H, s), 7.68 (2H, s), 7.61 (1H, d, J = 8.64 Hz), 8.09 (1H, s).

Chem.

[0053] Example 7, Synthesis of Intermediate III-12-Cyano-3-phenylacrylic Acid Dissolve 0.425 g (5 mmol) of cyanoacetic acid and 0.477 g (4.5 mmol) of benzaldehyde in 20 mL of toluene, add 0.058 g (0.75 mmol) of ammonium acetate, heat under reflux, react for 6 - 7 h, and detect whether the reaction is complete by TLC. After cooling to room temperature, perform suction filtration, and vacuum dry the filter cake to obtain 0.643 g of the white solid of intermediate III - 1, with a yield of 82.4%.

[0054] ESI-MS, 174.0 [M + H] + The reaction formula is as follows.

Chemical formula

[0055] Example 8, Synthesis of I - 7 Referring to the manufacturing method of compound I - 1, use 2 - cyano - 3 - phenylacrylic acid as the reaction reagent instead of 3 - phenylacrylic acid, with other conditions unchanged, to produce 0.026 g of the target compound I - 7 as a pale yellow solid, with a yield of 30.7%.

[0056] ESI-MS: 618.4 [M + H] + , 640.4 [M + Na] + , 616.4 [M - H] - . 1 1H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.93 (3H, s), 1.06 (3H, s), 1.13 (3H, s), 1.18 (3H, s), 1.27 (3H, s), 1.34 (3H, s), 1.49 (3H, s), 6.01 (1H, s), 7.35 (3H, m), 7.48 (2H, m), 8.06 (1H, s), 8.21 (1H, s).

Chemical formula

[0057] Example 9, Synthesis of Intermediate III - 2 - Cyano - 3-(4 - methylphenyl)acrylic Acid Referring to the synthesis method of Intermediate 2 - Cyano - 3 - phenylacrylic Acid III - 1, using 4 - methylbenzaldehyde instead of benzaldehyde, with other conditions unchanged, 0.727 g of Intermediate III - 2 was produced, and the yield was 86.3%.

[0058] ESI - MS, 188.0[M + H] + The reaction formula is as follows.

Chemical formula

[0059] Example 10, Synthesis of I - 8 Referring to the production method of Compound I - 1, using 2 - Cyano - 3-(4 - methylphenyl)acrylic Acid as the reaction reagent instead of 3 - phenylacrylic Acid, with other conditions unchanged, 0.031 g of the target compound I - 8, a pale yellow solid, was produced, and the yield was 35.9%.

[0060] ESI - MS:632.4 [M + H] + , 654.4 [M + Na] + , 630.4 [M - H] - . 1 H - NMR (300 MHz, CDCl3, TMS), δ ppm: 0.93 (3H, s), 1.06 (3H, s), 1.13 (3H, s), 1.18 (3H, s), 1.27 (3H, s), 1.34 (3H, s), 1.49 (3H, s), 2.43 (3H, s), 6.01 (1H, s), 6.99 (2H, d, J = 8.48 Hz), 7.92 (2H, d, J = 8.61 Hz), 8.06 (1H, s), 8.21 (1H, s).

Chemical formula

[0061] Example 11, Synthesis of Intermediate III-3 2-Cyano-3-(4-methoxyphenyl)acrylic Acid Referring to the synthesis method of Intermediate III-1 2-Cyano-3-phenylacrylic Acid, using 4-methoxybenzaldehyde instead of benzaldehyde, with other conditions unchanged, 0.794 g of Intermediate III-3 was produced, and the yield was 86.8%.

[0062] ESI-MS, 204.0 [M+H] + The reaction formula is as follows.

Chemical formula

[0063] Example 12, Synthesis of I-9 Referring to the production method of Compound I-1, using 2-Cyano-3-(4-methoxyphenyl)acrylic Acid as the reaction reagent instead of 3-phenylacrylic Acid, with other conditions unchanged, 0.030 g of the target compound I-9 yellow solid was produced, and the yield was 33.7%.

[0064] ESI-MS: 648.4 [M+H] + , 670.4 [M+Na] + . 1 1H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.93 (3H, s), 1.06 (3H, s), 1.12 (3H, s), 1.17 (3H, s), 1.27 (3H, s), 1.34 (3H, s), 1.49 (3H, s), 3.11 (3H, s), 6.01 (1H, s), 6.99 (2H, d, J = 8.49 Hz), 7.92 (2H, d, J = 8.67 Hz), 8.06 (1H, s), 8.21 (1H, s).

Chemical formula

[0065] Example 13, Synthesis of Intermediate III-4 2-Cyano-3-(4-trifluoromethylphenyl)acrylic Acid Referring to the synthesis method of Intermediate III-1 2-Cyano-3-phenylacrylic Acid, using 4-trifluoromethylbenzaldehyde instead of benzaldehyde, with other conditions unchanged, 0.872 g of Intermediate III-4 was produced, and the yield was 80.4%.

[0066] ESI-MS, 242.0 [M+H] + The reaction formula is as follows.

Chemical formula

[0067] Example 14, Synthesis of I-10 Referring to the production method of Compound I-1, using 2-Cyano-3-(4-trifluoromethylphenyl)acrylic Acid as the reaction reagent instead of 3-phenylacrylic Acid, with other conditions unchanged, 0.031 g of the target compound I-10, a white solid, was produced, and the yield was 32.9%.

[0068] ESI-MS: 686.4 [M+H] + , 684.4 [M-H] - . 1 1H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.91 (3H, s), 1.02 (3H, s), 1.08 (3H, s), 1.17 (3H, s), 1.27 (3H, s), 1.44 (3H, s), 1.50 (3H, s), 6.05 (1H, s), 7.68 (2H, d, J = 8.07 Hz), 7.88 (2H, d, J = 8.01 Hz), 8.09 (1H, s).

Chemical formula

[0069] Example 15, Synthesis of Intermediate III-5 2-Cyano-3-(4-fluorophenyl)acrylic Acid Referring to the synthesis method of Intermediate 2-cyano-3-phenylacrylic acid III-1, using 4-fluorobenzaldehyde instead of benzaldehyde, with other conditions unchanged, 0.683 g of Intermediate III-5 was produced, and the yield was 79.4%.

[0070] ESI-MS, 192.0 [M+H] + The reaction formula is as follows.

Chemical formula

[0071] Example 16, Synthesis of I-11 Referring to the production method of Compound I-1, using 2-cyano-3-(4-fluorophenyl)acrylic acid as the reaction reagent instead of 3-phenylacrylic acid, with other conditions unchanged, 0.022 g of the target compound I-11, a pale yellow solid, was produced, and the yield was 25.4%.

[0072] ESI-MS: 636.4 [M+H] + , 658.4 [M+Na] + , 635.4 [M-H] - . 1 H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.94 (3H, s), 1.07 (3H, s), 1.13 (3H, s), 1.18 (3H, s), 1.27 (3H, s), 1.44 (3H, s), 1.50 (3H, s), 6.05 (1H, s), 7.21 (4H, m), 8.06 (1H, s), 8.26 (1H, s).

Chemical formula

[0073] Example 17, Synthesis of Intermediate III-6 2-Cyano-3-(4-chlorophenyl)acrylic Acid Referring to the synthesis method of the intermediate 2-cyano-3-phenylacrylic acid III-1, 4-chlorobenzaldehyde was used instead of benzaldehyde, and other conditions remained unchanged. Intermediate III-6 with a weight of 0.712 g was produced, and the yield was 76.2%.

[0074] ESI-MS, 208.0 [M+H] + The reaction formula is as follows.

Chemical formula

[0075] Example 18, Synthesis of I-12 Referring to the production method of compound I-1, 2-cyano-3-(4-chlorophenyl)acrylic acid was used as the reaction reagent instead of 3-phenylacrylic acid, and other conditions remained unchanged. 0.033 g of the target compound I-12, a yellowish-white solid, was produced, and the yield was 37.9%.

[0076] ESI-MS: 652.3 [M+H] + , 674.3 [M+Na] + , 650.3 [M-H] - . 1 H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.92 (3H, s), 1.02 (3H, s), 1.08 (3H, s), 1.17 (3H, s), 1.27 (3H, s), 1.44 (3H, s), 1.50 (3H, s), 6.05 (1H, s), 7.42 (2H, d, J = 8.47 Hz), 7.82 (2H, d, J = 8.51 Hz), 8.12 (1H, s).

Chemical formula

[0077] Example 19, Synthesis of Intermediate III-7 3-cyano-3-phenylacrylic acid Dissolve 0.735 g (5 mmol) of phenylacetonitrile in 50 mL of methanol, add 0.464 g (5 mmol) of glyoxylic acid, add 1.035 g (7.5 mmol) of potassium carbonate, heat to reflux, and indicate the reaction end point by the result of TLC. After suction filtration, wash the filter cake with dichloromethane, dissolve the filter cake in water, add dilute hydrochloric acid to adjust to pH = 4, and a white solid precipitates. Extract with ethyl acetate (30 mL × 2), combine the organic layers, and dry over anhydrous sodium sulfate. After suction filtration, rotary dry and dry in a constant temperature drying oven to obtain 0.679 g of intermediate III-7 white solid, and the yield reaches 78.5%.

[0078] ESI-MS, 174.0 [M+H] + The reaction formula is as follows.

Chemical formula

[0079] Example 20, Synthesis of I-13 Referring to the manufacturing method of compound I-1, using intermediate III-7 as the reaction reagent instead of 3-phenylacrylic acid, with other conditions unchanged, 0.030 g of the target compound I-13 light yellowish-white solid is manufactured, and the yield is 36.2%.

[0080] ESI-MS: 618.4 [M+H] + , 640.4 [M+Na] + . 1 1H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.91 (3H, s), 1.04 (3H, s), 1.06 (3H, s), 1.18 (3H, s), 1.34 (3H, s), 1.42 (3H, s), 1.47 (3H, s), 6.00 (1H, s), 7.03 (1H, s), 7.47 (2H, m), 7.69 (3H, m), 8.04 (1H, s).

Chemical formula

[0081] Example 21, Synthesis of Intermediate III-83-Cyano-3-(4-methylphenyl)acrylic Acid Referring to the synthesis method of Intermediate 3-cyano-3-phenylacrylic acid III-7, 4-methylphenylacetonitrile was used as the reaction reagent instead of phenylacetonitrile, and other conditions remained unchanged. 0.733 g of Intermediate III-8 was produced with a yield of 78.2%.

[0082] ESI-MS, 188.0 [M+H] + The reaction formula is as follows.

Chemical formula

[0083] Example 22, Synthesis of I-14 Referring to the manufacturing method of Compound I-1, Intermediate III-8 was used as the reaction reagent instead of 3-phenylacrylic acid, and other conditions remained unchanged. 0.035 g of the target compound I-14, a yellowish-white solid, was produced with a yield of 40.7%.

[0084] ESI-MS: 632.4 [M+H] + , 654.4 [M+Na] + . 1 H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.90 (3H, s), 1.03 (3H, s), 1.06 (3H, s), 1.18 (3H, s), 1.27 (3H, s), 1.42 (3H, s), 1.47 (3H, s), 2.40 (3H, s), 6.00 (1H, s), 7.01 (1H, s), 7.26 (2H, d, J = 8.16 Hz), 7.68 (2H, d, J = 7.7 Hz), 8.04 (1H, s).

Chemical formula

[0085] Example 23, Synthesis of Intermediate III-93-Cyano-3-(4-methoxyphenyl)acrylic Acid Referring to the synthesis method of Intermediate 3-Cyano-3-phenylacrylic Acid III-7, 4-Methoxyphenylacetonitrile was used as a reaction reagent instead of Phenylacetonitrile, and other conditions remained unchanged. 0.757 g of Intermediate III-9 was produced with a yield of 74.5%.

[0086] ESI-MS, 204.0 [M+H] + The reaction formula is as follows.

Chemical Structure

[0087] Example 24, Synthesis of I-15 Referring to the production method of Compound I-1, Intermediate III-9 was used as a reaction reagent instead of 3-Phenylacrylic Acid, and other conditions remained unchanged. 0.031 g of the target compound I-15, a light tan solid, was produced with a yield of 35.4%.

[0088] ESI-MS: 648.4 [M+H] + , 670.4 [M+Na] + . 1 H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.89 (3H, s), 0.99 (3H, s), 1.04 (3H, s), 1.17 (3H, s), 1.27 (3H, s), 1.40 (3H, s), 1.45 (3H, s), 3.85 (3H, s), 6.00 (1H, s), 6.94 (2H, d, J = 8.76 Hz), 7.03 (1H, s), 7.65 (2H, d, J = 8.85 Hz), 8.06 (1H, s).

Chemical Structure

[0089] Example 25, Synthesis of Intermediate III-103-Cyano-3-(4-trifluoromethylphenyl)acrylic Acid Referring to the synthesis method of Intermediate 3-Cyano-3-phenylacrylic Acid III-7, 4-Trifluoromethylphenylacetonitrile was used as the reaction reagent instead of phenylacetonitrile, and other conditions remained unchanged. 0.833 g of Intermediate III-10 was produced, and the yield was 69.1%.

[0090] ESI-MS, 242.0 [M+H] + The reaction formula is as follows.

Chemical Structure

[0091] Example 26, Synthesis of I-16 Referring to the production method of Compound I-1, Intermediate III-10 was used as the reaction reagent instead of 3-Phenylacrylic Acid, and other conditions remained unchanged. 0.056 g of the target compound I-16, a light tan solid, was produced, and the yield was 60.2%.

[0092] ESI-MS: 686.4 [M+H] + , 708.4 [M+Na] + . 1 H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.89 (3H, s), 1.00 (3H, s), 1.06 (3H, s), 1.18 (3H, s), 1.28 (3H, s), 1.42 (3H, s), 1.47 (3H, s), 6.04 (1H, s), 7.15 (1H, s), 7.73 (2H, d, J = 8.13 Hz), 7.82 (2H, d, J = 7.89 Hz), 8.09 (1H, s).

Chemical Structure

[0093] Example 27, Synthesis of Intermediate III-113-Cyano-3-(4-fluorophenyl)acrylic Acid Referring to the synthesis method of Intermediate 3-Cyano-3-phenylacrylic Acid III-7, 4-Fluorophenylacetonitrile was used as the reaction reagent instead of phenylacetonitrile. Other conditions remained unchanged, and 0.956 g of Intermediate III-11 was produced with a yield of 81.9%.

[0094] ESI-MS, 192.0 [M+H] + The reaction formula is as follows.

Chemical Structure

[0095] Example 28, Synthesis of I-17 Referring to the production method of Compound I-1, Intermediate III-11 was used as the reaction reagent instead of 3-Phenylacrylic Acid. Other conditions remained unchanged, and 0.023 g of the target compound I-17, a yellowish-white solid, was produced with a yield of 27.0%.

[0096] ESI-MS: 636.4 [M+H] + , 658.4 [M+Na] + . 1 H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.90 (3H, s), 1.01 (3H, s), 1.06 (3H, s), 1.18 (3H, s), 1.34 (3H, s), 1.43 (3H, s), 1.49 (3H, s), 6.04 (1H, s), 6.96 (1H, s), 7.15 (2H, d, J = 8.49 Hz), 7.70 (2H, d, J = 8.76 Hz), 8.09 (1H, s).

Chemical Structure

[0097] Example 29, Synthesis of Intermediate III-12 3-Cyano-3-(4-chlorophenyl)acrylic Acid

[0098] Referring to the synthesis method of Intermediate 3-Cyano-3-phenylacrylic Acid III-7, 4-Chlorophenylacetonitrile was used as the reaction reagent instead of phenylacetonitrile, and other conditions remained unchanged. 0.843 g of Intermediate III-12 was produced, and the yield was 81.2%.

[0099] ESI-MS, 208.0 [M+H] +

[0100] The reaction formula is as follows.

Chemical formula

[0101] Example 30, Synthesis of I-18 Referring to the production method of Compound I-1, Intermediate III-12 was used as the reaction reagent instead of 3-Phenylacrylic Acid, and other conditions remained unchanged. 0.027 g of the target compound I-18, a light tan solid, was produced, and the yield was 30.3%.

[0102] ESI-MS: 652.3 [M+H] + , 674.3 [M+Na] + . 1 H-NMR (300 MHz, CDCl3, TMS), δ ppm: 0.88 (3H, s), 0.96 (3H, s), 1.04 (3H, s), 1.17 (3H, s), 1.26 (3H, s), 1.40 (3H, s), 1.45 (3H, s), 6.02 (1H, s), 7.23 (1H, s), 7.40 (2H, d, J = 8.40 Hz), 7.64 (2H, d, J = 8.43 Hz), 8.09 (1H, s).

Chemical formula

[0103] Example 31, Pharmacological Experiment of Compound The anti-tumor activity test was carried out on the compound of the present invention by adopting the tetramethyl azole blue colorimetric method (MTT method), and methyl bercolone (CDDO-Me) was selected as the positive control drug.

[0104] Equipment, clean bench (SW-CJ-1FD, AIRTECH, Sujing Antai), constant temperature CO2 incubator (3111, Thermo, USA), inverted biological microscope (IX71, OLYMPUS, Japan), enzyme-linked immunosorbent detector (Model680, BIO-RAD, USA), plate shaking table (Kylin-belllabInstruments), high-pressure sterilizer (YXO.SG41.280, Shanghai Huaxian), centrifuge (SIGMA).

[0105] Reagents, DMEM (GIBCO), fetal bovine serum (GIBCO), trypsin (SIGMA), DMSO (SIGMA).

[0106] Cell lines, human non-small cell lung cancer cell line A549, human hepatocellular carcinoma cell line HepG2, human breast cancer cell line MCF-7, human renal tubular epithelial cell HK-2, rat embryonic cardiomyocyte H9C2 (all provided by Jiangsu KGI Biotechnology Co., Ltd.).

[0107] Method: Resuscitate the cryopreserved cell line, place it in a constant temperature 37°C CO2 incubator for culture, change the liquid once a day, and it can be plated when the cell line is in the exponential growth phase and in good condition. Add 1 mL of 0.25% trypsin digestion solution, digest for 1 - 2 min, observe the cell state under a microscope, and the digestion solution can be aspirated and removed when the adherent cells become round and shrink. Add 1 - 2 mL of DMEM medium containing 10% fetal bovine serum to prepare a cell suspension, perform cell counting, and calculate the required amount of cell suspension according to the number of 5×10 4 cells per well and the total number of wells. Inoculate this cell suspension into a 96-well plate, 100 μL / well, seal the periphery with PBS, and place it in a constant temperature 37°C CO2 incubator for 24 h of culture.

[0108] Prepare the test drug, positive control drug CDDO-Me, and blank control group DMSO in DMEM medium, and adjust their final concentrations to 5 μM / well. Each drug has three replicate wells and is cultured for 48 hours. Add MTT reagent to the 96-well plate at 10 μL / well and continue to incubate for 4 h. Aspirate and remove the medium in the plate, add 100 μL of DMSO to each well, and shake on a plate shaking table for 10 min to dissolve the crystals. Use an enzyme-linked immunosorbent detector to detect the absorbance value of each well at a wavelength of 570 nm, and calculate the cell inhibition rate using the following formula. The average value of the results of three primary screenings is the final inhibition rate. For compounds with a primary screening inhibition rate greater than 60%, perform a concentration gradient screening (5-fold dilution) to calculate the IC 50 value (calculated by graphpad software), and the results of three repeated experiments are the final IC 50 value of the measured compound.

[0109] Cell inhibition rate % = [(OD value of blank control - OD value of administration group) / OD value of blank control group] × 100%

[0110] Table 1 Inhibitory activities (IC50, μM) of representative compounds of the present invention against the proliferation of some tumor cells and cardiomyocytes

Table 2

[0111] As can be seen from Table 1, the inhibitory activities of most compounds of the present invention against cardiomyocytes H9C2 are greater than or equivalent to those of CDDO-Me. Compounds I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, I-12, I-13, I-14, I-16, I-18 all showed lower toxicity to human renal tubular epithelial cells HK-2. In particular, the inhibitory activities of compounds I-7 and I-12 against normal cardiomyocytes H9C2 are small (IC 50The values were 3.176±1.74 μM and 3.143±1.53 μM respectively), which was about 1 / 10 of CDDO-Me (IC50 was 0.308±0.01 μM), and showed low cardiotoxicity.

[0112] Example 32 Inhibitory effect of the compound of the present invention on tumors in tumor-bearing mice

[0113] 1) Experimental animals: 72 female ICR mice at 6 - 8 weeks old, experimental cells: S180 ascites tumor cells. Eight animals were set as the normal group, and the remaining mice were modeled by the following method.

[0114] 2) Amplification and subculture of S180 ascites tumor cells: Take S180 ascites tumor cells at -80 °C, dissolve them in a 37 °C water bath, place them in a 12 mL centrifuge tube, add an appropriate amount of physiological saline, centrifuge at 1200 rpm for 5 min, discard the supernatant, add 300 μL of physiological saline to resuspend the cells, inject them into the abdominal cavity of ICR mice. When the abdomen of the mice becomes enlarged, take these cells and inject them into the abdominal cavity of another ICR mouse. When the abdomen of the mouse becomes enlarged, it can be used for inoculation.

[0115] 3) Take second-generation S180 cells inoculated with S180 tumor cells under the armpit of the mice, centrifuge them, and add physiological saline to make the concentration 1×10 7 cells / mL, and subcutaneously inject 100 μL of cells under the armpit. A mouse tumor-bearing model was established.

[0116] The model mice were divided into a tumor-bearing model group, a compound I-7 5 mg / kg group, a compound I-7 15 mg / kg group, a compound I-12 5 mg / kg group, a compound I-12 15 mg / kg group, a compound I-16 5 mg / kg group, a compound I-16 15 mg / kg group, and a CDDO-Me 15 mg / kg group.

[0117] Each group had 8 mice. 4) Administration: When the tumor grows, administer it intragastrically.

[0118] Administer 5 mg / kg of compound I-7 to the compound I-7 5 mg / kg group. The compound I-7 group (15 mg / kg) was administered with 15 mg / kg of compound I-7. The compound I-12 group (5 mg / kg) was administered with 5 mg / kg of compound I-12. The compound I-12 group (15 mg / kg) was administered with 15 mg / kg of compound I-12. The compound I-16 group (5 mg / kg) was administered with 5 mg / kg of compound I-16. The compound I-16 group (15 mg / kg) was administered with 15 mg / kg of compound I-16. The CDDO-Me group (15 mg / kg) was administered with 15 mg / kg of compound CDDO-Me.

[0119] The normal group and the model group were administered with an equal volume of 0.5% sodium carboxymethyl cellulose.

[0120] Administration was carried out continuously for 7 days, and autopsy was performed on the second day after the last administration.

[0121] Detection indicators, body weight, spleen weight, and tumors were measured. The spleen was ground on ice to lyse red blood cells, and after incubating with FITC-CD11b, LY6C, and LY6G antibodies for 15 min, CD11b+LY6Chi+ and CD11b+LY6G+ were assayed by flow cytometry.

[0122] The experimental results are shown in Tables 2 and 3.

[0123] Table 2 Inhibitory effect of the compounds of the present invention on tumors [Table 3]

[0124] Table 3 Effects of the compounds of the present invention on M-MDSC, PMN-MDSC, and spleen index [Table 4]

[0125] MDSCs are a group of immature cells derived from the myeloid lineage, which can normally differentiate into mature macrophages, granulocytes, and dendritic cells. However, in pathological conditions, they aggregate and become activated. In tumors, MDSCs can suppress T cell immune responses and interact with other immune cells to jointly promote the formation of an immunosuppressive state in the tumor microenvironment. Additionally, MDSCs can further promote tumor growth and metastasis through non-immune pathways such as extracellular matrix destruction and angiogenesis promotion. Indoleamine 2,3-dioxygenase 1 (IDO1) is one of the important functional markers of MDSCs, which can catalyze the metabolism of tryptophan in the tumor microenvironment, causing the release of soluble canine threonine and its downstream metabolites, inducing immune tolerance in Tregs and antigen-presenting cells, thereby causing tumor immune escape. There have been reports that MDSCs are involved in the progression of pancreatic cancer, breast cancer, brain metastatic tumors, etc., and as potential therapeutic targets and highly reliable prognostic markers, MDSCs have great research value.

[0126] The research in the examples of the present invention indicates that the compounds of the present invention can suppress the expression of MDSCs, improve the tumor microenvironment, and have a therapeutic effect on lung cancer, liver cancer, pancreatic cancer, breast cancer, ascites tumors, brain metastatic tumors, etc.

[0127] It should also be noted that the compound selected in Example 32 is a representative example of the target compounds of the present invention and does not limit the present invention.

Claims

1. 【Fig. 1】 R １ 、R ２ 、R ３ are each H, C １－６ Alkyl, C １－６ Alkoxy, halogen, cyano, C substituted with halogen or cyano １－６ Alkyl, or H, C １－３ Alkyl, C １－３ Alkoxy, halogen, cyano, C substituted with halogen or cyano １－３ A compound of formula I or a pharmaceutically acceptable salt thereof, independently selected from the substituents of alkyl.

2. The aforementioned R １ , R ２ is independently selected from H or cyano respectively, and R ３ is H, C １－３ alkyl, alkoxy, halogen, C substituted with halogen １－３ alkyl, and the alkoxy contains 1 to 3 carbon atoms, and the halogen is selected from F, Cl, Br, I. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that

3. R １ is H or cyano, R ２ is H or cyano, R ３ is selected from H, methyl, methoxy, trifluoromethyl, Cl or F, a compound according to claim 1 or a pharmaceutically acceptable salt thereof.

4. Said R １ is H or cyano, and R ２ is H or cyano, and R ３ is Cl, H or trifluoromethyl, a compound according to claim 1 or a pharmaceutically acceptable salt thereof.

5. The compound is 3-phenyl-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 3-(4-methylphenyl)-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 3-(4-methoxyphenyl)-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 3-(4-trifluoromethylphenyl)-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 3-(4-fluorophenyl)-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 3-(4-chlorophenyl)-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 2-cyano-3-ylphenyl-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 2-cyano-3-(4-methylphenyl)-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 2-cyano-3-(4-methoxyphenyl)-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 2-cyano-3-(4-trifluoromethylphenyl)-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 2-cyano-3-(4-fluorophenyl)-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 2-cyano-3-(4-chlorophenyl)-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 3-phenyl-3-cyano-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 3-(4-methylphenyl)-3-cyano-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 3-(4-Methoxyphenyl)-3-cyano-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 3-(4-Trifluoromethylphenyl)-3-cyano-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, 3-(4-Fluorophenyl)-3-cyano-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide, and The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that it is selected from 3-(4-chlorophenyl)-3-cyano-N-(2-cyano-3,12-dioxoolean-1,9(11)-dien-19-yl)acrylamide.

6. A method for synthesizing the compound of formula I according to any one of claims 1 to 5, [Chemical 2] wherein the compound of formula II and formula III are subjected to a condensation reaction to produce the compound of formula I, Here, the R １ , R ２ , R ３ is limited as described in any one of claims 1 to 5, and the condensing agent is one or more selected from dicyclohexylcarbodiimide, N,N-diisopropylcarbodiimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide, benzotriazole-1-oxytri(dimethylamino)phosphonium hexafluorophosphate or benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate. A synthesis method characterized by this.

7. The synthesis method of the formula II is [Chemical Formula 3] The synthesis method according to claim 6, characterized in that CDDO is subjected to a Curtius rearrangement to obtain a C-17 aminoated derivative, namely formula II.

8. A pharmaceutical composition comprising the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable pharmaceutical adjuvants.

9. Use of the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 8 in the manufacture of a drug for treating tumors.

10. The tumor is selected from lung cancer, liver cancer, ascites tumor, brain metastasis tumor, colon cancer, pancreatic cancer, breast cancer, prostate cancer, brain cancer, ovarian cancer, cervical cancer, testicular cancer, kidney cancer, head and neck cancer, lymphoma, melanoma or leukemia, or is selected from lung cancer, liver cancer, breast cancer, ascites tumor, pancreatic cancer, brain metastasis tumor, or is selected from non-small cell lung cancer, liver cancer, breast cancer, ascites tumor. The use according to claim 9.

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