Chalcone derivative as well as pharmaceutical composition and application thereof

Abstract

The invention discloses a chalcone derivative, the structural formula of the chalcone derivative is as shown in a general formula (1), imgabs0 #. The invention also provides a pharmaceutical composition of the chalcone derivative and application of the pharmaceutical composition, and the newly synthesized compounds show good inhibitory activity through a test on MTHFD inhibitory activity. The invention has important significance for treating MTHFD related diseases, especially lung cancer, colorectal cancer, breast cancer and the like.

Description

Technical Field

[0001] The present invention relates to the field of compound technology, in particular to the field of medicinal chemistry, and specifically refers to chalcone derivatives, drug combinations thereof and their application in drugs for fighting cancer (including lung cancer, colorectal cancer, breast cancer, ovarian cancer, bladder cancer, prostate cancer, esophageal cancer, liver cancer, pancreatic cancer and inflammation). Background Art

[0002] Methylenetetrahydrofolate dehydrogenase (MTHFD) is a key enzyme in the folate cycle pathway in the one-carbon metabolic pathway and plays a key role in biological processes such as purine synthesis and oxidative stress. MTHFD contains four isoenzymes, namely MTHFD1, MTHFD2, MTHFD1L and MTHFD2L, of which MTHFD1 exists in the cytoplasm, and MTHFD2, MTHFD1L and MTHFD2L all exist in the mitochondria.

[0003] MTHFD1 is a trifunctional enzyme with 5,10-methylenetetrahydrofolate dehydrogenase, 5,10-methenyltetrahydrofolate cyclohydrolase and 10-formyltetrahydrofolate synthetase activities. MTHFD1 is highly expressed in normal tissues, and its derived methylenetetrahydrofolate is a one-carbon source for cellular methylation reactions, which is beneficial for homocysteine ​​remethylation. MTHFD1L is a monofunctional enzyme with 10-formyl synthetase activity. It plays an indispensable role in the production of tetrahydrofolate and plays a very important role in maintaining the folate circulation pathway. MTHFD1 is highly expressed mainly in the neural tube and the developing brain. Therefore, the loss of MTHFD1L is closely related to neural tube abnormalities and craniosynthesis.

[0004] MTHFD2 is a bifunctional enzyme with dehydrogenase / cyclohydrolase activity. It was first extracted from ascites tumor cells by Scrimgeour et al. in 1960 as a nicotinamide adenine dinucleotide (NAD)-dependent methylenetetrahydrofolate dehydrogenase. It was not until 1985 that Mejia detected and confirmed its activity from differentiated cells. A large number of studies have shown that MTHFD2 is highly expressed in a variety of cancers, such as lung cancer, colorectal cancer, ovarian cancer, bladder cancer, prostate cancer, esophageal cancer, liver cancer, and pancreatic cancer, and is associated with poor prognosis of patients. At the same time, MTHFD2 also plays a key role in the synthesis of purines, the generation of ROS, the effects of epigenetic modification and transcriptional regulation, which provides a basis for the development of drugs targeting MTHFD2. MTHFD2L is also a bifunctional enzyme with dehydrogenase / cyclohydrolase activity, and has 60-65% homology with MTHFD2. However, unlike MTHFD2, which is only expressed in embryonic or undifferentiated adult tissues, MTHFD2L is expressed in adult mammals, with the highest expression in brain and lung tissues. Therefore, MTHFD2L can fill the deficiency of one-carbon metabolism caused by the lack of MTHFD2 in adult mammals. However, MTHFD2L is not highly expressed in cancer and has been less studied than MTHFD2.

[0005] According to the 2024 report of the National Cancer Center, the incidence of malignant tumors in my country in 2022 is estimated to be 1.0606 million lung cancer cases and 517,100 colorectal cancer cases. In 2022, the number of deaths from lung cancer in my country was 733,300, and the number of deaths from colorectal cancer was about 240,000. At present, the incidence and mortality of malignant tumors in my country continue to rise, and the annual medical expenses caused by malignant tumors exceed 220 billion. Among men, the incidence and mortality of lung cancer and colorectal cancer are both on the rise, which has attracted attention from all walks of life. At present, the incidence of lung cancer in my country is increasing by 26.9% each year. If effective control measures are not taken in time, it is estimated that by 2025, the number of lung cancer patients in my country will reach 1 million, making it the world's first "lung cancer power". Ovarian cancer refers to a malignant tumor that grows on the ovaries, of which about 90% are primary ovarian cancers, and the rest are primary cancers in other parts of the body that have metastasized to the ovaries. Due to the lack of symptoms in the early stages of ovarian cancer, even if there are symptoms, they are non-specific, and the role of screening is limited. Therefore, early diagnosis is difficult, and most of them are in the late stage when they are treated, and the efficacy of late cases is poor. Therefore, although the incidence of ovarian cancer is lower than that of cervical cancer and endometrial cancer, the mortality rate exceeds the sum of cervical cancer and endometrial cancer, ranking first among gynecological cancers and a major disease threatening women's health. Deepak Nagrath, associate professor of biomedical engineering at the University of Michigan North Campus Research Center (NCRC), and Xiongbin Lu, professor at the Indiana University School of Medicine, developed a CLIM (Collateral Lethal Gene Identification by Metabolic Flux) method to identify collateral lethal genes and successfully revealed a collateral lethal gene (MTHFD2) in ovarian cancer through CLIM. The researchers confirmed the indirect lethality of the UQCR11–MTHFD2 gene pair in mice. The study showed that by inhibiting the expression of MTHFD2 protein, ovarian tumors in mice due to the loss of UQCR11 could be completely alleviated, and inhibiting MTHFD2 basically shut down the energy source of cancer cells. Based on the high expression of MTHFD2 in tumors, inhibiting the metabolic process of MTHFD2 has been shown by more and more studies to inhibit the malignant progression of cancer cells, leading to the death of various cancer cells and improving patient survival. These studies suggest that MTHFD2 may be a new drug target for tumor treatment.

[0006] In addition, as mentioned above, the MTHFD family plays an important role in regulating biological processes such as inflammatory response, cell differentiation, cell proliferation, apoptosis and gene transcription. At the same time, MTHFD2 is the most potential target for tumor treatment compared to other MTHFD family genes, and can be designed for a variety of pathological conditions, including a wide range of cancers and inflammation. At the same time, there are currently no drugs on the market that inhibit the MTHFD pathway to treat cancer and other diseases related to this pathway, such as inflammatory diseases, and the known MTHFD inhibitors have poor selectivity or most of the synthetic steps are complicated. Therefore, the development of new MTHFD inhibitor compounds is of great significance for the treatment of the above-mentioned diseases, especially lung cancer, colorectal cancer and breast cancer. Summary of the invention

[0007] The purpose of the present invention is to overcome the deficiencies in the above-mentioned prior art and provide chalcone derivatives, drug combinations thereof and applications thereof that can be used for MTHFD-related diseases.

[0008] In order to achieve the above object, the first aspect of the present invention provides a chalcone derivative, the main feature of which is that the structural formula of the chalcone derivative is as shown in general formula (1):

[0009]

[0010] In the formula, X is selected from O or S;

[0011] R 1 , R 2 , R 3 , R 4 , R 5 and A are independently selected from: H, hydroxyl, amino, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted C 2-10 Alkenyl, substituted or unsubstituted C 2-6 Alkynyl, substituted or unsubstituted C 3-6 Cycloalkyl, halogen, nitrile, nitro, carboxyl, substituted or unsubstituted sulfonate, substituted or unsubstituted amide, substituted or unsubstituted ester; or, R 2 and R 3 or R 3 and R 4 Together with the carbon atoms to which they are attached, they form a 5- or 6-membered ring structure containing 1-3 heteroatoms selected from O, N or S or containing an alkenyl group;

[0012] R 6 , R 7 , R 9 and R 10 are independently selected from: H, hydroxyl, amino, substituted or unsubstituted C1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted amide, substituted or unsubstituted ester, halogen, amino, nitrile, nitro, carboxyl;

[0013] R 8 for Q is selected from: H, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 2-6 Alkenyl, substituted or unsubstituted C 2-6 Alkynyl, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted C 5-10 aryl or heteroaryl, substituted or unsubstituted sulfonate, substituted or unsubstituted ester, substituted or unsubstituted amide; R 11 is selected from: H, hydroxy, halogen, cyano, nitro, a substituted or unsubstituted 5-7 membered heterocyclic ring containing 1-2 heteroatoms selected from N, O or S, a substituted C 1-10 Alkyl, substituted or unsubstituted C 5-10 aryl or heteroaryl, substituted or unsubstituted amide, substituted or unsubstituted ester;

[0014] The carbon at position 2 and the carbon at position 3 are connected by a carbon-carbon single bond or a double bond;

[0015] When A is a hydroxyl group or an amino group, it can form a ring with the 2-carbon to form chalcone derivatives: dihydroflavonoids, flavones, flavonols and isoflavones.

[0016] Replace C 1-10 Alkyl, C 1-10 Alkoxy, C 2-10 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, sulfonate, amide, ester, C 5-10 The substituents of the aryl or heteroaryl are independently selected from one or more of cyano, halogen, hydroxyl, amino, nitro, carboxyl, ester, oxo, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted sulfonate, substituted or unsubstituted amide, substituted or unsubstituted ester, and substituted or unsubstituted aryl.

[0017] Preferably, R 2 and R 4 Independently selected from: H,

[0018] The 2' carbon and 3' carbon are connected by a carbon-carbon single bond or double bond; when the 2' carbon and 3' carbon are connected by a carbon-carbon single bond, B is selected from the following group: H, hydroxyl, amino, substituted or unsubstituted C 1-5 Alkyl, substituted or unsubstituted C 1-5 Alkoxy, substituted or unsubstituted C 2-5 Alkenyl, substituted or unsubstituted C 2-5 Alkynyl, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted amide, substituted or unsubstituted ester, halogen, nitrile, nitro, carboxyl;

[0019] Preferably, the chalcone derivative compound is selected from the following compounds:

[0020]

[0021]

[0022]

[0023]

[0024]

[0025] The second aspect of the present invention provides the use of the chalcone derivatives in the preparation of drugs for MTHFD inhibitors.

[0026] The third aspect of the present invention provides the use of chalcone derivatives in the preparation of drugs for MTHFD inhibitors, wherein the chalcone derivatives are selected from:

[0027]

[0028]

[0029] The fourth aspect of the present invention provides a pharmaceutical composition, the main feature of which is that the pharmaceutical composition comprises the chalcone derivative or its tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier.

[0030] The fifth aspect of the present invention provides the use of the pharmaceutical composition in treating and / or preventing MTHFD-related diseases.

[0031] Preferably, the MTHFD-related disease is cancer.

[0032] Preferably, the cancer includes lung cancer, colorectal cancer, breast cancer, ovarian cancer, bladder cancer, prostate cancer, esophageal cancer, liver cancer, pancreatic cancer and inflammation.

[0033] As used herein, “a” or “an” or “a kind” means at least one / kind / kind or one / kind / kind or more than one / kind / kind.

[0034] As used herein, "alkyl" refers to various saturated straight-chain, side-chain or cyclic hydrocarbon groups, and particularly includes small alkyl groups containing ten or less carbon atoms.

[0035] The "aryl group" used in the present invention is a cyclic conjugated aromatic system and may contain one or more non-carbon atoms (other heteroatoms other than carbon, such as nitrogen) in the ring, such as phenyl, pyridyl, pyrazinyl, etc.

[0036] Specifically, the various substituents defined herein are selected from one or more of cyano, halogen, hydroxyl, amino, nitro, carboxyl, ester, oxo, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted sulfonate, substituted or unsubstituted amide, substituted or unsubstituted ester, and substituted or unsubstituted aryl. The various substituents defined herein also include groups formed by further substitutions, wherein these new substituents may also contain other groups. For example, hydrogen atoms on alkyl and aryl groups are replaced by amino, halogen or other groups to become groups in the above definitions.

[0037] A series of new compounds are synthesized by using the chalcone derivatives, the drug combination and the application thereof of the present invention. Many of these new compounds show excellent activity through the inhibitory activity test on MTHFD, and are very valuable for treating MTHFD-related diseases. The lack of existing drug types and efficacy for treating MTHFD-related diseases is made up, which is very meaningful. DETAILED DESCRIPTION

[0038] In order to further understand the present invention, preferred embodiments of the present invention are described below in conjunction with examples, but it should be understood that these descriptions are only for further illustrating the features and advantages of the present invention, rather than limiting the claims of the invention.

[0039] All raw materials of the present invention have no particular limitation on their sources, and can be purchased from the market or prepared according to conventional methods known to those skilled in the art.

[0040] In order to further illustrate the present invention, the chalcone derivatives provided by the present invention are described in detail below in conjunction with the examples, and the protection scope of the present invention is not limited by the following examples.

[0041] Example 1

[0042] Preparation of chalcone MTHFD inhibitors M1-M56

[0043]

[0044] Take a dry and clean reaction bottle, dissolve S1 (168 mg, 1.0 mmol) in 2-3 mL of dry ethyl acetate, add S2 (239 mg, 1.6 mmol), stir at room temperature for 10 min, then add KOH (84 mg, 1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in turn, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound S3 (59.7%) by column chromatography (ethyl acetate / petroleum ether=1:8).

[0045] Take a dry and clean reaction bottle, dissolve the above synthesized compound S3 (1.0 eq) in 5-10 mL DCM, add DIPEA (3.0 eq), stir under ice bath for 10 min, add dropwise to dissolve in MOMBr (2.2 eq), react at room temperature for 1-24 h, add 10 mL of water to quench the reaction after TLC detection, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine solution (10 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound S4 (60-90%) by column chromatography (ethyl acetate / petroleum ether=1:15).

[0046] Take a dry and clean reaction bottle, dissolve the above-synthesized compound S4 (1.0 eq) in 5-10 mL of ethanol aqueous solution, add S5 (1.0 eq), stir at room temperature for 10 min, then add KOH (7.0 eq), react at room temperature for 1-24 h, add 10 mL of water after the reaction is completed by TLC detection, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine solution (10 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound S6 (60-90%) by column chromatography (ethyl acetate / petroleum ether=1:5).

[0047] Take a dry and clean reaction bottle, dissolve the above synthesized compound S6 (1.0eq) in 5-10mL methanol, add 3M hydrochloric acid (0.1mL), react at reflux temperature for 1-24h, add 10mL water after TLC detection, extract with ethyl acetate three times (10mL*3), combine the organic phases and wash with saturated saline solution (10mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure and obtain compounds M1-M56 (50-70%) by column chromatography (ethyl acetate / petroleum ether=2:1). The structural formula and characterization information of compounds M1-M56 are shown in Table 1 below.

[0048] Table 1 Structural formula and characterization information of compounds M1-M56

[0049]

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056] Example 2

[0057] Preparation of chalcone MTHFD inhibitors M57-M63

[0058]

[0059] Take a dry and clean reaction bottle, dissolve S7 (1.0 mmol) in 2-3 mL of dry ethyl acetate, add S2 (239 mg, 1.6 mmol), stir at room temperature for 10 min, then add KOH (84 mg, 1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in turn, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound S8 (5-47%) by column chromatography (ethyl acetate / petroleum ether=1:8).

[0060] Take a dry and clean reaction bottle, dissolve the above-synthesized compound S8 (1.0 eq) in 5-10 mL of ethanol aqueous solution, add S5 (1.0 eq), stir at room temperature for 10 min, then add KOH (7.0 eq), react at room temperature for 1-24 h, add 10 mL of water after the reaction is completed by TLC detection, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine solution (10 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compounds M57-M63 (60-79%) by column chromatography (ethyl acetate / petroleum ether = 1:5).

[0061] The structural formula and characterization information of compounds M57-M63 are shown in Table 2 below.

[0062] Table 2 Structural formula and characterization information of compounds M57-M63

[0063]

[0064]

[0065] Example 3

[0066] Preparation of chalcone MTHFD inhibitors M64-M67

[0067]

[0068] Take a dry and clean reaction bottle, dissolve S9 (168 mg, 1.0 mmol) in 2-3 mL of dry ethyl acetate, add S2 (239 mg, 1.6 mmol), stir at room temperature for 10 min, then add KOH (84 mg, 1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound S10 (5-47%) by column chromatography (ethyl acetate / petroleum ether=1:8).

[0069] Take a dry and clean reaction bottle, dissolve the above-synthesized compound S10 (1.0 eq) in 5-10 mL of ethanol aqueous solution, add S5 (1.0 eq), stir at room temperature for 10 min, then add KOH (7.0 eq), react at room temperature for 1-24 h, add 10 mL of water after the reaction is completed by TLC detection, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine solution (10 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compounds M64-M67 (60-90%) by column chromatography (ethyl acetate / petroleum ether = 1:5).

[0070] The structural formula and characterization information of compounds M64-M67 are shown in Table 3 below.

[0071] Table 3 Structural formula and characterization information of compounds M64-M67

[0072]

[0073] Example 4

[0074] Preparation of dihydroflavonoid MTHFD inhibitors M68-M77

[0075]

[0076] Take a dry and clean reaction bottle, take the similar raw material chalcone (0.5 mmol) in Example 1 and dissolve it in 3 mL of ethanol, add sodium acetate (6.0 eq), and react at reflux temperature for 1-24 h. After the reaction is completed by TLC detection, add 10 mL of water, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (10 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compounds M68-M77 (32-76%) by column chromatography (ethyl acetate / petroleum ether=1:4).

[0077] The structural formula and characterization information of compounds M68-M77 are shown in Table 4 below.

[0078] Table 4 Structural formula and characterization information of compounds M68-M77

[0079]

[0080]

[0081] Example 5

[0082] Preparation of chalcone MTHFD inhibitors M78-M79

[0083]

[0084] Take a dry and clean reaction bottle, dissolve S14 (168 mg, 1.0 mmol) in 2-3 mL of dry ethyl acetate, add S2 (239 mg, 1.6 mmol), stir at room temperature for 10 min, then add KOH (84 mg, 1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in turn, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compounds M78-M79 (11-17%) by column chromatography (ethyl acetate / petroleum ether = 1:8).

[0085] The structural formula and characterization information of compounds M78-M79 are shown in Table 5 below.

[0086] Table 5 Structural formula and characterization information of compounds M78-M79

[0087]

[0088] Example 6

[0089] Preparation of dihydroflavonoid MTHFD inhibitors M80-M81

[0090]

[0091] Take a dry and clean reaction bottle, dissolve S15 (168 mg, 1.0 mmol) in 2-3 mL of dry ethyl acetate, add S2 (239 mg, 1.6 mmol), stir at room temperature for 10 min, then add KOH (84 mg, 1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed, and extract with ethyl acetate three times (10 mL*3). After combining the organic phases, wash with saturated brine (6 mL) in turn, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compounds M80-M81 (9-14%) by column chromatography (ethyl acetate / petroleum ether = 1:8).

[0092] The structural formula and characterization information of compounds M80-M81 are shown in Table 6 below.

[0093] Table 6 Structural formula and characterization information of compounds M80-M81

[0094]

[0095] Example 7

[0096] Preparation of flavonol MTHFD inhibitor M82

[0097]

[0098] Take a dry and clean reaction bottle, dissolve S16 (168 mg, 1.0 mmol) in 2-3 mL of dry ethyl acetate, add S2 (239 mg, 1.6 mmol), stir at room temperature for 10 min, then add KOH (84 mg, 1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in turn, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound M82 (12%) by column chromatography (ethyl acetate / petroleum ether = 1:8).

[0099] The structural formula and characterization information of compound M82 are shown in Table 7 below.

[0100] Table 7 Structural formula and characterization information of compound M82

[0101]

[0102] Example 8

[0103] Preparation of flavonol MTHFD inhibitor M83

[0104]

[0105] Take a dry and clean reaction bottle, dissolve S17 (168 mg, 1.0 mmol) in 2-3 mL of dry ethyl acetate, add S18 (239 mg, 1.6 mmol), stir at room temperature for 10 min, then add KOH (84 mg, 1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in turn, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound M83 (18%) by column chromatography (ethyl acetate / petroleum ether = 1:8).

[0106] The structural formula and characterization information of compound M83 are shown in Table 8 below.

[0107] Table 8 Structural formula and characterization information of compound M83

[0108]

[0109] Example 9

[0110] Preparation of chalcone MTHFD inhibitor M84

[0111]

[0112] Take a dry and clean reaction bottle, dissolve S1 (168 mg, 1.0 mmol) in 2-3 mL of dry ethyl acetate, add S2 (239 mg, 1.6 mmol), stir at room temperature for 10 min, then add KOH (84 mg, 1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in turn, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound S19 (8.7%) by column chromatography (ethyl acetate / petroleum ether = 1:15).

[0113] Take a dry and clean reaction bottle, dissolve the above-synthesized compound S19 (1.0 eq) in 5-10 mL of ethanol aqueous solution, add S20 (1.0 eq), stir at room temperature for 10 min, then add KOH (7.0 eq), react at room temperature for 1-24 h, add 10 mL of water after the reaction is completed by TLC, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine solution (10 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound M84 (67%) by column chromatography (ethyl acetate / petroleum ether = 1:5).

[0114] The structural formula and characterization information of compound M84 are shown in Table 9 below.

[0115] Table 9 Structural formula and characterization information of compound M84

[0116]

[0117] Example 10

[0118] Preparation of chalcone MTHFD inhibitors M85-M87

[0119]

[0120] Take a dry and clean reaction bottle, dissolve S21 (1.0 mmol) in 2-3 mL of dry ethyl acetate, add S2 (1.6 mmol), stir at room temperature for 10 min, then add KOH (84 mg, 1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in turn, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound S22 (8.7%) by column chromatography (ethyl acetate / petroleum ether = 1:15).

[0121] Take a dry and clean reaction bottle, dissolve the above-synthesized compound S22 (1.0 eq) in 5-10 mL of ethanol aqueous solution, add S23 (1.0 eq), stir at room temperature for 10 min, then add KOH (7.0 eq), react at room temperature for 1-24 h, add 10 mL of water after the reaction is completed by TLC, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (10 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound S24 (40-70%) by column chromatography (ethyl acetate / petroleum ether=1:5).

[0122] Take a dry and clean reaction bottle, dissolve the above-synthesized compound S24 (1.0 eq) in 5-10 mL of methanol, add 3 M hydrochloric acid (0.1 mL), react at reflux temperature for 1-24 h, add 10 mL of water after the reaction is completed by TLC detection, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (10 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compounds M85-M87 (50-70%) by column chromatography (ethyl acetate / petroleum ether=1:1).

[0123] The structural formula and characterization information of compounds M85-M87 are shown in Table 10 below.

[0124] Table 10 Structural formula and characterization information of compounds M85-M87

[0125]

[0126]

[0127] Embodiment 11

[0128] Preparation of chalcone MTHFD inhibitors M88-M100

[0129]

[0130] Take a dry and clean reaction bottle, dissolve M78 (1.0 mmol) in 2-3 mL of dry THF, add S25 (1.5 mmol), stir at room temperature for 10 min, then add NaH (2.4 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in turn, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compounds M88-M100 (20-50%) by column chromatography (ethyl acetate / petroleum ether = 1:15).

[0131] The structural formula and characterization information of compounds M88-M100 are shown in Table 11 below.

[0132] Table 11 Structural formula and characterization information of compounds M88-M100

[0133]

[0134]

[0135]

[0136] Example 12

[0137] Preparation of chalcone MTHFD inhibitor M101

[0138]

[0139] Take a dry and clean reaction bottle, add M9 (1.0 mmol), S26 (1.2 mmol), DCC (1.5 mmol) and HOBt (1.2 mmol) in sequence, and then add ultra-dry DMF (3 mL). React at room temperature overnight. After the reaction is completed by TLC monitoring, add 5 mL of water, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in sequence, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound M101 (2.1%) by column chromatography (dichloromethane / methanol = 10:1).

[0140] The structural formula and characterization information of compound M101 are shown in Table 12 below.

[0141] Table 12 Structural formula and characterization information of compound M101

[0142]

[0143] Embodiment 13

[0144] Preparation of chalcone MTHFD inhibitors M102-M104

[0145]

[0146] Take a dry and clean reaction bottle, add S6 (1.0 mmol), S27 (1.2 mmol), DCC (1.5 mmol) and HOBt (1.2 mmol) in sequence, then add ultra-dry DMF (3 mL), react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in sequence, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound S28 (30-60%) by column chromatography (dichloromethane / methanol = 10:1).

[0147] Take a dry and clean reaction bottle, dissolve the above synthesized compound S28 (1.0 eq) in 5-10 mL of methanol, add 3 M hydrochloric acid (0.1 mL), react at reflux temperature for 1-24 h, add 10 mL of water after the reaction is completed by TLC detection, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine solution (10 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compounds M102-M104 (5-30%) by column chromatography (dichloromethane / methanol=10:1).

[0148] The structural formula and characterization information of compounds M102-M104 are shown in Table 13 below.

[0149] Table 13 Structural formula and characterization information of compounds M102-M104

[0150]

[0151]

[0152] Embodiment 14

[0153] Preparation of chalcone MTHFD inhibitors M105-M107

[0154]

[0155] Take a dry and clean reaction bottle, dissolve S21 (1.0 mmol) in 2-3 mL of dry ethyl acetate, add S2 (1.6 mmol), stir at room temperature for 10 min, then add KOH (84 mg, 1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in turn, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound S22 (8.7%) by column chromatography (ethyl acetate / petroleum ether = 1:15).

[0156] Take a dry and clean reaction bottle, dissolve the above-synthesized compound S22 (1.0 eq) in 5-10 mL of ethanol aqueous solution, add S29 (1.0 eq), stir at room temperature for 10 min, then add KOH (7.0 eq), react at room temperature for 1-24 h, add 10 mL of water after the reaction is completed by TLC detection, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine solution (10 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compounds M105-M107 (40-70%) by column chromatography (ethyl acetate / petroleum ether = 1:5).

[0157] The structural formula and characterization information of compounds M105-M107 are shown in Table 14 below.

[0158] Table 14 Structural formula and characterization information of compounds M105-M107

[0159]

[0160] Embodiment 15

[0161] Preparation of chalcone MTHFD inhibitor M108

[0162]

[0163] Take a dry and clean reaction bottle, dissolve S5 (1.0 mmol) in 2-3 mL of dry ethyl acetate, add S2 (239 mg, 1.6 mmol), stir at room temperature for 10 min, then add KOH (1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed, and extract with ethyl acetate three times (10 mL*3). After combining the organic phases, wash with saturated brine (6 mL) in turn, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound S30 (58%) by column chromatography (ethyl acetate / petroleum ether = 1:10).

[0164] Take a dry and clean reaction bottle, dissolve the above-synthesized compound S30 (1.0 eq) in 5-10 mL of ethanol, add S21 (1.0 eq), stir at room temperature for 10 min, then add KOH (7.0 eq), react at room temperature for 1-24 h, add 10 mL of water after the reaction is completed by TLC, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (10 mL), dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound M108 (37%) by column chromatography (ethyl acetate / petroleum ether = 1:3).

[0165] The structural formula and characterization information of compound M108 are shown in Table 15 below.

[0166] Table 15 Structural formula and characterization information of compound M108

[0167]

[0168] Example 16

[0169] Preparation of chalcone MTHFD inhibitor M109

[0170]

[0171] Take a dry and clean reaction bottle, add M106 (1.0 mmol), S26 (1.2 mmol), DCC (1.5 mmol) and HOBt (1.2 mmol) in sequence, then add ultra-dry DMF (3 mL), react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in sequence, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound M109 (9.4%) by column chromatography (dichloromethane / methanol = 10:1).

[0172] The structural formula and characterization information of compound M109 are shown in Table 16 below.

[0173] Table 16 Structural formula and characterization information of compound M109

[0174]

[0175] Embodiment 17

[0176] Preparation of chalcone MTHFD inhibitors M110-M111

[0177]

[0178] Take a dry and clean reaction bottle, add M1 or M23 (1.0 mmol), boron trifluoride ether complex (1.2 mmol) and ultra-dry THF (3 mL) in sequence, react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in sequence, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compounds M110-M111 (30-50%) by column chromatography (dichloromethane / methanol = 20:1).

[0179] The structural formula and characterization information of compounds M110-M111 are shown in Table 17 below.

[0180] Table 17 Structural formula and characterization information of compounds M110-M111

[0181]

[0182] Embodiment 18

[0183] Preparation of chalcone MTHFD inhibitors M112-M113

[0184]

[0185] Take a dry and clean reaction bottle, add M1 or M11 (1.0 mmol), 10% palladium carbon catalyst (10 wt%) and methanol (3 mL) in sequence, replace three times and then pass hydrogen, react overnight at room temperature, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in sequence, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and then obtain compounds M112-M113 (43-58%) by column chromatography (dichloromethane / methanol=20:1).

[0186] The structural formula and characterization information of compounds M112-M113 are shown in Table 18 below.

[0187] Table 18 Structural formula and characterization information of compounds M112-M113

[0188]

[0189]

[0190] Embodiment 19

[0191] Preparation of chalcone MTHFD inhibitors M114-M115

[0192]

[0193] Take a dry and clean reaction bottle, dissolve S31 (1.0 mmol) in 2-3 mL of dry ethyl acetate, add S2 (239 mg, 1.6 mmol), stir at room temperature for 10 min, then add KOH (1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed under TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in turn, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compounds M114-M115 (58%) by column chromatography.

[0194] The structural formula and characterization information of compounds M114-M115 are shown in Table 19 below.

[0195] Table 19 Structural formula and characterization information of compounds M114-M115

[0196]

[0197] Embodiment 20

[0198] Preparation of chalcone MTHFD inhibitor M116

[0199]

[0200] Take a dry and clean reaction bottle, dissolve S15 (1.0 mmol) in 2-3 mL of dry ethanol, add S2 (1.6 mmol), stir at room temperature for 10 min, then add KOH (1.5 mmol), react at room temperature overnight, add 5 mL of water after the reaction is completed by TLC monitoring, extract with ethyl acetate three times (10 mL*3), combine the organic phases, wash with saturated brine (6 mL) in sequence, dry over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain compound M116 (37%) by column chromatography.

[0201] The structural formula and characterization information of compound M116 are shown in Table 20 below.

[0202] Table 20 Structural formula and characterization information of compound M116

[0203]

[0204] Embodiment 21

[0205] Inhibitory activity of chalcone derivatives on MTHFD

[0206] (1) Reagents and materials: 3-Methylpropanesulfonic acid (MOPS), tripotassium phosphate (K3PO4), and magnesium chloride (MgCl2) were purchased from Shanghai McLean Biochemical Technology Co., Ltd. Formaldehyde (HCHO) and β-mercaptoethanol were purchased from Merck-Sigma Co., Ltd., Germany. Tetrahydrofolate (THF), nicotinamide adenine dinucleotide phosphate (NADP), and nicotinamide adenine dinucleotide (NAD) were purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0207] (2) Experimental instrument: ELISA reader (Bio Tek, USA)

[0208] (3) Experimental method: MTHFD2 has the functions of dehydrogenase and cyclohydrolase. Based on its dehydrogenase activity, we designed a kinetic simulation method to screen MTHFD2 inhibitors by detecting the absorbance of the product. The specific principle is that MTHFD2 can catalyze the conversion of 5,10-methylenetetrahydrofolate into 5,10-methylenetetrahydrofolate, and NAD + After obtaining hydrogen ions, it is converted into NADH. Since the generated product 5,10-methylenetetrahydrofolate has specific absorption at 350nm, the enzymatic activity efficiency of MTHFD2 can be reflected by measuring the change in the absorption value at 350nm, thereby judging the strength of the compound's inhibitory activity on proteins. In general, the greater the rate of change in the absorption value, the stronger the enzyme activity, and the weaker the inhibitory activity of the corresponding compound.

[0209] The specific process is as follows: Add the purified MTHFD2 protein to the enzyme activity test buffer (25mM MOPS, 5mMK3PO4, 5mM MgCl2, 2.5mM HCHO, 36mMβ-mercaptoethanol) to a final concentration of 1μg / mL of MTHFD2 protein. Add coenzyme NAD to the solution containing MTHFD2 protein to a concentration of 600μM. After mixing, draw 100μL of the mixed solution into a 96-well plate. At the same time, add the corresponding concentration of compounds to the 96-well plate and incubate at room temperature for 5min, then add the substrate THF to start the reaction, where the final concentration of THF is 200μM. Use the kinetic detection module of the microplate reader, set the reading every 30s, the total detection time is 6min, and read the absorption value at 350nm. Finally, the data processing of the inhibitor is to calculate the average rate (Vmean) of the derived DMSO group and compound group using the following formula:

[0210] Inhibitor (%) = (Vmean (DMSO) - Vmean (compound)) / Vmean (compound) * 100

[0211] (4) Experimental results

[0212]

[0213] All the compounds synthesized above and the natural products described in the above table have a certain inhibitory effect on MTHFD1 and MTHFD2. The inhibition rate of all compounds (at concentration = 10 μM) is graded by activity (mainly based on MTHFD2):

[0214] A: Compound inhibition rate (%) ≥ 70%; B: 70%> Compound inhibition rate (%) ≥ 50%; C: 50%> Compound inhibition rate (%) ≥ 30%; D: 30%> Compound inhibition rate (%)

[0215] The results of the inhibitory activity of each compound on MTHFD are shown in Table 21 below.

[0216] Table 21 Inhibitory activity of synthetic compounds on MTHFD1 and MTHFD2 proteins

[0217]

[0218]

[0219] LY345899: Positive control compound, a folic acid analog, is an inhibitor of MTHFD1 and MTHFD2, with inhibition rates of 97.21% and 89.67% at 10 μM, respectively. Other MOM-protected chalcone and M127 also have a certain inhibitory effect.

[0220] Embodiment 22

[0221] Effects of MTHFD2 inhibitors on cancer cell proliferation

[0222] (1) Cells: Human lung cancer cells H1299, H2030, H441; human colon cancer cells HCT116, LS174T; human breast cancer cells MCF7, MDA-MB-231

[0223] (2) Reagents: MTT was purchased from Beijing Lanjieke Technology Co., Ltd., DMSO was purchased from Shanghai McLean Biochemical Technology Co., Ltd.

[0224] (3) Experimental instrument: ELISA reader (Bio Tek, USA)

[0225] (4) Experimental method: This cell proliferation experiment uses the MTT test method. The specific principle is that in living cells, succinate dehydrogenase in the mitochondria can reduce MTT to blue-purple crystalline formazan, and formazan is insoluble in water but soluble in DMSO, and has specific absorption at 490nm.

[0226] The specific process is as follows: 3000 cells were plated in each well of a 96-well plate. After the cells were completely attached to the wall, they were treated with different concentrations of MTHFD2 inhibitor for 72 hours, and three groups were paralleled in each experimental group. After the compound treatment was completed, 10% MTT solution (5 mg / mL) was added to the cells and the cells were placed for another 4 hours. The supernatant solution was gently sucked off with a spray gun, and 100 μL of DMSO solution was added to dissolve the formazan. After oscillation, the absorbance at 490 nm was detected with an ELISA reader.

[0227] (5) Experimental results:

[0228] The following experimental data are the IC values ​​of MTHFD2 inhibitors on the proliferation of human colon cancer cells. 50 The results are shown in Table 22 below.

[0229] Table 22 IC50 values ​​of each compound on the proliferation of human colon cancer cells

[0230]

[0231] The following experimental data are the IC values ​​of MTHFD2 inhibitors on human breast cancer cell proliferation. 50 The results are shown in Table 23 below.

[0232] Table 23 IC50 values ​​of each compound on the proliferation of human breast cancer cells

[0233]

[0234] The following experimental data are the inhibitory effects of 30 μM MTHFD2 inhibitor on lung cancer cell proliferation, and the results are shown in Table 24 below.

[0235] Table 24 Inhibition rate of each compound against lung cancer cell proliferation inhibition rate (30 μM)

[0236]

[0237]

[0238] The experimental results showed that all the tested compounds had an inhibitory effect on the proliferation of different cancer cells.

[0239] In this specification, the present invention has been described with reference to specific embodiments thereof. However, it is apparent that various modifications and variations may be made without departing from the spirit and scope of the present invention. Therefore, the description should be regarded as illustrative rather than restrictive.

Claims

1. A chalcone derivative, or its tautomer, mesomer, racemate, enantiomer, diastereomer, mixture thereof or pharmaceutically acceptable salt thereof, characterized in that: The structural formula of the chalcone derivatives is shown in general formula (1): In the formula, X is selected from O or S; R 1 , R 2 , R 3 , R 4 , R 5 and A are independently selected from: H, hydroxyl, amino, substituted or unsubstituted C 1-10 Alkyl, substituted or unsubstituted C 1-10 Alkoxy, substituted or unsubstituted C 2-10 Alkenyl, substituted or unsubstituted C 2-6 Alkynyl, substituted or unsubstituted C 3-6 Cycloalkyl, halogen, nitrile, nitro, carboxyl, substituted or unsubstituted sulfonate, substituted or unsubstituted amide, substituted or unsubstituted ester; or, R 2 and R 3 or R 3 and R 4 Together with the carbon atoms to which they are attached, they form a 5- or 6-membered ring structure containing 1-3 heteroatoms selected from O, N or S or containing an alkenyl group; R 6 , R 7 , R 9 and R 10 are independently selected from: H, hydroxyl, amino, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 2-10 alkenyl, substituted or unsubstituted amide, substituted or unsubstituted ester, halogen, amino, nitrile, nitro, carboxyl; R 8 for Q is selected from: H, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 1-6 Alkoxy, substituted or unsubstituted C 2-6 Alkenyl, substituted or unsubstituted C 2-6 Alkynyl, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted C 5-10 aryl or heteroaryl, substituted or unsubstituted sulfonate, substituted or unsubstituted ester, substituted or unsubstituted amide; R 11 is selected from: H, hydroxy, halogen, cyano, nitro, a substituted or unsubstituted 5-7 membered heterocyclic ring containing 1-2 heteroatoms selected from N, O or S, a substituted C 1-10 Alkyl, substituted or unsubstituted C 5-10 aryl or heteroaryl, substituted or unsubstituted amide, substituted or unsubstituted ester; The carbon at position 2 and the carbon at position 3 are connected by a carbon-carbon single bond or a double bond; When A is a hydroxyl group or an amino group, it can form a ring with the 2-carbon to form a chalcone derivative.

2. The chalcone derivative according to claim 1, characterized in that: Replace C 1-10 Alkyl, C 1-10 Alkoxy, C 2-10 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, sulfonate, amide, ester, C 5-10 The substituents of the aryl or heteroaryl are independently selected from one or more of cyano, halogen, hydroxyl, amino, nitro, carboxyl, ester, oxo, substituted or unsubstituted alkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted sulfonate, substituted or unsubstituted amide, substituted or unsubstituted ester, and substituted or unsubstituted aryl.

3. The chalcone derivative according to claim 1 or 2, characterized in that: R 2 and R 4 Independently selected from: H, The 2' carbon and 3' carbon are connected by a carbon-carbon single bond or double bond; when the 2' carbon and 3' carbon are connected by a carbon-carbon single bond, B is selected from: H, hydroxyl, amino, substituted or unsubstituted C 1-5 Alkyl, substituted or unsubstituted C 1-5 Alkoxy, substituted or unsubstituted C 2-5 Alkenyl, substituted or unsubstituted C 2-5 Alkynyl, substituted or unsubstituted C 3-6 Cycloalkyl, substituted or unsubstituted amide, substituted or unsubstituted ester, halogen, nitrile, nitro, carboxyl; when the 2' carbon and 3' carbon are connected by a carbon-carbon double bond, B does not exist.

4. The chalcone derivative according to any one of claims 1 to 3, characterized in that The chalcone derivatives are selected from the following compounds:

5. Use of the chalcone derivative according to any one of claims 1 to 4 in the preparation of a drug for an MTHFD inhibitor.

6. Use of chalcone derivatives in the preparation of drugs for MTHFD inhibitors, characterized in that: The chalcone derivatives are selected from:

7. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the chalcone derivative according to any one of claims 1 to 6 or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof or a pharmaceutically acceptable salt thereof, and optionally a pharmaceutically acceptable carrier.

8. Use of the pharmaceutical composition according to claim 7 in treating and / or preventing MTHFD-related diseases.

9. The use according to claim 8, characterized in that The MTHFD-related disease is cancer.

10. The use according to claim 9, characterized in that: The cancers include lung cancer, colorectal cancer, breast cancer, ovarian cancer, bladder cancer, prostate cancer, esophageal cancer, liver cancer, pancreatic cancer and inflammation.

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