Compound containing thioether skeleton structure and application of compound in preparation of phosphoglycerate dehydrogenase inhibitor

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a compound containing a thioether skeleton structure and application of the compound in preparation of a phosphoglycerate dehydrogenase inhibitor. According to the invention, a series of novel PHGDH small-molecule inhibitors containing thioether skeletons are obtained through screening of a compound library and structural modification, and the structural types and indications of the PHGDH inhibitors are enriched. The invention has the following technical effects: the invention provides a compound as a PHGDH inhibitor, tests show that the compound directly generates an inhibition effect on PHGDH, active small molecules with novel structures and good physicochemical properties are found, and the small molecules show good activity in various experiments and are expected to be developed into specific drugs for treatment.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and in particular relates to a compound containing a thioether skeleton structure and application thereof in the preparation of a phosphoglycerate dehydrogenase inhibitor. Background Art

[0002] Among the nutrients required for mammalian cell proliferation, serine is the third largest metabolite after glucose and glutamine, and ranks second among all amino acids. Serine can promote the generation of energy required to support normal cell proliferation and differentiation metabolism and the synthesis of many biomacromolecules, and can provide building blocks for molecules involved in various biochemical pathways, including amino acids, lipids, nucleotides, and cofactors. Cells can obtain serine from the external environment through transporters, or through the serine biosynthesis pathway, using the glycolysis intermediate 3-phosphoglycerate (3-PG) for biosynthesis. SSP is the main way to provide intracellular serine, and studies have shown that increasing the intake of exogenous serine cannot make up for the functional defects of SSP. Phosphoglycerate dehydrogenase (PHGDH) catalyzes the oxidation of 3-PG in SSP and is the only rate-limiting enzyme in the serine synthesis pathway. Since the uptake of exogenous serine is not enough to make up for the lack of serine in cells caused by SSP dysfunction, the normal physiological function of PHGDH is particularly critical for the homeostasis of serine in cells. In recent years, pharmacological studies on PHGDH have mostly focused on its high expression. Currently, this target has been found to be upregulated in diseases such as breast cancer, melanoma, liver cancer, lung cancer, colon cancer, leukemia and liver damage.

[0003] Currently, there are few studies on small molecule compounds targeting PHGDH. Although existing PHGDH inhibitors such as NCT-503, CBR-5884 series, SENKYUNOLIDEA, etc. have shown certain potential in the treatment of tumors and liver damage, their widespread application in clinical practice is still limited due to problems such as poor selectivity, drug resistance, poor pharmacokinetics and side effects.

[0004] The present invention provides a class of thioether compounds, which have a significant inhibitory effect on the serine synthesis pathway regulated by PHGDH, and this effect has not been fully revealed in existing PHGDH inhibitors. By optimizing the molecular structure of thioether inhibitors, the compounds of the present invention show potential application value as anti-tumor drugs and liver injury treatment, providing new treatment methods for related diseases. Summary of the invention

[0005] Compared with existing PHGDH inhibitors, the thioether inhibitors of the present invention exhibit a unique mechanism of action. By targeting the specific binding site of PHGDH, the compounds of the present invention can not only show a high degree of inhibition on the catalytic activity of PHGDH, but also effectively reduce the expression of PHGDH in cells.

[0006] In order to achieve the above-mentioned object of the invention, the present invention adopts the following technical solutions:

[0007] A compound containing a thioether skeleton structure of general formula (I) or a pharmaceutically acceptable salt thereof,

[0008]

[0009] Wherein, R is a substituted or unsubstituted aromatic ring or aromatic heterocycle, wherein the aromatic ring is a five-membered or six-membered aromatic ring, the aromatic heterocycle is a monocyclic aromatic heterocycle or a polycyclic condensed heterocycle, and the substituent groups include monosubstituted or polysubstituted phenyl, alkyl, alkoxy, amino, hydroxyl, nitro, halogen, trifluoromethyl, carbonyl, sulfonamide, and sulfenyl imide groups.

[0010] Preferably, R is a substituted or unsubstituted aromatic heterocycle, the aromatic heterocycle is selected from any one of tetrazole, thiazole, quinoline, thiadiazole, benzothiazole, thiazolylpyridine, benzoxazole, thiazoline, indole, purine, pyridine, pyrimidine, triazole, and oxazole, and the substituent group includes a monosubstituted or polysubstituted phenyl, methyl, amino, nitro or hydroxyl group.

[0011] More preferably, R is selected from any one of the following groups: phenyltetrazole, methyltetrazole, thiazole, quinoline, 1,3,4-thiadiazole, methylbenzothiazole, phenylthiazole, methylthiazolopyridine, nitrothiazole, benzothiazole, benzoxazole, thiazoline, indole, purine, pyridine, pyrimidine, methylbenzothiazole, methylbenzoxazole, triazole, hydroxytriazole, hydroxymethylpyrimidine, diphenyloxazole, nitrobenzothiazole, and aminothiadiazole.

[0012] The compound may be any one of the following compound formulas, but is not limited to the following molecules:

[0013]

[0014]

[0015] The pharmaceutically acceptable salt of the compound of general formula (I) described in the present invention refers to an acid addition salt formed by the compound of general formula (I) and a pharmaceutically acceptable acid or a base addition salt formed by the compound of general formula (I) and a pharmaceutically acceptable base, wherein the acid includes: hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or ferulic acid; the base addition salt includes: sodium salt, potassium salt, ammonium salt, calcium salt, aluminum salt, magnesium salt or other metal salt, ethylenediamine, ethanolamine or other common base addition salts.

[0016] The compound of general formula (I) can be synthesized by the following route.

[0017] The substituent R is as defined above.

[0018] The compound of the general formula (I) can be prepared by a substitution reaction between a halogen atom and a thiol group, wherein the reactant is an aromatic molecule containing a halogen or thiol group, or can be prepared by tautomerism to form a thioketone structure raw material of a thiol group.

[0019] Preferably, the molar ratio of the raw material compound 1-bromo-4-nitrothiazole to R-SH in the reaction is 0.8 to 1:2.

[0020] Preferably, the reaction temperature is room temperature and the reaction time is 0 to 6 hours.

[0021] The pharmaceutically acceptable salt of the compound of general formula (I) can be prepared by reacting with an equal chemical equivalent or excess amount of an acid (inorganic acid or organic acid) in a suitable solvent or solvent combination. The acid includes but is not limited to hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or ferulic acid. The solvent includes but is not limited to methanol, ethanol, dichloromethane, acetone, ethyl acetate, toluene or tetrahydrofuran, or any mixed solvents thereof.

[0022] More preferably,

[0023] When the compound of general formula (Ⅰ) is 5-nitro-2-((1-phenyl-1H-tetrazol-5-yl)thio)thiazole (LBX-1), the synthesis method is as follows: 1-phenyl-1H-tetrazol-5-thiol and sodium ethoxide are added to ethanol, stirred at room temperature for 30 minutes, and then 1-bromo-4-nitrothiazole is added and reacted at room temperature for 2 hours.

[0024] When the compound of general formula (I) is 2-((1-methyl-1H-tetrazol-5-yl)thio)-5-nitrothiazole (LBX-2), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazol-5-thiol is replaced by the raw material 1-methyl-1H-tetrazol-5-thiol.

[0025] When the compound of general formula (I) is 5-nitro-2-(thiazol-2-ylthio)thiazole (LBX-3), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material thiazole-2-thiol.

[0026] When the compound of general formula (I) is 5-nitro-2-(quinolin-4-ylthio)thiazole (LBX-4), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material quinoline-4-thiol.

[0027] When the compound of general formula (I) is 2-((5-nitrothiazol-2-yl)thio)-1,3,4-thiadiazole (LBX-5), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 1,3,4-thiadiazole-2-thiol.

[0028] When the compound of general formula (I) is 4-methyl-2-((5-nitrosothiazol-2-yl)thio)benzo[d]thiazole (LBX-6), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 4-methylbenzo[d]thiazole-2-thiol.

[0029] When the compound of general formula (I) is 5-nitro-2-((4-phenylthiazol-2-yl)thio)thiazole (LBX-7), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 4-phenylthiazole-2-thiol.

[0030] When the compound of general formula (I) is 6-methyl-2-((5-nitrosothiazol-2-yl)thio)thiazolo[4,5-b]pyridine (LBX-8), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 6-methylthiazolo[4,5-b]pyridine-2-thiol.

[0031] When the compound of general formula (I) is bis(5-nitrosothiazol-2-yl)sulfane (LBX-9), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 5-nitrothiazole-2-thiol.

[0032] When the compound of general formula (I) is 2-((5-nitrosothiazol-2-yl)thio)benzo[d]thiazole (LBX-10), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material benzo[d]thiazole-2-thiol.

[0033] When the compound of general formula (I) is 2-((5-nitrothiazol-2-yl)thio)benzo[d]oxazole (LBX-11), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material benzo[d]oxazole-2-thiol.

[0034] When the compound of general formula (I) is 2-((4,5-dihydrothiazol-2-yl)thio)-5-nitrothiazole (LBX-12), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazole-5-thiol is replaced by the raw material 4,5-dihydrothiazol-2-thiol.

[0035] When the compound of general formula (I) is 2-((1H-indol-2-yl)thio)-5-nitrothiazole (LBX-13), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 1H-indole-2-thiol.

[0036] When the compound of general formula (I) is 2-((9H-purine-6-yl)thio)-5-nitrothiazole (LBX-14), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 9H-purine-6-thiol.

[0037] When the compound of general formula (I) is 5-nitro-2-(pyridin-2-ylthio)thiazole (LBX-15), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material pyridine-2-thiol.

[0038] When the compound of general formula (I) is 5-nitro-2-(pyrimidin-2-ylthio)thiazole (LBX-16), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material pyrimidine-2-thiol.

[0039] When the compound of general formula (I) is 5-methyl-2-((5-nitrothiazol-2-yl)thio)benzo[d]thiazole (LBX-17), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 5-methylbenzo[d]thiazole-2-thiol.

[0040] When the compound of general formula (I) is 4-methyl-2-((5-nitrothiazol-2-yl)thio)benzo[d]oxazole (LBX-18), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 4-methylbenzo[d]oxazole-2-thiol.

[0041] When the compound of general formula (I) is 2-((1H-1,2,4-triazol-3-yl)thio)-5-nitrothiazole (LBX-19), the synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 1H-1,2,4-triazole-3-thiol.

[0042] When the compound of general formula (I) is 5-((5-nitrothiazol-2-yl)thio)-4H-1,2,4-triazole-3-ol (LBX-20), the synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 5-mercapto-4H-1,2,4-triazole-3-ol.

[0043] When the compound of general formula (I) is 6-methyl-2-((5-nitrothiazol-2-yl)thio)pyrimidin-4-ol (LBX-21), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 2-mercapto-6-methylpyrimidin-4-ol.

[0044] When the compound of general formula (I) is 2-((5-nitrothiazol-2-yl)thio)-4,5-diphenyloxazole (LBX-22), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 4,5-diphenyloxazole-2-thiol.

[0045] When the compound of general formula (I) is 6-nitro-2-((5-nitrothiazol-2-yl)thio)benzo[d]thiazole (LBX-23), the synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 6-nitrobenzo[d]thiazole-2-thiol.

[0046] When the compound of general formula (I) is 5-((5-nitrothiazol-2-yl)thio)-1,3,4-thiadiazole-2-amine (LBX-24), the synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 5-amino-1,3,4-thiadiazole-2-thiol.

[0047] The compounds of the present invention can be administered clinically by oral administration, injection or the like.

[0048] Generally, when the compounds of the present invention are used for treatment, the dosage range for human use is 1-500 mg / day. The dosage may also exceed this range depending on the dosage form and the severity of the disease.

[0049] The present invention also provides the use of the compound represented by the general formula (I) in the preparation of PHGDH inhibitors.

[0050] The present invention also provides the use of the compound represented by the general formula (I) in preparing therapeutic drugs for PHGDH-mediated related diseases.

[0051] The PHGDH-mediated related diseases of the present invention include cancer and liver damage diseases. Among them, cancer is preferably but not limited to breast cancer, melanoma, liver cancer, lung cancer, colon cancer, and leukemia; liver damage diseases are preferably but not limited to drug and poison liver damage, alcohol and non-alcoholic liver damage, and hepatitis virus liver damage.

[0052] Unless otherwise stated, the following terms used in the specification and claims have the meanings discussed below:

[0053] The term "aromatic ring" refers to a monocyclic or fused polycyclic or biphenyl group containing 1-12 carbon atoms, with a complete conjugated π electron system. Non-limiting examples of aromatic rings are phenyl, naphthyl and biphenyl, and the aromatic rings may be substituted or unsubstituted.

[0054] The term "aromatic heterocycle" refers to a monocyclic system or fused polycyclic ring containing 1-12 atoms, the system containing one, two, three or four ring heteroatoms that are N, O or S, the remaining ring atoms being C, and having a complete conjugated π electron system. Non-limiting examples of unsubstituted aromatic heterocycles include pyrrole, furan, thiophene, imidazole, oxazole, thiazole, thiadiazole, pyrazole, pyridine, pyrimidine, tetrazole and triazine. The aromatic heterocycle can be substituted or unsubstituted.

[0055] The term "alkyl" refers to a saturated aliphatic hydrocarbon group of 1 to 20 carbon atoms, including straight chain and branched groups. The alkyl group may be substituted or unsubstituted. When it is a substituted alkyl group, the substituent is preferably one or more.

[0056] The term "alkoxy" refers to -O-(unsubstituted alkyl) and -O-(unsubstituted cycloalkyl). Representative examples include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy, and the like.

[0057] The term "carbonyl" means Group.

[0058] The term "sulfonamide" means Group, wherein Ar represents an aromatic ring.

[0059] The term "sulfenyl imine" means It is an amide derivative of sulfonic acid

[0060] The term "trifluoromethyl" refers to a -CF3 group.

[0061] The term "nitro" refers to a -NO2 group.

[0062] The term "amino" refers to a -NH2 group.

[0063] The term "hydroxy" refers to an -OH group.

[0064] The term "halogen" means fluorine, chlorine, bromine or iodine, preferably fluorine, chlorine or bromine.

[0065] The present invention has the following technical effects: The present invention provides a class of compounds as PHGDH inhibitors, and experiments show that they directly inhibit PHGDH, and an active small molecule with a novel structure and good physical and chemical properties is discovered. The small molecule has shown good activity in various experiments and is expected to be developed into a specific drug treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0066] Figure 1 It is the experimental result of Example 4, wherein: Figure 1 A shows the cell cycle distribution of MCF-7 cells under the treatment of different concentrations of LBX-5. Figure 1 B is Figure 1 A is the statistical result of the bar chart of data. Figure 1 C: Western Blot analysis of the expression of PHGDH, Cyclin D1 and CDK4.

[0067] Figure 2 It is the experimental result of Example 5, wherein: Figure 2 A is the change of tumor volume between the LBX-5 (0.3 mg / kg) administration group and the blank solvent control group. Figure 2 B is the comparison of the changes in body weight of mice between the LBX-5 (0.3 mg / kg) administration group and the blank solvent control group.

[0068] Figure 3 It is the experimental result of Example 6, wherein: Figure 3 A shows the effect of LBX-5 (3 mg / kg) on ​​serum alanine aminotransferase (ALT) levels in a CCl4-induced acute liver injury mouse model. Figure 3 B shows the effect of LBX-5 on serum aspartate aminotransferase (AST) levels in the CCl4-induced acute liver injury mouse model. DETAILED DESCRIPTION

[0069] In order to further illustrate the present invention, a series of examples are given below. These examples are purely illustrative and are only used to specifically describe the present invention, and should not be understood as limiting the present invention.

[0070] Example 1

[0071] 5-Nitro-2-((1-phenyl-1H-tetrazol-5-yl)thio)thiazole (LBX-1)

[0072] 1-Phenyl-1H-tetrazolyl-5-thiol (0.78 g, 4.35 mmol), sodium ethoxide (0.36 g, 5.22 mmol) and ethanol (15 mL) were added to a single-necked bottle, stirred at room temperature for 30 min, and 1-bromo-4-nitrothiazole (1.00 g, 4.78 mmol) was added, and the reaction was allowed to react at room temperature for 2 h. After the reaction was completed by TLC monitoring, the reaction solution was concentrated under reduced pressure, extracted with ethyl acetate and water three times, the organic layers were combined, dried over anhydrous sodium sulfate, and then filtered, the filtrate was distilled under reduced pressure to remove the solvent, and the filtrate was purified by silica gel column chromatography (V 石油醚 :V 乙酸乙酯 =5:1) to obtain 1.13 g of a light yellow solid with a yield of 85%. 1 HNMR (300MHz, DMSO-d6): δ=8.68(s,1H,Ar-H),7.73-7.63(m,6H,Ar-H)ppm.MS(ASAP),[M+H] + calculated for C 10 H6N6O2S2 + 307.0,found306.9.

[0073] 2-((1-Methyl-1H-tetrazol-5-yl)thio)-5-nitrothiazole (LBX-2)

[0074] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 1-methyl-1H-tetrazolyl-5-thiol, the feed amount is 0.51 g, and finally 0.85 g of white solid is obtained with a yield of 80%. 1 H NMR (300MHz, DMSO-d6): δ=8.71(s,1H,Ar-H),4.10(s,3H,CH3)ppm.MS(ASAP),[M+H] + calculated forC5H4N6O2S2 + 246.0, found 245.8.

[0075] 5-Nitro-2-(thiazol-2-ylthio)thiazole (LBX-3)

[0076] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material thiazole-2-thiol, the feed amount is 0.51 g, and finally 0.97 g of white solid is obtained with a yield of 91%. 1 H NMR (300MHz, DMSO-d6): δ = 8.79 (s, 1H, Ar-H), 8.15 (s, 1H, Ar-H), ppm.MS (ASAP), [M+H]+ calculated for C6H3N3O2S3 + 245.3, found 245.1.

[0077] 5-Nitro-2-(quinolin-4-ylthio)thiazole (LBX-4)

[0078] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material quinoline-4-thiol, the feed amount is 0.70 g, and finally 1.02 g of yellow solid is obtained with a yield of 81%. 1 H NMR (300MHz, DMSO-d6): δ = 9.07-9.05 (d, 1H, J = 4.44Hz, Ar-H), 8.73 (s, 1H, Ar-H), 8.29-8.27 (d, 1H, J = 9.99Hz, Ar-H), 8.22-8.19 (d, 1H, J = 7. 74Hz,Ar-H),8.07-8.05(d,1H,J=4.50Hz,Ar-H),7.96-7.90(t,1H,J=6.90Hz,Ar-H),7.81-7.75(t,1H,J=7.02Hz,Ar-H)ppm.MS(ASAP),[M+H] + calculated for C 12 H7N3O2S2 + 290.0, found 289.9.

[0079] 2-((5-nitrothiazol-2-yl)thio)-1,3,4-thiadiazole (LBX-5)

[0080] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 1,3,4-thiadiazole-2-thiol, the feed amount is 0.51 g, and finally 1.02 g of white solid is obtained with a yield of 81%. 1 HNMR (300MHz, DMSO-d6): δ=9.86(s,1H,Ar-H),8.85(s,1H,Ar-H)ppm.MS(ASAP),[M+H] + calculated forC5H2N4O2S3 + 246.9, found 246.7.

[0081] 4-Methyl-2-((5-nitrosothiazol-2-yl)thio)benzo[d]thiazole (LBX-6)

[0082] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 4-methylbenzo[d]thiazole-2-thiol, the feed amount is 0.79 g, and finally 1.17 g of yellow solid is obtained with a yield of 83%. 1 H NMR (300MHz, DMSO-d6): δ=8.88(s,1H,Ar-H),8.02-7.99(t,1H,J=5.67Hz,Ar-H),7.45-7.40(m,2H,Ar-H),2.77(s,3H,CH3)ppm.MS(ASAP),[M+H] + calculated for C 11 H7N3O2S3 + 310.0,found309.9.

[0083] 5-Nitro-2-((4-phenylthiazol-2-yl)thio)thiazole (LBX-7)

[0084] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 4-phenylthiazole-2-thiol, the feed amount is 0.84 g, and finally 1.22 g of white solid is obtained with a yield of 87%. 1 HNMR (300MHz, DMSO-d6): δ=8.83(s,1H,Ar-H),8.48(s,1H,Ar-H),8.03-8.01(d,2H,J=7.08Hz,Ar-H),7.54-7.39(m,3H,Ar-H)ppm.MS(ASAP),[M+H] + calculated for C 12 H7N3O2S3 + 322.0, found 321.8.

[0085] 6-Methyl-2-((5-nitrosothiazol-2-yl)thio)thiazolo[4,5-b]pyridine(LBX-8)

[0086] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 6-methylthiazolo[4,5-b]pyridine-2-thiol, the feed amount is 0.79 g, and finally 1.23 g of white solid is obtained with a yield of 91%. 1 H NMR (300MHz, DMSO-d6): δ=8.91(s,1H,Ar-H),8.58(s,1H,Ar-H),8.47(s,1H,Ar-H),2.47(s,3H,CH3)ppm.MS(ASAP),[M+H] +calculated for C 10 H6N4O2S3 + 295.9, found 295.7.

[0087] Bis(5-nitrosothiazol-2-yl)sulfane(LBX-9)

[0088] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 5-nitrothiazole-2-thiol, the feed amount is 0.71 g, and finally 0.89 g of yellow solid is obtained with a yield of 71%. 1 HNMR (300MHz, DMSO-d6): δ=8.97(s,2H,Ar-H)ppm.MS(ASAP),[M+H] + calculated for C6H2N4O4S3 + 290.9,found290.8.

[0089] 2-((5-Nitrosothiazol-2-yl)thio)benzo[d]thiazole (LBX-10)

[0090] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material benzo[d]thiazole-2-thiol, the feed amount is 0.73 g, and finally 1.13 g of yellow solid is obtained with a yield of 88%. 1 HNMR (300MHz, DMSO-d6): δ=8.86(s,1H,Ar-H),8.20-8.10(m,2H,Ar-H),7.63-7.50(m,2H,Ar-H)ppm.MS(ASAP),[M+H] + calculated for C 10 H5N3O2S3 + 296.0, found 295.9.

[0091] 2-((5-Nitrothiazol-2-yl)thio)benzo[d]oxazole (LBX-11)

[0092] The synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material benzo[d]oxazole-2-thiol, the feed amount is 0.51 g, and finally 0.60 g of yellow solid is obtained, with a yield of 64%. 1 H NMR (300MHz, DMSO-d6): δ=8.87(s,1H,Ar-H),7.92-7.81(m,2H,Ar-H),7.54-7.46(m,2H,Ar-H)ppm.MS(ASAP),[M+H]+ calculated for C 10 H6N3O3S2 + 280.0,found 280.1.

[0093] 2-((4,5-dihydrothiazol-2-yl)thio)-5-nitrothiazole (LBX-12)

[0094] The synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 4,5-dihydrothiazole-2-thiol, the feed amount is 0.62 g, and finally 0.75 g of white solid is obtained, with a yield of 58%. 1 H NMR (300MHz, DMSO-d6): δ = 8.83 (s, 1H, Ar-H), 4.45-4.40 (t, 2H, J = 8.19Hz, CH2), 3.71-3.65 (t, 2H, J = 8.13Hz, CH2) ppm.MS (ASAP), [M+H] + calculated for C6H6N3O2S3 + 248.0, found 247.9.

[0095] 2-((1H-indol-2-yl)thio)-5-nitrothiazole (LBX-13)

[0096] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 1H-indole-2-thiol, the feed amount is 0.83 g, and finally 1.00 g of white solid is obtained with a yield of 65%. 1 HNMR (300MHz, DMSO-d6): δ = 8.82 (s, 1H, Ar-H), 7.65-7.63 (d, 1H, J = 5.64Hz, Ar-H), 7.52-7. 51(d,2H,J=3.21Hz,Ar-H),7.12-6.88(m,2H,Ar-H),6.68(s,1H,Ar-H)ppm.MS(ASAP),[M+H] + calculated for C 11 H8N3O2S2 + 278.0, found 278.2.

[0097] 2-((9H-purin-6-yl)thio)-5-nitrothiazole (LBX-14)

[0098] The synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 9H-purine-6-thiol, the feed amount is 0.77 g, and finally 1.05 g of white solid is obtained, with a yield of 74%. 1 HNMR (300MHz, DMSO-d6): δ=9.06(s,1H,Ar-H),8.90(s,1H,Ar-H),8.68(s,1H,Ar-H)ppm.MS(ASAP),[M+H] + calculated for C 10 H6N3O3S2 + 280.0,found 280.1.

[0099] 5-Nitro-2-(pyridin-2-ylthio)thiazole (LBX-15)

[0100] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material pyridine-2-thiol, the feed amount is 0.74 g, and finally 0.62 g of off-white solid is obtained, with a yield of 39%. 1 H NMR (300MHz, DMSO-d6): δ = 8.84 (s, 1H, Ar-H), 8.81-8.79 (m, 1H, Ar-H), 8.01-7.95 (m, 1H, Ar-H),7.81-7.79(d,1H,J=8.04Hz,Ar-H),7.53-7.48(m,1H,Ar-H)ppm.MS(ASAP),[M+H] + calculated for C8H6N3O2S2 + 240.0, found 239.8.

[0101] 5-Nitro-2-(pyrimidin-2-ylthio)thiazole (LBX-16)

[0102] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material pyrimidine-2-thiol, the feed amount is 0.68 g, and finally 1.27 g of yellow solid is obtained with a yield of 87%. 1 H NMR (300MHz, DMSO-d6): δ = 9.01-8.99 (d, 2H, J = 4.98Hz, Ar-H), 8.90 (s, 1H, Ar-H), 7.62-7.59 (t, 1H, J = 4.92Hz, Ar-H) ppm.MS (ASAP), [M+H] + calculated for C7H5N4O2S2 +241.0, found 241.2.

[0103] 5-Methyl-2-((5-nitrothiazol-2-yl)thio)benzo[d]thiazole (LBX-17)

[0104] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 5-methylbenzo[d]thiazole-2-thiol, the feed amount is 0.48 g, and finally 0.66 g of yellow solid is obtained with a yield of 81%. 1 H NMR (300MHz, DMSO-d6): δ=8.74(s,1H,Ar-H),7.86-7.83(m,2H,Ar-H),7.31-7.29(d,1H,J=5.92Hz,Ar-H),2.47(s,3H,CH3)ppm.MS(ASAP),[M+H] + calculated for C 11 H8N3O2S3 + 310.0,found310.2.

[0105] 4-Methyl-2-((5-nitrothiazol-2-yl)thio)benzo[d]oxazole (LBX-18)

[0106] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 4-methylbenzo[d]oxazole-2-thiol, the feed amount is 0.50 g, and finally 0.67 g of white solid is obtained, with a yield of 76%. 1 H NMR (300MHz, DMSO-d6): δ = 8.71 (s, 1H, Ar-H), 7.67-7.65 (d, 1H, J = 4.67Hz, Ar-H), 7.45-7.44 (d, 1H,J=3.74Hz,Ar-H),7.33-7.31(t,1H,J=5.51Hz,Ar-H),2.28(s,3H,CH3)ppm.MS(ASAP),[M+H] + calculated for C 11 H8N3O3S2 + 294.0, found 293.9.

[0107] 2-((1H-1,2,4-triazol-3-yl)thio)-5-nitrothiazole (LBX-19)

[0108] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 1H-1,2,4-triazole-3-thiol, the feed amount is 0.61 g, and finally 1.34 g of white solid is obtained with a yield of 82%. 1 H NMR (300MHz, DMSO-d6): δ = 8.98 (s, 1H, Ar-H), 8.77 (s, 1H, Ar-H) ppm. MS (ASAP), [M+H] + calculated forC5H4N5O2S2 + 230.0, found 230.3.

[0109] 5-((5-Nitrothiazol-2-yl)thio)-4H-1,2,4-triazol-3-ol (LBX-20)

[0110] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 5-mercapto-4H-1,2,4-triazole-3-ol, the feed amount is 0.66 g, and finally 1.22 g of off-white solid is obtained, with a yield of 88%. 1 H NMR (300MHz, DMSO-d6): δ = 8.78 (s, 1H, Ar-H) ppm.MS (ASAP), [M+H] + calculated for C5H4N5O3S2 + 246.0, found 245.8.

[0111] 6-Methyl-2-((5-nitrothiazol-2-yl)thio)pyrimidin-4-ol (LBX-21)

[0112] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 2-mercapto-6-methylpyrimidin-4-ol, the feed amount is 1.02 g, and finally 0.99 g of yellow solid is obtained with a yield of 51%. 1 H NMR (300MHz, DMSO-d6): δ=8.87(s,1H,Ar-H),6.56(s,1H,Ar-H),2.47(s,3H,CH3)ppm.MS(ASAP),[M+H] + calculated for C8H7N4O3S2 + 271.0, found 271.1.

[0113] 2-((5-Nitrothiazol-2-yl)thio)-4,5-diphenyloxazole (LBX-22)

[0114] The synthesis method is the same as LBX-1, the only difference is that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 4,5-diphenyloxazole-2-thiol, the feed amount is 0.48 g, and finally 0.53 g of white solid is obtained, with a yield of 73%. 1 H NMR (300MHz, DMSO-d6): δ=8.99(s,1H,Ar-H),7.81-7.73(m,5H,Ar-H),7.65-7.59(m,5H,Ar-H)ppm.MS(ASAP),[M+H] + calculated for C 18 H 12 N3O3S2 + 382.0, found 382.4.

[0115] 6-Nitro-2-((5-nitrothiazol-2-yl)thio)benzo[d]thiazole (LBX-23)

[0116] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 6-nitrobenz[d]thiazole-2-thiol, the feed amount is 0.45 g, and finally 0.33 g of yellow solid is obtained, with a yield of 46%. 1 H NMR (300MHz, DMSO-d6): δ = 9.19-9.18 (d, 1H, J = 2.37Hz, Ar-H), 8.91 (s, 1H, Ar-H), 8.37 -8.33(d,1H,J=6.63Hz,Ar-H),8.25-8.22(d,1H,J=8.97Hz,Ar-H)ppm.MS(ASAP),[M+H] + calculated for C 10 H5N4O4S3 + 340.9, found 340.7.

[0117] 5-((5-nitrothiazol-2-yl)thio)-1,3,4-thiadiazol-2-amine (LBX-24)

[0118] The synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazolyl-5-thiol is replaced by the raw material 5-amino-1,3,4-thiadiazole-2-thiol, the feed amount is 0.85 g, and finally 1.38 g of yellow solid is obtained with a yield of 83%. 1 H NMR (300MHz, DMSO-d6): δ = 8.92 (s, 1H, Ar-H) ppm.MS (ASAP), [M+H] + calculated for C5H4N5O2S3+ 262.0, found 261.9.

[0119] Example 2

[0120] Enzyme inhibition activity test of PHGDH small molecule inhibitors

[0121] In this example, a high-throughput screening system targeting PHGDH was established by coupling the resazurin reduction reaction. + , 56.25mM N2H4-H2SO4 pH 9.0, 125mM Tris-HCl pH 7.5, 2.5mM EDTA, 0.0125% Tween20; dilute the PHGDH protein stock solution with protein buffer, add 9μL of PHGDH protein with a final concentration of 5nM to the 384-well black board, and add 9μL of protein buffer to the blank control well; prepare compound solutions with different concentration gradients with DMSO, set three replicates for each concentration, add 1μL of compound solution to each well of the compound well in the plate, add 1μLDMSO to the negative control well and the blank control well, and place the plate on a horizontal shaker at 30rpm and incubate at room temperature for 30 minutes; prepare substrate buffer 175μM 3-PG, 25μM resazurin, 5μg / mL diaphorase, 56.25mM N2H4-H2SO4 pH9.0, 125mM Tris-HCl pH 7.5, 2.5mM EDTA, 0.0125% Tween20; add 20 μL substrate buffer to each well of the plate in the dark, incubate at room temperature in the dark for 3 hours on a horizontal shaker at 50 rpm; briefly centrifuge the 384-well plate at 1000 rpm for 30 seconds at room temperature, set the excitation wavelength to 550 nm, the emission wavelength to 590 nm, read the fluorescence signal value using a Molecular Devices multi-function microplate reader, and calculate the inhibition rate at the corresponding concentration; use GraphPad Prism 8.0 software to fit the PHGDH inhibitory activity IC of the test compound 50 The results are shown in Table 1.

[0122] Table 1 IC of some compounds of the present invention against PHGDH 50 scope

[0123] serial number <![CDATA[PHGDHIC 50 (μM)]]> serial number <![CDATA[PHGDHIC 50 (μM)]]> LBX-1 +++ LBX-13 ++ LBX-2 +++ LBX-14 ++ LBX-3 +++ LBX-15 ++ LBX-4 +++ LBX-16 ++ LBX-5 +++ LBX-17 ++ LBX-6 +++ LBX-18 + LBX-7 +++ LBX-19 + LBX-8 +++ LBX-20 + LBX-9 +++ LBX-21 + LBX-10 +++ LBX-22 + LBX-11 ++ LBX-23 + LBX-12 ++

[0124] Note: The inhibitory activity of the compounds in Table 2 against PHGDH is indicated by 0 to 3 plus signs. "+++": IC 50 <1μM, indicating that the compound has strong PHGDH inhibitory activity. "++": 1μM≤IC 50 <5μM, indicating that the compound has moderate PHGDH inhibitory activity. "+" indicates 5μM≤IC50 <10 μM, indicating that the compound has weak PHGDH inhibitory activity.

[0125] As can be seen from Table 2, the inhibitory activity of the compounds increases with GI 50 The "+++" group (IC 50 <1μM) showed significant inhibitory effects and are suitable for therapeutic scenarios requiring potent inhibition; "++" group (1μM≤IC 50 <5μM) showed a moderate inhibitory effect and was suitable for moderate inhibition requirements; the "+" group (5μM≤IC 50 <10 μM) indicates weak inhibitory activity and may need to be used in combination with other drugs to enhance the effect.

[0126] Example 3

[0127] Cell survival assay to test the inhibitory effect of compounds on the proliferation of tumor cell lines

[0128] In this example, human breast cancer cell MCF-7, liver cancer cell H22 and colon cancer cell CT26 were selected for the experiment. The cell lines were purchased from the cell bank of the Chinese Academy of Sciences. The MTT method was used to detect the inhibitory effect of the compound on cell proliferation. The cells were all cultured at 2*10 ^ 4mL -1 The cells were cultured at a density of 100 μg / mL in a 96-well transparent plate and treated with a DMSO solution of the compound or the same volume of DMSO. After 72 hours, 5 mg / mL MTT solution was added and incubated at 37°C. After 4 hours, the culture medium was discarded, and the blue-purple crystals at the bottom of the plate were dissolved with DMSO. The OD of each well was measured using a Thermo microplate reader. 490 , reflecting the number of living cells. Half-inhibitory concentration GI 50 The values ​​were obtained by fitting with GraphPad Prism 8.0 software, and the results are shown in Table 2.

[0129] Table 2 Proliferation inhibition activity GI of some compounds of the present invention on tumor cell lines 50 value

[0130]

[0131] Note: The inhibitory activity of the compounds in Table 2 on cell proliferation is represented by 0 to 3 plus signs. "+++": IC 50 <1μM, indicating that the compound has a strong inhibitory activity on cell proliferation. "++": 1μM≤IC 50 <5μM indicates that the compound has moderate inhibitory activity on cell proliferation. "+" indicates 5μM≤IC 50 <10 μM, indicating that the compound has weak inhibitory activity on cell proliferation.

[0132] As can be seen from the above table, some compounds of the present invention exhibited good proliferation inhibition effects on various tumor cell lines.

[0133] Example 4

[0134] In order to further clarify the effect of LBX-5 on the cell cycle of human breast cancer cells MCF7, the cell cycle distribution was detected by flow cytometry and the expression of cyclin D1 and cyclin-dependent kinase 4 (CDK4) was detected by immunohistochemistry. The results showed that with the increase of LBX-5 concentration, the proportion of MCF-7 cells in the G1 phase increased significantly ( Figure 1 A-1B), and LBX-5 can downregulate PHGDH, Cyclin D1 and CDK4 in MCF-7 cells in a concentration-dependent manner ( Figure 1 C). These results verified that LBX-5 could significantly inhibit the proliferation of MCF-7 cells by regulating the cell cycle process, especially by inhibiting the expression of G1 phase-related proteins. This finding suggests that LBX-5 has a potential cell cycle regulatory effect and may exert its anti-tumor activity by intervening in the PHGDH, Cyclin D1 and CDK4 signaling pathways.

[0135] Example 5

[0136] In vivo pharmacodynamics of PHGDH small molecule inhibitors in a hepatoma cell H22 xenograft mouse model

[0137] In this example, a xenograft tumor mouse model of liver cancer cells H22 was constructed to verify the in vivo anti-tumor activity of the representative compound LBX-5. Specifically, liver cancer cells H22 were inoculated into the abdomen of BALB / c mice to form a xenograft tumor model. The construction method of this model refers to the technical solution described in the document [DOI:10.2147 / DDDT.S461594]. After the model was established, LBX-5 was injected and the tumor growth of the mice was monitored to evaluate the inhibitory effect of LBX-5 on tumors.

[0138] When the tumor grows to 60 mm 3 -80mm 3 The mice were randomly divided into groups and given blank solvent or LBX-5 (0.3 mg / kg) by tail vein injection every 4 days for 12 days. The results showed that LBX-5 (0.3 mg / kg) could inhibit tumor growth to a certain extent. Compared with the blank solvent control group, the LBX-5 group did not lose significant weight ( Figure 2 A, 2B).

[0139] Example 6

[0140] In vivo pharmacodynamics of a small molecule inhibitor of PHGDH in a mouse model of acute liver injury induced by CCl4

[0141] In this example, a mouse model of acute liver injury induced by carbon tetrachloride (CCl4) was constructed to verify the liver protective effect of the representative compound LBX-5. Acute liver injury was induced by intraperitoneal injection of CCl4 solution (0.3%, 10 mL / kg), and the mice were divided into a solvent control group and an LBX-5 administration group. After the induction of liver injury, the mice in the LBX-5 administration group were immediately treated with LBX-5 (3 mg / kg). Venous blood samples were collected at 0, 6, 12, 24 and 48 hours after CCl4 treatment for blood biochemical analysis. The results showed that LBX-5 significantly inhibited CCl4-induced acute liver injury, as manifested by a significant decrease in the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) ( Figure 3 A, 3B).

[0142] The above are only embodiments of the present invention, and are not intended to limit the patent scope of the present invention. Any equivalent transformations made using the contents of the present invention specification, or directly or indirectly applied in other related technical fields, are also included in the patent protection scope of the present invention.

Claims

1. A compound of the general formula (I) containing a thioether skeleton structure or a pharmaceutically acceptable salt thereof, in, R is a substituted or unsubstituted aromatic ring or aromatic heterocycle, wherein the aromatic ring is a five-membered or six-membered aromatic ring, the aromatic heterocycle is a monocyclic aromatic heterocycle or a polycyclic condensed heterocycle, and the substituent groups include monosubstituted or polysubstituted phenyl, alkyl, alkoxy, amino, hydroxyl, nitro, halogen, trifluoromethyl, carbonyl, sulfonamide, and sulfenyl imide groups.

2. The compound of general formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The substituent R is a substituted or unsubstituted aromatic heterocycle, and the aromatic heterocycle is selected from any one of tetrazole, thiazole, quinoline, thiadiazole, benzothiazole, thiazolyl pyridine, benzoxazole, thiazoline, indole, purine, pyridine, pyrimidine, triazole, and oxazole, and the substituent group includes a monosubstituted or polysubstituted phenyl, methyl, amino, nitro, or hydroxyl group.

3. The compound of general formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The substituent R is selected from any one of the following groups: phenyltetrazole, methyltetrazole, thiazole, quinoline, 1,3,4-thiadiazole, methylbenzothiazole, phenylthiazole, methylthiazolopyridine, nitrothiazole, benzothiazole, benzoxazole, thiazoline, indole, purine, pyridine, pyrimidine, methylbenzothiazole, methylbenzoxazole, triazole, hydroxytriazole, hydroxymethylpyrimidine, diphenyloxazole, nitrobenzothiazole, and aminothiadiazole.

4. The compound of general formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound is any one of the following compounds:

5. The compound of general formula (I) or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The pharmaceutically acceptable salt refers to an acid addition salt formed by the compound of general formula (I) with a pharmaceutically acceptable acid or a base addition salt formed with a pharmaceutically acceptable base, wherein the acid is hydrogen chloride, hydrogen bromide, sulfuric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, phosphoric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid or ferulic acid; the base addition salt is sodium salt, potassium salt, ammonium salt, calcium salt, aluminum salt, magnesium salt, ethylenediamine salt or ethanolamine salt.

6. A pharmaceutical composition comprising a pharmaceutically effective amount of an active ingredient and a pharmaceutically acceptable excipient, characterized in that: The active ingredient comprises one or more of the compounds of general formula (I) or pharmaceutically acceptable salts thereof according to any one of claims 1 to 5.

7. Use of the compound of general formula (I) or a pharmaceutically acceptable salt thereof as claimed in claims 1 to 5 or the composition as claimed in claim 6 in the preparation of a PHGDH inhibitor.

8. Use of the compound of general formula (I) as claimed in claims 1 to 5 or a pharmaceutically acceptable salt thereof or the composition as claimed in claim 6 in the preparation of therapeutic drugs for PHGDH-mediated related diseases.

9. The use according to claim 8, characterized in that: The PHGDH-mediated related diseases include cancer and liver damage diseases.

10. The use according to claim 9, characterized in that: The cancers described are breast cancer, melanoma, liver cancer, lung cancer, colon cancer, and leukemia; the liver damage diseases described include drug and toxic liver damage, alcohol and non-alcoholic liver damage, and hepatitis virus liver damage.

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