Compounds of the formula (i) wherein r, s, t, u, v, w, x, y, z, a, b, c, d, e, g, h, j, k, m, n, o, p, q, r, s, t, u, v, w, x, y, z

Abstract

The application belongs to the technical field of medicines, and particularly relates to a kind of compounds containing sulfide skeleton structure and application thereof in preparation of phosphoglycolate dehydrogenase inhibitors.A series of novel PHGDH small molecule inhibitors containing sulfide skeleton are obtained by screening compound library and structural modification, which enriches the structure type and indications of PHGDH inhibitors.The application has the following technical effects: the application provides a kind of compounds as PHGDH inhibitors, tests show that it directly produces inhibitory effect on PHGDH, and finds an active small molecule with novel structure and good physicochemical properties, which shows good activity in various experiments and is expected to be developed into a specific drug treatment.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a kind of compound containing sulfide skeleton structure and its application in preparing phosphoglycerate dehydrogenase inhibitor. BACKGROUND

[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 biological macromolecules, and can provide building blocks for molecules involved in various biochemical pathways, including amino acids, lipids, nucleotides and cofactors. Cells can obtain serine by transport proteins from the external environment, or by the serine biosynthesis pathway, using glycolysis intermediate 3-phosphoglycerate (3-PG) for biosynthesis. SSP is the main pathway for providing intracellular serine, and studies have shown that increasing the uptake of exogenous serine cannot compensate 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. Due to the insufficient uptake of exogenous serine to compensate for the lack of serine caused by SSP dysfunction, the normal physiological function of PHGDH is particularly critical for the intracellular serine homeostasis balance. In recent years, pharmacological research on PHGDH has focused on its high expression, and the target has been found to be up-regulated in breast cancer, melanoma, liver cancer, lung cancer, colon cancer, leukemia and liver injury, etc.

[0003] Currently, there are few studies on small molecule compounds targeting PHGDH. Existing PHGDH inhibitors such as NCT-503, CBR-5884 series, SENKYUNOLIDEA, etc. Although they show certain potential in tumor treatment and liver injury treatment, due to poor selectivity, drug resistance, poor pharmacokinetics and side effects, their widespread use in clinical practice is still limited.

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

[0005] Compared with existing PHGDH inhibitors, the sulfide inhibitors described in the present application exhibit a unique mechanism of action in function. By targeting the specific binding site of PHGDH, the compounds of the present application not only exhibit high inhibition of PHGDH catalytic activity, but also effectively reduce the expression of PHGDH in cells.

[0006] In order to achieve the above-mentioned purposes of the present application, the following technical solutions are adopted in the present application:

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

[0008]

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

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

[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 can 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) refers to the acid addition salt formed by the compound of general formula (I) and a pharmaceutically acceptable acid, or the 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; and 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 salt.

[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 general formula (I) can be synthesized by using the substitution reaction of halogen atom and mercapto group, and the reactant is an aromatic molecule containing halogen or mercapto group, or by forming the thione structure of mercapto group through tautomerism.

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

[0020] Preferably, the temperature of the reaction is room temperature, and the reaction time is 0-6h.

[0021] The pharmaceutically acceptable salt of the compound of general formula (I) can be prepared by reacting with an equivalent or excess amount of 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 solvent.

[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-tetrazole-5-thiol and sodium ethoxide are added to ethanol, stirred at room temperature for 30 min, then 1-bromo-4-nitrothiazole is added, and reacted at room temperature for 2h.

[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 that of LBX-1, except that the raw material 1-phenyl-1H-tetrazole-5-thiol is replaced by the raw material 1-methyl-1H-tetrazole-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 difference is only that the raw material 1-phenyl-1H-tetrazole-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 difference is only that the raw material 1-phenyl-1H-tetrazole-5-thiol is replaced by the raw material quinolin-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 difference is only that the raw material 1-phenyl-1H-tetrazole-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-nitrothiazol-2-yl)thio)benzo[d]thiazole (LBX-6), the synthesis method is the same as LBX-1, the difference is only that the raw material 1-phenyl-1H-tetrazole-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 difference is only that the raw material 1-phenyl-1H-tetrazole-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-nitrothiazol-2-yl)thio)thiazolo[4,5-b]pyridine (LBX-8), the synthesis method is the same as LBX-1, the difference is only that the raw material 1-phenyl-1H-tetrazole-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-nitrothiazol-2-yl) sulfane (LBX-9), the synthesis method is the same as LBX-1, the difference is only that the raw material 1-phenyl-1H-tetrazole-5-thiol is replaced by the raw material 5-nitrothiazole-2-thiol.

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

[0036] When the compound of general formula (I) is 2-((9H-purin-6-yl)thio)-5-nitrothiazole (LBX-14), the synthesis method is the same as LBX-1, except that the raw material 1-phenyl-1H-tetrazol-5-thiol is replaced by the raw material 9H-purin-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, except that the raw material 1-phenyl-1H-tetrazol-5-thiol is replaced by the raw material pyridin-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, except that the raw material 1-phenyl-1H-tetrazol-5-thiol is replaced by the raw material pyrimidin-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, except that the raw material 1-phenyl-1H-tetrazol-5-thiol is replaced by the raw material 5-methylbenzo[d]thiazol-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, except that the raw material 1-phenyl-1H-tetrazol-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 starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 1H-1,2,4-triazol-3-thiol.

[0042] When the compound of general formula (Ⅰ) is 5-((5-nitrothiazol-2-yl)thio)-4H-1,2,4-triazol-3-ol (LBX-20), the synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 5-mercapto-4H-1,2,4-triazol-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, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting 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, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting 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, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting 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 starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 5-amino-1,3,4-thiadiazole-2-thiol.

[0047] The compounds described in this invention can be administered clinically via oral administration, injection, or other methods.

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

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

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

[0051] The PHGDH-mediated diseases described in this invention include cancer and liver damage. Preferred cancers include, but are not limited to, breast cancer, melanoma, liver cancer, lung cancer, colon cancer, and leukemia. Liver damage diseases are preferred but not limited to drug-induced and toxic liver damage, alcoholic and non-alcoholic liver damage, and hepatitis virus-induced 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, possessing a fully conjugated π-electron system. Non-limiting examples of aromatic rings include phenyl, naphthyl, and biphenyl, and aromatic rings can be substituted or unsubstituted.

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

[0055] The term "alkyl" refers to a saturated aliphatic hydrocarbon group of 1-20 carbon atoms, including straight-chain and branched groups. Alkyl groups can be substituted or unsubstituted. When substituted, 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, cyclopropoxy, cyclobutoxy, cyclopentoxy, and cyclohexyloxy.

[0057] The term "carbonyl" means Group.

[0058] The term "sulfonamide" indicates Group, where Ar represents an aromatic ring.

[0059] The term "sulfinimide" indicates It is an amide derivative of sulfonic acid.

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

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

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

[0063] The term "hydroxyl group" refers to the -OH group.

[0064] The term "halogen" refers to 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. Experiments show that they directly inhibit PHGDH. A novel active small molecule with good physicochemical properties has been discovered. This small molecule has shown good activity in various experiments and is expected to be developed into a specific drug for treatment. Attached Figure Description

[0066] Figure 1 The experimental results are for Example 4, where: Figure 1 A represents the cell cycle distribution of MCF-7 cells under different concentrations of LBX-5 treatment. Figure 1 B is Figure 1 The bar chart statistics of data A. Figure 1 C represents the expression of PHGDH, Cyclin D1, and CDK4 detected by Western blotting.

[0067] Figure 2 The experimental results are for Example 5, wherein: Figure 2 A represents the change in tumor volume between the LBX-5 (0.3 mg / kg) administration group and the blank solvent control group. Figure 2 B represents a comparison of mouse body weight changes between the LBX-5 (0.3 mg / kg) administration group and the blank solvent control group.

[0068] Figure 3 The experimental results are for Example 6, where: Figure 3 A represents the effect of LBX-5 (3 mg / kg) on ​​serum alanine aminotransferase (ALT) levels in a mouse model of CCl4-induced acute liver injury. Figure 3 B represents the effect of LBX-5 on serum aspartate aminotransferase (AST) levels in a mouse model of CCl4-induced acute liver injury. Detailed Implementation

[0069] To further illustrate the present invention, a series of embodiments are given below. These embodiments are purely illustrative and are only used to specifically describe the present invention. They should not be construed as limiting the present invention.

[0070] Example 1

[0071] 5-Nitro-2-((1-Phenylacetazol-5-yl)thio)thiazole (LBX-1)

[0072] 1-Phenylacetazol-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 flask and stirred at room temperature for 30 min. Then, 1-bromo-4-nitrosothiazole (1.00 g, 4.78 mmol) was added, and the mixture was reacted at room temperature for 2 h. After the reaction was completed by TLC monitoring, the reaction mixture was concentrated under reduced pressure, extracted three times with ethyl acetate and water, and the organic layers were combined. After drying with anhydrous sodium sulfate, the mixture was filtered, and the solvent was removed by distillation under reduced pressure. The filtrate was then subjected to silica gel column chromatography (V... 石油醚 V 乙酸乙酯 The mixture was purified at a ratio of 5:1 to obtain 1.13 g of a pale 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, found 306.9.

[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 starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 1-methyl-1H-tetrazole-5-thiol. The feed amount is 0.51g, and the final result is 0.85g of white solid 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 for C5H4N6O2S2 + 246.0, found 245.8.

[0075] 5-Nitro-2-(thiazolyl-2-thio)thiazole (LBX-3)

[0076] The synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material thiazol-2-thiol. The feed amount is 0.51g, and the final result is 0.97g of white solid 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 starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material quinoline-4-thiol. The feed amount is 0.70 g, and the final yield is 1.02 g of yellow solid, 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-nitrothiazolyl-2-yl)thio)-1,3,4-thiadiazole (LBX-5)

[0080] The synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 1,3,4-thiadiazole-2-thiol. The feed amount is 0.51g, and the final result is 1.02g of white solid 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 for C5H2N4O2S3 + 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 starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 4-methylbenzo[d]thiazol-2-thiol. The feed amount is 0.79g, and the final yield is 1.17g of yellow solid, 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, found 309.9.

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

[0084] The synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 4-phenylthiazol-2-thiol. The feed amount is 0.84g, and the final yield is 1.22g of white solid, 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)thiazo[4,5-b]pyridine (LBX-8)

[0086] The synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 6-methylthiazo[4,5-b]pyridine-2-thiol. The amount of material fed is 0.79 g, and 1.23 g of white solid is finally 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)thione (LBX-9)

[0088] The synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 5-nitrothiazol-2-thiol. The feed amount is 0.71g, and the final yield is 0.89g of yellow solid, 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, found 290.8.

[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 starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material benzo[d]thiazol-2-thiol. The feed amount is 0.73g, and the final yield is 1.13g of yellow solid, 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-nitrothiazolyl-2-yl)thio)benzo[d]oxazole (LBX-11)

[0092] The synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material benzo[d]oxazol-2-thiol. The feed amount is 0.51g, and the final yield is 0.60g of yellow solid, 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-dihydrothiazolyl-2-yl)thio)-5-nitrothiazolyl (LBX-12)

[0094] The synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 4,5-dihydrothiazol-2-thiol. The feed amount is 0.62g, and the final result is 0.75g of white solid, 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 starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 1H-indole-2-thiol. The feed amount is 0.83g, and the final result is 1.00g of white solid 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, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 9H-purine-6-thiol. The amount of material fed is 0.77g, and 1.05g of white solid is finally 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 starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material pyridine-2-thiol. The feed amount is 0.74g, and the final product is 0.62g of off-white solid 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 starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with pyrimidine-2-thiol. The amount of material fed is 0.68g, and 1.27g of yellow solid is finally 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-nitrothiazolyl-2-yl)thio)benzo[d]thiazole (LBX-17)

[0104] The synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 5-methylbenzo[d]thiazol-2-thiol. The feed amount is 0.48g, and the final yield is 0.66g of yellow solid, 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, found 310.2.

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

[0106] The synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 4-methylbenzo[d]oxazol-2-thiol. The feed amount is 0.50 g, and the final yield is 0.67 g of white solid, 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 starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 1H-1,2,4-triazol-3-thiol. The feed amount is 0.61g, and the final yield is 1.34g of white solid, 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 for C5H4N5O2S2 + 230.0, found 230.3.

[0109] 5-((5-nitrothiazolyl-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 starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 5-mercapto-4H-1,2,4-triazol-3-ol. The amount of material fed is 0.66g, and 1.22g of off-white solid is finally 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-nitrothiazolyl)thio)pyrimidin-4-ol (LBX-21)

[0112] The synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 2-mercapto-6-methylpyrimidin-4-ol. The amount of material fed is 1.02 g, and 0.99 g of yellow solid is finally 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-nitrothiazolyl-2-yl)thio)-4,5-diphenyloxazole (LBX-22)

[0114] The synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 4,5-diphenyloxazole-2-thiol. The feed amount is 0.48g, and the final result is 0.53g of white solid 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-nitrothiazolyl-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-tetrazole-5-thiol is replaced with the raw material 6-nitrobenz[d]thiazol-2-thiol. The feed amount is 0.45g, and the final yield is 0.33g of yellow solid, 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-nitrothiazolyl)thio)-1,3,4-thiadiazole-2-amine (LBX-24)

[0118] The synthesis method is the same as LBX-1, except that the starting material 1-phenyl-1H-tetrazole-5-thiol is replaced with the starting material 5-amino-1,3,4-thiadiazole-2-thiol. The feed amount is 0.85g, and the final yield is 1.38g of yellow solid, 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 inhibitory activity assay of PHGDH small molecule inhibitor

[0121] This embodiment utilizes a coupled rezinosa reduction reaction to establish a high-throughput screening system targeting PHGDH. A protein buffer of 20 μM NAD was prepared. + 56.25mM N2H4-H2SO4 pH 9.0, 125mM Tris-HCl pH 7.5, 2.5mM EDTA, 0.0125% Tween 20; Dilute the PHGDH protein stock solution with protein buffer, add 9μL of PHGDH protein to a 384-well black plate to a final concentration of 5nM, and add 9μL of protein buffer to the blank control wells; Prepare compound solutions of different concentration gradients with DMSO, with three replicates for each concentration, add 1μL of compound solution to each compound well, and add 1μL of DMSO to the negative control and blank control wells. Incubate the plate at room temperature for 30 minutes on a horizontal shaker at 30 rpm; Prepare substrate buffer in the dark: 175μM 3-PG, 25μM resazurin, 5μg / mL myocardial flavin, 56.25mM N2H4-H2SO4 pH 9.0, 125mM Tris-HCl pH 7.5, 2.5mM EDTA, 0.0125% Tween 20. EDTA, 0.0125% Tween 20; add 20 μL of substrate buffer to each well of the plate in the dark, and incubate on a horizontal shaker at 50 rpm at room temperature in the dark for 3 hours; briefly centrifuge the 384-well plate at 1000 rpm at room temperature for 30 seconds, set the excitation wavelength to 550 nm and the emission wavelength to 590 nm, read the fluorescence signal values ​​using a Molecular Devices multi-functional microplate reader, and calculate the inhibition rate at the corresponding concentration; use GraphPad Prism 8.0 software to fit and obtain the IC50 of the PHGDH inhibitory activity of the test compound. 50 Values. The results are shown in Table 1.

[0122] Table 1. IC50 of some compounds of the present invention against PHGDH enzyme activity. 50 scope

[0123] Number PHGDH IC 50 (μM) Number PHGDH IC 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. "+++": IC50 50 <1 μM indicates that the compound has extremely strong PHGDH inhibitory activity. "++": 1 μM ≤ IC 50 <5 μM indicates that the compound has moderate PHGDH inhibitory activity. "+" indicates 5 μM ≤ IC50.50 <10μM indicates 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 value decreases and then increases. "+++" group (IC) 50 <1μM) showed significant inhibitory effects, suitable for therapeutic scenarios requiring potent inhibition; "++" group (1μM≤IC) 50 <5μM) showed moderate inhibition, suitable for moderate inhibition needs; "+" group (5μM≤IC) showed moderate inhibition. 50 <10μM indicates weaker inhibitory activity, which may require combination with other drugs to enhance the effect.

[0126] Example 3

[0127] Cell survival assays were conducted to test the inhibitory effect of the compounds on the proliferation of tumor cell lines.

[0128] This embodiment uses human breast cancer cell line MCF-7, liver cancer cell line H22, and colon cancer cell line CT26 for experiments. All cell lines were purchased from the Cell Bank of the Chinese Academy of Sciences. The MTT assay was used to detect the inhibitory effect of the compounds on cell proliferation. Cells were cultured at 2*103 ^ 4mL -1 Cells were cultured at a density of [insert density here] in 96-well clear plates and treated with either DMSO solution of the compound or an equal volume of DMSO. After 72 h, 5 mg / mL LMT solution was added, and the plates were incubated at 37 °C. After 4 h, the culture medium was aspirated, and the blue-purple crystals at the bottom of the plate were dissolved in DMSO. The OD of each well was measured using a Thermo microplate reader. 490 This reflects the number of living cells. Half-proliferation inhibitory concentration (GI) 50 The values ​​were obtained by fitting using GraphPad Prism 8.0 software, and the results are shown in Table 2.

[0129] Table 2. Inhibitory activity of some compounds of the present invention against tumor cell lines. 50 value

[0130]

[0131] Note: The inhibitory activity of compounds on cell proliferation in Table 2 is indicated by 0 to 3 plus signs. "+++": IC50 50 <1μM indicates that the compound has extremely strong inhibitory activity against cell proliferation. "++": 1μM≤IC 50 <5 μM indicates that the compound has moderate inhibitory activity against cell proliferation. "+" indicates 5 μM ≤ IC50. 50 <10μM indicates that the compound has weak inhibitory activity against cell proliferation.

[0132] As shown in the table above, some of the compounds of this invention exhibited good inhibitory effects on the proliferation of various tumor cell lines.

[0133] Example 4

[0134] To further elucidate the effect of LBX-5 on the cell cycle of human breast cancer cells MCF7, 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 increasing LBX-5 concentration, the proportion of MCF-7 cells in the G1 phase significantly increased. 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 validate that LBX-5 can significantly inhibit the proliferation of MCF-7 cells by regulating cell cycle progression, particularly by suppressing the expression of G1 phase-related proteins. This finding suggests that LBX-5 has potential cell cycle regulatory functions and may exert its anti-tumor activity by intervening in the PHGDH, Cyclin D1, and CDK4 signaling pathways.

[0135] Example 5

[0136] In vivo pharmacodynamic study of PHGDH small molecule inhibitor in a mouse model of H22 hepatocellular carcinoma xenograft tumor

[0137] In this embodiment, a xenograft mouse model of hepatocellular carcinoma (H22) cells was constructed to verify the in vivo antitumor activity of the representative compound LBX-5. Specifically, H22 hepatocellular carcinoma cells were inoculated into the abdomen of BALB / c mice to form a xenograft tumor model. The method for constructing this model was based on the technical scheme described in the literature [DOI:10.2147 / DDDT.S461594]. After the model was established, the inhibitory effect of LBX-5 on tumors was evaluated by injecting LBX-5 and monitoring tumor growth in the mice.

[0138] When the tumor grows to 60mm 3 -80mm 3 Mice were randomly assigned to groups and administered either a blank solvent or LBX-5 (0.3 mg / kg) via tail vein injection every 4 days for 12 days. Results showed that LBX-5 (0.3 mg / kg) could inhibit tumor growth to some extent. Compared with the blank solvent control group, the LBX-5 administration group did not show significant weight loss. Figure 2 A, 2B).

[0139] Example 6

[0140] In vivo pharmacodynamic study of PHGDH small molecule inhibitor in a mouse model of CCl4-induced acute liver injury

[0141] In this embodiment, a carbon tetrachloride (CCl4)-induced acute liver injury mouse model was constructed to verify the hepatoprotective effect of the representative compound LBX-5. Acute liver injury was induced by intraperitoneal injection of CCl4 solution (0.3%, 10 mL / kg), and mice were divided into a solvent control group and an LBX-5 treatment group. Immediately after liver injury induction, mice in the LBX-5 treatment group were 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 evidenced by a significant decrease in alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Figure 3 A, 3B).

[0142] The above are merely embodiments of the present invention and do not limit the scope of the patent. Any equivalent modifications made based on the content of this specification, or direct or indirect applications in other related technical fields, are similarly included within the scope of patent protection of the present invention.

Claims

1. The use of a compound containing a thioether skeleton or a pharmaceutically acceptable salt thereof as a PHGDH inhibitor in the preparation of a medicament for treating liver injury, characterized in that, The compound is the following compound: 。 2. The application according to claim 1, wherein the liver injury disease includes drug and toxin liver injury, alcoholic and non-alcoholic liver injury, and hepatitis virus liver injury.

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