A diphenyltriazine compound, its preparation method and application

Abstract

The present invention belongs to the technical field of chemical drugs, and particularly relates to a diphenyltriazine compound, a preparation method thereof, and an application thereof. The diphenyltriazine compound does not have a PGI2 skeleton, has strong selectivity and affinity for the PGI2 receptor, and has good target selectivity compared with PGI2 analogs. Through experimental verification, the in vivo efficacy of this compound is superior to that of selexipag, and its toxicity is far less than that of selexipag. Therefore, this compound can be used as a pulmonary hypertension treatment drug with greater clinical value and development prospects.

Description

[0001] Cross - reference to related applications

[0002] This application claims the priority of Chinese Patent Application CN202410169645.1 with the filing date of February 6, 2024, and this application incorporates the full text of the above - mentioned Chinese patent application by reference. Technical field

[0003] The present invention belongs to the technical field of chemical drugs, and particularly relates to a diphenyltriazine compound, a preparation method thereof, and an application thereof. Background art

[0004] Pulmonary arterial hypertension (PAH) is a disease mainly characterized by vasospasm, intimal hyperplasia, and remodeling of pulmonary arterioles. The vascular hyperplasia and remodeling of pulmonary arterioles lead to a progressive increase in pulmonary vascular resistance, ultimately causing right - heart failure and even death. PAH has been ranked as the third most common cardiovascular disease, second only to hypertension and coronary heart disease in terms of prevalence, and has become a major public health problem seriously threatening human physical and mental health and has been included in the scope of detection of major chronic diseases by the World Health Organization.

[0005] PGI2 is a substance produced from arachidonic acid via prostaglandin H2 (PGH2) in vivo. Deficiency of PGI2 can cause pulmonary arterial hypertension. Currently, the marketed PGI2 receptor agonists include epoprostenol, beraprost, iloprost, etc., all of which are PGI2 analogs. However, due to the very short biological half - life of PGI2 and poor selectivity for the target, it is difficult to separate the intended effect from other effects, so adverse reactions are likely to occur. Selexipag is currently the only PGI2 agonist that does not have a PGI2 skeleton but has good selectivity for the PGI2 receptor and definite efficacy, and has been approved for marketing in many countries for the treatment of adult pulmonary arterial hypertension. Its specific therapeutic effect is stronger and more long - acting compared to other drugs with similar mechanisms, but its price is high, which undoubtedly increases a great economic burden for patients with pulmonary arterial hypertension who need long - term treatment. In addition, although Selexipag has relatively good specificity, it still has obvious adverse reactions, such as headache, flushing, nausea, vomiting, etc. For patients who need to take drugs for a long time, the cumulative damage to the body caused by the drug will reduce their health level and quality of life to varying degrees. Summary of the invention

[0006] Aiming at the above problems, the present invention provides a diphenyltriazine compound, a preparation method thereof, and an application thereof. This compound has good affinity for the PGI2 receptor and very low adverse reactions, avoiding the disadvantages of current PGI2 analogs and Selexipag.

[0007] To achieve the above - mentioned invention object, the present invention adopts the following technical solutions:

[0008] In a first aspect, the present invention provides a diphenyltriazine compound or a pharmaceutically acceptable salt, stable isotope derivative, isomer, and mixture thereof, and its structural formula is as shown in Formula I:

[0009]

[0010] This compound does not have a PGI2 skeleton, has strong selectivity and affinity for the PGI2 receptor, and has good target selectivity compared with PGI2 analogs. It has been experimentally verified that the in vivo efficacy of this compound is better than that of selexipag, and its toxicity is much less than that of selexipag. Therefore, this compound can be used as a pulmonary hypertension treatment drug with greater clinical value and development prospects.

[0011] In a second aspect, the present invention also provides a preparation method of the above diphenyltriazine compound, which specifically includes the following operations:

[0012] Mix a compound with a structural formula as shown in Formula II, methylsulfonamide, 4-dimethylaminopyridine (DMAP), and carbodiimide (EDCI) in dichloromethane, heat to 35-45 °C and react for 2-3 h, cool to 20-40 °C, wash the obtained reaction solution with purified water, then wash with a hydrochloric acid solution, and then dehydrate and remove the solvent to obtain the product.

[0013]

[0014] Combined with the second aspect, the equivalent ratio of the methylsulfonamide to the compound with a structural formula as shown in Formula II is ≥1. Preferably, the equivalent ratio of the methylsulfonamide to the compound with a structural formula as shown in Formula II is ≥1.3.

[0015] Combined with the second aspect, the equivalent ratio of the 4-dimethylaminopyridine to the compound with a structural formula as shown in Formula II is ≥1. Preferably, the equivalent ratio of the 4-dimethylaminopyridine to the compound with a structural formula as shown in Formula II is ≥1.1.

[0016] Combined with the second aspect, the equivalent ratio of the carbodiimide to the compound with a structural formula as shown in Formula II is ≥1. Preferably, the equivalent ratio of the carbodiimide to the compound with a structural formula as shown in Formula II is greater than ≥1.1.

[0017] Combined with the second aspect, the amount of dichloromethane used is such that it can ensure that all reactants are dissolved therein, and the present invention does not limit this.

[0018] Combined with the second aspect, the concentration of the hydrochloric acid solution is 1.8-2.2 M.

[0019] In combination with the second aspect, the dehydration can be carried out by drying with anhydrous sodium sulfate. Anhydrous sodium sulfate can also be replaced with other desiccants that can meet the requirement of dehydrating the organic phase and have no influence on the reactants.

[0020] In combination with the second aspect, the compound with the structural formula shown in Formula II can be prepared by the following method. The specific steps include:

[0021] S1. Mix glacial acetic acid, benzil, semicarbazide hydrochloride and purified water, heat up to 100 - 110 °C and keep the reaction for 2 - 2.5 h, then cool down to 30 - 40 °C, add purified water, stir and react at 20 - 30 °C for 0.5 - 1 h, separate the solid and liquid, place the obtained solid phase in ethyl acetate, reflux and react for 2 - 3 h, cool down and then separate the solid and liquid to obtain Intermediate II-1;

[0022] S2. Mix phosphorus oxychloride and Intermediate II-1, heat up to 80 - 85 °C, stir until dissolved and clear, keep the reaction for 1 - 1.5 h, remove the solvent, add a mixed solvent of toluene and isopropanol to the obtained product, stir and disperse, separate the solid and liquid to obtain Intermediate II-2;

[0023] S3. Add Intermediate II-2 and 4-(isopropylamino)butanol to the reaction flask in sequence, heat up to 140 - 150 °C, keep the reaction for 12 - 15 h, cool down to room temperature, pour the reaction solution into water, extract with ethyl acetate, wash the organic phase with saturated sodium chloride aqueous solution and then dry it, remove the solvent, and purify the obtained product by silica gel column to obtain Intermediate II-3;

[0024] S4. Mix Intermediate II-3 with dichloromethane, cool down to 0 - 10 °C, add Dess-Martin reagent, react at 0 - 10 °C for 12 - 15 h, wash the obtained reaction solution with saturated sodium bicarbonate solution, then wash with saturated sodium chloride aqueous solution, dry the organic phase, remove the solvent, and purify the obtained product by silica gel column to obtain Intermediate II-4;

[0025] S5. Mix triethyl phosphonoacetate with tetrahydrofuran, cool down to 0 - 10 °C, add sodium hydride and react for 1 - 1.5 h, add Intermediate II-4 at 0 - 5 °C, raise the temperature to room temperature and react for 2 - 3 h, then add water dropwise to the reaction solution to quench the reaction, then concentrate to remove at least 80% of tetrahydrofuran, add ethyl acetate to the residue for extraction, wash the organic phase with saturated sodium chloride aqueous solution and then dry it, remove the solvent, and purify the obtained product by silica gel column to obtain Intermediate II-5;

[0026] S6. Mix Intermediate II-5, absolute ethanol and palladium carbon, displace with hydrogen, react at room temperature for 5 - 8 h, remove palladium carbon, then remove the solvent from the obtained reaction solution to obtain Intermediate II-6;

[0027] S7. Mix the intermediate II-6, tetrahydrofuran, sodium hydroxide and water, reflux for 2 - 4 h, concentrate to remove tetrahydrofuran, add purified water and ethyl acetate, stir evenly, let stand for liquid separation, retain the aqueous phase, adjust the pH of the aqueous phase to 3 - 5 with hydrochloric acid, extract with methyl tert-butyl ether, dry the organic phase and remove the solvent to obtain the compound with the structure shown in Formula II;

[0028] Preferably, in step S1 of this preparation method, the equivalent ratio of semicarbazide hydrochloride to benzil is greater than 1. Preferably, the equivalent ratio of semicarbazide hydrochloride to benzil is ≥1.4.

[0029] Preferably, in step S1 of this preparation method, the volume ratio of glacial acetic acid to purified water is 2 - 3:1.

[0030] Preferably, in step S1 of this preparation method, the amounts of glacial acetic acid and purified water are such that all reactants can be dissolved therein, and the present invention does not limit this.

[0031] Preferably, in step S2 of this preparation method, the mass ratio of phosphorus oxychloride to intermediate II-1 is ≥6.5.

[0032] Preferably, in step S2 of this preparation method, the volume ratio of toluene to isopropanol in the mixed solvent of toluene and isopropanol is 1:2.5 - 3.5.

[0033] Preferably, in step S3 of this preparation method, the equivalent ratio of 4-(isopropylamino)butanol to intermediate II-2 is 3 - 5:1.

[0034] Preferably, in step S3 of this preparation method, purification is carried out using a 200-mesh silica gel column, and the eluent is a dichloromethane-methanol mixed solution with a volume ratio of 30:1.

[0035] Preferably, in step S4 of this preparation method, the equivalent ratio of Dess-Martin reagent to intermediate II-3 is 1.5 - 3:1.

[0036] Preferably, in step S4 of this preparation method, purification is carried out using a 200-mesh silica gel column, and the eluent is a n-hexane-ethyl acetate mixed solution with a volume ratio of 2:1.

[0037] Preferably, in step S5 of this preparation method, the equivalent ratio of triethyl phosphonoacetate to intermediate II-4 is 1 - 2:1.

[0038] Preferably, in step S5 of this preparation method, the equivalent ratio of sodium hydride to intermediate II-4 is 1 - 2:1.

[0039] Preferably, in step S5 of the preparation method, purification is carried out using a 200-mesh silica gel column, and the eluent is a mixture of n-hexane and ethyl acetate with a volume ratio of 5:1.

[0040] Preferably, in step S5 of the preparation method, the mass of the palladium-carbon is 5% to 20% of the mass of the intermediate II-5.

[0041] In a third aspect, the present invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier, excipient or diluent, and the above-mentioned diphenyltriazine compound or a pharmaceutically acceptable salt, stable isotope derivative, isomer and mixture thereof as an active ingredient.

[0042] In combination with the third aspect, the dosage form of the pharmaceutical composition is a pharmaceutically acceptable dosage form.

[0043] Exemplarily, the above dosage forms include conventional dosage forms such as tablets, granules, capsules, powders or injections.

[0044] In a fourth aspect, the present invention also provides the use of the above-mentioned diphenyltriazine compound or a pharmaceutically acceptable salt, stable isotope derivative, isomer and mixture thereof or the above-mentioned pharmaceutical composition in the preparation of a drug for treating or preventing pulmonary hypertension, pulmonary arterial hypertension, chronic thromboembolic pulmonary hypertension, Fontan disease and pulmonary hypertension associated with Fontan disease, sarcoidosis and pulmonary hypertension associated with sarcoidosis.

[0045] Preferably, the above application is, for example, the application in the preparation of a drug for treating or preventing pulmonary hypertension, pulmonary arterial hypertension or chronic thromboembolic pulmonary hypertension.

[0046] In a fifth aspect, the present invention also provides the use of the above-mentioned diphenyltriazine compound or a pharmaceutically acceptable salt, stable isotope derivative, isomer and mixture thereof or the above-mentioned pharmaceutical composition in the preparation of a drug for treating or preventing peripheral circulatory disorders, connective tissue diseases, chronic kidney diseases including glomerulonephritis and diabetic nephropathy at any stage, diseases involving organ or tissue fibrosis, respiratory diseases, anti-ulcers, finger ulcers, diabetic gangrene, or diabetic foot ulcers. BRIEF DESCRIPTION OF THE DRAWINGS

[0047] Figure 1 For Example 1 of the present invention 1 1H-NMR spectrum;

[0048] Figure 2 For Example 1 of the present invention 13 13C-NMR spectrum;

[0049] Figure 3 For the HPLC spectrum of Example 1 of the present invention;

[0050] Figure 4EC of compound A against IP, EP1, EP2, EP3, EP4, DP, FP, and TP targets in Example 2 of the present invention 50 Figure

[0051] Figure 5 EC of Selexipag against IP, EP1, EP2, EP3, EP4, DP, FP, and TP targets in Example 2 of the present invention 50 Figure

[0052] Figure 6 Effects of compound A and Selexipag on the proliferation of human pulmonary artery smooth muscle cells in Example 2 of the present invention

[0053] Figure 7 Effects of compound A and Selexipag on the tension of rat pulmonary artery rings in Example 2 of the present invention

[0054] Figure 8 Effects of intragastric administration of compound A and Selexipag on the survival rate (%) of rats with pulmonary hypertension in Example 2 of the present invention (Note: Survival rate (%) = Number of surviving animals in the group / Total number of animals in the group × 100%)

[0055] Figure 9 Effects of intragastric administration of compound A and Selexipag on the right ventricular pressure (mmHg) of rats with pulmonary hypertension in Example 2 of the present invention (*** indicates P ≤ 0.001 compared with the model control group);

[0056] Figure 10 Effects of intragastric administration of compound A and Selexipag on the right ventricular hypertrophy index (%) of rats with pulmonary hypertension in Example 2 of the present invention (* indicates P ≤ 0.05 compared with the model control group, *** indicates P ≤ 0.001 compared with the model control group);

[0057] Figure 11 Comparison of the percentage of the medial thickness of pulmonary arterioles in each group of animals in Example 2 of the present invention (*** indicates P ≤ 0.001 compared with the model control group);

[0058] Figure 12 Pathological pictures of the lungs of rats in each group in Example 2 of the present invention. Among them, Figure A is the normal control group, lung, HE staining, 200×; Figure B is the model control group, lung, HE staining, 200×; Figure C is the low-dose test article group, lung, HE staining, 200×; Figure D is the medium-dose test article group, lung, HE staining, 200×; Figure E is the high-dose test article group, lung, HE staining, 200×; Figure F is the positive control group, lung, HE staining, 200×. Detailed implementation manners

[0059] In order to make the objectives, technical solutions and advantages of the present invention clearer and more understandable, the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0060] Term

[0061] In the preparation method of the compounds of the present application, "eq" represents equivalent, which means equivalent in chemistry. This concept is very crucial in chemical stoichiometry and chemical reactions, especially when it comes to the molar ratio of substances. For example, if 0.3 mol (1 eq) of A is used in a certain reaction, and the usage amount of B is 6 times that of A, that is, 6 eq., then the usage amount of B is 1.8 mol. This representation method helps to be more convenient and accurate when calculating the molar ratio of substances in chemical reactions.

[0062] In the present application document, both "eq" and "equivalent" are calculated according to the molar ratio, and each step can adopt its own independent equivalent standard for the convenience of calculation and experiment.

[0063] PGI2 receptor agonists are commonly used drugs for the treatment of PAH at present, but they have a short biological half-life, poor target selectivity, and are prone to adverse reactions. Selexipag does not have a PGI2 skeleton and has relatively good target selectivity, but there are still certain adverse reactions. The embodiments of the present invention provide a diphenyltriazine compound, and its structural formula is shown in Formula I:

[0064]

[0065] This compound does not have a PGI2 skeleton, has strong target selectivity, and the in vivo pharmacodynamic experimental results prove that the efficacy of this compound is better than that of selexipag, and the toxicological experimental results prove that the toxicity of this compound is much less than that of selexipag, and it has higher clinical value and broader development prospects in the treatment of pulmonary hypertension.

[0066] "Isomer" as defined herein refers to a compound having the same molecular formula but different in the nature or order of bonding of its atoms or the spatial arrangement of its atoms. Isomers with different spatial arrangements of their atoms are called "stereoisomers". Stereoisomers include optical isomers, geometric isomers and conformational isomers.

[0067] The compounds of the present invention can exist in the form of optical isomers. According to the configuration of the substituents around the chiral carbon atom, these optical isomers are in the "R" or "S" configuration. Optical isomers include enantiomers and diastereoisomers. Methods for preparing and separating optical isomers are known in the art.

[0068] The compounds of the present invention may also exist as geometric isomers. The present invention contemplates various geometric isomers and mixtures thereof resulting from the distribution of substituents around carbon-carbon double bonds, carbon-nitrogen double bonds, cycloalkyl or heterocyclic groups. Substituents around carbon-carbon double bonds or carbon-nitrogen bonds are designated as the Z or E configuration, and substituents around cycloalkyl or heterocycles are designated as the cis or trans configuration.

[0069] "Isotope" includes all isotopes of atoms that occur in the compounds of the present invention. Isotopes include those atoms having the same atomic number but different mass numbers. Examples of isotopes suitable for incorporation into the compounds of the present invention are hydrogen, carbon, nitrogen, oxygen, fluorine and chlorine, for example but not limited to 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 35 S, 18 F and 36 Cl. Isotopically labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described in the appended examples using appropriate isotopically labeled reagents in place of non-isotopically labeled reagents. Such compounds have various potential uses, such as standards and reagents in the determination of biological activity. In the case of stable isotopes, such compounds have the potential to advantageously alter biological, pharmacological or pharmacokinetic properties.

[0070] "Pharmaceutically acceptable salt" or "pharmacologically acceptable salt" refers to a salt formed from a pharmaceutically acceptable base or acid, including inorganic or organic bases or acids. In the case where the compounds of the present invention contain one or more acidic or basic groups, the present invention also encompasses their corresponding pharmaceutically acceptable salts. Thus, the compounds of the present invention containing acidic groups may exist in the form of salts and can be used according to the present invention, for example, as alkali metal salts, alkaline earth metal salts or as ammonium salts, exemplary including sodium salts, potassium salts, calcium salts, magnesium salts or salts with ammonia or organic amines, such as ethylamine, ethanolamine, triethanolamine or amino acids. The compounds of the present invention containing basic groups may exist in the form of salts and can be used according to the present invention in the form of their addition salts with inorganic or organic acids. Examples of suitable acids include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, aminosulfonic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid and other acids known to those skilled in the art. If the compounds of the present invention contain both acidic and basic groups in the molecule, the present invention also includes inner salts or inner ammonium salts in addition to the salt forms mentioned above. Each salt can be obtained by conventional methods known to those skilled in the art, for example, by contacting these with organic or inorganic acids or bases in a solvent or dispersant or by anion exchange or cation exchange with other salts.

[0071] "Pharmaceutical composition" refers to a composition containing one or more of the compounds described herein or their pharmaceutically acceptable salts, prodrugs, stable isotope derivatives, isomers and mixtures thereof, as well as other components such as pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration to an organism, promote absorption of the active ingredient and thereby exert biological activity.

[0072] The following will illustrate the solutions of the present invention through specific examples.

[0073] Example 1

[0074] This example provides a diphenyltriazine compound, its preparation method and structural characterization.

[0075] Preparation method:

[0076] 1. Preparation of the compound shown in Formula II

[0077] The intermediate shown in Formula II (chemical name: 6-((5,6-diphenyl-1,2,4-triazin-3-yl)(isopropyl)amino)hexanoic acid), its synthesis route is as follows:

[0078]

[0079] The specific synthesis method is as follows.

[0080] (1) Add glacial acetic acid (2.5 L), benzil (500 g, 1 eq), semicarbazide hydrochloride (889 g, 1.5 eq), and purified water (1 L) to the reaction flask in sequence. Stir evenly, heat up to 100 - 105 °C and hold for reaction for 2.5 h. Then cool down to 30 - 40 °C, add purified water, stir and react at 20 - 30 °C for 1 h. Filter, place the filter cake in ethyl acetate, heat up to reflux and stir for 3 h. After cooling to room temperature, filter again. Vacuum dry the filter cake to obtain 502 g of intermediate II-1 (chemical name: 5,6-diphenyl-1,2,4-triazin-3-ol).

[0081] (2) Add phosphorus oxychloride (2 L) and intermediate II-1 (500 g, 1 eq) to the reaction flask in sequence. Heat up to 80 - 85 °C, stir until dissolved and clear, and hold for reaction for 1.5 h. After concentrating to remove phosphorus oxychloride, add a mixed solvent of toluene (500 mL) and isopropanol (1500 mL). Stir and disperse at room temperature to obtain a solid. Filter and vacuum dry to obtain 350 g of intermediate II-2 (chemical name: 3-chloro-5,6-diphenyl-1,2,4-triazine).

[0082] (3) Add intermediate II-2 (100 g, 1 eq) and 4-(isopropylamino)butanol (171.5 g, 3.5 eq) to the reaction flask in sequence. Heat up to 140 - 150 °C and hold for reaction for 14 h. Cool down to room temperature, pour the reaction solution into water, add ethyl acetate for extraction. Wash the organic phase three times with saturated sodium chloride aqueous solution. Dry the organic phase with anhydrous sodium sulfate, concentrate to dryness. Purify the residue by silica gel column (200-mesh silica gel, eluent dichloromethane:methanol = 30:1) to obtain 71 g of intermediate II-3 (chemical name: 4-((5,6-diphenyl-1,2,4-triazin-3-yl)(isopropyl)amino)butan-1-ol).

[0083] (4) Add intermediate II-3 (70 g, 1 eq) and dichloromethane (700 ml) to the reaction flask. Cool down to 0 - 10 °C, and add Dess-Martin reagent (163.8 g, 2 eq) in batches. Hold for reaction at 0 - 10 °C for 14 h. Stop the reaction, wash the reaction solution twice with saturated sodium bicarbonate solution, then wash with saturated sodium chloride aqueous solution. Dry the organic phase with anhydrous sodium sulfate, concentrate to dryness. Purify the residue by silica gel column (200-mesh silica gel, eluent n-hexane:ethyl acetate = 2:1) to obtain 41.3 g of intermediate II-4 (chemical name: 4-((5,6-diphenyl-1,2,4-triazin-3-yl)(isopropyl)amino)butanal).

[0084] (5) Triethyl phosphonoacetate (29.8 g, 1.2 eq) and tetrahydrofuran (400 ml) were added to a reaction flask. The temperature was lowered to 0 - 10 °C, and sodium hydride (4.4 g, 1 eq) was added. The mixture was stirred for 1 h. Intermediate II-4 (40 g, 1 eq) was added at 0 - 5 °C. After addition, the temperature was raised to room temperature and the reaction was carried out for 2.5 h. 50 ml of water was added dropwise to the reaction solution to quench the reaction. Most of the tetrahydrofuran (more than 80%) was removed by concentration. 400 ml of ethyl acetate was added to the residue for extraction. The organic phase was washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated to dryness, and the residue was purified by silica gel column (200-mesh silica gel, eluent n-hexane:ethyl acetate = 5:1) to obtain 21.3 g of Intermediate II-5 (chemical name: ethyl 6-((5,6-diphenyl-1,2,4-triazin-3-yl)(isopropyl)amino)-2-hexenoate).

[0085] (6) 21 g of Intermediate II-5, 2.1 g of 10% palladium on carbon, and absolute ethanol (210 ml) were added to a reaction flask. The flask was purged with hydrogen, and the reaction was carried out at room temperature for 7 h. The palladium on carbon was removed by filtration, and the filtrate was concentrated to dryness to obtain 20.2 g of Intermediate II-6 (chemical name: ethyl 6-((5,6-diphenyl-1,2,4-triazin-3-yl)(isopropyl)amino)hexanoate).

[0086] (7) Intermediate II-6 (20 g, 1 eq), tetrahydrofuran (200 ml), purified water (20 ml), and sodium hydroxide (7.4 g, 4 eq) were successively added to a reaction flask. The temperature was raised to reflux and the reaction was carried out for 2 h. Tetrahydrofuran was removed by concentration. Purified water (100 ml) and ethyl acetate (100 ml) were added, and the mixture was stirred evenly and allowed to stand for liquid separation. The aqueous phase was retained. Hydrochloric acid was added to the aqueous phase to adjust the pH to 3 - 5, and methyl tert-butyl ether was added for extraction. The organic phase was dried over anhydrous sodium sulfate and then concentrated under reduced pressure to dryness to obtain 16.8 g of the compound shown in Formula II; 1 1H-NMR (500 MHz, CDCl3): δ: 7.510 - 7.493 (m, 2H), 7.475 - 7.448 (m, 2H), 7.396 - 7.364 (m, 1H), 7.317 - 7.269 (m, 5H), 5.141 - 5.070 (m, 1H), 3.592 (m, 2H), 2.407 - 2.362 (m, 2H), 1.779 - 1.707 (m, 4H), 1.506 - 1.445 (m, 2H), 1.322 - 1.308 (m, 6H), ppm.

[0087] 2. Preparation of the compound shown in Formula I

[0088] To the reaction flask, add successively the compound shown by the structural formula as shown in Formula II (1 g, 1 eq), dichloromethane (20 ml), methylsulfonamide (353 mg, 1.5 eq), DMAP (362 mg, 1.2 eq), and EDCI (569 mg, 1.2 eq). Heat the mixture to 35 - 45 °C and react for 3 h. Then cool it to room temperature. Wash the resulting reaction solution once with purified water, once with 2M hydrochloric acid, and then once again with purified water. Dry the organic phase over anhydrous sodium sulfate and concentrate it to dryness to obtain 650 mg of the compound shown by Formula I (chemical name: 6 - ((5,6 - diphenyl - 1,2,4 - triazin - 3 - yl)(isopropyl)amino)-N-(methylsulfonyl)hexanamide)).

[0089] The structural characterization spectra of the compound shown by Formula I are shown in Figure 1 、 Figure 2 and Figure 3 , and the specific data are as follows:

[0090] 1 1H - NMR (500 MHz, CDCl3): δ: 10.144 (br, 1H), 7.480 - 7.497 (m, 2H), 7.447 - 7.466 (m, 2H), 7.398 (m, 1H), 7.262 - 7.321 (m, 5H), 5.075 (m, 1H), 3.622 (m, 2H), 3.236 (s, 3H), 2.370 - 2.399 (t, 2H), 1.854 - 1.716 (m, 4H), 1.460 - 1.487 (m, 2H), 1.292 - 1.306 (m, 6H), ppm.

[0091] 13 13C NMR: (500 MHz, CDCl3): δ: 20.41, 23.89, 25.71, 36.13, 41.36, 41.67, 46.66, 77.25, 128.21, 128.30, 129.16, 129.69, 130.22, 136.14, 136.59, 147.75, 158.84 ppm.

[0092] High - resolution mass spectrometry: [M + 1] + Measured value (m / z): 482.2210.

[0093] HPLC spectrum: The retention time of the main peak is 5.520 minutes.

[0094] The results of elemental analysis are shown in Table 1 as follows:

[0095] Table 1 Results of elemental analysis

[0096]

[0097] Based on the above structural characterization results, it can be determined that the structural formula of the compound is as shown in Formula I:

[0098]

[0099] Example 2

[0100] This example provides the pharmacodynamic experiments and results of the compound shown in Formula I (hereinafter referred to as "Compound A").

[0101] I. Evaluation of the effects of Compound A on IP, EP1, EP2, EP3, EP4, DP, FP, and TP targets

[0102] In this experiment, cell lines stably expressing IP, EP1, EP2, EP3, EP4, DP, FP, and TP receptors were used. By using the methods of HTRF cAMP and HTRF IP1, according to the changes in cell signal intensity, the functional activities of Compound A and Selexipag on the 8 targets of IP, EP1, EP2, EP3, EP4, DP, FP, and TP were studied, and the corresponding concentration-effect curve EC 50 values were calculated. See Figure 4 .

[0103] The results showed that: Compound A had an agonist effect on the IP target under the initial detection condition of 10000 nM, and the absolute EC 50 value was 0.48 nM; there was no obvious agonist effect on the EP1, EP2, EP3, EP4, DP, FP, and TP targets under the initial detection condition of 10000 nM, and the absolute EC 50 values were all greater than 10000 nM.

[0104] As a comparison, the effects of Selexipag on IP, EP1, EP2, EP3, EP4, DP, FP, and TP targets were detected. The results showed that: under the initial detection condition of 10000 nM, Selexipag had an agonist effect on the IP, EP2, and DP targets, and the absolute EC 50 values were 0.27 nM, 9754.34 nM, and 2736.01 nM respectively; there was no obvious agonist effect on the EP1, EP3, EP4, FP, and TP targets under the initial detection condition of 10000 nM, and the absolute EC 50 values were all greater than 10000 nM. See Figure 5 .

[0105] The above results indicate that: The agonist activity of Compound A towards the IP receptor is weaker than that of Selexipag, but its target selectivity is stronger than that of Selexipag. It is speculated that the adverse reactions such as headache, flushing, nausea, vomiting, etc. caused by Compound A's over-activation of the IP receptor are less severe, and the adverse reactions such as muscle pain caused by the activation of other prostacyclin receptors are also less severe.

[0106] II. Effects of Compound A on the Proliferation of Human Pulmonary Artery Smooth Muscle Cells

[0107] Human pulmonary artery smooth muscle cells were cultured in a specialized smooth muscle medium and placed statically in a cell culture incubator at a temperature of 37°C, a CO2 concentration of 5%, and saturated humidity. 6×10 3 cells were seeded onto a 96-well plate and cultured in the 96-well plate (final volume: 100 μl). After 24 hours, they were starved with serum-free medium for 24 hours. Then, Compound A and Selexipag were serially diluted (10000, 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, 0.1 nM) in a medium containing 20 ng / ml hPDGF-BB at the final concentration. The medium in the cell culture wells was removed, and the serially diluted test substances were added to the cell culture wells, with a volume of 100 μl. The CTG method was used to study the inhibitory effects of the test substances Compound A and Selexipag on the proliferation of human pulmonary artery smooth muscle cells and to fit their half-maximal inhibitory concentration IC 50 , so as to evaluate the potential of the test substances for the treatment of pulmonary hypertension.

[0108] The results showed (see Appendix Figure 6 ) that: The IC 50 values of Compound A and Selexipag for the inhibition of PDGF-induced proliferation of pulmonary artery smooth muscle cells were 9.65 and 18.69 μM, respectively. The inhibitory effect of Compound A was positively correlated with the compound concentration at a concentration of 0.3 - 10 μM.

[0109] In summary, Compound A and Selexipag showed effective inhibition of PDGF-induced proliferation of pulmonary artery smooth muscle cells. The IC 50 value of the inhibitory effect of Compound A < Selexipag, indicating that the inhibitory effect of Compound A is stronger than that of Selexipag.

[0110] III. Effects of Compound A on the Tension of Rat Pulmonary Artery Rings

[0111] After anesthetizing the SD rats, the heart and lungs were located and dissected free, then immersed in oxygenated perfusion fluid. After washing away the blood, the dissected heart and lungs were placed into clean, oxygenated incubation fluid. The pulmonary artery was located, and the pulmonary artery vessel was dissected free under a microscope, taking care not to pull on the pulmonary artery during this process. The blood remaining on the vascular endothelium was cleared, and vascular rings with an inner diameter of about 2 mm to 3 mm were prepared and hung on the hook of a force transducer (taking care not to damage the endothelial cells), and the pulmonary artery branches were immersed in the liquid in the incubation bath. The basal tension of the pulmonary artery branches was adjusted to about 0.8 - 1 g through micromanipulation and stabilized for 1 - 2 hours. After the basal tension of the pulmonary artery rings was stabilized, high K + After the incubation fluid induced an increase and stabilization of the pulmonary artery ring tension, incubation fluid containing test compounds at different concentrations, Compound A (1 μM, 5 μM, 10 μM, 20 μM, 60 μM) and Selexipag (5 μM, 10 μM, 20 μM, 40 μM, 60 μM), N = 5, was added. The changes in the pulmonary artery ring tension were recorded. After observing the maximum drug efficacy, the pulmonary artery rings were washed with basal incubation fluid to restore the tension to the basal level.

[0112] The results showed (see Appendix Figure 7 ): Both Compound A and Selexipag could inhibit the contraction of rat pulmonary artery vessels induced by high K + to varying degrees at the five tested concentrations. The inhibitory effects, IC 50 , were 11.15 μM and 23.97 μM respectively. The order of the IC 50 of each compound, which could reflect the inhibitory efficiency of the compound from high to low, was: Compound A > Selexipag. In summary, both Compound A and Selexipag could effectively inhibit the contraction of pulmonary artery vessels induced by high K + , and the inhibitory effect of Compound A was greater than that of Selexipag.

[0113] IV. Pharmacodynamic Test of Compound A on Rats with Monocrotaline-Induced Pulmonary Hypertension Model

[0114] Sixty-five rats were divided into 6 groups, with 10 rats in the normal control group and 11 rats in each of the remaining 5 groups. The 5 model groups were used as the model control group, the low-dose test article group, the medium-dose test article group, the high-dose test article group, and the positive control group respectively. Among them, monocrotaline, 60 mg / kg, 5 ml / kg was used for model preparation. The animals in the normal control group were intraperitoneally injected with sodium chloride injection (5 ml / kg). Approximately 2 hours after the animals were given the modeling agent, the animals in each group started to be administered drugs.

[0115] The normal control group and the model control group were given 0.5% CMC-Na by gavage. The low-dose test article group was given 0.15 mg / kg of Compound A by gavage, the medium-dose test article group was given 0.5 mg / kg of Compound A, and the high-dose test article group was given 1.5 mg / kg of Compound A by gavage. The positive control group was given 1 mg / kg of Selexipag by gavage, twice a day for 19 consecutive days.

[0116] The body weight changes of the rats were recorded and the survival rate was observed. The mean pulmonary artery pressure of the rats was measured by the right heart catheterization method. HE staining was used to observe the pathological changes of the right ventricular tissue and lung tissue, and the percentage of the medial thickness and the right heart hypertrophy index were calculated.

[0117] The results showed that:

[0118] (1) Effect of Compound A on the body weight of rats with pulmonary hypertension

[0119] After the animals were given the modeling reagent, their body weight was significantly lower than that of the animals in the normal control group. With the extension of the test period, the average body weight of the animals in each dosing group increased steadily, and the body weight of the animals in the model control group decreased more significantly. As shown in Table 2.

[0120] Table 2 Effect of gavage administration of Compound A and Selexipag on the body weight (g) of rats with pulmonary hypertension

[0121]

[0122] Note: Compared with the model control group, "*" indicates P≤0.05, "**" indicates P≤0.01, and "***" indicates P≤0.001.

[0123] (2) Compound A reduces the mortality rate of rats with pulmonary hypertension

[0124] During the test, the survival rate of the animals in the model control group was 82%, and no animals died in the other groups, and the survival rate was 100%. As Figure 8 shown.

[0125] (3) Compound A dose-dependently reduces the right ventricular systolic pressure, right heart hypertrophy index and medial thickness of the pulmonary arterioles in rats

[0126] Compared with the right ventricular systolic pressure (RVSP, mmHg) of the animals in the model control group, the low-dose test article group (0.15 mg / kg), the medium-dose test article group (0.5 mg / kg), the high-dose test article group (1.5 mg / kg), and the positive drug control group (1 mg / kg) were all significantly reduced, and the maximum reduction amplitudes were approximately: 15%, 33%, 43%, 33% respectively, and all had statistical differences (P≤0.001). As Figure 9 and Table 3 shown.

[0127] Table 3 Effects of intragastric administration of Compound A and Selexipag on pulmonary artery blood pressure (mmHg) in rats with pulmonary hypertension

[0128]

[0129] Note: Compared with the model control group, "***" indicates P≤0.001.

[0130] Compared with the animals in the model control group, Compound A significantly reduced the right ventricular hypertrophy index in rats at the doses of 0.15 mg / kg, 0.5 mg / kg, and 1.5 mg / kg, and the maximum decreases were 17%, 27%, and 29% respectively. At the dose of 1 mg / kg, Selexipag reduced the right ventricular hypertrophy index in rats by 26%. As Figure 10 shown in and Table 4.

[0131] Table 4 Effects of intragastric administration of Compound A and Selexipag on right ventricular hypertrophy index (%) in rats with pulmonary hypertension

[0132]

[0133] Note: Compared with the model control group, "*" indicates P≤0.05 and "***" indicates P≤0.001.

[0134] Compared with the model control group, the percentage of intimal thickness was significantly decreased in the groups of Compound A at the doses of 0.5 mg / kg and 1.5 mg / kg and in the positive control group (P≤0.001); there was no significant change in the percentage of intimal thickness in the 0.15 mg / kg group of Compound A compared with the model control group. Compared with the 1.5 mg / kg group of Compound A, the percentage of intimal thickness in the positive control group was slightly decreased. As Figure 11 shown in and Table 5.

[0135] Table 5 Effects of intragastric administration of Compound A and Selexipag on the percentage of pulmonary artery medial thickness (%) in rats with pulmonary hypertension

[0136]

[0137] Note: Compared with the model control group, "***" indicates P≤0.001.

[0138] (4) Compound A alleviates pathological damage of lung tissue in rats with pulmonary hypertension

[0139] In the model control group and the drug administration group, under microscopic observation, some lesions of the lungs and bronchi caused by the pulmonary hypertension model were visible, such as alveolar foamy macrophage infiltration, alveolar hemorrhage, alveolar fibrinoid substance exudation, congestion, and thickening of the media of small pulmonary arteries / luminal stenosis, etc. Considering the lung lesions comprehensively, compared with the model control group, the incidence and degree of lung lesions in the animals given Compound A and the positive control were reduced to a certain extent. In addition, compared with the high, medium, and low doses of the test article group, the degree of lesions in the positive control group was reduced to a certain extent, as Figure 12 shown.

[0140] From the above results, it can be seen that under the conditions of this experiment, when Compound A was administered by gavage at doses of 0.15 mg / kg, 0.5 mg / kg, and 1.5 mg / kg, and Selexipag was administered by gavage at a dose of 1 mg / kg, twice a day for 19 consecutive days, a total of 38 times, it could significantly reduce the pulmonary artery pressure, right ventricular hypertrophy index, and percentage of pulmonary artery media thickness in rats with monocrotaline-induced pulmonary hypertension model, improve the survival rate, and had a certain ameliorating effect on lung tissue lesions, showing a dose-response relationship. The effect of Compound A at a dose of 0.5 mg / kg was close to that of the positive control Selexipag (1 mg / kg).

[0141] In summary, the diphenyltriazine compounds or their pharmaceutically acceptable salts, stable isotope derivatives, isomers, and mixtures thereof in this application have significant effects in the treatment or prevention of diseases such as pulmonary hypertension, chronic thromboembolic pulmonary hypertension, etc. In addition, they have significant effects in Fontan disease and pulmonary hypertension associated with Fontan disease, sarcoidosis and pulmonary hypertension associated with sarcoidosis.

[0142] Based on the same mechanism, the diphenyltriazine compounds or their pharmaceutically acceptable salts, stable isotope derivatives, isomers, and mixtures thereof in this application have therapeutic or prophylactic effects on peripheral circulatory disorders (such as chronic arterial occlusion, intermittent claudication, peripheral embolism, vibration syndrome, Raynaud's disease).

[0143] Based on the same above-mentioned mechanism, in connective tissue diseases, such as systemic lupus erythematosus, scleroderma, mixed connective tissue disease, vasculitis syndrome, it has therapeutic or prophylactic effects. In arteriosclerosis and thrombosis, it has therapeutic or prophylactic effects, such as acute cerebral thrombosis, pulmonary embolism, etc. In thrombocytopenia caused by dialysis and diseases involving organ or tissue fibrosis, it has prophylactic or therapeutic effects, such as kidney diseases such as tubulointerstitial nephritis. In respiratory diseases, such as interstitial pneumonia, (idiopathic) pulmonary fibrosis, chronic obstructive pulmonary disease, digestive system diseases (such as liver cirrhosis, viral hepatitis, chronic pancreatitis, and scirrhous gastric cancer), as well as in the prevention or treatment of ulcers, finger ulcers, diabetic gangrene, or diabetic foot ulcers, etc., it has prophylactic or therapeutic effects.

[0144] Example 3

[0145] This example provides the toxicological experiments and results of the compound shown in Formula I (hereinafter referred to as "Compound A").

[0146] I. Acute toxicity test of Compound A by oral gavage in rats

[0147] Twenty-four healthy SD rats, half male and half female, were randomly divided into 4 groups according to body weight. The first group was the vehicle group, and the second to fourth groups were the low, medium, and high dose groups of Compound A, with the dosing doses being 100, 300, and 1000 mg / kg respectively. All animals were given a single oral gavage on Day 1, and the dosing volume was 10 ml / kg. The animals were observed for 7 days after dosing.

[0148] One female rat in the fourth group was near death on Day 2 and was then euthanized.

[0149] In the detailed clinical observation, it was found that the above-mentioned female animals in the high-dose (1000 mg / kg) group of Compound A showed loss of righting reflex, cold skin when touched, disheveled hair, and near death on Day 2. The above-mentioned detailed observation symptoms were considered to be toxic reactions caused by Compound A; two female rats in the medium-dose (300 mg / kg) group of Compound A showed symptoms of red nasal secretions on Day 2. The nasal secretions only appeared on Day 2 and then returned to normal, without obvious dose correlation, and were considered to be related to Compound A but not toxic reactions. Compared with the body weight during the adaptation period, the body weights of female animals in the high-dose (1000 mg / kg) group of Compound A decreased on Day 2, which was considered to be a toxic reaction caused by Compound A. There were no obvious abnormalities in the food intake and gross anatomy of animals in each dose group. After a single oral gavage of Compound A to SD rats, the MTD was less than 1000 mg / kg.

[0150] The above toxic symptoms were basically the same as those of rats given Selexipag, but Compound A did not cause serious toxic symptoms such as blackening or shedding of the rat tail. The non-lethal dose of Selexipag in rats was 250 mg / kg, and the approximate lethal dose was 500 mg / kg. Combining with the pharmacodynamic data, the safe dose range of Compound A in rats was greater than that of Selexipag, and the tolerance dose of Compound A in rats was higher than that of Selexipag.

[0151] II. 4-week repeated-dose toxicity test of Compound A in rats

[0152] 160 healthy SD rats (SPF grade), half male and half female, were randomly divided into 8 groups according to gender and body weight. Groups 1 - 4 were the toxicity study groups, with 15 animals of each gender in each group; Groups 5 - 8 were the toxicokinetics study groups, with 5 animals of each gender in each group. All animals were orally gavaged with different doses of Compound A once a day for 4 consecutive weeks. The dosing doses were 0 mg / kg (vehicle), 50 mg / kg, 150 mg / kg, and 500 mg / kg respectively, and the dosing volume was 10 ml / kg for all.

[0153] During the experiment, the behavior and physical signs of the animals were observed and recorded; in Groups 1 - 4, body weight and food intake were measured once a week during the dosing period and the recovery period, and fundus examinations were performed at the end of the dosing and the end of the recovery period; at the end of the dosing and the end of the recovery period, clinical pathology examinations (including: hematology, coagulation, serum biochemistry, and urine analysis) were carried out on the animals scheduled for dissection. All the animals scheduled for sample collection were fasted overnight (≥10 h) before collecting clinical pathology samples, and about 12 h of urine was collected one day in advance for urine analysis; at the end of the dosing and the end of the recovery period, gross dissection examinations, organ weighing, and histopathological examinations were performed on the animals scheduled for dissection.

[0154] No animal deaths were observed during the experiment. The NOAEL was 50 mg / kg. The AUC of Compound A in male and female animals after repeated dosing at the NOAEL dose for 28 days (0-24) was 3784 h×ng / ml (male) and 7016 h×ng / ml (female) respectively. The NOAEL of Selexipag was 6 mg / kg. The AUC of Selexipag in male and female animals after repeated dosing at the NOAEL dose for 28 days (0-24) was 10 h×ng / ml (male) and 40 h×ng / ml (female) respectively.

[0155] Compound A at ≥500 mg / kg could cause salivation and an increase in thyroid volume in animals. Compound A at ≥150 mg / kg could cause a decrease in animal body weight, a decrease in RBC, HGB, and HCT, an increase in RET / RET%, an increase in ALT, an increase in urine volume, an increase in liver weight, and histopathology of hypertrophy of endocrine cells in the pars distalis of the pituitary (male), hypertrophy of thyroid follicular cells, and hypertrophy of liver cells. All changes recovered or showed a recovery trend at the end of the recovery period. Among the above changes, salivation only occurred on some detection days in some animals and disappeared after drug withdrawal. The change in body weight was small in the medium-dose (150 mg / kg) group. The decrease in RBC, HGB, and HCT was small, and the increase in RET / RET% was considered a compensatory effect. The increase in urine volume but no obvious abnormalities were found in other indicators were all considered non-harmful reactions related to Compound A. No toxicity reactions related to Compound A were observed in the food intake, ophthalmic examination, and coagulation examination of each group of animals.

[0156] III. Genotoxicity Test of Compound A

[0157] Genotoxicity experiment procedure: The test compound A and Selexipag were diluted in a serial dilution with the highest concentration of 1000 μg / well to obtain 8 concentrations. The 6-well plate incorporation method was used, with 2 wells treated in parallel. At the same time, negative (DMSO) and positive controls were set, and parallel tests were carried out under the conditions of the presence or absence of a metabolic activation system (±S9). After culturing for 48 - 72 hours, the precipitation of the test article and the growth of background bacteria plaques were observed, and the number of revertant colonies in each well was counted. The test results are shown in Tables 6 - 9.

[0158] Positive result determination

[0159] Results that meet one or both of the following criteria can be determined as positive:

[0160] 1) In at least one strain, with or without metabolic activation, the number of revertant colonies shows a dose-dependent increase, and the number of revertant colonies is 2 times or more than that of the negative control group.

[0161] 2) With or without metabolic activation, a significant increase in the number of revertant colonies is observed in one or more dose groups, and it can be repeated, and the number of revertant colonies is 2 times or more than that of the negative control group.

[0162] After the test sample is detected by 2 test strains, as long as one test strain is positive under the condition of adding or not adding the S9 mixture, the test article can be determined as a mutagen.

[0163] Negative result determination

[0164] If the test results show that there is no dose-dependent increasing relationship in the number of revertant colonies of each test strain, and the peak values of the number of revertant colonies in each dose group of all strains do not exceed 2 times that of the negative control group, then the test article can be determined as a non-mutagen.

[0165] The results showed that: Under the ±S9 conditions, for the TA98 and TA100 strains, when the final concentration of Selexipag ≥ 250 μg / well, non-interfering precipitation was observed; non-interfering precipitation was only observed when the final concentration of compound A was 1000 μg / well. Under the ±S9 conditions, for the TA98 and TA100 strains, when the final concentration of Selexipag ≥ 250 μg / well, a decrease in background bacteria plaques was observed; no abnormal background bacteria plaques were observed at each concentration of compound A. For the TA98 and TA100 strains, under the ±S9 conditions, the number of revertant colonies in each concentration group of compound A and Selexipag did not reach 2 times that of the negative control group, and there was no concentration-effect relationship, so the test results were considered negative.

[0166] Therefore, compound A and Selexipag are not genotoxic.

[0167] Table 6. Results of the preliminary screening test of compound A in the bacterial reverse mutation 6-well plate (TA98 strain)

[0168]

[0169]

[0170] Remarks:

[0171] Background bacterial plaque: T0 is normal.

[0172] Solubility of the test article / control: P0 is normal / without precipitation; P1 has non-interfering precipitation under the microscope.

[0173] *: The number of reverse mutation colonies in the positive control group is more than 3 times that of the negative control group.

[0174] Table 7. Results of the preliminary screening test of compound A in the bacterial reverse mutation 6-well plate (TA100 strain)

[0175]

[0176]

[0177] Remarks:

[0178] Background bacterial plaque: T0 is normal.

[0179] Solubility of the test article / control: P0 is normal / without precipitation; P1 has non-interfering precipitation under the microscope.

[0180] *: The number of reverse mutation colonies in the positive control group is more than 3 times that of the negative control group.

[0181] Table 8. Results of the preliminary screening test of Selexipag in the bacterial reverse mutation 6-well plate (TA98 strain)

[0182]

[0183]

[0184] Remarks:

[0185] NA: Not counted.

[0186] Background bacterial plaque: T0 is normal; T1 shows a slight reduction in the background bacterial plaque; T3 shows a severe reduction in the background bacterial plaque; T4 shows the disappearance of the background bacterial plaque.

[0187] Solubility of the test article / control: P0 is normal / without precipitation; P1 has non-interfering precipitation under the microscope.

[0188] a : The positive drugs without and with S9 were sodium azide (0.4 μg / well) and 2-aminoanthracene (0.6 μg / well), respectively. *: The number of revertant colonies in the positive control group was more than three times that in the negative control group.

[0189] Table 9 Results of the preliminary screening test of Selexipag bacterial reverse mutation in 6-well plates (TA100 strain)

[0190]

[0191] Remarks:

[0192] NA: Not counted.

[0193] Background plaque: Normal at T0; Slightly reduced background plaque at T1; Severely reduced background plaque at T3; Disappearance of background plaque at T4.

[0194] Solubility of the test article / control article: Normal / without precipitation at P0; Non-interfering precipitation under the microscope at P1.

[0195] a : The positive drugs without and with S9 were sodium azide (0.4 μg / well) and 2-aminoanthracene (0.6 μg / well), respectively. *: The number of revertant colonies in the positive control group was more than three times that in the negative control group.

[0196] In summary, the NOAEL of compound A in rats is 100 times the pharmacodynamic dose, and the NOAEL of Selexipag is 6 times the pharmacodynamic dose, indicating that the safety range of compound A is greater than that of Selexipag. In addition, compared with Selexipag, repeated administration of compound A for 4 weeks did not cause serious adverse reactions. The above results indicate that the safety of compound A is superior to that of Selexipag.

[0197] Example 4

[0198] This example provides a tablet for treating pulmonary hypertension, and its preparation method is: using the compound shown in Formula I as the active ingredient, and preparing the tablet with tablet excipients by a conventional process.

[0199] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, or improvements made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims

1. A diphenyltriazine compound or a pharmaceutically acceptable salt thereof, characterized in that, Its structural formula is shown in Formula I: Formula I.

2. The preparation method of the diphenyltriazine compound according to claim 1, characterized in that, Specifically, it includes the following steps: Mix a compound with a structural formula shown in Formula II, methylsulfonamide, 4-dimethylaminopyridine, and carbodiimide in dichloromethane, heat to 35 - 45 °C and react for 2 - 3 h, cool to 20 - 40 °C, wash the obtained reaction solution with purified water, then wash with hydrochloric acid solution, and then dehydrate and remove the solvent to obtain the product; Formula II.

3. The preparation method according to claim 2, characterized in that, The equivalent ratio of the methylsulfonamide to the compound with a structural formula shown in Formula II ≥ 1; and / or The equivalent ratio of the 4-dimethylaminopyridine to the compound with a structural formula shown in Formula II ≥ 1; and / or The equivalent ratio of the carbodiimide to the compound with a structural formula shown in Formula II ≥ 1.

4. The preparation method according to claim 2, characterized in that, The preparation method of the compound with a structural formula shown in Formula II specifically includes the following steps: S1. Mix glacial acetic acid, benzil, semicarbazide hydrochloride, and purified water, heat to 100 - 110 °C and keep the reaction for 2 - 2.5 h, then cool to 30 - 40 °C, add purified water, stir and react at 20 - 30 °C for 0.5 - 1 h, perform solid-liquid separation, place the obtained solid phase in ethyl acetate, reflux and react for 2 - 3 h, cool and perform solid-liquid separation to obtain Intermediate II-1; S2. Mix phosphorus oxychloride and Intermediate II-1, heat to 80 - 85 °C, stir until dissolved and clear, keep the reaction for 1 - 1.5 h, after removing the solvent, add a mixed solvent of toluene and isopropanol to the obtained product, stir and disperse, perform solid-liquid separation to obtain Intermediate II-2; S3. Sequentially add Intermediate II-2 and 4-(isopropylamino)butanol to the reaction flask, heat to 140 - 150 °C, keep the reaction for 12 - 15 h, cool to room temperature, pour the reaction solution into water, add ethyl acetate for extraction, wash the organic phase with saturated sodium chloride aqueous solution and then dry, remove the solvent, and purify the obtained product by silica gel column to obtain Intermediate II-3; S4. Mix Intermediate II-3 with dichloromethane, cool to 0 - 10 °C, add Dess-Martin reagent, react at 0 - 10 °C for 12 - 15 h, wash the obtained reaction solution with saturated sodium bicarbonate solution, then wash with saturated sodium chloride aqueous solution, dry the organic phase, remove the solvent, and purify the obtained product by silica gel column to obtain Intermediate II-4; S5. Mix triethyl phosphonoacetate with tetrahydrofuran, cool to 0 - 10 °C, add sodium hydride and react for 1 - 1.5 h, add Intermediate II-4 at 0 - 5 °C, raise the temperature to room temperature and react for 2 - 3 h, then add water dropwise to the reaction solution to quench the reaction, then concentrate to remove at least 80% of tetrahydrofuran, add ethyl acetate for extraction to the residue, wash the organic phase with saturated sodium chloride aqueous solution and then dry, remove the solvent, and purify the obtained product by silica gel column to obtain Intermediate II-5; S6. Mix Intermediate II-5, absolute ethanol, and palladium carbon, displace with hydrogen, react at room temperature for 5 - 8 h, remove palladium carbon, then remove the solvent from the obtained reaction solution to obtain Intermediate II-6; S7. Mix the intermediate II-6, tetrahydrofuran, sodium hydroxide and water, reflux and react for 2 - 4 h, concentrate to remove tetrahydrofuran, add purified water and ethyl acetate, stir evenly, let stand for liquid separation, retain the aqueous phase, adjust the pH of the aqueous phase to 3 - 5 with hydrochloric acid, extract with methyl tert-butyl ether, and remove the solvent after drying the organic phase to obtain the compound with the structure shown in Formula II.

5. The preparation method according to claim 4, characterized in that, In step S1, the equivalent ratio of semicarbazide hydrochloride to benzil is greater than 1; and / or The volume ratio of glacial acetic acid to purified water is 2 - 3:1; and / or In step S2, the mass ratio of phosphorus oxychloride to intermediate II-1 is ≥6.5; and / or In step S2, the volume ratio of toluene to isopropanol in the mixed solvent of toluene and isopropanol is 1:2.5 - 3.5; and / or In step S3, the equivalent ratio of 4-(isopropylamino)butanol to intermediate II-2 is 3 - 5; and / or In step S3, purify with a 200-mesh silica gel column, and the eluent is a dichloromethane-methanol mixed solution with a volume ratio of 30:1; and / or In step S4, the equivalent ratio of Dess-Martin reagent to intermediate II-3 is 1.5 - 3:1; and / or In step S4, purify with a 200-mesh silica gel column, and the eluent is a n-hexane-ethyl acetate mixed solution with a volume ratio of 2:1; and / or In step S5, the equivalent ratio of triethyl phosphonoacetate to intermediate II-4 is 1 - 2:1; and / or In step S5, the equivalent ratio of sodium hydride to intermediate II-4 is 1 - 2:1; and / or In step S5, purify with a 200-mesh silica gel column, and the eluent is a n-hexane-ethyl acetate mixed solution with a volume ratio of 5:1; and / or In step S6, the mass of the palladium-carbon is 5% - 20% of the mass of the intermediate II-5.

6. A pharmaceutical composition, characterized in that, Comprising a pharmaceutically acceptable carrier, excipient or diluent, and the compound according to claim 1 as the active ingredient or a pharmaceutically acceptable salt thereof.

7. The pharmaceutical composition according to claim 6, wherein The dosage form of the pharmaceutical composition is a pharmaceutically acceptable dosage form.

8. The pharmaceutical composition according to claim 7, characterized in that, The dosage form includes tablets, granules, capsules, powders or injections.

9. Use of the diphenyltriazine compound according to claim 1 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to any one of claims 6 - 8 in the preparation of a medicament for treating or preventing pulmonary hypertension.

10. The application according to claim 9, wherein, The pulmonary hypertension is chronic thromboembolic pulmonary hypertension.

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