Pgi2 receptor agonist compounds, pharmaceutical compositions and uses

Abstract

The application belongs to the technical field of chemical medicines, and provides a PGI2 receptor agonist compound shown in a general formula I or a pharmaceutically acceptable salt, a stable isotope derivative, an isomer and a mixture thereof. In the formula, R1 and R2 are independently selected from H, C1-C3 alkyl or a halogen atom; Z is selected from CR7 or an N atom, wherein R7 is selected from H or C1-C3 alkyl, a halogen atom or a halogenated C1-C3 alkyl; R3 is selected from C1-C3 alkyl, C 3‑ C6 monocyclic cycloalkyl; R4 is selected from H, C1-C3 alkoxy or a 3-6-membered heteroaliphatic ring group; R5 represents OH, OR6 or NHSO2R6, wherein R6 represents C 1‑4 alkyl or C 1‑4 alkyl substituted by a halogen; and represents a single bond or a double bond. The compound has good affinity to PGI2 receptors, and has very low adverse reactions, and avoids the defects existing in current PGI2 analogues and seipag.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to Chinese patent application CN202311824286.0, filed on December 27, 2023, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This invention belongs to the field of chemical pharmaceutical technology, specifically relating to a PGI2 receptor agonist compound, a pharmaceutical composition, and its application. Background Technology

[0004] Pulmonary arterial hypertension (PAH) is a disease characterized by vasospasm, intimal hyperplasia, and remodeling of the pulmonary arterioles. Vascular hyperplasia and remodeling of the pulmonary arterioles lead to a progressive increase in pulmonary vascular resistance, eventually causing right ventricular failure and death. PAH is now ranked third among common cardiovascular diseases, second only to hypertension and coronary heart disease in prevalence. It has become a serious public health issue threatening human physical and mental health and has been included in the World Health Organization's global monitoring of major chronic diseases.

[0005] PGI2 is a substance produced in the body from arachidonic acid via prostaglandin H2 (PGH2). PGI2 deficiency can cause pulmonary hypertension. Currently, marketed PGI2 receptor agonists include eprostol, beraprost, and iloprost, all of which are PGI2 analogs. However, due to the very short biological half-life of PGI2 and its poor selectivity for targets, its target effect is difficult to separate from other effects, thus easily leading to adverse reactions. Selexipag is currently the only PGI2 agonist that does not possess a PGI2 backbone but has good selectivity for PGI2 receptors and definite efficacy. It has been approved for marketing in many countries for the treatment of pulmonary hypertension in adults. Its specific therapeutic effect is stronger and longer-lasting than other drugs with similar mechanisms, but its high price undoubtedly adds a significant financial burden to patients with pulmonary hypertension who require long-term treatment. In addition, although selexipag has relatively good specificity, it still has significant adverse reactions, such as headache, facial flushing, nausea, and vomiting. For patients who need long-term medication, the cumulative damage to the body caused by the drug can reduce their health and quality of life to varying degrees. Summary of the Invention

[0006] To address the above problems, the present invention provides a PGI2 receptor agonist compound, a pharmaceutical composition, and an application. This compound has good affinity for the PGI2 receptor and very low adverse reactions, thus avoiding the shortcomings of current PGI2 analogs and selexiparg.

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

[0008] In a first aspect, the present application provides a PGI2 receptor agonist compound of the general formula I:

[0009]

[0010] wherein R1 and R2 are independently selected from H, C1-C3 alkyl or halogen;

[0011] Z is selected from CR7 or N atom, wherein R7 is selected from H or C1-C3 alkyl, halogen or halogenated C1-C3 alkyl;

[0012] R3 is selected from C2-C4 alkyl, C 3- C6 monocyclic cycloalkyl;

[0013] R4 is selected from H, C1-C3 alkoxy or 3-6 membered heteroalicyclic group;

[0014] R5 represents OH, OR6 or NHSO2R6, wherein R6 represents C 1-4 alkyl or C 1-4 alkyl substituted with halogen; carboxyl forms an acid, ester or sulfonamide with R5;

[0015] represents a single or double bond.

[0016] One embodiment of the present application relates to the compound of the general formula I or its pharmaceutically acceptable salts, stable isotope derivatives, isomers and mixtures thereof, wherein Z is N atom; or Z is C atom and R7 is selected from H or CH3.

[0017] In one embodiment, R1 and R2 are independently selected from H, CH3 or F; Z is N atom; R3 is selected from C2-C4 alkyl, C represents a double bond, R4 is H, and accordingly, its structural formula is shown in formula II:

[0018]

[0019] In formula II, R1 and R2 are selected from H, CH3 or F; Z is N atom, or Z is CR7, R7 is selected from H or CH3, more preferably H atom; R3 is selected from isopropyl, ethyl or cyclopropane, more preferably isopropyl; R5 is selected from OH or NHSO2R6, R6 is selected from C 1-4 alkyl or C 1-4 alkyl substituted with halogen; R6 is more preferably methyl.

[0020] One embodiment of the present application relates to the compound of the general formula I or its pharmaceutically acceptable salts, stable isotope derivatives, isomers and mixtures thereof, wherein represents a single bond, and its structural formula is shown in formula III:

[0021]

[0022] In formula III, R4 is selected from H, C1-C3 alkoxy or 3-6 membered heteroalicyclic group, preferably R4 is H, methoxy, ethoxy or More preferably, R4 is H, methoxy or ethoxy.

[0023] One embodiment of the present application relates to the compound of general formula I or general formula III or pharmaceutically acceptable salts, stable isotope derivatives, isomers and mixtures thereof, wherein R1 and R2 are selected from H, methyl or F; R3 is selected from isopropyl, ethyl or cyclopropane group, preferably R3 is isopropyl; R5 is preferably OH or NHSO2CH3.

[0024] One embodiment of the present application relates to the compound of general formula (I) as described above, wherein the compound is selected from, but not limited to:

[0025]

[0026]

[0027] or a prodrug, stable isotope derivative, pharmaceutically acceptable salt, isomer and mixture thereof of the above structure.

[0028] The compound of the present application does not have the PGI2 skeleton, has strong selectivity and affinity to PGI2 receptor, has good target selectivity compared with PGI2 analogs. It is verified by experiments that the compound has better in vivo efficacy than selexipag, and its toxicity is less than selexipag, so the compound can be used as a more clinically valuable and promising drug for treating pulmonary arterial hypertension.

[0029] The second aspect of the present application relates to a pharmaceutical composition comprising a pharmaceutically acceptable carrier, excipient or diluent, and the above-mentioned compound or pharmaceutically acceptable salts, prodrugs, stable isotope derivatives, isomers and mixtures thereof as an active ingredient.

[0030] The above-mentioned pharmaceutical composition can be any dosage form, optionally, conventional dosage forms such as tablets, granules, capsules, powders or injections.

[0031] Another aspect of the present application relates to the use of the above-mentioned compound or pharmaceutically acceptable salts, stable isotope derivatives, isomers and mixtures thereof, and the above-mentioned pharmaceutical composition in the preparation of a drug for treating pulmonary arterial hypertension. BRIEF DESCRIPTION OF DRAWINGS

[0032] Figure 1Figure showing the effect of compound in the example and Selexipag on pulmonary arterial systolic pressure by gavage administration (*** indicates P < 0.001 compared with the model control group, ** indicates P < 0.01 compared with the model control group). DETAILED DESCRIPTION

[0033] In order to make the objects, technical solutions and advantages of the present application clearer, the present application will be further described in detail below with specific examples. It should be understood that the specific examples described herein are only used to explain the present application and are not used to limit the present application.

[0034] TERMS

[0035] In the method for preparing the compound of the present application, "eq" means equivalent, which means equivalent in chemistry. This concept is very important in stoichiometry and chemical reactions, especially when the molar ratio of substances is involved. For example, if 0.3 mol (1 eq) of A is used in a certain reaction, and 6 eq of B is used, which is 6 times the amount of A, then the amount of B used is 1.8 mol. This representation method helps to calculate the molar ratio of substances in chemical reactions more conveniently and accurately. The "eq" in the present application is calculated according to the molar ratio.

[0036] As used herein, the expression "C x-y " represents the range of the number of carbon atoms, wherein x and y are both integers, for example, C 3-6 Cycloalkyl represents a cycloalkyl group having 3-6 carbon atoms.

[0037] "Alkyl" represents a saturated straight-chain or branched-chain hydrocarbon group having a certain number of carbon atoms, for example, 1-6 carbon atoms or 1-4 carbon atoms. Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, and 2-ethylbutyl, and the like.

[0038] Heteroalicyclic group represents a monocyclic or fused ring having one or more N, O or S heteroatoms. Typically, 3-8 membered heterocyclyl having one or more N, O or S heteroatoms, preferably 3-6 membered heterocyclyl having one or more N, O or S heteroatoms, such as propylene oxide, butylene oxide, piperazino, morpholino, piperidino and derivatives thereof.

[0039] For example, morpholino refers to a group having the following chemical structure: Piperidino refers to a group of the chemical structure:

[0040] "Halogen" means fluorine, chlorine, bromine or iodine.

[0041] "Isomers" as defined herein refer to compounds that have the same molecular formula but differ in the nature or sequence of bonding of their atoms or in the arrangement of their atoms in space. It is understood that isomers have the same number and kind of atoms, but their atoms are not in the same arrangement or order. Isomers that differ in the arrangement of their atoms in space are termed "stereoisomers." Stereoisomers include optical isomers, geometric isomers, and conformational isomers.

[0042] The compounds of the present application can exist in optically active or racemic forms. These optically active forms are "R" or "S" as determined by the Cahn- Ingold-Prelog convention. The optically active forms can be obtained by chiral chromatographic separation techniques or by chiral synthesis techniques. Optical isomers can also be separated using other techniques such as fractional crystallization, distribution between two immiscible solvents, selective absorption, or selective hydrolysis. Methods for the resolution of racemic compounds are known in the art and are described in, for example, Burgmaier et al. (1980) Synthesis 139; and Nohira et al. (1982) YAKUGAKU ZASSHI 102: 58-74.

[0043] The compounds of the present application can also exist as geometric isomers. The present application contemplates various geometric isomers resulting from the configuration of substituents around a carbon-carbon double bond, carbon-nitrogen double bond, a cycloalkyl, or a heterocyclic group, and mixtures thereof. The substituents around a carbon-carbon double bond or carbon-nitrogen bond are designated as being in the "Z" or "E" configuration, and the substituents around a cycloalkyl or heterocycle are designated as being in the "cis" or "trans" configuration.

[0044] "Isotopes" include all isotopes of atoms occurring in the present compounds. Isotopes include those atoms having the same atomic number but different mass atoms, e.g., 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 application can generally be prepared by conventional techniques known to those skilled in the art, or by processes analogous to those described in the accompanying Examples using an appropriate isotopically-labeled reagent in place of the non- isotopically labeled reagent. Such compounds are within the scope of the present application. Such compounds have a variety of potential uses, for example as standards and reagents in determining biological activity. In the case of stable isotopes, such compounds have the potential to alter the biological, pharmacological, or pharmacokinetic properties of the compound.

[0045] "Prodrug" means that the compounds of the present application can be administered in the form of a prodrug. Prodrugs are derivatives of the biologically active compounds of the present application that are modified to improve their delivery to the site of action in the body. Examples of prodrugs are compounds wherein amino groups are acylated, alkylated or phosphorylated, such as methylacylamino, alanyl-amino, pivaloyloxymethylamino, or wherein hydroxy groups are acylated, alkylated, phosphorylated or converted into borates, such as acyloxy, fumaryloxy, alanyloxy, or wherein carboxy groups are esterified or amidated. These compounds can be prepared from the compounds of the present application according to methods known in the art.

[0046] "Pharmaceutically acceptable salt" or "pharmaceutically acceptable salts" means salts of a compound of the present application derived from pharmaceutically acceptable bases or acids, including inorganic bases or acids and organic bases or acids. In cases where the compounds of the present application contain one or more acidic or basic groups, the present application also comprises the corresponding pharmaceutically acceptable salts thereof. Thus, compounds of the present application containing an acidic group can exist in the form of salts and can be used according to the present application, 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. Compounds of the present application containing a basic group can exist in the form of salts and can be used according to the present application 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, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid and other acids known to the person skilled in the art. If the compounds of the present application contain both acidic and basic groups in the molecule, the present application comprises, in addition to the mentioned salt forms, internal salts or betaines. The respective salts can be obtained by customary methods known to the person skilled in the art, for example by contacting these with organic or inorganic acids or bases in solvents or dispersants or by ion exchange with other salt anions or cations.

[0047] "Pharmaceutical composition" means a composition comprising one or more compounds described herein or pharmaceutically acceptable salts, prodrugs, stable isotopic derivatives, isomers, and mixtures thereof, and other components, such as pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of the active ingredient to an organism for the purpose of absorption and thereby exerting a biological activity.

[0048] Thus, when in the present application reference is made to "compounds", "compounds of the present application" or "compounds described by the present application", all forms of the described compounds are meant to be included, such as pharmaceutically acceptable salts, prodrugs, stable isotopic derivatives, isomers, and mixtures thereof.

[0049] The present application also provides a process for preparing the compounds. The compounds of the present application of general formula (I) can be prepared by the following exemplary methods and examples, but these methods and examples should not be considered in any way to be limiting to the scope of the present application. The compounds of the present application can also be synthesized by synthetic techniques known to those skilled in the art, or by a combination of methods known in the art and those described herein. The products resulting from each step of the reaction are obtained using separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, chromatography, and the like. The starting materials and chemical reagents required for the synthesis can be routinely synthesized according to the literature or purchased.

[0050] Reference Example 1

[0051] Preparation of 6-isopropylamino-3-ethoxy-hexanoic acid ethyl ester (denoted as SM1) was carried out according to the following scheme:

[0052]

[0053] Preparation of SM1-B: Into a reaction flask was added SM1-A (4-isopropylamino-1-n-butanol, 200 g, 1 eq), Boc anhydride (349.3 g, 1.05 eq), tetrahydrofuran (1200 ml) sequentially, and the reaction was carried out at room temperature for 2-2.5 h, and then concentrated to obtain 353 g of compound SM1-B.

[0054] Preparation of SM1-C: Into a reaction flask was added compound SM1-B (300 g, 1 eq), dichloromethane (1500 ml), and then cooled in an ice bath, and then added Dess-Martin reagent (660 g, 1.2 eq) in batches, and the reaction was carried out at 0-5 °C for 2-3 h, and then the reaction solution was washed with saturated sodium bicarbonate solution, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated to dryness, and then the residue was purified by silica gel column to obtain 172 g of compound SM1-C.

[0055] Preparation of SM1-D: Into a reaction flask was added triethyl phosphonoacetate (199 g, 1.2 eq), tetrahydrofuran (1300 ml), and then cooled in an ice bath, and then added sodium hydride (35.6 g, 1.2 eq) in batches, and stirred for 1-2 h, and then added compound SM1-C (170 g, 1 eq) in batches at 0-5 °C, and the reaction was carried out at room temperature for 2-3 h, and then the reaction solution was quenched by dropwise addition of water, and then extracted with ethyl acetate, and then the organic phase was washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, concentrated to dryness, and then the residue was purified by silica gel column to obtain 135 g of compound SM1-D.

[0056] SM1-E: Into a reaction flask was added compound SM1-D (30 g, 1 eq), anhydrous ethanol (150 ml), and ice bath cooling. Sodium hydride (6 g, 1.5 eq) was added, and the reaction was allowed to warm to room temperature for 2-3 h. The reaction was cooled to 0-10 °C, and 100 ml of water was added to quench the reaction. Most of the ethanol was removed by concentration, and the reaction was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the reaction was concentrated to dryness. The reaction was purified by silica gel column to give 26.3 g of compound SM1-E.

[0057] SM1: Into a reaction flask was added SM1-E (25 g, 1 eq), dichloromethane, and hydrochloric acid / ethanol solution (1.1 eq) was added dropwise at room temperature. After the dropwise addition was completed, the reaction was concentrated to dryness to give 19.4 g of SM1.

[0058] Example 1

[0059] Synthesis of 6-{N-[5,6-bis(4-methylphenyl)pyrazin-2-yl]-N-isopropylamino}-2-hexenoic acid, which is referred to as SYN-001, corresponds to structural formula (1).

[0060] The synthesis route is as follows:

[0061]

[0062] STEP 1: Into a reaction flask was added compound 1A (4,4'-dimethylbenzil 300 g, 1 eq), 2-aminoacetamide hydrochloride (209 g, 1.5 eq), and methanol (2.0 L). The reaction was warmed to reflux, and sodium hydroxide aqueous solution (sodium hydroxide 101 g, 2 eq, purified water 200 ml) was added dropwise. The reaction was stirred at elevated temperature for 4-5 h, and the reaction was cooled to room temperature. The reaction was adjusted to pH 6-7 with hydrochloric acid, and the reaction was filtered. The filter cake was rinsed with 300 ml of methanol, and the reaction was dried under vacuum to give 320 g of compound 1B.

[0063] STEP 2: Into a reaction flask was added compound 1B (100 g, 1 eq) and phosphorus oxychloride (277.4 g, 5 eq). The reaction was warmed to 100-105 °C, and the reaction was stirred at elevated temperature for 3-5 h. The reaction was concentrated under reduced pressure to remove residual phosphorus oxychloride. 200 ml of toluene was added, and the reaction was further concentrated to remove residual phosphorus oxychloride. 100 ml of dichloromethane was added to the reaction, and the reaction was stirred to dissolve. The reaction was added dropwise to 1000 ml of isopropyl alcohol. After the dropwise addition was completed, the reaction was stirred at 0-5 °C for 1-3 h. The reaction was filtered, and the filter cake was dried under vacuum to give 90.7 g of compound 1C.

[0064] STEP3: Into a reaction flask, was added compound 1C (100 g, 1 eq), 4- (isopropylamino)butanol (245 g, 5.5 eq) sequentially, and the temperature was raised to 160-170 °C, and the reaction was maintained for 20-30 h. The temperature was then lowered to 60-80 °C, and the reaction mixture was poured into water (1.0 L), and extracted with ethyl acetate. The organic phase was washed with saturated ammonium chloride solution, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was purified by silica gel column chromatography to obtain 74 g of compound 1D.

[0065] STEP4: Into a reaction flask, was added compound 1D (70 g, 1 eq), dichloromethane (700 ml), and the temperature was lowered in an ice bath. Then, Dess-Martin reagent (152 g, 2 eq) was added portionwise, and the reaction was maintained for 3-5 h. The reaction mixture was washed with saturated sodium bicarbonate solution, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was purified by silica gel column chromatography to obtain 21.5 g of compound 1E.

[0066] STEP5: Into a reaction flask, was added triethyl phosphonoacetate (9 g, 1.2 eq), tetrahydrofuran (130 ml), and the temperature was lowered in an ice bath. Then, sodium hydride (1.34 g, 1 eq) was added portionwise, and stirred for 1 h. Compound 1E (13 g, 1 eq) was added portionwise at 0-5 °C, and the temperature was raised to room temperature. The reaction was maintained for 2-3 h, and 50 ml of water was added dropwise to quench the reaction. The reaction mixture was extracted with ethyl acetate, and the organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated to dryness. The residue was purified by silica gel column chromatography to obtain 13 g of compound 1F.

[0067] STEP6: Into a reaction flask, was added compound 1F (3 g, 1 eq), tetrahydrofuran (30 ml), and purified water (3 ml). Then, sodium hydroxide (1.05 g, 4 eq) was added, and the temperature was raised to reflux. The reaction was maintained for 12-15 h, and the temperature was lowered to room temperature. Then, 50 ml of water was added, and the pH was adjusted to 5-6 with hydrochloric acid. The reaction mixture was extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure, and the residue was purified by silica gel column chromatography to obtain 1.3 g of compound SYN-001.

[0068] 1 H NMR (500 MHz, CDC13): δ: 8.019 (s, 1H), 7.366-7.350 (m, 2H), 7.282 (s, 2H), 7.158-7.057 (m, 5H), 5.925-5.894 (d, 1H), 4.714-4.688 (m, 1H), 3.460-3.429 (m, 2H), 2.371-2.332 (m, 8H), 1.918-1.857 (m, 2H), 1.291-1.277 (m, 6H) ppm.

[0069] Example 2

[0070] 6-[N-(5,6-diphenylpyrazin-2-yl)-N-isopropylamino]-2-hexenoic acid, compound noted as SYN-002, corresponding to structural formula (2).

[0071] The preparation route is as follows:

[0072]

[0073] The preparation method of 6-[N-(5,6-diphenylpyrazin-2-yl)-N-isopropylamino]-2-hexenoic acid is the same as that of 6-{N-[5,6-bis(4-methylphenyl)pyrazin-2-yl]-N- isopropylamino}-2-hexenoic acid (numbered as SYN-001) in Example 1, except that the starting material is benzil.

[0074] 1 H NMR (500 MHz, CDC13): δ: 8.050 (s, 1H), 7.449-7.430 (m, 2H), 7.366-7.347 (m, 2H), 7.285-7.247 (m, 6H), 7.232-7.211 (m, 1H), 5.920-5.889 (m, 1H), 4.741-4.715 (m, 1H), 3.454-3.422 (m, 2H), 2.354-2.340 (m, 2H), 1.898-1.867 (m, 2H), 1.285-1.272 (m, 6H) ppm.

[0075] Example 3

[0076] 6-{N-[5,6-bis(4-methylphenyl)-1,2,4-triazin-2-yl]-N-isopropylamino}hexanoic acid, compound noted as SYN-003, corresponding to structural formula (3).

[0077] The preparation route is as follows:

[0078]

[0079] STEP 1: Add glacial acetic acid (2.5 L), 4,4'-dimethylbenzil (compound 6A; 500 g, 1 eq), semicarbazide hydrochloride (351 g, 1.5 eq), and purified water (1 L) into a reaction bottle in sequence, stir uniformly, and warm to 100-110 °C. Keep the reaction at 100-110 °C for 2-3 h, cool to 30-40 °C, add purified water, stir at room temperature for 0.5-1.5 h, filter, add the filter cake into 3 L of ethyl acetate, warm to reflux and stir for 2-3 h, cool to room temperature, filter, and vacuum dry to obtain 523 g of intermediate 6B.

[0080] STEP 2: To the reaction flask was added phosphorous oxychloride (2 L) followed by 6B (500 g, 1 eq) and the temperature was raised to 80-85 °C. The solution was stirred until clear. The reaction was maintained for 1-1.5 h and the phosphorous oxychloride was removed by concentration. Toluene (1 L) was added and the remaining phosphorous oxychloride was removed by concentration. Methyl tert-butyl ether was added to the residue and stirred for 1 h. The mixture was filtered and dried under vacuum to give 320 g of intermediate 6C.

[0081] STEP 3: To the reaction flask was added intermediate 6C (100 g, 1 eq) followed by 4-(isopropylamino)butanol (155.3 g, 3.5 eq) and the temperature was raised to 140-160 °C. The reaction was maintained for 14-20 h and the temperature was lowered to room temperature. The reaction mixture was poured into water and extracted with ethyl acetate. The organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, concentrated to dryness and purified on a silica gel column to give 71 g of intermediate 6D.

[0082] STEP 4: To the reaction flask was added intermediate 6D (70 g, 1 eq) followed by dichloromethane (700 ml) and the temperature was lowered to 0-10 °C. Dess-Martin reagent (152 g, 2 eq) was added portionwise and the reaction was maintained for 3-5 h at 0-10 °C. The reaction was stopped and the reaction mixture was washed twice with saturated sodium bicarbonate solution and once with saturated aqueous sodium chloride solution. The organic phase was dried over anhydrous sodium sulfate, concentrated to dryness and purified on a silica gel column to give 41.3 g of intermediate 6E.

[0083] STEP 5: To the reaction flask was added triethyl phosphonoacetate (27.7 g, 1.2 eq) followed by tetrahydrofuran (400 ml) and the temperature was lowered to 0-10 °C. Sodium hydride (4.1 g, 1 eq) was added and stirred for 1-2 h. Intermediate 6E (40 g, 1 eq) was added at 0-5 °C and the reaction was maintained for 2-3 h at room temperature. The reaction was quenched by the dropwise addition of 100 ml of water. Most of the tetrahydrofuran (80% or more) was removed by concentration. The residue was extracted with 200 ml of ethyl acetate and the organic phase was washed with saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, concentrated to dryness and purified on a silica gel column to give 21.3 g of intermediate 6F.

[0084] STEP 6: To the reaction flask was added 21 g of intermediate 6F, 2.1 g of 10% palladium on carbon and anhydrous ethanol (210 ml). The reaction was maintained for 5-7 h under hydrogen atmosphere at room temperature. The palladium on carbon was removed by filtration and the filtrate was concentrated to dryness to give 20.2 g of intermediate 6G.

[0085] STEP 7: Into the reaction flask, sequentially added intermediate 6G (20 g, 1 eq), tetrahydrofuran (200 ml), purified water (20 ml), sodium hydroxide (6.9 g, 4 eq), warmed to reflux, reacted for 2-3 h, concentrated to remove tetrahydrofuran, added purified water (100 ml) and ethyl acetate (100 ml), stirred uniformly, allowed to separate into layers, retained the aqueous phase, added hydrochloric acid to the aqueous phase to adjust the pH to 3-5, added methyl tert-butyl ether to extract, dried the organic phase over anhydrous sodium sulfate, concentrated the organic phase under reduced pressure to dryness, to obtain 16.8 g of compound SYN-003.

[0086] 1H NMR (500 MHz, CDC13): δ: 7.455-7.439 (m, 2H), 7.396-7.380 (m, 2H), 7.146-7.129 (m, 4H), 5.137 (m, IH), 3.599 (m, 2H), 2.462-2.378 (m, 8H), 1.795-1.735 (m, 4H), 1.527-1.454 (m, 2H), 1.365-1.238 (m, 6H) ppm.

[0087] Example 4

[0088] Example 4

[0089]

[0090] Into the reaction flask, sequentially added compound IF, named 6-{N-[5,6-bis(4- methylphenyl)pyrazin]-2-yl-N-isopropylamino}-2-hexenoic acid ethyl ester (the preparation method of this compound is the same as "compound IF" in Example 1) (3 g, 1 eq), anhydrous ethanol (30 ml), cooled in an ice bath, added sodium hydride (0.79 g, 3 eq), warmed to reflux, reacted for 12-15 h, cooled to room temperature, added hydrochloric acid to adjust the pH to 5-6, added 30 ml of water, extracted with ethyl acetate, dried the organic phase over anhydrous sodium sulfate, concentrated to dryness, purified by silica gel column to obtain 1.2 g of compound SYN-004. 1H NMR (500 MHz, CDC13): δ: 8.013 (s, 1H), 7.378-7.366 (m, 2H), 7.350 (s, 1H), 7.090-7.065 (m, 5H), 4.803-4.776 (m, 1H), 3.814-3.792 (m, 1H), 3.594-3.536 (m, 2H), 3.449-3.398 (m, 2H), 2.628-2.461 (m, 2H), 2.348-2.337 (m, 6H), 1.802-1.642 (m, 4H), 1.281-1.268 (m, 6H), 1.212-1.185 (t, 3H) ppm.

[0091] Example 5

[0092] Prepare 3-ethoxy-6-{N-[5,6-di(4-methylphenyl)-1,2,4-triazinyl]-2-yl-N- isopropylamino}hexanoic acid, compound named as SYN-005, corresponding to structural formula (5).

[0093] Prepare the following route:

[0094]

[0095] STEP 1: Into a reaction flask, add compound 6C (30 g, 1 eq, prepared by the same method as compound 6C in Example 3, named as 5,6-di(4-methylphenyl)-3-chloro-1,2,4-triazine), SM1 (37.2 g, 1.3 eq, prepared by the method described in Reference Example), potassium carbonate (42 g, 3 eq), DMF (1.5 L), and heat to 80-85 °C, and keep the reaction for 2-3 h. Add water, extract with ethyl acetate, and concentrate to obtain 43.9 g of compound 7D.

[0096] STEP 2: Into a reaction flask, add compound 7D (10 g, 1 eq), tetrahydrofuran (100 ml), and purified water (10 ml), and add sodium hydroxide (2.4 g, 3 eq). Heat to reflux, and keep the reaction for 12-15 h. Cool to room temperature, adjust the pH to 5-6 with hydrochloric acid, add 100 ml of water, extract with ethyl acetate, dry the organic phase with anhydrous sodium sulfate, concentrate to dryness, and purify by silica gel column to obtain 4.8 g of compound SYN-005.

[0097] 1H NMR (500 MHz, CDC13): δ: 7.451 ~ 7.425 (m, 2H), 7.381 ~ 7.365 (m, 2H), 7.133 ~ 7.117 (m, 4H), 3.835 ~ 3.811 (m, 1H), 3.605 ~ 3.654 (m, 4H), 2.601 ~ 2.573 (m, 2H), 2.384 ~ 2.365 (m, 7H), 1.699 ~ 1.688 (m, 4H), 1.328 ~ 1.315 (m, 6H), 1.210 ~ 1.182 (t, 3H) ppm.

[0098] Example 6

[0099] Prepare 3-methoxy-6-{N-[5,6-bis(4-methylphenyl)pyrazin]-2-yl-N-isopropylamino} hexanoic acid, compound is recorded as SYN-006, corresponding to structural formula (6).

[0100]

[0101] STEP1: Into a reaction bottle, sequentially add compound 1F (3 g, 1 eq, preparation method see compound 1F in Example 1), anhydrous methanol (30 ml), ice bath cooling, sodium hydride (0.79 g, 3 eq), and then warm to room temperature for 3-5 h. Add 50 ml water to quench, add ethyl acetate to extract, dry the organic phase with anhydrous sodium sulfate, concentrate to dryness, and purify by silica gel column to obtain 2.1 g of compound 4G.

[0102] STEP2: Into a reaction bottle, sequentially add compound 4G (2 g, 1 eq), tetrahydrofuran (30 ml), and purified water 3 ml. Add sodium hydroxide (0.65 g, 4 eq), and then warm to reflux for 12-15 h. Cool to room temperature, adjust pH to 5-6 with hydrochloric acid, add 30 ml water, extract with ethyl acetate, dry the organic phase with anhydrous sodium sulfate, concentrate to dryness, and purify by silica gel column to obtain 0.9 g of compound SYN-006.

[0103] 1 H NMR (500 MHz, CDC13): δ: 8.021 (s, 1H), 7.368 ~ 7.352 (m, 2H), 7.278 ~ 7.258 (m, 2H), 7.091 ~ 7.067 (m, 4H), 4.812 (m, 1H), 3.740 ~ 3.716 (m, 1H), 3.428 ~ 3.413 (m, 5H), 2.642 ~ 2.487 (m, 2H), 2.349 ~ 2.338 (m, 6H), 1.784 ~ 1.671 (m, 4H), 1.293 ~ 1.269 (m, 6H) ppm.

[0104] Example 7

[0105] 6-[N-(5,6-diphenyl-l,2,4-triazin-2-yl)-N-isopropylamino]hexanoic acid, compound noted as SYN-007, corresponding to formula (7) was prepared.

[0106] The preparation of 6-[N-(5,6-diphenyl-l,2,4-triazin-2-yl)-N-isopropylamino]hexanoic acid was carried out in the same way as the preparation of 6-{N-[5,6-bis(4-methylphenyl)-l,2,4-triazin-2-yl]-N- isopropylamino}hexanoic acid (noted as SYN-003) in Example 3, except that the starting material was benzil.

[0107] 1 H NMR (500 MHz, CDC13): δ: 7.510-7.493 (m, 2H), 7.475-7.448 (m, 2H), 7.396-7.364 (m, IH), 7.317-7.269 (m, 5H), 5.141-5.070 (m, IH), 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.

[0108] Example 8

[0109] 3-ethoxy-6-[N-(5,6-diphenylpyrazin-2-yl)-N-isopropylamino]hexanoic acid, compound noted as SYN-008, corresponding to formula (8) was prepared.

[0110]

[0111] To the reaction flask was added compound 8F (3 g, 1 eq, prepared in Example 2), anhydrous ethanol (30 ml), ice bath cooling, sodium hydride (0.84 g, 3 eq), elevated to reflux reaction for 12-15 h, reduced to room temperature, pH adjusted to 5-6 with hydrochloric acid, 100 ml water was added, extracted with ethyl acetate, the organic phase was dried with anhydrous sodium sulfate, concentrated to dryness, purified by silica gel column to obtain 1.2 g of compound SYN-008.

[0112] 1H NMR (500 MHz, CDC13): δ: 8.062 (s, 1H), 7.413-7.385 (m, 2H), 7.326-7.298 (m, 2H), 7.143-7.084 (m, 1H), 6.981-6.933 (m, 4H), 5.919-5.888 (d, 1H), 4.709-4.683 (m, 1H), 3.455-3.424 (m, 2H), 2.361-2.319 (m, 2H), 1.886-1.840 (m, 2H), 1.297-1.274 (m, 6H) ppm.

[0113] Example 9

[0114] Example 9

[0115] Example 9

[0116]

[0117] Example 9

[0118] 1 H NMR (500 MHz, CDC13): δ: 8.062 (s, 1H), 7.413-7.385 (m, 2H), 7.326-7.298 (m, 2H), 7.143-7.084 (m, 1H), 6.981-6.933 (m, 4H), 5.919-5.888 (d, 1H), 4.709-4.683 (m, 1H), 3.455-3.424 (m, 2H), 2.361-2.319 (m, 2H), 1.886-1.840 (m, 2H), 1.297-1.274 (m, 6H) ppm.

[0119] Example 10

[0120] Example 10

[0121]

[0122] STEP 1 : Into a reaction flask was added compound 12F (6 g, 1 eq, see Example 9 for preparation of compound 12F), 60 ml of absolute ethanol, 0.6 g of 10% wet Pd-C, and the reaction was stirred under H2at room temperature for 3-4 h. The Pd-C was removed by filtration and the filtrate was concentrated to give 5.8 g of compound 13G.

[0123] STEP 2: Into a reaction flask was added compound 13G (5 g, 1 eq), 50 ml of tetrahydrofuran, 5 ml of purified water, and sodium hydroxide (1.3 g, 3 eq). The reaction was heated to reflux for 12-15 h, cooled to room temperature, and the pH was adjusted to 5-6 with hydrochloric acid. The reaction was diluted with 100 ml of water and extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, concentrated to dryness, and purified by silica gel column chromatography to give 2.3 g of compound SYN-010.

[0124] 1 H NMR (500 MHz, CDC13): δ: 8.125 (s, 1H), 7.423-7.388 (m, 2H), 7.333-7.298 (m, 2H), 7.194-7.100 (m, 4H), 4.768-4.735 (m, 1H), 3.424 (m, 2H), 2.258-2.220 (m, 2H), 1.658-1.554 (m, 4H), 1.425-1.351 (m, 2H), 1.239-1.164 (m, 6H) ppm.

[0125] Example 11

[0126] Example 11

[0127]

[0128] STEP 1 : Into a reaction flask was added compound 12F (6 g, 1 eq, see Example 9 for preparation of compound 12F), 60 ml of absolute ethanol, 0.6 g of 10% wet Pd-C, and the reaction was stirred under H2at room temperature for 3-4 h. The Pd-C was removed by filtration and the filtrate was concentrated to give 5.8 g of compound 13G.

[0129] STEP 2: To the reaction flask was added compound 5G (2 g, 1 eq), tetrahydrofuran (30 ml), purified water 3 ml, sodium hydroxide (0.44 g, 3 eq) was added, warmed to reflux, reacted for 12-15 h, cooled to room temperature, pH was adjusted to 5-6 using hydrochloric acid, 60 ml of water was added, extracted using ethyl acetate, the organic layer was dried over anhydrous sodium sulphate, concentrated to dryness, purified over silica gel column to get 1.2 g of compound SYN-017.

[0130] 1 H NMR (500 MHz, CDC13): δ: 8.013 (s, 1H), 7.339-7.323 (m, 2H), 7.282 (m, 2H), 7.086-7.052 (m, 4H), 4.580-4.554 (m, 1H), 3.748 (m, 4H), 3.460-3.431 (m, 2H), 2.889-2.869 (m, 1H), 2.782-2.759 (m, 2H), 2.555-2.495 (m, 3H), 2.387-2.323 (m, 7H), 1.825-1.605 (m, 3H), 1.332-1.253 (m, 7H) ppm.

[0131] Example 12

[0132] Example 12

[0133]

[0134] To the reaction flask was added compound SYN-004 (1 g, 1 eq), dichloromethane (20 ml), methyl sulfonamide (300 mg, 1.5 eq), DMAP (308 mg, 1.2 eq), EDCI (484 mg, 1.2 eq), refluxed for 2-3 h, cooled to room temperature, the reaction mass was washed with purified water, the organic layer was dried over anhydrous sodium sulphate, concentrated to dryness, purified over silica gel column to get 650 mg of compound SYN-044.

[0135] 1H NMR (500 MHz, CDC13): δ: 8.013 (s, IH), 7.292 ~ 7.272 (m, 2H), 7.208 ~ 7.188 (m, 2H), 7.129 ~ 7.066 (m, 4H), 4.769 ~ 4.737 (m, IH), 3.771 ~ 3.741 (m, IH), 3.493 ~ 3.459 (m, 2H), 3.183 (s, 3H), 2.434 ~ 2.290 (m, 10H), 1.694 ~ 1.525 (m, 4H), 1.246 ~ 1.229 (m, 6H), 1.092 ~ 1.057 (t, 3H) ppm.

[0136] Example 13

[0137] To prepare 6-{N-[5,6-bis(4-fluorophenyl)pyrazin-2-yl]-N-isopropylamino}-N-(p- methylsulfonyl)hexanamide, compound SYN-046, corresponding to structural formula (13).

[0138]

[0139] To a reaction flask was added compound SYN-010 (1.1 g, 1 eq), dichloromethane (20 ml), methylsulfonamide (357 mg, 1.5 eq), DMAP (367 mg, 1.2 eq), EDCI (574 mg, 1.2 eq) sequentially, refluxed for 2 ~ 3 hours, reduced to room temperature, washed the reaction solution with purified water, dried the organic phase with anhydrous sodium sulfate, concentrated to dryness, purified by silica gel column to obtain 0.6 g of compound SYN-046.

[0140] 1 H NMR (500 MHz, CDC13): δ: 8.013 (s, IH), 7.292 ~ 7.272 (m, 2H), 7.208 ~ 7.188 (m, 2H), 7.129 ~ 7.066 (m, 4H), 4.769 ~ 4.737 (m, IH), 3.771 ~ 3.741 (m, IH), 3.493 ~ 3.459 (m, 2H), 3.183 (s, 3H), 2.434 ~ 2.290 (m, 10H), 1.694 ~ 1.525 (m, 4H), 1.246 ~ 1.229 (m, 6H), 1.092 ~ 1.057 (t, 3H) ppm.

[0141] Example 14

[0142] To prepare {N-[5,6-bis(4-methylphenyl)-1,2,4-triazin-2-yl]-N-isopropylamino}-2- hexenoic acid, compound SYN-018, corresponding to structural formula (14).

[0143]

[0144] Into the reaction bottle, compound 6F (5 g, 1 eq, see preparation method in Example 3), tetrahydrofuran (50 ml), purified water 5 ml, sodium hydroxide (0.87 g, 2 eq) were added in turn, warmed to reflux, reacted for 12-15 h, cooled to room temperature, pH was adjusted to 5-6 with hydrochloric acid, 100 ml of water was added, extracted with ethyl acetate, the organic phase was dried with anhydrous sodium sulfate, concentrated to dryness, and purified by silica gel column to obtain 3.5 g of compound SYN-018.

[0145] 1 H NMR (500 MHz, CDCl3): δ: 8.062 (s, 1H), 7.368-7.351

[0146] (m, 2H), 7.283-7.268 (m, 1H), 7.143-7.050 (m, 5H), 5.981-5.899 (d, 1H), 4.719-4.690 (m, 1H), 3.453-3.422 (m, 2H), 2.371-2.330 (m, 8H), 1.916-1.855

[0147] (m, 2H), 1.293-1.278 (m, 6H) ppm.

[0148] Experimental Example

[0149] I. Target selectivity

[0150] Experimental process: Using HEK cells stably expressing prostacyclin receptors, using HTRF cAMP and HTRF IP1 methods, the functional activity of compounds numbered SYN-001, SYN-002, SYN-003, SYN-004, SYN-005, SYN-006, SYN-007, SYN-008, SYN-010, SYN-013, SYN-017, SYN-044, SYN-046 and SYN-018 on the following 8 targets IP, EP1, EP2, EP3, EP4, DP, FP and TP was studied, and Selexipag (commercial product Selapag) was used as a comparison.

[0151] The experimental results are shown in Table 1.

[0152] Table 1 Target selectivity

[0153]

[0154]

[0155] The above results show that the compounds numbered SYN-001, SYN-002, SYN-003, SYN-004, SYN-005, SYN-006, SYN-007, SYN-008, SYN-010, SYN-013, SYN-017, SYN-044, SYN-046 and SYN-018 all have good agonistic activity on IP receptors, and the effect is equivalent to that of Selexipag. Except that SYN-001, SYN-002 and SYN-003 have slight agonistic activity on EP1, EP2, EP3, EP4, DP, FP and TP target points, the rest of the compounds have no agonistic activity on other receptors except IP receptors, and the selectivity is high.

[0156] The chemical structural formula (Formula IV) of the active ingredient of Selexipag is as follows.

[0157]

[0158] The chemical name is: 2-{4-[(5,6-diphenylpyrazin-2-yl)(2-propyl)amino]butoxy}-N- (methylsulfonyl)acetamide.

[0159] ACT-333679 is a metabolite of Selexipag, and the chemical structural formula is as follows.

[0160]

[0161] The chemical name is: 2-(4-((5,6-diphenylpyrazin-2-yl)(isopropyl)amino) butoxy) acetic acid.

[0162] The structure of the compound in the embodiments of the present application is different from the butoxy acetamide structure in the branched chain of the active ingredient of Selexipag. The main structure of the branched chain in the embodiments of the present application is hexanoic acid or hexenoic acid, or hexanoic acid sulfonamide. The compounds numbered SYN-001, SYN-002, SYN-003, SYN-004, SYN-005, SYN-006, SYN-007, SYN-008, SYN-010, SYN-013, SYN-017, SYN-044, SYN-046 and SYN-018 all have good agonistic activity on IP receptors. They have application prospects in the preparation of drugs for treating pulmonary arterial hypertension.

[0163] II. Drug efficacy test on pulmonary arterial hypertension rats

[0164] Experimental process: The model was prepared by subcutaneous injection of 40 mg / kg of Lycorine. The normal control group of animals was subcutaneously injected with sodium chloride injection. The animals were given the modeling agent about 2 hours before the administration, twice a day, and the administration was continuously given for 19 days.

[0165] Specifically, the compounds numbered SYN-003, SYN-004, SYN-006, SYN-007, SYN-008, SYN-010, SYN-013, and SYN-017 were prepared into suspensions by suspending in 10wt% DMSO + 90wt% NaCl, and were administered by gavage at an active ingredient of 3mg / kg body weight, with a dosing volume of 5mL / kg.

[0166] After 19 days of continuous administration, the mean pulmonary arterial pressure of the rats was determined by right heart catheterization. The experimental results are shown in Figure 1 , wherein Model is 10wt% DMSO + 90wt% NaCl, administered by gavage at 5ml / kg body weight, and Control is Selexipag, administered by gavage at 3mg / kg body weight.

[0167] The results show that the compounds numbered SYN-003, SYN-004, SYN-006, SYN-007, SYN-008, SYN-010, SYN-013, and SYN-017 all significantly reduced the pulmonary arterial pressure of rats, and SYN-004, SYN-007, and SYN-010 had the most obvious depressor effect.

[0168] III. Survival rate test of pulmonary arterial hypertension rats

[0169] Experimental process: The model was prepared by subcutaneous injection of 40mg / kg of monocrotaline. The normal control group animals were subcutaneously injected with sodium chloride injection. The animals were administered the modeling agent about 2 hours before the administration, twice a day, for 45 consecutive days. The administration method was as follows: the compounds numbered SYN-004, SYN-007, and SYN-010 were prepared into suspensions by suspending in 10wt% DMSO + 90wt% NaCl, and were administered by gavage at an active ingredient of 1mg / kg body weight. Selexipag was used as a positive drug group, and was administered by gavage at an active ingredient of 1mg / kg body weight. The model group was administered with normal saline by gavage.

[0170] The survival of the rats was recorded. The experimental results are shown in Table 2.

[0171] Table 2. Effect of SYN-004, SYN-007, and SYN-010 on the survival rate of pulmonary arterial hypertension rats

[0172] Group Number of animals (D0) / per group Number of remaining animals (D45) / per group Survival rate (%) Normal control group 10 10 100 Model group 10 3 30 Selexipag group 10 7 70 SYN-004 group 10 7 70 SYN-007 group 10 8 80 SYN-010 group 10 7 70

[0173] The results show that Selexipag, SYN-004, SYN-007, and SYN-010 can significantly improve the survival rate of pulmonary arterial hypertension rats.

[0174] IV. Toxicity comparison

[0175] Acute toxicity experiment process: rats were randomly divided into groups according to body weight, 10 rats / group, respectively, single gavage 250, 500, 1000, 2000 mg / kg compound, the volume of administration was 10 ml / kg, and the observation was carried out for 7 days after administration.

[0176] The experimental results are shown in Table 3.

[0177] Table 3 Acute toxicity of SYN-003, SYN-004, SYN-005, SYN-007 and SYN-010 in rats

[0178]

[0179]

[0180] The results show that the oral lethal dose of Selexipag in rats is about 500 mg / kg, and the non-lethal dose is 250 mg / kg. SYN-003, SYN-004, SYN-005, SYN-007 and SYN-010 did not show death at 1000 mg / kg, but showed death at 2000 mg / kg, indicating that the toxicity is less than Selexipag.

[0181] Genetic toxicity experiment process: the test samples SYN-004, SYN-007, SYN-010 and ACT-333679 were diluted by 8 times with the highest concentration of 1000 μg / well. The 6-well plate incorporation method was used, 2 wells were treated in parallel, and negative (DMSO) and positive controls were set up, and parallel tests were carried out under the conditions with or without metabolic activation system (± S9). After 48-72 hours of culture, whether the test sample was precipitated and the growth of background plaque was observed, the number of revertant colonies per well was counted, and the test results are shown in Tables 4-11.

[0182] Positive result determination

[0183] Results meeting one or two of the following criteria can be determined as positive:

[0184] 1) At least in one strain, under the condition with or without metabolic activation, the number of revertant colonies showed dose-dependent increase, and the number of revertant colonies was 2 times or more than that of the negative control group.

[0185] 2) Under the condition with or without metabolic activation, the number of revertant colonies in one or more dose groups showed significant increase and could be repeated, and the number of revertant colonies was 2 times or more than that of the negative control group.

[0186] If the test sample is detected by 2 test strains, and as long as one of the test strains is positive, no matter under the condition of adding S9 mixture or not, the test sample is determined as mutagen.

[0187] Negative result determination

[0188] If the test results show that the number of revertant colonies of each test strain does not have a dose-dependent increasing relationship, and the peak value of the number of revertant colonies of each dose group of all strains does not exceed 2 times of the negative control group, the test sample is determined as non-mutagenic.

[0189] The results show that under ± S9 conditions, for TA98 and TA100 strains, ACT-333679 final concentration ≥ 250 μg / well, non-interfering precipitate was observed; SYN-004, SYN-007 and SYN-010 were observed to precipitate at a final concentration of 1000 μg / well. Under ± S9 conditions, for TA98 and TA100 strains, ACT-333679 final concentration ≥ 250 μg / well, background plaque reduction was observed; SYN-010 final concentration was 1000 μg / well, background plaque reduction was observed; SYN-004, SYN-007 each concentration did not observe abnormal background plaque. For TA98 and TA100 strains, under ± S9 conditions, the number of revertant colonies of SYN-004, SYN-007 and SYN-010 and ACT-333679 at each concentration group did not reach 2 times of the negative control group, and there was no concentration-effect relationship, so the test results were negative.

[0190] Therefore, SYN-004, SYN-007, SYN-010 and ACT-333679 do not have genetic toxicity.

[0191] Table 4 SYN-004 bacterial revertant mutation 6-well plate primary screening test results (TA98 strain)

[0192]

[0193] Note:

[0194] Background plaque: T0: normal;

[0195] Test sample / control solubility: P0: normal / no precipitation, P1: non-interfering precipitate under a microscope;

[0196] *: The number of revertant colonies of the positive control group is more than 3 times of the negative control group.

[0197] Table 5 SYN-004 bacterial revertant mutation 6-well plate primary screening test results (TA100 strain)

[0198]

[0199] Notes:

[0200] Background plaque: T0: normal;

[0201] Test article / control article solubility: P0: normal / no precipitate, PI: non-interfering precipitate under microscope;

[0202] *: The revertant colony counts of the positive control group are more than 3 times of the negative control group.

[0203] Table 6 Results of SYN-007 bacterial reverse mutation 6-well plate primary screening test (TA98 strain)

[0204]

[0205]

[0206] Notes:

[0207] Background plaque: T0: normal;

[0208] Test article / control article solubility: P0: normal / no precipitate, PI: non-interfering precipitate under microscope;

[0209] *: The revertant colony counts of the positive control group are more than 3 times of the negative control group.

[0210] Table 7 Results of SYN-007 bacterial reverse mutation 6-well plate primary screening test (TA100 strain)

[0211]

[0212]

[0213] Notes:

[0214] Background plaque: T0: normal;

[0215] Test article / control article solubility: P0: normal / no precipitate, PI: non-interfering precipitate under microscope;

[0216] *: The revertant colony counts of the positive control group are more than 3 times of the negative control group.

[0217] Table 8 Results of SYN-010 bacterial reverse mutation 6-well plate primary screening test (TA98 strain)

[0218]

[0219] Notes:

[0220] Background plaque: T0: normal, Tl: slight decrease in background plaque;

[0221] Test article / control article solubility: P0: normal / no precipitate, PI: non-interfering precipitate under microscope;

[0222] *: The number of revertant colonies of the positive control group is more than 3 times of that of the negative control group.

[0223] Table 9 Results of the bacterial reversion 6-well plate primary screening test of SYN-010 (TA100 strain)

[0224]

[0225]

[0226] Note:

[0227] Background plaque: T0: normal, Tl: mild reduction of background plaque;

[0228] Test article / control article solubility: P0: normal / no precipitate, PI: non-interfering precipitate under microscope;

[0229] *: The number of revertant colonies of the positive control group is more than 3 times of that of the negative control group.

[0230] Table 10 Results of the bacterial reversion 6-well plate primary screening test of ACT-333679 (TA98 strain)

[0231]

[0232]

[0233] Note:

[0234] NA: Not counted;

[0235] Background plaque: T0: normal, Tl: mild reduction of background plaque, T3: severe reduction of background plaque, T4: disappearance of background plaque;

[0236] Test article / control article solubility: P0: normal / no precipitate, PI: non-interfering precipitate under microscope;

[0237] a : The positive drugs without and with S9 are sodium azide (0.4 μg / well) and 2-aminoanthracene (0.6 μg / well), respectively;

[0238] *: The number of revertant colonies of the positive control group is more than 3 times of that of the negative control group.

[0239] Table 11 Results of the bacterial reversion 6-well plate primary screening test of ACT-333679 (TA100 strain)

[0240]

[0241] Remarks:

[0242] NA: Not counted;

[0243] Background plaque: T0: normal, T1: mild reduction of background plaque, T3: severe reduction of background plaque, T4: disappearance of background plaque;

[0244] Test article / control article solubility: P0: normal / no precipitation, P1: non-interfering precipitate under microscope;

[0245] a : The positive drugs were sodium azide (0.4 μg / well) and 2-aminoanthracene (0.6 μg / well) when S9 was added or not added, respectively;

[0246] *: The number of revertant colonies of the positive control group was more than 3 times of that of the negative control group.

[0247] In summary, a series of new compounds with good affinity to PGI2 receptor and low adverse reactions are synthesized in the application, and the application prospect is wide in the preparation of drugs for treating pulmonary arterial hypertension.

[0248] The above only describes the preferred embodiments of the present application and is not intended to limit the present application. Any modification, equivalent replacement or improvement made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A PGI2 receptor agonist compound of Formula I or pharmaceutically acceptable salts, deuterated isotope derivatives, stereoisomers and mixtures thereof, ###0001### Formula I wherein, R1 and R2 are each independently selected from H, C1-C3 alkyl or halogen atom; Z represents N atom; R3 is selected from isopropyl; R4 is selected from H or C1-C3 alkoxy; wherein, R1 and R2 are each independently selected from H, C1-C3 alkyl or halogen atom; Z represents N atom; R3 is selected from isopropyl; R4 is selected from H or C1-C3 alkoxy; wherein, R1 and R2 are each independently selected from H, C1-C3 alkyl or halogen atom; Z represents N atom; R3 is selected from isopropyl; R4 is selected from H or C1-C3 alkoxy; wherein, R1 and R2 are each independently selected from H, C1-C3 alkyl or halogen atom; Z represents N atom; R3 is selected from isopropyl; R4 is selected from H or C1-C3 alkoxy; wherein, R1 and R2 are each independently selected from H, C1-C3 alkyl or halogen atom; Z represents N atom; R3 is selected from isopropyl; R4 is selected from H or C1-C3 alkoxy; wherein, R1 and R2 are each independently selected from H, C1-C3 alkyl or halogen atom; Z represents N atom; R3 is selected from isopropyl; R4 is selected from H or C1-C3 alkoxy; wherein, R1 and R2 are each independently selected from H, C1-C3 alkyl or halogen atom; Z represents N atom; R3 is selected from isopropyl; R4 is selected from H or C1-C3 alkoxy; wherein, R1 and R2 are each independently selected from H, C1-C3 alkyl or halogen atom; Z represents N atom; R3 is selected from isopropyl; R4 is selected from H or C1-C3 alkoxy; wherein, R1 and R2 are each independently selected from H, C1-C3 alkyl or halogen atom; Z represents N atom; R3 is selected from isopropyl; R4 is selected from H or C1-C3 alkoxy; wherein, R1 and R2 are each independently selected from H, C1-C3 alkyl or halogen atom; Z represents N atom; R3 is selected from isopropyl; R4 is selected from H or C1-C3 alkoxy; wherein, R1 and R2 are each independently selected from H, C1-C3 alkyl or halogen atom; Z represents N atom; R3 is selected from isopropyl; R4 is selected from H or C1-C3 alkoxy; wherein, R1 and R2 are each independently selected from H, C1-C3 alkyl or halogen atom; Z represents N atom; R3 is selected from isopropyl; R4 is selected from H or C1-C3 alkoxy; wherein, R1 and R2 are each independently selected I ​ ​ ​ ​ R5represents OH or NHSO2R6, wherein R6represents C 1-4 alkyl or C 1-4 alkyl.

2. The PGI2 receptor agonist compound or pharmaceutically acceptable salt, deuterated isotope derivative, stereoisomer thereof and mixture thereof according to claim 1, wherein ​ ​ 3. The PGI2 receptor agonist compound or pharmaceutically acceptable salt, deuterated isotope derivative, stereoisomer thereof and mixture thereof according to claim 2, wherein ​ ​ 4. The PGI2 receptor agonist compound or pharmaceutically acceptable salt, deuterated isotope derivative, stereoisomer thereof and mixtures thereof of any one of claims 1-3, wherein ​ ​ 5. The compound of claim 1, or pharmaceutically acceptable salts, deuterated isotopes derivatives, stereoisomers thereof, and mixtures thereof, wherein, ​ （3） （5） （7）。 6. A pharmaceutical composition, characterized by, ​ 7. The pharmaceutical composition of claim 6, wherein ​ 8. The pharmaceutical composition of claim 7, wherein, ​ ​

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