A hydroxyl-substituted beraprost derivative, a synthetic method and application thereof

Abstract

The application provides a hydroxyl-substituted beraprost derivative, a synthesis method and application thereof, and belongs to the technical field of drugs.The compound is a coupling compound containing beraprost and a nitric oxide donor.The compound solves the defects of short elimination half-life of beraprost sodium, multiple single-day administration, saturation ceiling effect of curative effect, short metabolic half-life of NO which is a gas and is rapidly decomposed in a solution, and the like, the new compound reduces the original beraprost administration dose and administration frequency, and simultaneously utilizes the relaxation of smooth muscle caused by the release of NO molecules in the body of the compound, so that the two drugs play a synergistic effect through double action, and the effectiveness and safety of the drug are improved.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to a hydroxylated beraprost derivative, its synthesis method, and its application. Background Technology

[0002] Pulmonary hypertension (PH, pulmonary arterial hypertension, PAH) is a group of diseases characterized by increased pulmonary vascular resistance and right ventricular failure. Patients diagnosed with PAH have a short survival rate and high mortality rate, making it a malignant disease.

[0003] Currently, there are three main classes of drugs used clinically to treat pulmonary hypertension: 1) endothelin receptor antagonists (such as bosentan, ambesentan, and macitentan); 2) nitric oxide pathway, such as phospholipase 5 inhibitors (such as nitric oxide, cildira, and fetadalafil) and guanylate cyclase agonists (such as riociguat); and 3) prostaglandin pathway, such as prostacyclin analogs (such as beprostaglandin sodium, eprostol, enoprostol, and treprostinil) and prostacyclin receptor agonists (such as selexizone). The mechanism of action of these drugs ultimately involves relaxing endothelial vascular smooth muscle through the final pathway of the second messengers cAMP and cGMP.

[0004] Among these drugs, prostaglandin analogs are the most effective and classic. Oral beraprost sodium (beraprost sudiam) is currently the main formulation used clinically in China, Japan, South Korea, Indonesia and other countries. However, due to its pharmacokinetic defects, it needs to be administered multiple times a day. Therefore, some researchers have made improvements and innovations to the formulation (such as the 60mcg extended-release tablet Careload from Toray Industries, Inc. in Japan, which was successfully launched in Japan) and structure (the beraprost optical matrix Esuberaprost was developed by UT Pharmaceuticals in the United States, but its phase III clinical trial failed and was terminated).

[0005] Nitric oxide (NO) plays an important role in the relaxation of pulmonary vascular smooth muscle, but due to its gaseous nature, rapid metabolism, and difficulty in administration, inhaled nitric oxide (iNO), although approved for marketing, is difficult to implement in actual clinical treatment of PAH. Therefore, using a nitric oxide donor (NOdonor) approach is a novel approach to drug development. For example, long-acting inhalers of liposomal aerosols prepared using NO donors are used to treat PAH (Nahar K, et al., Pharma Res. 2016). Valeant Pharma's latanoprost nitrate, composed of latanoprost acid butanediol mononitrate, has a dual mechanism of action in glaucoma treatment: latanoprost acid (a marketed drug) acts on the uveal-scleral pathway, promoting aqueous humor outflow; butanediol mononitrate releases nitric oxide (NO), which, through the trabecular meshwork and Schlemm's canal, further promotes aqueous humor outflow. This dual-pronged approach has been validated in clinical trials: compared to latanoprost alone, latanoprost nitrate significantly reduces intraocular pressure, demonstrating better clinical advantages. This product was approved by the FDA in 2017 (brand name VYZULTA).

[0006] Besides its use in treating pulmonary hypertension, beta-prostaglandins have been explored for treatment of metastatic malignant tumors (developed by United Therap, USA), atherosclerosis (developed by Kaken Pharma, Japan), hypertension (developed by Kaken Pharma and United Therap, respectively), diabetic neuropathy (developed by Kaken Pharma, Japan), as well as nephritis and renal failure, vascular dementia (CN 112691109A), and alcoholic fatty liver disease (HK1219665A). NO donor drugs have also been used to develop treatments for various diseases, such as anti-inflammatory and cardiovascular diseases (Megson IL & Webb DJ, Expert Opin Investig Drugs, 2002; Knox CD et al., MK5108, J AmHeartAssoc, 2016).

[0007] However, beraprost sodium currently available on the market has disadvantages such as a short elimination half-life, multiple daily dosings, a saturation cap effect on efficacy, and the fact that NO is a gas that rapidly decomposes and metabolizes in solution with a short half-life. Summary of the Invention

[0008] The purpose of this invention is to address the shortcomings of beraprost sodium, such as its short elimination half-life, frequent daily dosing, and saturation-ceiling effect, as well as the short half-life of NO due to its rapid decomposition and metabolism in solution. This invention provides a new series of compounds combining beraprost sodium with an NO donor. These compounds, combining beraprost sodium with an NO donor, can be used in the treatment of various diseases, including pulmonary hypertension, acute respiratory distress syndrome, arterial occlusive disease, organ fibrosis, kidney disease, eye diseases (such as diabetic retinopathy), osteoporosis, thromboangiitis obliterans, and myocardial infarction.

[0009] The technical solution of this invention is implemented as follows:

[0010] This invention provides a hydroxylated beraprost derivative having the structure shown in Formula I:

[0011] R1 and R2 are selected from H, respectively. Furthermore, R1 and R2 are not both H; where R3 is a straight-chain or branched C1-C chain. 10 Alkyl, C 5-7 cycloalkyl or -C1-C 10 alkyl-aryl ring-, R4 and R5 are straight-chain or branched C1-C 10 Alkyl; wherein C1-C 10 Alkyl, C 5-7 The cycloalkyl or aromatic ring may be substituted with one or more of the following substituents: halogen atom, hydroxyl group, carboxyl group, cyano group, or -(C1-C) group. 10 Alkyl)-ONO2.

[0012] As a further improvement of the present invention, the compound comprises any of the following structures:

[0013]

[0014]

[0015] This invention further protects a method for synthesizing the above-mentioned hydroxylated beraprost derivative, comprising:

[0016]

[0017] S1. Starting material A is used to synthesize carboxylic acid nitrate intermediate B via a substitution reaction;

[0018] S2. Raw material C is synthesized via esterification to obtain intermediate D;

[0019] S3. Intermediate C and intermediate B undergo esterification, and three types of substituted intermediates E, F or G are obtained by purification and separation.

[0020] S4. Intermediates E, F, or G undergo deprotection reactions to yield products H, I, or J.

[0021] The reagents for the S1 substitution reaction include, but are not limited to, silver nitrate and concentrated nitric acid, and the solvents used include, but are not limited to, acetonitrile, dichloromethane, chloroform, acetone, ethyl acetate, toluene, dioxane, etc.

[0022] The condensing agents used in the S2 esterification reaction include, but are not limited to: (1-ethyl-3-(3-dimethylpropylamine)carbodiimide), dicyclohexylcarbodiimide, trifluoromethanesulfonic anhydride, and p-toluenesulfonic acid. The solvents used include, but are not limited to: acetonitrile, dichloromethane, chloroform, acetone, ethyl acetate, toluene, and dioxane.

[0023] The condensing agents used in the S3 esterification reaction include, but are not limited to: (1-ethyl-3-(3-dimethylpropylamine)carbodiimide), dicyclohexylcarbodiimide, trifluoromethanesulfonic anhydride, and p-toluenesulfonic acid. The solvents used include, but are not limited to: acetonitrile, dichloromethane, chloroform, acetone, ethyl acetate, toluene, and dioxane.

[0024] The reagents used in the S4 deprotection reaction include, but are not limited to: trifluoroacetic acid, tetrabutylammonium fluoride, tetrabutylammonium iodide, hydrofluoric acid, hydrochloric acid, potassium hydroxide, sodium hydroxide, palladium on carbon, etc., and the solvents used include, but are not limited to: acetonitrile, dichloromethane, chloroform, acetone, ethyl acetate, toluene, dioxane, methanol, ethanol, etc.

[0025] The present invention further protects the use of the above-mentioned hydroxylated beraprostine derivative as a procyclosporine analog and nitric oxide coupling.

[0026] The present invention further protects the use of the above-mentioned hydroxylated beraprost derivative in the preparation of a therapeutic drug for pulmonary hypertension, acute respiratory distress syndrome, arterial occlusive disease, organ fibrosis, nephropathy, diabetic retinopathy, osteoporosis, thromboangiitis obliterans, and myocardial infarction.

[0027] The present invention has the following beneficial effects: The present invention is a drug developed by combining a series of compounds of beraprost sodium and NO donor. After entering the body, the compounds decompose into beraprost prostaglandins and produce nitric oxide (NO), which can produce dual pharmacological effects. On the one hand, beraprost prostaglandins can specifically bind to prostaglandin receptors and exert a vasodilatory effect on vascular smooth muscle through the cAMP pathway. On the other hand, the NO molecules released by these compounds in the body can also exert a vasodilatory effect through the cGMP pathway of endothelial cells, thus achieving a synergistic therapeutic effect.

[0028] These compounds can be used in the treatment of a variety of diseases, including pulmonary hypertension, acute respiratory distress syndrome, arterial occlusive disease, organ fibrosis, kidney disease, eye diseases (such as diabetic retinopathy), osteoporosis, thromboangiitis obliterans, and myocardial infarction.

[0029] This invention provides a class of drugs combining beraprost sodium and a NO donor, which overcomes the shortcomings of both beraprost sodium (short elimination half-life, frequent daily dosing, and saturation capping effect) and NO (rapid decomposition and metabolism in solution with a short half-life). The new compound reduces the dosage and frequency of administration of the original beraprost sodium, and at the same time utilizes the smooth muscle relaxant effect caused by the release of NO molecules in vivo. Through this dual action, the synergistic effect of the two drugs is achieved, improving the efficacy and safety of the drug. Attached Figure Description

[0030] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0031] Figure 1 This is a comparison chart of NO release from different groups of compounds in the experimental cases. Detailed Implementation

[0032] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0033] Example 1: Preparation of Compound 1

[0034]

[0035] The synthesis route is as follows:

[0036]

[0037] The synthesis method is as follows:

[0038] 2-Bromoacetic acid (5 mmol) was dissolved in acetonitrile, and silver nitrate (8 mmol) was added under stirring. The reaction was carried out at 60 °C for 4 hours. The reaction was monitored by TLC until complete. The solvent was evaporated, dichloromethane was added, and the mixture was washed once with water and once with saturated brine. The mixture was then subjected to column chromatography to obtain intermediate 1-B.

[0039] Beraprost (100 mg) was dissolved in 10 mL of dichloromethane. 2,4-Dimethylbenzyl alcohol (65 mg), DMAP (25 mg), and EDCI.HCl (66 mg) were added in sequence. The mixture was stirred at room temperature for 14 h. The reaction was confirmed to be complete by TLC. The reaction mixture was diluted with ethyl acetate, washed once with 0.1 M hydrochloric acid aqueous solution, and once with saturated brine. The mixture was purified by column chromatography to obtain intermediate 1-D.

[0040] Intermediate 1-D (0.1 mmol), intermediate 1-B (0.15 mmol), EDCI.HCl (0.2 mmol), and HOBT (0.2 mmol) were dissolved in dichloromethane, stirred at room temperature for 4 hours, and the reaction was monitored by TLC until complete. The solvent was evaporated and purified by column chromatography to obtain intermediate 1-E.

[0041] Intermediate 1-E (0.1 mmol) was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added and stirred. After reacting at room temperature for 2 hours, the solvent was evaporated and the product was purified by HPLC to obtain compound 1 in 23% yield. 1 H NMR(500MHz,Chloroform-d)δ7.11–7.00(m,2H),6.96(dq,J=7.7,1.0Hz,1H),5.84(dd,J=15.6,7.0Hz,1H),5. 72(dd,J=15.6,6.4Hz,1H),5.08(td,J=5.3,4.1Hz,1H),5.00(dt,J=5.1,4.3Hz,1H),4.74–4.57(m,3H),4.17(q d,J=6.4,1.1Hz,1H),3.47(dd,J=5.7,4.2Hz,1H),3.00–2.87(m,1H),2.87–2.68(m,4H),2.42(dt,J=14.6,4.2 Hz,1H),2.38–2.27(m,3H),2.27–2.13(m,2H),2.01–1.81(m,3H),1.65(t,J=2.0Hz,3H),1.02(d,J=6.8Hz,3H).

[0042] Example 2: Preparation of Compound 2

[0043]

[0044] The synthesis route is as follows:

[0045]

[0046] Intermediate 2-B can be prepared by referring to the synthesis method of intermediate 1-B.

[0047] Intermediate 1-D (0.1 mmol), intermediate 2-B (0.15 mmol), EDCI.HCl (0.2 mmol), and HOBT (0.2 mmol) were dissolved in dichloromethane, stirred at room temperature for 6 hours, and the reaction was monitored by TLC until complete. The solvent was evaporated and purified by HPLC to obtain intermediate 2-C.

[0048] Compound 2 can be prepared by replacing the raw material with 2-C according to the synthesis method of Example 1. 1 H NMR(500MHz,Chloroform-d)δ7.10–7.00(m,2H),6.94(ddt,J=6.2,2.9,1.0Hz,1H),5.95–5.78(m,2H),5. 12(td,J=6.1,1.0Hz,1H),5.02–4.89(m,1H),4.32(t,J=5.8Hz,2H),4.10(qd,J=5.5,4.7Hz,1H),3.72(d, J=5.3Hz,1H),3.40(dd,J=5.7,4.2Hz,1H),2.76(ddddd,J=6.5,5.1,4.2,3.2,1.1Hz,3H),2.44–2.28(m,4 H), 2.27–2.19 (m, 2H), 2.18–1.98 (m, 3H), 1.98–1.72 (m, 6H), 1.65 (t, J = 2.0Hz, 3H), 1.03 (d, J = 6.8Hz, 3H).

[0049] Example 3: Preparation of Compound 3

[0050]

[0051] Compound 3 can be prepared by replacing the raw material with 4-bromobutyric acid according to the synthesis method of Example 1. 1HNMR(500MHz,Chloroform-d)δ7.14–6.98(m,2H),6.96(dq,J=7.7,1.0Hz,1H),5.84(dd,J=15.6,7.0Hz,1H),5.72(dd,J=15.6, 6.4Hz,1H),5.09(td,J=5.2,4.1Hz,1H),5.02(dt,J=5.1,4.3Hz,1H),4.69(d,J=6.2Hz,1H),4.43(qt,J=10.4,6.1Hz,2H),4.17( qd,J=6.4,1.1Hz,1H),3.47(dd,J=5.7,4.2Hz,1H),2.94(dtd,J=6.6,5.4,1.0Hz,1H),2.78(td,J=6.4,1.0Hz,2H),2.61–2.47(m ,2H),2.21(dp,J=6.2,2.0Hz,2H),2.07(tt,J=7.0,6.0Hz,2H),2.02–1.84(m,3H),1.65(t,J=2.0Hz,3H),1.02(d,J=6.8Hz,3H).

[0052] Example 4: Preparation of Compound 4

[0053]

[0054] Compound 4 can be prepared by replacing the raw material with 4-bromobutyric acid according to the synthesis method of Example 2. 1 HNMR(500MHz,Chloroform-d)δ7.21–6.99(m,2H),6.94(dq,J=7.7,1.0Hz,1H),5.96–5.71(m,2H),5.09 (td,J=6.1,0.9Hz,1H),4.99–4.87(m,1H),4.54–4.37(m,2H),4.10(qd,J=5.5,4.7Hz,1H),3.72(d,J=5 .3Hz,1H),3.40(dd,J=5.7,4.2Hz,1H),2.88–2.66(m,3H),2.50(t,J=7.0Hz,2H),2.35(t,J=7.0Hz,2H) ,2.29–2.17(m,2H),2.17–2.01(m,5H),2.01–1.85(m,2H),1.65(t,J=2.0Hz,3H),1.03(d,J=6.7Hz,3H).

[0055] Example 5: Preparation of Compound 5

[0056]

[0057] Compound 5 can be prepared by replacing the raw material with 6-bromohexanoic acid, referring to the synthesis method of Example 2. 1 HNMR(500MHz,Chloroform-d)δ7.14–7.01(m,2H),6.94(ddt,J=5.6,3.5,1.1Hz,1H),5.98–5.71(m,2H),5.12(td ,J=6.1,1.0Hz,1H),4.94(dt,J=5.0,4.3Hz,1H),4.29(t,J=6.0Hz,2H),4.10(qd,J=5.6,4.7Hz,1H),3.72(d,J=5. 3Hz, 1H), 3.40 (dd, J=5.7, 4.2Hz, 1H), 2.93–2.64 (m, 3H), 2.32 (td, J=6.9, 4.5Hz, 4H), 2.27–2.17 (m, 2H), 2.17–1. 98(m,3H),1.98–1.86(m,2H),1.86–1.73(m,2H),1.65(t,J=2.0Hz,3H),1.62–1.44(m,4H),1.03(d,J=6.8Hz,3H).

[0058] Example 6: Preparation of Compound 6

[0059]

[0060] Compound 6 can be prepared by replacing the starting material with 6-bromohexanoic acid, referring to the synthesis method of Example 1. 1 H NMR(500MHz,Chloroform-d)δ7.11–6.99(m,2H),6.96(ddt,J=7.5,1.8,0.9Hz,1H),5.83(dd,J=15.7,6.9Hz,1H ),5.72(dd,J=15.6,6.4Hz,1H),5.03(ddt,J=14.8,5.1,4.1Hz,2H),4.69(d,J=6.2Hz,1H),4.32(t,J=6.0Hz,2H) ,4.17(qd,J=6.4,1.1Hz,1H),3.47(dd,J=5.7,4.2Hz,1H),2.94(dtd,J=6.6,5.5,1.0Hz,1H),2.87–2.69(m,4H), 2.48–2.27(m,4H),2.21(dp,J=6.2,2.0Hz,2H),2.06–1.71(m,7H),1.65(t,J=2.0Hz,3H),1.02(d,J=6.8Hz,3H).

[0061] Example 7: Preparation of Compound 7

[0062]

[0063] Compound 7 can be prepared by replacing the raw material with 7-bromoheptanoic acid according to the synthesis method of Example 2. 1 H NMR(500MHz,Chloroform-d)δ7.15–7.01(m,2H),6.95(ddt,J=5.5,3.3,0.9Hz,1H),5.94–5.73(m,2H),5.12(td, J=6.1,1.0Hz,1H),4.94(dt,J=5.0,4.3Hz,1H),4.29(t,J=6.1Hz,2H),4.10(qd,J=5.6,4.7Hz,1H),3.72(d,J=5.3 Hz,1H),3.40(dd,J=5.5,4.2Hz,1H),2.86–2.63(m,3H),2.32(td,J=7.0,2.7Hz,4H),2.31–2.18(m,3H),2.18–2. 02(m,3H),1.97–1.84(m,2H),1.84–1.72(m,2H),1.65(t,J=2.0Hz,3H),1.55–1.27(m,6H),1.03(d,J=6.8Hz,3H).

[0064] Example 8: Preparation of Compound 8

[0065]

[0066] Compound 8 can be prepared by replacing the raw material with 7-bromoheptanoic acid according to the synthesis method of Example 1. 1H NMR(500MHz,Chloroform-d)δ7.25–6.99(m,2H),6.96(ddt,J=7.0,2.2,1.1Hz,1H),5.83(dd,J=15.6,6.9Hz,1H),5.73(dd,J=15 .6,6.4Hz,1H),5.16–4.94(m,2H),4.76(d,J=6.2Hz,1H),4.29(t,J=6.1Hz,2H),4.17(qd,J=6.4,1.1Hz,1H),3.47(dd,J=5.7,4. 2Hz,1H),2.94(dtd,J=6.6,5.5,1.0Hz,1H),2.78(td,J=6.4,1.0Hz,2H),2.40(dt,J=14.5,4.2Hz,1H),2.37–2.26(m,5H),2.21( dp,J=6.2,2.0Hz,2H),2.02–1.85(m,3H),1.81(p,J=6.3Hz,2H),1.65(t,J=2.0Hz,3H),1.63–1.41(m,4H),1.02(d,J=6.8Hz,3H).

[0067] Example 9: Preparation of Compound 9

[0068]

[0069] The synthesis route is as follows:

[0070]

[0071] Intermediate 1-D (0.1 mmol), intermediate 1-B (0.3 mmol), EDCI.HCl (0.4 mmol), and HOBT (0.4 mmol) were dissolved in dichloromethane and stirred at room temperature for 6 hours. The reaction was monitored by TLC until complete. The solvent was evaporated and purified by HPLC to obtain intermediate 3-A. Compound 9 can be prepared by referring to the synthesis method in Example 1. 1H NMR(500MHz,Chloroform-d)δ7.15–7.00(m,2H),6.97(ddt,J=6.2,2.9,1.1Hz,1H),5.99–5.72( m,2H),5.20–5.04(m,2H),4.99(dt,J=5.1,4.2Hz,1H),4.63(q,J=7.2Hz,4H),3.46(dd,J=5.6,4 .3Hz,1H),2.98–2.88(m,1H),2.88–2.70(m,6H),2.41(dt,J=14.4,4.2Hz,1H),2.35–2.28(m,3H ), 2.24(dp,J=6.1,2.0Hz,2H),2.10–1.86(m,3H),1.65(t,J=2.0Hz,3H),1.03(d,J=6.8Hz,3H).

[0072] Example 10: Preparation of Compound 10

[0073]

[0074] Compound 10 can be prepared by replacing the raw materials with 8-bromooctanoic acid according to the synthesis method of Example 2. 1 H NMR(300MHz,Chloroform-d)δ7.16–7.02(m,2H),6.95(ddt,J=6.0,2.7,0.9Hz,1H),5.93–5.61(m,4 H),4.76–4.42(m,3H),4.29–4.04(m,2H),3.92(d,J=5.5Hz,1H),3.43(dd,J=5.5,4.2Hz,1H),2.96–

[0075] 2.63(m,5H),2.45(td,J=7.0,0.9Hz,2H),2.30–2.06(m,4H),2.06–1.84(m,3H),1.57(t,J=2.0Hz,3H),1.00(d,J=6.2Hz,3H).

[0076] Example 11: Preparation of Compound 11

[0077]

[0078] Compound 11 can be prepared by replacing the raw materials with 9-bromononanoic acid according to the synthesis method of Example 2. 1H NMR(500MHz,Chloroform-d)δ7.19–7.02(m,2H),6.95(ddt,J=7.1,2.0,0.9Hz,1H),5.97–5.71(m,2H),5.13(td,J=6. 1,1.0Hz,1H),4.94(dt,J=5.0,4.3Hz,1H),4.29(t,J=6.1Hz,2H),4.10(qd,J=5.6,4.7Hz,1H),3.72(d,J=5.3Hz,1H),3 .39(dd,J=5.6,4.3Hz,1H),2.87–2.63(m,3H),2.32(dt,J=15.5,7.1Hz,4H),2.26–2.17(m,2H),2.17–1.98(m,3H),1.9 7–1.87(m,2H),1.86–1.73(m,2H),1.65(t,J=2.0Hz,3H),1.56–1.41(m,4H),1.41–1.18(m,6H),1.03(d,J=6.7Hz,3H).

[0079] Example 12: Preparation of Compound 12

[0080]

[0081] Compound 12 can be prepared by replacing the raw materials with 7-bromoheptanoic acid according to the synthesis method of Example 1. 1 HNMR(500MHz,Chloroform-d)δ7.13–7.01(m,2H),6.97(ddt,J=6.8,2.1,0.9Hz,1H),5.83(dd,J=15.6,6.9Hz,1H),5.73(dd,J=15.6 ,6.4Hz,1H),5.03(ddt,J=11.3,5.1,4.3Hz,2H),4.76(d,J=6.2Hz,1H),4.29(t,J=6.1Hz,2H),4.17(qd,J=6.4,1.1Hz,1H),3.47(dd ,J=5.7,4.2Hz,1H),2.94(dtd,J=6.6,5.4,1.0Hz,1H),2.78(td,J=6.4,1.0Hz,2H),2.40(dt,J=14.5,4.2Hz,1H),2.36–2.24(m,5H) ,2.21(dp,J=6.2,2.0Hz,2H),2.02–1.85(m,3H),1.85–1.71(m,2H),1.65(t,J=2.0Hz,3H),1.55–1.31(m,6H),1.02(d,J=6.8Hz,3H).

[0082] Example 13: Preparation of Compound 13

[0083]

[0084] Compound 13 can be prepared by replacing the raw materials with 10-bromodecanoic acid according to the synthesis method of Example 2. 1 H NMR(500MHz,Chloroform-d)δ7.18–7.01(m,2H),6.95(ddt,J=7.1,2.0,0.9Hz,1H),5.94–5.75(m,2H),5.13(td,J=6.1,1.0 Hz,1H),4.94(dt,J=5.0,4.3Hz,1H),4.29(t,J=6.1Hz,2H),4.10(qd,J=5.5,4.7Hz,1H),3.95(d,J=5.3Hz,1H),3.39(dd,J=5 .6,4.3Hz,1H),2.86–2.68(m,3H),2.32(dt,J=12.1,7.1Hz,4H),2.27–2.18(m,2H),2.15–1.99(m,3H),1.95–1.86(m,2H),1 .82–1.73(m,2H),1.65(t,J=2.0Hz,3H),1.49(p,J=6.9Hz,2H),1.46–1.38(m,2H),1.37–1.19(m,8H),1.03(d,J=6.8Hz,3H).

[0085] Example 14: Preparation of Compound 14

[0086]

[0087] Compound 14 can be prepared by replacing the raw material with 6-bromohexanoic acid according to the synthesis method of Example 2. 1H NMR(500MHz,Chloroform-d)δ7.15–7.02(m,2H),6.97(ddt,J=6.6,2.0,1.0Hz,1H),5.92–5.73(m,2H) ,5.25–5.04(m,2H),4.99(dt,J=5.1,4.2Hz,1H),4.29(t,J=6.1Hz,4H),3.45(dd,J=5.5,4.2Hz,1H),2 .92(dtd,J=6.6,5.4,1.0Hz,1H),2.86–2.63(m,2H),2.40(dt,J=14.5,4.2Hz,1H),2.35–2.18(m,9H), 2.08–1.85(m,3H),1.84–1.70(m,4H),1.64(t,J=2.0Hz,3H),1.62–1.42(m,8H),1.03(d,J=6.8Hz,3H).

[0088] Example 15: Preparation of Compound 15

[0089]

[0090] Compound 15 can be prepared by replacing the raw materials with 4-bromocyclohexane-1-carboxylic acid, referring to the synthesis method of Example 2. 1 HNMR(500MHz,Chloroform-d)δ7.16–7.02(m,2H),6.95(ddt,J=5.5,3.3,0.9Hz,1H),5.92–5.75(m,2H ),5.13(td,J=6.1,0.9Hz,1H),4.94(dt,J=5.1,4.3Hz,1H),4.67(p,J=5.4Hz,1H),4.10(qd,J=5.6,4. 7Hz,1H),3.72(d,J=5.3Hz,1H),3.39(dd,J=5.6,4.3Hz,1H),2.88–2.64(m,3H),2.40–2.29(m,3H),2. 29–2.18(m,2H),2.15–2.02(m,3H),1.99–1.72(m,10H),1.65(t,J=2.0Hz,3H),1.03(d,J=6.7Hz,3H).

[0091] Example 16: Preparation of Compound 16

[0092]

[0093] Compound 16 can be prepared by replacing the raw materials with 3-bromo-2-methylpropionic acid according to the synthesis method of Example 1. 1HNMR(500MHz,Chloroform-d)δ7.17–6.99(m,2H),6.96(dq,J=7.7,1.0Hz,1H),5.84(dd,J=15.6,7.0Hz,1H),5.72(dd,J=15.6 ,6.4Hz,1H),5.10(td,J=5.3,4.1Hz,1H),4.97(dt,J=5.1,4.2Hz,1H),4.69(d,J=6.2Hz,1H),4.38(d,J=6.3Hz,2H),4.17(qd, J=6.4,1.1Hz,1H),3.47(dd,J=5.7,4.2Hz,1H),3.19–2.82(m,2H),2.78(td,J=6.4,1.0Hz,2H),2.43(dt,J=14.5,4.2Hz,1H), 2.39–2.29(m,3H),2.26–2.12(m,2H),2.02–1.82(m,3H),1.65(t,J=2.0Hz,3H),1.27(d,J=7.3Hz,3H),1.02(d,J=6.8Hz,3H).

[0094] Example 17: Preparation of Compound 17

[0095]

[0096] Compound 17 can be prepared by replacing the raw materials with 2,2-dibromoacetic acid according to the synthesis method of Example 2. 1 H NMR(500MHz,Chloroform-d)δ7.21–7.00(m,2H),6.95(ddt,J=7.1,2.0,0.9Hz,1H),5.97–5.67(m,2H),5.17(t d,J=6.1,1.0Hz,1H),4.94(dt,J=5.0,4.3Hz,1H),4.64(d,J=6.4Hz,4H),4.10(qd,J=5.5,4.6Hz,1H),3.72(d, J=5.3Hz,1H),3.41(dd,J=5.5,4.2Hz,1H),3.12(p,J=6.5Hz,1H),2.91–2.61(m,3H),2.33(t,J=7.1Hz,2H),2. 24(dp,J=6.0,2.0Hz,2H),2.18–1.97(m,3H),1.97–1.77(m,2H),1.65(t,J=2.0Hz,3H),1.03(d,J=6.8Hz,3H).

[0097] Example 18: Preparation of Compound 18

[0098]

[0099] Compound 18 can be prepared by replacing the raw materials with 4-bromo-3-methylbutyric acid according to the synthesis method of Example 2. 1 HNMR(500MHz,Chloroform-d)δ7.15–6.99(m,2H),6.94(ddt,J=6.2,2.9,1.0Hz,1H),5.96–5.71(m ,2H),5.10(td,J=6.1,1.0Hz,1H),5.02–4.86(m,1H),4.32–4.16(m,2H),4.10(qd,J=5.5,4.7Hz,1H ),3.72(d,J=5.3Hz,1H),3.40(dd,J=5.7,4.2Hz,1H),2.87–2.66(m,3H),2.43–2.17(m,7H),2.17–2 .01(m,3H),1.97–1.85(m,2H),1.65(t,J=2.0Hz,3H),1.24(d,J=6.7Hz,3H),1.03(d,J=6.7Hz,3H).

[0100] Example 19: Preparation of Compound 19

[0101]

[0102] The synthesis route is as follows:

[0103]

[0104] The synthesis of intermediate 4-A can be found in Molecules, 2012, 17, 7556-7568.

[0105] Intermediate 1-D (0.1 mmol), intermediate 4-A, DMAP, and TEA were dissolved in 2 mL of anhydrous dichloromethane and stirred at room temperature for four hours. Then, 3 mL of dichloromethane was added to the reaction solution for dilution. The solution was washed twice with 10% hydrochloric acid and once with saturated saline solution. After filtration, the filtrate was concentrated and purified by HPLC to obtain intermediate 4-B.

[0106] Compound 19 can be prepared by referring to the synthesis method of Example 2. 1HNMR(500MHz,Chloroform-d)δ7.91–7.80(m,2H),7.67–7.48(m,3H),7.15–7.01(m,2H),6.95(ddt,J=7 .3,1.9,0.9Hz,1H),6.12–5.99(m,2H),5.89–5.70(m,2H),5.14(td,J=6.1,1.0Hz,1H),4.94(dt,J=5.0, 4.3Hz,1H),4.16–3.96(m,2H),3.57(s,2H),3.43(dd,J=5.7,4.2Hz,1H),2.87–2.64(m,3H),2.33(t,J=7 .1Hz,2H),2.24(dp,J=5.7,1.9Hz,2H),2.18–1.84(m,5H),1.65(t,J=2.0Hz,3H),1.03(d,J=6.7Hz,3H).

[0107] Example 20: Preparation of Compound 20

[0108]

[0109] Compound 20 can be prepared by referring to the synthesis method of Example 19. 1 HNMR(500MHz,Chloroform-d)δ7.76–7.47(m,5H),7.18–7.04(m,2H),6.95(ddt,J=7.3,1.9,1.0Hz,1H),6.0 4–5.66(m,2H),5.13(td,J=6.0,1.1Hz,1H),4.94(dt,J=5.0,4.3Hz,1H),4.70–4.58(m,2H),4.54(td,J=6.2, 0.8Hz,2H),4.21–3.92(m,2H),3.54(s,2H),3.43(dd,J=5.7,4.2Hz,1H),2.93–2.69(m,3H),2.33(t,J=7.0Hz ,2H),2.27–2.17(m,2H),2.17–2.05(m,2H),2.05–1.85(m,3H),1.65(t,J=2.0Hz,3H),1.03(d,J=6.7Hz,3H).

[0110] Example 21: Preparation of compound 21

[0111]

[0112] Compound 21 can be prepared by referring to the synthesis method of Example 19. 1H NMR(500MHz,Chloroform-d)δ7.92–7.75(m,2H),7.68–7.43(m,3H),7.15–7.01(m,2H),6.95(ddt,J=7.3,1. 9,1.0Hz,1H),6.01–5.58(m,2H),5.14(td,J=6.1,1.0Hz,1H),4.94(dt,J=5.0,4.3Hz,1H),4.38(t,J=6.1Hz ,2H),4.21(t,J=6.1Hz,2H),4.16–3.96(m,2H),3.54(s,2H),3.43(dd,J=5.7,4.2Hz,1H),2.88–2.64(m,3H) ,2.33(t,J=7.1Hz,2H),2.29–2.19(m,2H),2.19–1.86(m,7H),1.65(t,J=2.0Hz,3H),1.03(d,J=6.7Hz,3H).

[0113] Experimental Example 1: In vitro NO release test

[0114] Test materials: The blank solution was a mixture of DMSO and PBS; Griess reagent was prepared as follows: sulfonamide (4.0 g), N-(1-naphthyl)ethylenediamine dihydrochloride (0.2 g) and 10 mL of 85% H3PO4 were dissolved in 90 mL of distilled water and stirred until a clear solution was obtained; the test compound (compound 2, compound 5, compound 8, compound 10, compound 14 and compound 19 in this patent example) solution was prepared by accurately weighing the test compound, dissolving it in DMSO and diluting it with PBS to a concentration of 200 μM.

[0115] Experimental method and standard curve equation: 0-100 μmol / L sodium nitrite standard solutions were prepared using blank solutions, mixed with Griess reagent, and incubated in a constant temperature shaker at 37℃ for 30 min. The absorbance was measured at 540 nm, and the standard curve equation was obtained by subtracting the blank solution reading from the absorbance.

[0116] Experimental test: The test compound solution and L-cysteine ​​solution were mixed and incubated in a shaker at 37°C for 120 min. Then, the mixture was mixed with Griess reagent and incubated in a shaker at 37°C for another 30 min. The absorbance at 540 nm was measured, and the NO release was calculated according to the standard curve equation.

[0117] Data for some compounds, such as Figure 1 As shown, the compounds in the examples have good NO release effects.

[0118] Experimental Example 2: Antiplatelet Aggregation Effect

[0119] Experimental materials: ADP (adenosine diphosphate), adrenaline, collagen, and platelet aggregation function assay kit (CAT#5393).

[0120] Experimental Methods: Platelet-rich plasma was prepared using blood samples from healthy individuals. Bornl's turbidimetric assay was used for testing. 225 μL of platelet-rich plasma was added to a reaction vessel, followed by 25 μL of a solution of the test compound (the compound in this patent embodiment) prepared as a 1.8-1800 nM solution (25 mM Tris-acetate and 120 mM NaCl). After co-incubation at 37°C for 2 min, ADP was added to a final concentration of 2 μM to induce platelet aggregation. The inhibition rate of the compound against ADP-induced platelet aggregation was evaluated.

[0121] Experimental results: see Table 1.

[0122] Table 1. Compounds in the examples inhibit ADP-induced platelet aggregation.

[0123]

[0124]

[0125] As shown in the table above, the compounds in the examples have a good effect on inhibiting ADP-induced platelet aggregation.

[0126] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A hydroxylated beraprost derivative, characterized in that, It has the structure shown in Equation I: Formula I; Among them, R1 and R2 are selected from H, or Furthermore, R1 and R2 are not both H; where R3 is a straight-chain or branched C1-C 10 Alkyl, C 5-7 Cycloalkyl groups, where R4 and R5 are straight-chain or branched C1-C atoms. 10 Alkyl; wherein C1-C 10 Alkyl, C 5-7 Cycloalkyl groups may be substituted with one or more of the following substituents: halogen atom, hydroxyl group, carboxyl group, cyano group, or -(C1-C2) group. 10 Alkyl)-ONO2.

2. The hydroxy-substituted beraprost derivative according to claim 1, wherein, The compound comprises any of the following structures: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 3. A method for synthesizing the hydroxy-substituted beraprost derivative according to claim 1 or 2, characterized by, Includes the following steps: wherein, S1. Starting material A is used to synthesize carboxylic acid nitrate intermediate B via a substitution reaction; S2. Intermediate D is synthesized from raw material C via esterification. S3. Intermediate D and intermediate B undergo esterification, and three types of substituted intermediates E, F or G are obtained by purification and separation; S4. Intermediates E, F, or G undergo deprotection reactions to yield products H, I, or J.

4. The preparation method according to claim 3, characterized in that, The reagents for the S1 substitution reaction are selected from silver nitrate or concentrated nitric acid, and the solvents used are selected from acetonitrile, dichloromethane, chloroform, acetone, ethyl acetate, toluene, or dioxane. The condensing agent used in the S2 esterification reaction is selected from (1-ethyl-3(3-dimethylpropylamine)carbodiimide), dicyclohexylcarbodiimide, trifluoromethanesulfonic anhydride or p-toluenesulfonic acid, and the solvent used is selected from acetonitrile, dichloromethane, chloroform, acetone, ethyl acetate, toluene or dioxane. The condensing agent used in the S3 esterification reaction is selected from (1-ethyl-3(3-dimethylpropylamine)carbodiimide), dicyclohexylcarbodiimide, trifluoromethanesulfonic anhydride or p-toluenesulfonic acid, and the solvent used is selected from acetonitrile, dichloromethane, chloroform, acetone, ethyl acetate, toluene or dioxane. The reagents used in the S4 deprotection reaction are selected from trifluoroacetic acid, tetrabutylammonium fluoride, tetrabutylammonium iodide, hydrofluoric acid, hydrochloric acid, potassium hydroxide, sodium hydroxide, or palladium on carbon, and the solvents used are selected from acetonitrile, dichloromethane, chloroform, acetone, ethyl acetate or toluene, dioxane, methanol, or ethanol.

5. The use of a hydroxylated beraprost derivative as described in claim 1 or 2 in the preparation of a therapeutic medicament for pulmonary hypertension, arterial occlusive disease, nephropathy, and thromboangiitis obliterans.

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