A nitric oxide donor-type beraprost derivative, and pharmaceutical compositions and uses thereof

Abstract

The present application discloses a series of new compounds, which contain beraprost and a nitric oxide donor, as shown in the following formula I, and relates to the series of compounds, pharmaceutical compositions and uses thereof. The present application provides a kind of beraprost sodium and NO donor combination drug, which solves the short clearance half-life of beraprost sodium, the high frequency of single day administration, the saturation ceiling effect of curative effect, and the short half-life of NO gas in solution, and other defects. The new compound reduces the original dose and frequency of beraprost administration, and simultaneously utilizes the relaxation of smooth muscle caused by the release of NO molecules in the body, so as to exert the synergistic effect of the two drugs through double action, and improve the effectiveness and safety of the drug.

Description

[0001] This application is a divisional application of the application filed on November 18, 2021, with application number 202111366683.9 and invention title "A nitrogen oxide donor type beraprost derivative and its pharmaceutical composition and use". Technical Field

[0002] This invention belongs to the field of biomedicine, specifically relating to a nitrogen oxide donor type beraprost derivative or its pharmaceutically acceptable salt, its pharmaceutical composition and uses. Background Technology

[0003] Pulmonary hypertension (PH, including pulmonary arterial hypertension, PAH) is a group of diseases characterized by increased pulmonary vascular resistance and right ventricular failure. Patients diagnosed with PH have a short survival rate and high mortality rate, making it a malignant disease.

[0004] Currently, the main drugs used in clinical practice to treat pulmonary hypertension include endothelin receptor antagonists (such as bosentan), phosphoesterase 5 inhibitors (such as sildenafil), guanylate cyclase agonists (such as riociguat), prostacyclin analogs (such as beprostaglandin), and prostacyclin receptor agonists (such as celecoxib). The mechanism of action of these drugs ultimately involves the nitric oxide (NO) and cGMP pathways to dilate endothelial blood vessels.

[0005] Among these drugs, prostaglandin analogs are the most effective and classic. Beraprost sudiam is the main oral formulation used in clinical practice. However, due to its pharmacokinetic defects, it needs to be administered multiple times a day. Therefore, researchers have made improvements to the formulation of beraprost (such as the extended-release tablet Careload, which has been successfully launched in Japan) and its structure (the phase III clinical trial of the beraprost optical matrix Esuberaprost failed).

[0006] Because NO plays a crucial role in the entire pathway of action, but is difficult to administer due to its gaseous nature and rapid metabolism, the use of a NO donor model 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). ValeantPharma's latanoprost acid 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; while butanediol mononitrate releases nitric oxide (NO), which, through the trabecular meshwork and Schlemm's canal, further promotes aqueous humor outflow. This novel two-pronged approach has been validated in clinical trials: compared to latanoprost alone, latanoprost nitrate significantly reduces intraocular pressure, demonstrating superior clinical advantages. This product was approved by the FDA in 2017 (brand name VYZULTA). Therefore, modifying a prostaglandin with a NO donor modifier can enhance drug efficacy through two pharmacological pathways, representing a more convenient path for new drug development.

[0007] Besides its use in treating pulmonary hypertension, beta-prostaglandin has 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). Meanwhile, NO donor drugs are also being developed for the treatment of various diseases such as inflammation and cardiovascular diseases (Megson IL & Webb DJ, Expert Opin Investig Drugs, 2002; Knox CD et al., MK5108, J AmHeart Assoc, 2016). Therefore, both beta-prostaglandin sodium and NO donors hold the potential to be developed into a variety of therapeutic drugs.

[0008] This invention relates to a series of nitric oxide donor-type beraprost derivatives or their pharmaceutically acceptable salts. After entering the body, these compounds decompose into beraprostine and produce nitric oxide (NO), which can produce dual pharmacological effects. On the one hand, beraprostine can specifically bind to prostaglandin receptors and exert a vasodilatory effect on vascular smooth muscle. 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. The two mechanisms work synergistically to achieve a therapeutic effect.

[0009] These compounds can be used in the treatment of various diseases, including pulmonary hypertension, myocardial infarction, kidney disease, peripheral vascular diseases such as occlusive arteriosclerosis, ophthalmic diseases (such as diabetic retinopathy and glaucoma), osteoporosis, thromboangiitis obliterans, and thromboembolic diseases. Summary of the Invention

[0010] This application addresses 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. It provides a new series of compounds that combine beraprost sodium with a NO donor.

[0011] To achieve the above objectives, a nitric oxide donor-type beraprost derivative or a pharmaceutically acceptable salt thereof, as described in this invention, is provided as follows:

[0012]

[0013] n is 0, 1, 2, 3 or 4;

[0014] R is -X-ONO2, -OC(O)-X-ONO2, -OX-ONO2, or Where X is the C1-C of a straight chain or a branched chain. 10 Alkyl, cycloalkyl or -C1-C 10 alkyl-aromatic ring-; 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.

[0015] The cycloalkyl group is preferably C. 5-7 cycloalkyl, with an aromatic ring of C 5-10 Aromatic ring.

[0016] Furthermore, the compound comprises any of the following specific structures:

[0017]

[0018]

[0019] The pharmaceutically acceptable salts of the nitric oxide donor-type beraprost derivatives mentioned in this invention can be either acidic or basic. Acidic salts include, for example, hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, pyrosulfuric acid, phosphoric acid or nitric acid, or bisulfate, or acid addition salts formed with the following organic acids: for example, formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pyruvic acid, pectinic acid, persulfate, 3- Phenylacetic acid, picric acid, pentanoic acid, 2-hydroxyethanesulfonic acid, itaconic acid, aminosulfonic acid, trifluoromethanesulfonic acid, dodecyl sulfate, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucohepanoic acid, glycerophosphate, aspartic acid, sulfosalicylic acid, hemisulfonic acid, or thiocyanate. Basic salts such as sodium ions, potassium ions, N-methylglucosamine, dimethylglucosamine, ethylglucosamine, lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, meglumine, sarcosine, serine, trihydroxymethylaminomethane, aminopropylene glycol, 1-amino-2,3,4-butanetriol.

[0020] Another technical solution of this application provides a pharmaceutical composition containing the above-mentioned nitric oxide donor type beraprost derivative, which includes a compound with the structure shown in general formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0021] The carrier is any one or a mixture of two or more of the following: sustained-release agent, excipient, filler, binder, wetting agent, disintegrant, absorption promoter, adsorbent carrier, surfactant, and lubricant.

[0022] The pharmaceutical composition is preferably any one of a topical formulation, an oral formulation, and an injectable formulation.

[0023] The oral preparation is any one of granules, capsules, and tablets.

[0024] The pharmaceutical compositions of nitric oxide donor-type beraprost derivatives of the present invention include their use as procyclosporine analogs.

[0025] The pharmaceutical composition of nitric oxide donor-type beraprost derivatives described in this invention includes its use in the preparation of therapeutic drugs for various diseases such as pulmonary hypertension, myocardial infarction, kidney disease, occlusive arteriosclerosis and other peripheral vascular diseases, ophthalmic diseases (such as diabetic retinopathy, glaucoma, etc.), osteoporosis, thromboangiitis obliterans, and thromboembolic diseases.

[0026] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0027] 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

[0028] Figure 1 This indicates the therapeutic effect of compound 15 on hypoxic pulmonary hypertension in mice.

[0029] Figure 2 This represents bone mineral density data in mice after treatment with compound 15;

[0030] Figure 3 This indicates the effect of compound 15 on the proliferation of renal tubular epithelial cells in acute renal failure. Detailed Implementation

[0031] The present invention will be further described below with reference to the embodiments.

[0032] Example 1

[0033]

[0034] Synthesis route:

[0035]

[0036] Synthesis of Example 1

[0037] Concentrated sulfuric acid (13 mmol) was dissolved in dichloromethane, and fuming nitric acid (14 mmol) was slowly added dropwise at 0 °C. After reacting for 20 min, 2-bromoethanol (6 mmol) was added to the reaction solution. The reaction was continued at 0 °C for 4 hours. The reaction solution was then slowly poured into ice water and extracted twice with dichloromethane (50 mL). The organic phase was collected, washed once with water and once with saturated brine, and then evaporated to dryness to obtain the product 2-bromoethyl nitrate.

[0038] Beraprost (60 mg) was dissolved in 2 mL of anhydrous DMF. A solution of potassium iodide (75 mg), potassium carbonate (62 mg), and 2-bromoethyl nitrate (80 mg) in dichloromethane was added dropwise. The mixture was stirred at 50 °C for 2 h. The reaction was confirmed to be complete by TLC. The solvent was evaporated and purified by HPLC to obtain Example 1 with a yield of 67%. 1 HNMR(300MHz,DMSO-d)δ7.38–7.22(m,2H),7.00(d,J=5.8Hz,1H),5.85(d,J=9.5Hz ,2H),4.63–4.51(m,2H),4.38(d,J=12.5Hz,1H),4.23–4.15(m,2H),3.49–3.36(m, 2H),2.95–2.76(m,2H),2.61(dq,J=12.5,2.0Hz,1H),2.56–2.39(m,4H),2.28(d,J =13.0Hz,1H),2.01–1.99(m,1H),1.95–1.76(m,5H),1.03(d,J=5.5Hz,3H).ESI-MS m / z:510.2[M+Na] + .

[0039] Example 2

[0040]

[0041] Example 2 can be prepared by referring to the synthesis method of Example 1. 1 H NMR (300MHz, DMSO-d) δ7.35–7.24(m,2H),7.04(ddd,J=3.7,2.7,1.4Hz,1H),5.88(s,1H),5.82(s,1H),4.71–4.52(m,3H),4.35–4.10(m,6H),2. 86(qd,J=12.3,0.9Hz,2H),2.61(dq,J=12.5,2.0Hz,1H),2.51–2.37(m, 4H),2.36–2.15(m,3H),1.94–1.74(m,6H),1.00(d,J=5.7Hz,3H).ESI-MS m / z:524.2[M+Na] + .

[0042] Example 3

[0043]

[0044] Example 3 can be prepared by referring to the synthesis method of Example 1. 1H NMR(300MHz, Methanol-d4)δ7.39–7.22(m,2H),7.04(d,J=3.1Hz,1H),5.69–5.46 (m,2H),4.69–4.52(m,3H),4.31–4.09(m,5H),4.01(d,J=12.5Hz,1H),2.86(qd,J= 12.3,0.9Hz,2H),2.70(dq,J=12.5,2.0Hz,1H),2.55(dq,J=12.3,1.9Hz,1H),2.4 9–2.38(m,3H),2.28(d,J=13.0Hz,1H),2.09–1.78(m,10H),1.02(d,J=5.3Hz,3H).

[0045] Example 4

[0046]

[0047] Example 4 can be prepared by referring to the synthesis method of Example 1. 1 H NMR (300MHz, DMSO-d4) δ7.41–7.24(m,1H),7.07(d,J=7.4Hz,1H),5.85(d,J=7.5Hz,1H),4.71–4.51(m,2H),4.28–4.00(m,3H),2.94–2.77(m ,1H),2.66(dq,J=12.5,2.0Hz,1H),2.56–2.37(m,2H),2.28(d,J=13.0Hz,1H),1.94–1.78(m,5H),1.68–1.54(m,1H),1.02(d,J=5.5Hz,2H).

[0048] Example 5

[0049]

[0050] Example 5 can be prepared by referring to the synthesis method of Example 1. 1H NMR(300MHz,DMSO-d4)δ7.37(t,J=7.5Hz,1H),7.24(dd,J=7.5,2.0Hz,1H), 7.02(d,J=7.7Hz,1H),5.65–5.42(m,2H),4.71–4.52(m,3H),4.32–4.02(m,6 H),2.93–2.77(m,2H),2.66(dq,J=12.4,2.0Hz,1H),2.52–2.26(m,5H),1.9 5–1.81(m,5H),1.68–1.59(m,4H),1.53–1.35(m,4H),1.00(d,J=6.5Hz,3H).

[0051] Example 6

[0052]

[0053] Synthesis of Example 6

[0054] Beraprost (60 mg) was dissolved in 2 mL of anhydrous acetonitrile. Potassium iodide (75 mg) and potassium carbonate (62 mg) were added. After stirring at room temperature for 10 min, 2-chloromethyl ethyl nitrate (15 mg) was added. The mixture was reacted at 60 °C for 8 h. The reaction was stopped, the solvent was evaporated, dichloromethane was added, and the mixture was washed twice with water and once with saturated brine. The organic phase was concentrated and purified by HPLC to obtain Example 6, with a yield of 42%. 1 H NMR(300MHz,DMSO-d)δ7.11–7.01(m,2H),6.95(dq,J=7.7,1.2Hz,1H),5.90–5.6 8(m,2H),5.09(s,2H),4.66–4.56(m,2H),4.22–4.09(m,2H),3.87(t,J=6.2Hz,2 H),2.88–2.68(m,3H),2.46(t,J=7.1Hz,2H),2.21(dp,J=6.2,2.0Hz,2H),2.11( t,J=4.7Hz,2H),2.05–1.85(m,3H),1.65(t,J=2.0Hz,3H),1.02(d,J=6.8Hz,3H).

[0055] Example 7

[0056]

[0057] Example 7 can be prepared by referring to the synthesis method of Example 6. 1H NMR(300MHz,DMSO-d)δ7.28–7.08(m,2H),7.05(dq,J=7.7,1.2Hz,1H),5.89–5.58(m,2H) ,5.06(q,J=2.7Hz,2H),4.94(q,J=4.5Hz,1H),4.37(t,J=6.1Hz,2H),4.22–4.04(m,2H), 3.59(t,J=6.1Hz,2H),3.43(dd,J=5.5,4.2Hz,1H),2.92–2.66(m,3H),2.46(t,J=7.1Hz, 2H),2.27–2.16(m,2H),2.16–1.85(m,7H),1.65(t,J=2.0Hz,3H),1.01(d,J=6.5Hz,3H).

[0058] Example 8

[0059]

[0060] Compound 8 was prepared by referring to the synthesis method in Example 6. ESI-MS m / z: 513.3 [M+H] + . 1 H NMR(300MHz,DMSO-d)δ7.25–7.04(m,2H),7.00(dq,J=7.7,1.0Hz,1H),5.91–5.66(m,2H),4.94( q,J=4.5Hz,1H),4.57(t,J=6.2Hz,2H),4.30–4.04(m,4H),3.82(t,J=6.2Hz,2H),3.72(t,J=6.2 Hz,2H),3.43(dd,J=5.5,4.2Hz,1H),2.86–2.68(m,3H),2.41(t,J=7.0Hz,2H),2.21(dp,J=6.2, 2.0Hz,2H),2.18–2.07(m,2H),2.07–1.85(m,3H),1.66(t,J=2.0Hz,3H),1.01(d,J=6.9Hz,3H).

[0061] Example 9

[0062]

[0063] Example 9 can be prepared by referring to the synthesis method of Example 6. 1H NMR(300MHz,DMSO-d)δ7.16–7.02(m,2H),6.95(dq,J=7.7,1.2Hz,1H),5.98–5.66(m ,4H),5.11–4.87(m,3H),4.24–4.03(m,2H),3.43(dd,J=5.5,4.2Hz,1H),2.77(dqd, J=30.2,6.5,6.1,1.1Hz,3H),2.46(t,J=7.1Hz,2H),2.21(dp,J=6.2,2.0Hz,2H),2. 11(t,J=4.7Hz,2H),2.02–1.87(m,3H),1.55(t,J=2.0Hz,3H),1.02(d,J=6.8Hz,3H).

[0064] Example 10

[0065]

[0066] Example 10 can be prepared by referring to the synthesis method of Example 6. 1 H NMR(300MHz,Chloroform-d)δ7.16–7.02(m,2H),6.95(ddt,J=6.0,2.7,0.9Hz,1 H),5.93–5.61(m,4H),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),2.96–2.63(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).

[0067] Example 11

[0068]

[0069] Example 11 can be prepared by referring to the synthesis method of Example 1. 1H NMR(300MHz,DMSO-d)δ7.49–7.30(m,2H),7.18–7.02(m,4H),6.97(ddt,J=7.0,1 .9,1.0Hz,1H),5.78(qd,J=15.6,6.2Hz,2H),5.52–5.24(m,2H),4.80(d,J=6.2H z,1H),4.39–4.02(m,5H),3.42(dd,J=5.5,4.2Hz,1H),2.89–2.56(m,5H),2.40( t,J=7.1Hz,2H),2.26–1.85(m,9H),1.61(t,J=2.0Hz,3H),1.01(d,J=6.7Hz,3H).

[0070] Example 12

[0071]

[0072] Example 12 can be prepared by referring to the synthesis method of Example 1. 1 H NMR(300MHz,DMSO-d)δ7.13–7.02(m,2H),6.95(ddt,J=5.5,3.3,0.9Hz,1H) ,5.89–5.57(m,4H),4.94(dt,J=5.1,4.3Hz,1H),4.42(qt,J=10.4,6.1Hz,2H ),4.24–4.06(m,2H),3.43(dd,J=5.5,4.2Hz,1H),2.94–2.63(m,3H),2.63–2 .39(m,4H),2.26–1.79(m,9H),1.62(t,J=2.0Hz,3H),0.99(d,J=6.8Hz,3H).

[0073] Example 13

[0074]

[0075] Example 13 can be prepared by referring to the synthesis method of Example 1. 1H NMR(500MHz,Chloroform-d)δ7.51–7.30(m,2H),7.19–7.05(m,4H),6.97(ddt,J=7.0,2.0,1.0Hz,1H) ,5.78(qd,J=15.6,6.2Hz,2H),5.52–5.31(m,2H),4.94(dt,J=5.1,4.2Hz,1H),4.22–4.05(m,5H),3.4 2(dd,J=5.5,4.2Hz,1H),2.87–2.68(m,3H),2.62(tq,J=6.5,1.0Hz,2H),2.40(t,J=7.1Hz,2H),2.21( dp,J=5.9,2.0Hz,2H),2.16–2.01(m,2H),2.01–1.87(m,3H),1.81–1.60(m,7H),1.02(d,J=6.7Hz,3H).

[0076] Example 14

[0077]

[0078] Compound 14 can be prepared by referring to the synthesis method of Example 1. 1 H NMR(300MHz,DMSO-d)δ7.22–6.99(m,2H),6.97(ddt,J=7.3,1.8,0.9Hz,1H),5.78 (qd,J=15.6,6.2Hz,2H),4.94(dt,J=5.0,4.2Hz,1H),4.29–4.15(m,3H),4.15–4. 03(m,3H),3.42(dd,J=5.5,4.2Hz,1H),2.89–2.66(m,3H),2.40(t,J=7.0Hz,2H), 2.31–2.18(m,2H),2.18–1.84(m,6H),1.76–1.33(m,14H),1.04(d,J=6.1Hz,3H).

[0079] Example 15

[0080]

[0081] Compound 15 can be prepared by referring to the synthesis method of Example 1. 1H NMR(300MHz,DMSO-d)δ7.21–7.05(m,2H),6.97(ddt,J=5.6,3.5,1.1Hz,1H),5.78(qd,J=15 .6,6.2Hz,2H),4.94(dt,J=5.0,4.2Hz,1H),4.34–4.10(m,5H),3.75(dd,J=10.5,6.3Hz,1H ),3.42(dd,J=5.5,4.2Hz,1H),2.89–2.66(m,3H),2.41(t,J=7.1Hz,2H),2.21(dp,J=6.2,2 .0Hz,2H),2.17–1.82(m,7H),1.82–1.59(m,5H),1.59–1.39(m,6H),1.01(d,J=6.7Hz,3H).

[0082] Example 16

[0083]

[0084] Compound 16 can be prepared by referring to the synthesis method of Example 6. 1 H NMR(300MHz,DMSO-d)δ7.13–7.04(m,2H),6.97(ddt,J=5.7,3.5,1.1Hz,1H),5 .87–5.62(m,4H),4.94(dt,J=5.1,4.3Hz,1H),4.32(t,J=6.0Hz,2H),4.23–4. 05(m,2H),3.92(d,J=5.5Hz,1H),2.91–2.66(m,3H),2.51–2.32(m,4H),2.26– 2.17(m,2H),2.17–1.75(m,9H),1.62(t,J=2.0Hz,3H),0.99(d,J=6.1Hz,3H).

[0085] Example 17

[0086]

[0087] Compound 17 can be prepared by referring to the synthesis method of Example 6. 1H NMR(300MHz,DMSO-d)δ7.13–7.00(m,2H),6.96(ddt,J=6.2,2.9,1.1Hz,1H),5.8 9–5.63(m,2H),5.04–4.83(m,3H),4.69(d,J=6.2Hz,1H),4.48–4.28(m,4H),4.22 –4.03(m,2H),3.92(d,J=5.5Hz,1H),2.93–2.60(m,3H),2.40(t,J=7.1Hz,2H),2. 25–2.07(m,4H),2.07–1.86(m,3H),1.57(t,J=2.0Hz,3H),1.00(d,J=6.2Hz,3H).

[0088] Example 18

[0089]

[0090] Compound 18 can be prepared by referring to the synthesis method of Example 6. 1 H NMR(300MHz,DMSO-d)δ7.18–7.02(m,2H),6.97(ddt,J=5.6,3.5,1.1Hz,1H),5.78(q d,J=15.6,6.2Hz,2H),4.94(dt,J=5.0,4.2Hz,1H),4.71–4.52(m,2H),4.23–4.04(m, 6H),3.43(dd,J=5.5,4.2Hz,1H),2.92–2.66(m,5H),2.40(t,J=7.1Hz,2H),2.21(dp ,J=6.2,2.0Hz,2H),2.18–1.85(m,7H),1.60(t,J=2.0Hz,3H),1.04(d,J=6.8Hz,3H).

[0091] Example 19

[0092]

[0093] Compound 19 can be prepared by referring to the synthesis method of Example 6. 1H NMR(300MHz,DMSO-d)δ7.15–7.02(m,2H),6.97(ddt,J=7.0,1.9,1.0Hz,1H),5.78(qd,J=1 5.6, 6.2Hz, 2H), 4.94 (dt, J=5.1, 4.2Hz, 1H), 4.61 (t, J=7.1Hz, 2H), 4.26–4.04 (m, 6H), 3. 42(dd,J=5.5,4.2Hz,1H),2.89–2.67(m,5H),2.39(t,J=7.0Hz,2H),2.21(dp,J=6.2,2.0H z,2H),2.17–1.87(m,5H),1.87–1.73(m,4H),1.60(t,J=2.0Hz,3H),0.98(d,J=6.7Hz,3H).

[0094] Example 20

[0095]

[0096] Compound 20 can be prepared by referring to the synthesis method of Example 6. 1 H NMR(300MHz, DMSO-d)δ7.13–7.02(m,2H),6.96(ddt,J=5.5,3.3,1.0Hz,1H),5.78(qd,J= 15.6,6.2Hz,2H),4.94(dt,J=5.0,4.2Hz,1H),4.71–4.51(m,2H),4.40–4.26(m,4H),4.26 –4.05(m,2H),3.43(dd,J=5.5,4.2Hz,1H),2.91–2.63(m,5H),2.40(t,J=7.1Hz,2H),2.2 1(dp,J=6.2,2.0Hz,2H),2.20–1.85(m,5H),1.65(t,J=2.0Hz,3H),1.02(d,J=6.7Hz,3H).

[0097] Example 21

[0098]

[0099] Compound 21 can be prepared by referring to the synthesis method of Example 6. 1H NMR(300MHz,DMSO-d)δ7.20–7.10(m,2H),7.01(ddt,J=5.5,3.3,1.0Hz,1H),5.78(qd,J=15.6,6.2Hz ,2H),5.25(q,J=6.6Hz,1H),4.94(dt,J=5.1,4.3Hz,1H),4.52–4.41(m,2H),4.41–4.26(m,2H),4.26 –4.05(m,2H),3.43(dd,J=5.5,4.2Hz,1H),2.91–2.67(m,3H),2.40(t,J=7.0Hz,2H),2.21(dp,J=6.2 ,2.0Hz,2H),2.17–1.84(m,5H),1.65(t,J=2.0Hz,3H),1.47(d,J=6.6Hz,3H),1.00(d,J=6.2Hz,3H).

[0100] Example 22

[0101]

[0102] Compound 22 can be prepared by referring to the synthesis method of Example 6. 1 H NMR(300MHz,DMSO-d)δ7.21–7.08(m,2H),7.00(ddt,J=6.9,1.9,1.0Hz,1H),5.78(qd,J=1 5.6,6.2Hz,2H),4.94(dt,J=5.0,4.2Hz,1H),4.54–4.31(m,2H),4.22–4.03(m,6H),3.42( dd,J=5.5,4.2Hz,1H),2.91–2.62(m,3H),2.49(t,J=7.0Hz,2H),2.40(t,J=7.1Hz,2H),2. 21(dp,J=6.2,2.0Hz,2H),2.18–1.82(m,9H),1.55(t,J=2.0Hz,3H),1.00(d,J=6.2Hz,3H).

[0103] Example 23

[0104]

[0105] Compound 23 can be prepared by referring to the synthesis method of Example 6. 1H NMR(300MHz,DMSO-d)δ7.17–7.08(m,2H),7.00(ddt,J=5.6,3.5,1.1Hz,1H),5.78 (qd,J=15.6,6.2Hz,2H),5.09–4.87(m,3H),4.27–4.05(m,6H),3.43(dd,J=5.5,4. 2Hz,1H),2.86–2.67(m,3H),2.38(t,J=7.1Hz,2H),2.21(dp,J=6.2,2.0Hz,2H),2 .17–1.87(m,5H),1.87–1.72(m,4H),1.61(t,J=2.0Hz,3H),1.01(d,J=6.7Hz,3H).

[0106] Example 24

[0107]

[0108] Compound 24 can be prepared by referring to the synthesis method of Example 6. 1 H NMR(300MHz,DMSO-d)δ7.54–7.38(m,2H),7.30–7.15(m,2H),7.15–7.02(m,2H),6.97(ddt,J=7 .0,2.0,1.0Hz,1H),5.97–5.70(m,4H),5.50–5.26(m,2H),4.94(dt,J=5.1,4.2Hz,1H),4.80(d, J=6.2Hz,1H),4.26–4.04(m,3H),3.62(q,J=0.8Hz,2H),2.77(dqd,J=31.3,6.5,6.1,1.1Hz,3H ), 2.43(td,J=7.1,1.0Hz,2H),2.27–1.82(m,7H),1.60(t,J=2.0Hz,3H),1.04(d,J=6.6Hz,3H).

[0109] Example 25

[0110]

[0111] Compound 25 can be prepared by referring to the synthesis method of Example 6. 1H NMR (300MHz, DMSO-d) δ8.04(tt,J=2.2,1.0Hz,1H),7.92(ddd,J=7.7,2.2,1.1Hz,1H),7.57(ddq,J=7.9,2.2,1.1Hz,1H),7.47(t,J=7 .8Hz,1H),7.20–7.02(m,2H),6.97(ddt,J=7.0,2.0,1.0Hz,1H),5.89–5.70(m,2H),5.45(t,J=1.0Hz,2H),4.94(dt,J=5.0,4.2Hz,1H ),4.80(d,J=6.2Hz,1H),4.46(td,J=6.0,0.8Hz,2H),4.37(td,J=6.0,0.8Hz,2H),4.26–4.04(m,3H),3.42(dd,J=5.5,4.2Hz,1H),2. 93–2.62(m,3H),2.41(t,J=7.0Hz,2H),2.21(dp,J=5.9,2.0Hz,2H),2.16–1.84(m,5H),1.65(t,J=2.0Hz,3H),1.02(d,J=6.6Hz,3H).

[0112] Example 26

[0113]

[0114] Compound 26 can be prepared by referring to the synthesis method of Example 6. 1 H NMR(300MHz,DMSO-d)δ8.11–7.84(m,2H),7.65–7.37(m,2H),7.13–7.01(m,2H),6.97(ddt,J =7.0,2.0,1.0Hz,1H),5.92–5.62(m,2H),4.94(dt,J=5.0,4.2Hz,1H),4.80(d,J=6.2Hz,1H) ,4.32–3.95(m,7H),3.45(dd,J=5.6,4.3Hz,1H),2.89–2.67(m,3H),2.40(t,J=7.1Hz,2H),2 .21(dp,J=5.9,2.0Hz,2H),2.18–1.85(m,7H),1.61(t,J=2.0Hz,3H),1.00(d,J=6.8Hz,3H).

[0115] Example 27

[0116]

[0117] Compound 27 can be prepared by referring to the synthesis method of Example 6. 1 H NMR(300MHz,DMSO-d)δ8.05–7.89(m,2H),7.58–7.35(m,2H),7.17–7.04(m,2H),6.97(ddt, J=6.9,2.0,1.0Hz,1H),5.90–5.65(m,2H),4.94(dt,J=5.1,4.2Hz,1H),4.80(d,J=6.2Hz,1 H),4.30–4.02(m,7H),3.45(dd,J=5.6,4.3Hz,1H),2.88–2.66(m,3H),2.49–2.31(m,2H),2 .21(dp,J=5.9,2.0Hz,2H),2.18–1.73(m,9H),1.61(t,J=2.2Hz,3H),1.01(d,J=6.4Hz,3H).

[0118] Example 28

[0119]

[0120] Compound 28 can be prepared by referring to the synthesis method of Example 6. 1 H NMR(300MHz,DMSO-d)δ7.22–7.06(m,2H),7.00(dtt,J=5.7,3.5,1.0Hz,1H),5.7 8(qd,J=15.6,6.2Hz,2H),4.94(dt,J=5.1,4.3Hz,1H),4.62–4.40(m,4H),4.29–4 .05(m,4H),3.43(dd,J=5.5,4.2Hz,1H),2.87–2.67(m,3H),2.55–2.39(m,3H),2. 26–2.16(m,2H),2.16–1.85(m,5H),1.66(t,J=2.1Hz,3H),1.00(d,J=6.5Hz,3H).

[0121] Example 29

[0122]

[0123] Compound 29 can be prepared by referring to the synthesis method of Example 1. 1H NMR(300MHz,DMSO-d)δ7.19–7.07(m,2H),7.01(ddt,J=6.9,1.9,1.0Hz,1H),5.7 8(qd,J=15.6,6.2Hz,2H),4.94(dt,J=5.0,4.2Hz,1H),4.56–4.36(m,4H),4.29– 4.01(m,4H),3.92(d,J=5.5Hz,1H),2.91–2.65(m,3H),2.40(t,J=7.0Hz,2H),2. 33–2.17(m,3H),2.17–1.81(m,7H),1.60(t,J=1.9Hz,3H),0.99(d,J=6.2Hz,3H).

[0124] Example 30

[0125]

[0126] Compound 30 can be prepared by referring to the synthesis method of Example 6. 1 H NMR(300MHz, DMSO-d)δ7.21–7.07(m,2H),7.00(ddt,J=6.9,1.9,1.0Hz,1H),5.78(qd,J=15.6,6.2 Hz,2H),5.28–5.02(m,3H),4.94(dt,J=5.1,4.2Hz,1H),4.74(dd,J=21.4,5.9Hz,3H),4.23–4.05(m ,2H),3.93(dd,J=5.5,3.9Hz,3H),3.42(dd,J=5.5,4.2Hz,1H),2.87–2.67(m,3H),2.43(t,J=7.1Hz ,2H),2.21(dp,J=6.2,2.0Hz,2H),2.18–1.84(m,5H),1.60(t,J=2.2Hz,3H),1.00(d,J=6.6Hz,3H).

[0127] Example 31

[0128]

[0129] Compound 31 can be prepared by referring to the synthesis method of Example 6. 1H NMR(300MHz, DMSO-d)δ7.15–7.04(m,2H),6.97(ddt,J=6.9,1.9,1.0Hz,1H),5.78(qd,J=15.6 ,6.2Hz,2H),5.11(p,J=5.6Hz,1H),4.94(dt,J=5.0,4.2Hz,1H),4.68(d,J=5.6Hz,2H),4.27–4 .02(m,5H),3.84(d,J=5.5Hz,2H),3.56–3.29(m,3H),2.86–2.63(m,3H),2.40(t,J=7.1Hz,2H) ,2.21(dp,J=5.9,2.0Hz,2H),2.17–1.88(m,7H),1.61(t,J=2.2Hz,3H),0.99(d,J=6.6Hz,3H).

[0130] Example 32

[0131]

[0132] Synthesis method

[0133]

[0134] Beraprost (60 mg), chloromethylfuran nitroxide, DMAP, and TEA were dissolved in 2 mL of anhydrous dichloromethane. After stirring at room temperature for four hours, 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. The solution was filtered, concentrated, and purified by HPLC to obtain Example 32 with a yield of 40%. 1 H NMR(300MHz,DMSO-d)δ7.94–7.73(m,2H),7.69–7.50(m,3H),7.13–7.02(m,2H),6.97(ddt,J=7.0 ,2.0,1.0Hz,1H),6.11–5.95(m,2H),5.89–5.64(m,2H),4.94(dt,J=5.0,4.2Hz,1H),4.80(d,J=6 .2Hz,1H),4.22–4.03(m,3H),3.45(dd,J=5.6,4.3Hz,1H),2.88–2.65(m,3H),2.43(t,J=7.1Hz,2 H),2.21(dp,J=5.9,2.0Hz,2H),2.18–1.84(m,5H),1.61(t,J=2.1Hz,3H),1.00(d,J=6.2Hz,3H).

[0135] Example 33

[0136]

[0137] Compound 33 can be prepared by referring to the synthesis method of Example 32. 1 H NMR(300MHz,DMSO-d)δ8.04–7.90(m,2H),7.73–7.49(m,3H),7.17–7.02(m,2H),6.97(ddt,J =7.0,2.0,1.0Hz,1H),5.93–5.61(m,2H),4.94(dt,J=5.0,4.2Hz,1H),4.63(td,J=6.2,1.0H z,2H),4.43(t,J=6.2Hz,2H),4.27–4.00(m,3H),3.45(dd,J=5.6,4.3Hz,1H),2.88–2.62(m, 3H), 2.40 (t, J = 7.1Hz, 2H), 2.29–1.86 (m, 7H), 1.55 (t, J = 2.1Hz, 3H), 1.01 (d, J = 6.2Hz, 3H).

[0138] Example 34

[0139]

[0140] Compound 34 can be prepared by referring to the synthesis method of Example 32. 1 H NMR(300MHz,DMSO-d)δ8.12–7.91(m,2H),7.70–7.54(m,3H),7.17–7.04(m,2H),6.97(ddt ,J=7.0,2.0,1.0Hz,1H),5.89–5.63(m,2H),4.94(dt,J=5.0,4.2Hz,1H),4.80(d,J=6.2Hz, 1H),4.43(t,J=6.1Hz,2H),4.24–4.03(m,5H),3.45(dd,J=5.6,4.3Hz,1H),2.88–2.67(m,3 H),2.40(t,J=7.1Hz,2H),2.27–1.85(m,9H),1.57(t,J=1.9Hz,3H),0.99(d,J=6.6Hz,3H).

[0141] Example 35

[0142]

[0143] Compound 35 can be prepared by referring to the synthesis method of Example 32. 1H NMR(300MHz,DMSO-d)δ8.04–7.84(m,2H),7.70–7.52(m,3H),7.16–7.04(m,2H),6.97(ddt,J=7 .3,1.8,0.9Hz,1H),5.90–5.63(m,2H),4.94(dt,J=5.1,4.2Hz,1H),4.47–4.31(m,2H),4.24–4. 00(m,5H),3.45(dd,J=5.6,4.3Hz,1H),2.89–2.62(m,3H),2.48–2.34(m,2H),2.21(dp,J=5.9, 2.0Hz,2H),2.20–1.87(m,5H),1.87–1.72(m,4H),1.62(t,J=1.8Hz,3H),0.99(d,J=6.2Hz,3H).

[0144] Experimental Example 1: In vitro NO release test of the compound

[0145] The Griess process utilizes the instantaneous oxidation of released NO in aqueous solution to form NO2. - NO2 - It forms a complex with Griess reagent, which has strong UV absorption at 540 nm, thereby determining the NO release amount of the compound.

[0146] 1) Solution preparation: Blank solution: Mix 10 mL DMSO and 190 mL PBS; Griess reagent: Dissolve sulfonamide (4.0 g), N-(1-naphthyl)ethylenediamine dihydrochloride (0.2 g) and 10 mL 85% H3PO4 in 90 mL distilled water and stir until a clear solution is obtained; L-cysteine ​​solution: Accurately weigh L-cysteine ​​and add a certain amount of PBS to prepare a 200 μM solution; Test compound solution: Accurately weigh the test compound, dissolve and dilute it with DMSO to a concentration of 1 mM, and then dilute it with PBS to a concentration of 200 μM.

[0147] 2) Formulation of standard curve equation: Sodium nitrite standard solutions with concentrations of 0, 0.78, 1.56, 3.13, 6.25, 12.5, 25, 50, and 100 μmol / L were prepared using blank solution. 150 μL of each concentration was taken each time, and 50 μL of Griess reagent was added to each solution. After incubation in a shaker at 37℃ for 30 min, the absorbance of each tube was measured at 540 nm using an ELISA reader. The standard curve equation was obtained by subtracting the blank solution reading from the absorbance of each tube.

[0148] 3) Test of the test compound: Mix 2.5 mL each of the prepared test compound solution and L-cysteine ​​solution, and incubate in a shaker at 37°C for 120 min. Take 150 μL of the mixture every 15 min, add 50 μL of Griess reagent to each, mix well, and incubate in a shaker at 37°C for another 30 min. Measure the absorbance of each tube at 540 nm using a microplate reader. Subtract the blank solution reading from the absorbance and substitute the values ​​into the standard curve equation to obtain the NO release amount.

[0149] The test results show that the nitric oxide donor type beraprostine derivatives or their pharmaceutically acceptable salts have good NO release effects.

[0150] Table 1. NO release effect of compounds in the examples

[0151]

[0152]

[0153] Experimental Example 2: In vivo experiment of hypoxic pulmonary hypertension in rats

[0154] 1) Experimental instruments and materials: HX-200 animal ventilator; male SD rats used in the experiment were all purchased from Yangzhou University. All control groups were raised in normal conditions, while the drug intervention group and model group were raised in a low-pressure hypoxic chamber (pressure 50 kPa, oxygen concentration 10%).

[0155] 2) Experimental Procedure: Compound 15 was dissolved in DMSO / solutol / water (10 / 10 / 80) to prepare a clear solution. Starting from day 2 of hypoxia, the intervention group was administered compound 15 by gavage at a dose of 5 mg / kg. All rats were weighed weekly, and their survival status was recorded. Pulmonary artery pressure was measured after four weeks. Rats were anesthetized with chloral hydrate (100 g / L) (3 ml / kg), fixed in a supine position, and underwent tracheotomy. They were assisted with ventilation using a small animal ventilator (60 breaths / min, tidal volume 5 ml, I:E ratio 4:5). The left third rib was freed, and a catheter with one end connected to a tension transducer was inserted into the pulmonary artery. Mean pulmonary artery pressure (mPAP) was recorded using a BL-420E biomechanical experimental system. Pleural and peritoneal fluid were examined and collected. Finally, blood was drawn from the abdominal aorta to euthanize the rats.

[0156] 3) Experimental results: Compared with the control group, the mPAP of rats in the model group was significantly increased, while the mPAP of the intervention group treated with compound 15 was lower than that in the model group, demonstrating a good therapeutic effect on hypoxic pulmonary hypertension. Figure 1 The results indicate the therapeutic effect of nitric oxide donor-type beraprostine derivative compound 15 on hypoxic pulmonary hypertension in mice.

[0157] Experimental Case 3: Therapeutic Effect on Osteoporosis

[0158] 1) Experimental Materials; 1. Clean-grade C57BL / 6 strain, 8-10 weeks old, non-pregnant female mice were purchased from Yangzhou University. 2. Main reagents and instruments: Estradiol valerate tablets (Bayer); mouse osteocalcin (OC) enzyme-linked immunosorbent assay kit, alkaline phosphatase (ALP) test kit, and tartrate-resistant acid phosphatase (StrACP) test kit (all kits were purchased from Nanjing Jiancheng). Dual-energy X-ray absorptiometry (HOLOGIC); ECLIPSE 50i microscope (Nikon); MUTISKANMK3 microplate reader (Thermo); tissue sectioning equipment (including KD-TS3D1 automatic biological tissue dehydrator (Zhejiang Kedi), TB-718 automatic biological tissue embedding machine (Hubei Taiwei), R138 rotary microtome (Hubei Taiwei), TK-212 automatic constant temperature slide bleaching machine (Hubei Taiwei), TK-213 automatic constant temperature slide drying machine (Hubei Taiwei), etc.).

[0159] 2) Experimental methods: a. Animal grouping: Female mice were randomly divided into 4 groups, namely sham operation group, model group, positive drug group, and test drug group. b. Preparation of postmenopausal osteoporosis mouse model: After anesthetizing mice with chloral hydrate, the ovaries were removed in a supine position. In the sham operation group, only the same volume of adipose tissue near the ovary was removed. Vaginal smears were examined on days 4-8 after oophorectomy to determine whether the oophorectomy was complete. c. Administration method: The drug was administered on day 3 after surgery. The positive control drug (0.1 mL / 10 g of estradiol valerate was administered by gavage). The sham operation group and the model group were given the same volume of 0.9% sodium chloride solution, which was administered by gavage for 28 consecutive days. d. Results test: Blood and bone tissue samples were taken after day 28. The following indicators were tested: (1) Determination of bone mineral density: The bone mineral density of the lumbar vertebrae L4-6 was measured by dual-energy X-ray absorptiometry. (2) Observation of bone tissue morphology: Tibial bone was stained with HE to observe changes in bone tissue morphology. The main quantitative evaluation indicators were the bone volume / tissue volume (BV / TV), trabecular bone number (Tb.N), and trabecular separation (Tb.sp). (3) Determination of the levels of biomarkers ALP, StrACP, OC, and E2 in mouse serum: Blood was collected by enucleation and analyzed using a kit and enzyme-linked reaction adsorption method. All data were analyzed using SPSS 20.0 software.

[0160] 3) Experimental results: Compared with the model group, the test compound effectively increased the various indicators of lumbar vertebral bone mineral density in mice and significantly reduced osteocalcin content; the serum alkaline phosphatase and acid phosphatase levels were significantly reduced, indicating that the compound can improve the relevant indicators in estrogen deficiency-induced osteoporosis.

[0161] in, Figure 2 This table shows bone mineral density data in mice treated with compound 15, a nitric oxide donor-type beraprost derivative. Tables 2 and 3 show quantitative data on bone morphology and serum bone metabolism in mice treated with compound 15.

[0162] Table 2. Quantitative data on bone morphology

[0163]

[0164] Table 3. Bone metabolism markers in mouse serum

[0165]

[0166] Experimental Case 4: Tubular Protective Effect in Acute Renal Failure

[0167] 1) Experimental materials: Glycerol (Maclean's); CCK-8 assay kit, Annexin V-PE apoptosis assay kit, superoxide dismutase activity assay kit, malondialdehyde assay kit (Beyotime Biotechnology Co., Ltd.); Primary antibodies against cysteine-aspartic protease, caspase 3 and 9, B lymphoma-2 gene, Bcl-2-related X protein (BAX, anti-rabbit), TGF-β1, and smad3 (Abcam, USA). Protein gel imaging system, microplate reader, and flow cytometer (Beckman Coulter, CytoFLEX model). Clean-grade SD rats were purchased from Yangzhou University.

[0168] 2) Experimental Methods: An acute renal failure model was established in rats by glycerol injection into the hind limbs. Successful model establishment was indicated by elevated serum BUN and Cr levels, renal interstitial vascular necrosis, and inflammatory cell infiltration 24 hours later. Kidney tissue from the model animals was cultured in vitro. Renal tubular epithelial cells were isolated and identified, and co-cultured with the test drug (0-24 hours). Changes in various biochemical indicators and apoptosis were measured. All data were statistically analyzed.

[0169] 3) Experimental results: Compared with the control group, the OD of model animal cells after culture (3, 6, 12, 24 h) was significantly higher. 450The test compounds decreased MDA activity and BCL2 protein levels, while increasing SOD activity, apoptosis rate, and caspase 3, caspase 9, BAX, TGF-β1, and Smad3 protein levels. Compared to the model group, the test compounds induced OD after 6 hours of cell culture. 450 The compound increased and decreased apoptosis rate, SOD activity, and BCL2, TGF-β1, and Smad3 protein levels (P<0.05), while increasing MDA activity, caspase3, caspase9, and BAX protein levels. In conclusion, the tested compound can promote the proliferation of renal tubular epithelial cells in rats with acute renal failure, inhibit apoptosis, suppress oxidative stress, and alleviate renal tubular epithelial cell damage by inhibiting the expression of pro-apoptotic proteins.

[0170] in Figure 3 Tables 4 and 5 show the effects of compound 15 on renal tubular cell proliferation in mice with acute renal failure, including MDA, SOD, apoptosis rate, apoptosis proteins, TGF-β1, and Smad3.

[0171] Table 4. Effects of compound 15 on MDA, SOD and apoptosis rate in renal tubular epithelial cells during acute renal failure.

[0172]

[0173] Table 5. Effects of compound 15 on apoptosis proteins, TGF-β1, and Smad3 in renal tubular epithelial cells during acute renal failure.

[0174] Group caspase3 caspase9 BCL2 BAX TGF-β1 Smad3 control group 0.31±0.08 0.39±0.12 1.07±0.13 0.29±0.09 0.33±0.02 0.11±0.01 Model group 1.27±0.19 0.88±0.2 0.08±0.02 1.17±0.17 0.91±0.09 0.57±0.08 Group 15 of compounds at 200 μg / mL 0.28±0.06 0.48±0.1 0.56±0.11 0.32±0.12 0.24±0.05 0.19±0.07

[0175] Experimental Example 5: Antiplatelet Aggregation Effect

[0176] 1) Experimental materials: ADP, adrenaline, collagen (BaiLi Biotechnology)

[0177] 2) Experimental Methods: Blood samples from healthy subjects were mixed with 3.8% citric acid solution and centrifuged at 160 r / min to obtain platelet-rich plasma. For data calibration, the obtained platelet-rich plasma was further centrifuged at 2000 r / min to obtain anemic platelet-rich plasma, which was stored at -20°C for later use. Bornl's turbidimetric assay was used for testing. 225 μL of platelet-rich plasma was added to a reaction vessel, and the compound of the example was prepared into a 50 nM solution (25 mM Tris-acetate and 120 mM NaCl). 25 μL of the compound solution of the example was added, and after co-incubation at 37°C for 2 min, 5 μL of ADP (final concentration 2 μM) was added to induce platelet aggregation. To assess the degree of platelet aggregation, the maximum absorbance was taken from the data of the blank group 10 min after the addition of ADP, thereby calculating the inhibition rate of each compound of the example against ADP-induced platelet aggregation. As shown in Table 6 below, the compounds described in the embodiments of this application all have a good effect on inhibiting ADP-induced platelet aggregation.

[0178] Table 6. Compounds from the Examples Inhibit ADP-Induced Platelet Aggregation

[0179] compound Platelet aggregation inhibition rate (%) compound Platelet aggregation inhibition rate (%) 1 78 19 69 2 66 20 45 3 57 21 81 4 43 22 52 5 64 23 56 6 39 24 47 7 44 25 35 8 61 26 22 9 52 27 35 10 54 28 47 11 67 29 56 12 42 30 52 13 61 31 67 14 48 32 55 15 92 33 82 16 81 34 35 17 59 35 44 18 66

[0180] For those skilled in the art, this disclosure is not limited to the foregoing illustrative embodiments and can be embodied in other specific forms without departing from its essential attributes. Therefore, it is intended that all aspects be considered illustrative rather than restrictive, that references be made to the appended claims rather than the foregoing embodiments, that references be made only to the appended claims and not to the foregoing examples, and that all variations falling within the meaning and scope of claim equivalence are therefore intended to be included herein.

[0181] All patents, patent applications, and references listed in this specification are incorporated herein by reference in their entirety. In case of inconsistencies, this disclosure, including its definitions, will be considered more persuasive.

Claims

1. A nitric oxide donor-type beraprostine derivative or a pharmaceutically acceptable salt thereof, characterized in that, It is selected from the following structures: 。 2. The use of the nitric oxide donor type beraprost derivative or its pharmaceutically acceptable salt as described in claim 1 in the preparation of drugs for treating pulmonary hypertension, kidney disease, osteoporosis, and thromboembolic diseases.

3. A pharmaceutical composition, characterized in that... It comprises the nitric oxide donor type beraprost derivative of claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

4. The pharmaceutical composition according to claim 3, characterized in that, The carrier is any one or a mixture of two or more of the following: sustained-release agent, filler, binder, wetting agent, disintegrant, absorption promoter, adsorbent carrier, surfactant, and lubricant.

5. The pharmaceutical composition according to claim 4, characterized in that, The pharmaceutical composition is any one of a topical preparation, an oral preparation, and an injectable preparation.

6. The pharmaceutical composition according to claim 5, characterized in that, The oral preparation is any one of granules, capsules, and tablets.

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