WO2023131349A1 - Nitric oxide donor type treprostinil derivative, and pharmaceutical composition thereof and use thereof - Google Patents

Abstract

A treprostinil coupled nitric oxide (NO) donor drug as shown in formula I below, a pharmaceutical composition thereof and a use thereof.

Description

Nitric oxide donor type treprostinil derivative and its pharmaceutical composition and application technical field

The invention belongs to the field of biomedicine, and specifically relates to a coupled NO donor treprostinil derivative or a pharmaceutically acceptable salt thereof, its pharmaceutical composition and application.

Background technique

Pulmonary arterial hypertension (PAH) is a malignant disease characterized by elevated pulmonary vascular resistance and right ventricular failure. Prostaglandins in the body activate adenylyl cyclase by interacting with prostaglandin receptors on platelets or vascular smooth muscle, thereby increasing the concentration of intracellular cAMP, which can play a role in dilating blood vessels.

Prostacyclin (PGI2) drugs are one of the important types of targeted drugs for the treatment of PAH. Treprostinil, as a class of prostaglandin analogues, can specifically bind to prostaglandin receptors to relax vascular smooth muscle and reduce pulmonary artery Pressure and inhibition of pulmonary artery vascular remodeling and in situ thrombosis. Treprostinil is currently the only prostacyclin drug that is used in the treatment of PAH abroad and has multiple drug methods, such as subcutaneous, intravenous, inhalation or oral subcutaneous/intravenous administration. It has anti-platelet and vasodilator properties , with stable structure and longer action time than other prostacyclins, it is the preferred drug for the treatment of pulmonary arterial hypertension (Nika Skoro-Sajer, Drugs 2012; James C Coons, et al., Therapeutic Advances in Respiratory Disease, 2021; Steven D Nathan, et al., Lancent Respiratory Medicine, 2021).

Nitric oxide (Nitric oxide, NO) is a messenger molecule widely present in the human body and has various physiological activities. At physiological concentrations, it can activate soluble guanylate cyclase in smooth muscle cells to increase The level of cGMP, which makes blood vessels dilate, plays a role in regulating blood pressure. NO is also a commonly used drug for pulmonary hypertension and pulmonary embolism (Fernanda Blasina, et al., Pulmonary Pharmacology & Therapeutics, 2019; Jeffrey A. Kline, Am Heart J. 2017; ), especially in the treatment of children and neonates with pulmonary hypertension and the treatment of late hospitalized patients It has special advantages (Puthiyachirakkal M et al., Front Pediatr, 2013; Petros AJ and Pierce CM, Paediatr Anaesth, 2006), but because it is a gas, it must be administered in the hospital, and it needs to be administered by inhalation, but Due to the large individual differences among patients, it is difficult to control the dose, so the current clinical use is gradually reduced, and it is only used in hospitalized patients and critical situations.

Due to the complex pathogenesis of pulmonary arterial hypertension, two or three drugs with different mechanisms of action have been used in combination to improve efficacy (White RJ, et al., Am J Respir Crit Care Med, 2020; Verlinden NJ, et al., Pulmonary Circulation 2020). Since NO and treprostinil have their own special advantages and are effective through two second messenger cGMP and cAMP pathways respectively, clinically there is also the experience of using combined administration to improve the therapeutic effect in the future (Stacy Mandras, et al., J of Cardiovascular Pharmacol and Therap, 2021), but because the two drugs have to be administered in different ways (NO must be inhaled in the hospital, Quqian can be taken orally or injected or inhaled), so the drug is very inconvenient to use and the patient's compliance is low It is difficult to obtain the desired therapeutic effect.

The present invention is a series of innovative drugs coupled with Treprostinil and NO donors. This type of drug combines two characteristic components of Treprostinil and NO, which are decomposed into Treprostinil and NO donors in the body and then produced NO, on the one hand, provides treprostinil and prostaglandin receptor specific binding, and exerts the effect of dilating vascular smooth muscle; NO passes through different pathways of cGMPS and cAMP. Therefore, the two have a synergistic function, which strengthens the effect of dilating blood vessels. At the same time, it is more convenient to administer and solves the problem of patient compliance. Pulmonary hypertension, respiratory distress syndrome, chronic arterial occlusive disease and other diseases.

Contents of the invention

Purpose of the invention: In view of the defect that treprostinil is widely used clinically and has weak curative effect, a coupling NO donor type treprostinil derivative or a pharmaceutically acceptable salt thereof is provided. The coupled NO donor treprostinil derivative or its pharmaceutically acceptable salt can be used for the treatment of pulmonary arterial hypertension, respiratory distress syndrome, chronic arterial occlusive disease and the like.

Technical solution: In order to achieve the above purpose, as described in the present invention, a coupled NO donor treprostinil derivative or a pharmaceutically acceptable salt thereof shown in the following formula I:

n is 0-6, preferably 0, 1, 2, 3 or 4;

其中X为直链或支链的C₁-C₁₀烷基、C_5-7环烷基或-C₁-C₁₀烷基-芳环-；其中C₁-C₁₀烷基、C_5-7环烷基或芳环可被以下一个或者多个取代基取代：卤原子、羟基、羧基、氰基或-(C₁-C₁₀烷基)-ONO₂。R is -X-ONO ₂ , -OC(O)-X-ONO ₂ , -OX-ONO ₂ , or

Wherein X is straight chain or branched C ₁ -C ₁₀ alkyl, C _5-7 cycloalkyl or -C ₁ -C ₁₀ alkyl-aromatic ring-; wherein C ₁ -C ₁₀ alkyl, C _{5- 7} Cycloalkyl or aromatic ring can be substituted by one or more of the following substituents: halogen atom, hydroxyl, carboxyl, cyano or -(C ₁ -C ₁₀ alkyl)-ONO ₂ .

Further, the compound includes any one of the following structures:

The present invention also provides a pharmaceutical composition, which contains a therapeutically effective amount of coupled NO-donor treprostinil derivative or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

The carrier of the present invention can be any one or A mixture of two or more.

The pharmaceutical composition is preferably any one of external preparations, oral preparations and injection preparations.

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

The coupling NO donor treprostinil derivative or its pharmaceutically acceptable salt described in the present invention includes its application as a procyclin analogue.

The coupling NO donor type treprostinil derivative or its pharmaceutically acceptable salt described in the present invention includes its application in the preparation of medicines for treating pulmonary hypertension, respiratory distress syndrome, chronic arterial occlusive disease and the like.

Beneficial effect: compared with the prior art, the present invention has the following advantages:

The invention provides a kind of coupled NO donor type treprostinil derivatives or pharmaceutically acceptable salts thereof, which solves the defects of weak curative effect of treprostinil and inconvenient dosage of NO administration, etc. Drug effectiveness and patient compliance, and drug effectiveness control have also been improved.

Description of drawings

Fig. 1 is a comparison chart of hypoxic pulmonary arterial hypertension (mPAP) measured in vivo in rats.

Detailed ways

The present invention will be further described below in conjunction with embodiment.

Example 1

synthetic route:

Synthesis of Example 1

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

Dissolve Treprostinil (60mg) with 2mL of anhydrous DMF, drop potassium iodide (75mg), potassium carbonate (62mg) and 2-bromoethyl nitrate (80mg) in dichloromethane solution, move to 50°C The reaction was stirred for 2 h under the conditions, and the reaction was complete as detected by TLC. The solvent was spin-dried, and purified by HPLC to obtain Example 1 with a yield of 56%. ¹ H NMR (300MHz, DMSO-d) δ7.08(t, J=7.9Hz, 1H), 6.78(dq, J=7.8, 1.1Hz, 2H), 4.76–4.60(m, 4H), 4.47(td ,J=6.2,1.4Hz,2H),3.96(qd,J=5.7,4.8Hz,1H),3.58–3.48(m,1H),3.10–2.92(m,2H),2.87–2.67(m,2H ),2.24–1.96(m,3H),1.96–1.78(m,2H),1.73–1.50(m,4H),1.48–1.23(m,8H),0.96–0.79(m,3H).

Example 2

Referring to the synthetic method of Example 1, Example 2 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.12 (t, J = 7.2Hz, 1H), 6.92–6.64 (m, 2H), 4.41 (t, J = 6.1Hz, 2H), 4.29 (dd, J =6.4,5.7Hz,2H),3.95(qd,J=5.5,4.6Hz,1H),3.61–3.49(m,1H),2.96(d,J=7.2Hz,2H),2.88–2.67(m, 2H),2.29–1.98(m,5H),1.95–1.80(m,2H),1.70–1.53(m,4H),1.47–1.20(m,8H),0.98–0.79(m,3H).

Example 3

With reference to the synthetic method of Example 1, Example 3 can be prepared. ¹ H NMR (300MHz, Methanol-d ₄ ) δ7.13(t, J=7.0Hz, 1H), 6.78(dq, J=7.8, 1.1Hz, 2H), 4.67(s, 2H), 4.46–4.27( m,2H),4.23–4.09(m,2H),3.97(qd,J=5.5,4.6Hz,1H),3.64–3.46(m,1H),2.96(d,J=7.2Hz,2H),2.91 –2.67(m,2H),2.27–1.97(m,3H),1.97–1.78(m,6H),1.78–1.51(m,4H),1.44–1.20(m,8H),1.02–0.77(m, 3H).

Example 4

Referring to the synthetic method of Example 1, Example 4 can be prepared. ¹ H NMR (300MHz, DMSO-d ₄ )δ7.08(t, J=7.9Hz, 1H), 6.89–6.66(m, 2H), 4.67(s, 2H), 4.28(t, J=6.1Hz, 2H), 4.12(t, J=6.1Hz, 2H), 3.97 (qd,J=5.5,4.6Hz,1H),3.62–3.47(m,1H),2.87–2.68(m,2H),2.41(d,J=5.3Hz,1H),2.26–1.96(m,3H ),1.96–1.49(m,12H),1.49–1.17(m,8H),0.94–0.79(m,3H).

Example 5

Referring to the synthesis method of Example 1, Example 5 can be prepared. ¹ H NMR (300MHz, DMSO-d ₄ )δ7.08(t, J=7.9Hz, 1H), 6.89–6.71(m, 2H), 4.67(s, 2H), 4.29(t, J=6.1Hz, 2H), 4.12(td, J＝6.0, 1.0Hz, 2H), 3.97(qd, J＝5.5, 4.6Hz, 1H), 3.62–3.46(m, 1H), 2.89–2.67(m, 2H), 2.41 (d,J=5.3Hz,1H),2.29–1.98(m,3H),1.98–1.52(m,10H),1.52–1.25(m,12H),0.98–0.78(m,3H).

Example 6

Synthesis of Example 6

Referring to the synthesis method of Example 1, compound 6 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.61–7.39(m,2H),7.39–7.21(m,2H),7.07(t,J=7.9Hz,1H),6.87–6.68(m,2H) ,5.41(dt,J=2.9,1.0Hz,2H),5.19(t,J=1.1Hz,2H),3.97(qd,J=5.5,4.6Hz,1H),3.61–3.47(m,1H), 2.92–2.68(m,2H),2.25–1.96(m,3H),1.96–1.80(m,2H),1.77–1.49(m,4H),1.49–1.21(m,8H),1.00–0.75(m ,3H).

Example 7

With reference to the synthetic method of Example 1, Example 7 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.43–7.34(m,2H),7.09(t,J＝7.9Hz,1H),7.06–6.97(m,2H),6.81–6.72(m,2H) ,5.52–5.20(m,2H),4.67(s,2H),4.36–4.13(m,2H),3.97(qd,J＝5.5,4.6Hz,1H),3.64–3.49(m,1H),2.89 (tt,J=6.5,1.0Hz,2H),2.87–2.69(m,2H),2.22–1.97(m,3H),1.97–1.80(m,2H),1.76–1.51(m,4H),1.47 –1.24(m,8H),0.94–0.77(m,3H).

Example 8

Referring to the synthesis method of Example 1, compound 8 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.08(t, J=7.9Hz, 1H), 6.77(dd, J=8.2, 1.1Hz, 2H), 4.71–4.54(m, 3H), 4.37(t ,J=6.0Hz,2H),3.97(qd,J=5.5,4.6Hz,1H),3.62–3.47(m,1H),2.91–2.68(m,2H),2.26–1.96(m,3H), 1.96–1.77(m,6H),1.77–1.50(m,6H),1.50–1.39(m,4H),1.39–1.25(m,4H),1.07–0.67(m,6H).

Example 9

Referring to the synthesis method of Example 1, Example 9 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.08(t, J=7.9Hz, 1H), 6.83–6.60(m, 2H), 4.82–4.68(m, 2H), 4.65(s, 2H), 3.97 (qd,J=5.5,4.6Hz,1H),3.60 –3.44(m,1H),2.89–2.62(m,2H),2.29–1.99(m,3H),1.99–1.77(m,10H),1.77–1.48(m,4H),1.48–1.22(m, 8H),0.97–0.82(m,3H).

Example 10

Referring to the synthesis method of Example 1, Example 10 can be prepared. ¹ H NMR (300MHz, Chloroform-d) δ7.13–6.99(m,1H),6.85–6.70(m,2H),5.33(p,J=5.6Hz,1H),4.82–4.62(m,6H) ,3.97(qd,J＝5.5,4.6Hz,1H),3.65–3.46(m,1H),3.06–2.87(m,2H),2.87–2.69(m,2H),2.26–1.97(m,3H) ,1.97–1.78(m,2H),1.76–1.50(m,4H),1.50–1.25(m,8H),1.00–0.87(m,3H).

Example 11

Referring to the synthesis method of Example 1, Example 11 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.09(t, J=7.9Hz, 1H), 6.88–6.68(m, 2H), 4.68(d, J=1.0Hz, 2H), 4.42(t, J ＝6.6Hz, 2H), 4.35(d, J＝6.2Hz, 2H), 3.99(qd, J＝5.5, 4.6Hz, 1H), 3.60–3.49(m, 1H), 2.96(d, J＝7.2Hz ,2H),2.87–2.66(m,2H),2.38–2.16(m,2H),2.16–2.01(m,4H),2.01–1.78(m,2H),1.78–1.48(m,4H),1.48 –1.24(m,8H),1.02–0.77(m,3H).

Example 12

Referring to the synthesis method of Example 1, Example 12 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.08(t, J=7.9Hz, 1H), 6.78(dd, J=7.7, 1.4Hz, 2H), 5.28–5.01(m, 2H), 4.62(t ,J=6.2Hz,2H),3.97(qd,J=5.5,4.6Hz,1H),3.87(t,J=6.2Hz,2H),3.62–3.47(m,1H),2.87–2.63(m, 2H),2.26–2.16(m,1H),2.16–1.98(m,2H),1.96–1.79(m,2H),1.76–1.51(m,4H),1.47–1.21(m,8H),0.96– 0.80(m,3H).

Example 13

Referring to the synthesis method of Example 1, Example 13 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.08(t, J=7.9Hz, 1H), 6.85–6.70(m, 2H), 5.18–5.00(m, 2H), 4.35(t, J=6.1Hz ,2H),3.97(qd,J＝5.5,4.6Hz,1H),3.68–3.48(m,3H),2.89–2.67(m,2H),2.30–2.16(m,1H),2.16–1.96(m ,4H),1.96–1.77(m,2H),1.77–1.52(m,4H),1.49–1.24(m,8H),0.97–0.77(m,3H).

Example 14

Referring to the synthesis method of Example 1, compound 14 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.08(t, J=7.9Hz, 1H), 6.85–6.68(m, 2H), 4.33(td, J=6.0, 1.7Hz, 2H), 3.97(qd ,J＝5.5,4.6Hz,1H),3.60–3.50(m,1H),3.48–3.32(m,3H),2.91–2.67(m,2H),2.27–2.14(m,1H),2.14–1.97 (m,2H),1.97–1.75(m,6H),1.75–1.49(m,4H),1.49–1.25(m,8H),0.99–0.77(m,3H).

Example 15

Referring to the synthesis method of Example 1, Compound 15 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.08(t, J=7.9Hz, 1H), 6.77(dd, J=8.1, 1.4Hz, 2H), 4.57(t, J=6.2Hz, 2H), 4.37–4.21(m,2H),3.97(qd,J=5.5,4.6Hz,1H),3.81(t,J=6.2Hz,2H),3.72(t,J=6.1Hz,2H),3.61–3.49 (m,1H),3.06–2.91(m,2H),2.88–2.68(m,2H),2.24–1.96(m,3H),1.96–1.77(m,2H),1.76–1.52(m,4H) ,1.47–1.24(m,8H),0.98–0.82(m,3H).

Example 16

Referring to the synthesis method of Example 1, compound 16 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.08(t, J=7.9Hz, 1H), 6.89–6.70(m, 2H), 4.57(t, J=6.2Hz, 2H), 4.17(t, J ＝6.1Hz, 2H), 3.97(qd, J＝5.5, 4.6Hz, 1H), 3.76(t, J＝6.2Hz, 2H), 3.59–3.41(m, 3H), 2.87–2.67(m, 2H) ,2.25–1.79(m,7H),1.77–1.53(m,4H),1.49–1.21(m,8H),0.96–0.79(m,3H).

Example 17

Referring to the synthesis method of Example 1, compound 17 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.17–6.99(m,1H),6.89–6.68(m,2H),5.86(dd,J＝25.8,2.7Hz,2H),5.02(s,2H) ,4.70(s,2H),3.97(qd,J＝5.5,4.6Hz,1H),3.62–3.48(m,1H),2.89–2.66(m,2H),2.27–2.15(m,1H),2.15 –1.97(m,2H),1.97–1.78(m,2H),1.73–1.49(m,4H),1.48–1.24(m,8H),0.99–0.79(m,3H).

Example 18

Referring to the synthesis method of Example 1, compound 18 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.09(t, J=7.8Hz, 1H), 6.91–6.68(m, 2H), 5.93–5.63(m, 2H), 4.61(td, J=7.1, 1.8Hz, 2H), 3.97(qd, J=5.5, 4.6Hz, 1H), 3.62–3.46(m, 1H), 2.93–2.63(m, 4H), 2.27–2.15(m, 1H), 2.15–1.96 (m,2H),1.96–1.77(m,2H),1.77–1.49(m,4H),1.49–1.23(m,8H),0.98–0.70(m,3H).

Example 19

Referring to the synthesis method of Example 1, compound 19 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.09(t, J=7.8Hz, 1H), 6.90–6.73(m, 2H), 5.93–5.73(m, 2H), 4.50–4.34(m, 2H) ,3.97(qd,J＝5.5,4.6Hz,1H),3.61–3.47(m,1H),2.86–2.67(m,2H),2.61–2.38(m,3H),2.25–1.96(m,5H) ,1.96–1.77(m,2H),1.77–1.50(m,4H),1.48–1.23(m,8H),0.99–0.78(m,3H).

Example 20

Referring to the synthesis method of Example 1, compound 20 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.13–7.02(m,1H),6.85–6.71(m,2H),4.75–4.51(m,4H),4.50–4.21(m,4H),3.97( qd,J=5.5,4.6Hz,1H), 3.61–3.48(m,1H),3.06–2.88(m,2H),2.88–2.66(m,4H),2.27–1.96(m,3H),1.96–1.77(m,2H),1.77–1.48(m ,4H),1.48–1.19(m,9H),1.00–0.74(m,3H).

Example 21

Referring to the synthesis method of Example 1, compound 21 can be prepared. ¹ H NMR(300MHz,DMSO-d)δ7.07(t,J=7.9Hz,1H),6.88–6.65(m,2H),4.79–4.49(m,4H),4.31–4.04(m,4H) ,3.97(qd,J＝5.5,4.6Hz,1H),3.64–3.49(m,1H),2.87–2.66(m,4H),2.25–1.98(m,5H),1.98–1.77(m,2H) ,1.77–1.49(m,4H),1.49–1.20(m,8H),0.98–0.78(m,3H).

Example 22

Referring to the synthesis method of Example 1, compound 22 can be prepared. 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＝15.6, 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).

Example 23

Referring to the synthesis method of Example 1, compound 23 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ8.08–7.89(m,2H),7.53–7.37(m,2H),7.12(t,J=7.9Hz,1H),6.77(tt,J=6.7, 1.1Hz, 2H), 5.41(dt, J＝2.7, 1.0Hz, 2H), 4.66(s, 2H), 4.55–4.37(m, 4H), 4.04(qd, J＝5.5, 4.6Hz, 1H), 3.65–3.51(m,1H),3.05–2.88(m,2H),2.84–2.69(m,2H),2.31–2.16(m,1H),2.15–1.98(m,2H),1.97–1.79(m ,2H),1.79–1.53(m,4H),1.53–1.23(m,8H),0.94–0.80(m,3H).

Example 24

Referring to the synthesis method of Example 1, compound 24 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ8.03–7.89(m,2H),7.53–7.38(m,2H),7.11(t,J=7.9Hz,1H),6.88–6.64(m,2H) ,5.56–5.28(m,2H),4.66(s,2H),4.29–4.13(m,4H),4.04(qd,J＝5.5,4.6Hz,1H),3.60–3.50(m,1H),3.03 –2.85(m,2H),2.85–2.68(m,2H),2.26–1.96(m,5H),1.96–1.76(m,2H),1.76–1.51(m,4H),1.51–1.23(m, 8H),1.00–0.80(m,3H).

Example 25

Dissolve treprostinil (60mg), chloromethylfurazan nitrogen oxide, DMAP, and TEA in 2mL of anhydrous dichloromethane, and stir at room temperature for four hours, add 3mL of dichloromethane to the reaction solution for dilution, and successively add 10% Wash twice with hydrochloric acid and once with saturated brine, filter, concentrate the filtrate, and purify by HPLC to obtain Example 25 with a yield of 38%. ¹ H NMR (300MHz, DMSO-d) δ7.98–7.81(m,2H),7.71–7.52(m,3H),7.11(t,J=7.9Hz,1H),6.88–6.68(m,2H) ,6.12–5.92(m,2H),4.70(s,2H),4.04(qd,J＝5.5,4.6Hz,1H),3.61–3.49(m,1H),2.94(dd,J＝7.2,1.2Hz ,2H),2.86–2.66(m,2H),2.21(ddddd,J＝7.5,6.8,5.7,4.8,4.1Hz,1H),2.16–1.98(m,2H),1.98–1.78(m,2H) ,1.78–1.52(m,4H),1.52–1.22(m,8H),1.02–0.75(m,3H).

Example 26

Referring to the synthesis method of Example 25, Compound 26 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.72–7.55 (m, 5H), 7.19–7.05 (m, 1H), 6.77 (ddt, J=7.9, 3.3, 1.2Hz, 2H), 4.71–4.39 ( m,6H),4.04(qd,J＝5.5,4.6Hz,1H),3.62–3.49(m,1H),3.10–2.87(m,2H),2.87–2.67(m,2H),2.27–2.15( m,1H),2.15–1.97(m,2H),1.97–1.79(m,2H),1.79–1.53(m,4H),1.53–1.26(m,8H),0.96–0.79(m,3H).

Example 27

Referring to the synthesis method of Example 25, compound 27 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.95–7.75(m,2H),7.69–7.50(m,3H),7.10(t,J=7.9Hz,1H),6.87–6.71(m,2H) ,4.67(s,2H),4.38(t,J=6.1Hz,2H),4.20(td,J=6.1,1.1Hz,2H),4.04(qd,J=5.5,4.6Hz,1H),3.64– 3.49(m,1H),3.06–2.86(m,2H),2.86–2.67(m,2H),2.25–1.98(m,5H),1.98–1.77(m,2H),1.77–1.53(m,4H ),1.53–1.19(m,8H),0.97–0.75(m,3H).

Example 28

Referring to the synthesis method of Example 25, Compound 28 can be prepared. ¹ H NMR (300MHz, DMSO-d) δ7.96–7.77(m,2H),7.71–7.51(m,3H),7.10(t,J=7.9Hz,1H),6.88–6.71(m,2H) ,4.68(s,2H),4.32(t,J=6.0Hz,2H),4.15(t,J=6.0Hz,2H),4.04(qd,J=5.6,4.6Hz,1H),3.62–3.49( m,1H),3.05–2.88(m,2H),2.87–2.69(m,2H),2.25–1.97(m,3H),1.97–1.73(m,6H),1.73–1.52(m,4H), 1.52–1.21(m,8H),0.98–0.75(m,3H).

Test example 1

In vitro NO release test of compounds

Using the Griess method, the released NO is instantaneously oxidized in aqueous solution and called NO2 ^- , NO2 ^- and Griess reagent form a complex, which has a strong ultraviolet absorption at 540nm, so as to determine the NO release amount of the compound.

1. Solution preparation:

1) Blank solution: mix 10mL DMSO and 190mL PBS;

2) Griess reagent: sulfonamide (4.0 g), N-(1-naphthyl) ethylenediamine dihydrochloride (0.2 g) and 10 mL of 85% H ₃ PO ₄ were dissolved in 90 mL of distilled water, stirred until a clear solution;

3) L-cysteine solution: after accurately weighing L-cysteine, add a certain amount of PBS to prepare a solution weighing 200 μM;

4) Test compound solution: the test compound was accurately weighed, dissolved in DMSO and diluted to a concentration of 1 mM, and then diluted with PBS to make the concentration 200 μM.

Second, the formulation of the standard curve equation:

Use the blank solution to prepare sodium nitrite standard solution concentrations: 0, 0.78, 1.56, 3.13, 6.25, 12.5, 25, 50, 100 μmol/L, take 150 μL each time for each concentration, add 50 μL of Griess reagent and mix well, and mix well at 37 After incubating in a constant temperature shaker at ℃ for 30 minutes, measure the absorbance of each tube at 540 nm with a microplate reader, subtract the readings of the blank solution and return to the standard curve equation.

Three, test compound test:

Mix 2.5 mL each of the prepared test compound solution and L-cysteine solution, incubate in a constant temperature shaker at 37°C for 120 min, take 150 μL of the mixed solution for each 15 min, add 50 μL of Griess reagent and mix well. After incubating in a constant temperature shaker at 37°C for another 30 minutes, measure the absorbance of each tube at 540 nm with a microplate reader, subtract the readings of the blank solution and substitute the values into the standard curve equation to obtain the NO release.

Through testing, the data of some compounds of the present invention are shown in Table 1. The test results show that the nitric oxide-donating treprostinil derivatives or pharmaceutically acceptable salts thereof of the present invention have good NO releasing effects.

Table 1. Example compound NO release effect

Test example 2

In Vivo Experiment in Rats with Hypoxic Pulmonary Hypertension

(1) Experimental instruments and materials

HX-200 animal ventilator, SD male rats used in the experiment were purchased from Yangzhou University. All the control groups were reared under normal conditions, and the intervention group and the model group were reared in a hypobaric hypoxic chamber (air pressure 50kPa, oxygen concentration 10%).

(2) Experimental steps

Compound 5 was dissolved in DMSO/solutol/water (10/10/80) to make a clear solution, and the intervention group was given a dose of 5 mg/kg of compound 5 by intragastric administration from the second day of hypoxia, and all rats were weighed every week , recorded survival, and measured pulmonary artery pressure four weeks later. Rats were anesthetized with chloral hydrate (100g/L) (3mL/kg), fixed in the supine position, tracheotomy was performed, and a small animal ventilator was used to assist breathing (frequency 60 times/min, tidal volume 5mL, and respiratory ratio 4: 5). Free the third rib on the left side, send a catheter connected to a tension transducer at one end to the pulmonary artery, and record the mean pulmonary artery pressure (mPAP) through the BL-420E biological function experiment system. Examine and collect pleural fluid and ascites, and finally the rats are sacrificed by drawing blood from the abdominal aorta.

(3) Experimental results

Referring to Figure 1, compared with the control group, the mPAP of the rats in the model group was significantly increased, and the mPAP of the intervention group administered with Compound 5 was lower than that of the model group.

Test example 3, anti-platelet aggregation effect

1) Experimental materials: ADP, epinephrine, collagen (Baili Bio)

2) Experimental method: Blood samples provided by healthy subjects were mixed with 3.8% citric acid solution and centrifuged at 160 r/min to obtain platelet-rich plasma. To calibrate the data, the obtained platelet-rich plasma was further centrifuged at 2000r/min to obtain platelet-poor plasma, which was stored in a -20°C refrigerator for future use. Tests were performed using Bornl's nephelometric method. Add 225 μL of platelet-rich plasma to the cuvette, configure the compound of the example into a 50 nM solution (25 mM Tris-acetate and 120 mM NaCl), add 25 μL of the compound solution of the example, and incubate at 37°C for 2 minutes, then add 5 μL of ADP (final concentration of 2 μM) to induce platelet aggregation. In order to evaluate the degree of platelet aggregation, the maximum value of absorbance was taken after the addition of ADP in the blank group for 10 minutes, so as to calculate the inhibitory effect of each embodiment compound on the induction of ADP. Inhibition rate of platelet aggregation. From the data in Table 2 below, it can be seen that the compounds described in the Examples of the present application all have a good effect of inhibiting ADP-induced platelet aggregation.

Table 2. Example compounds inhibit ADP-induced platelet aggregation

It will be apparent to those skilled in the art that the present disclosure is not limited to the foregoing illustrative embodiments, but can be embodied in other specific forms without departing from its essential attributes. It is therefore intended to be considered in all respects as illustrative rather than restrictive, that reference is made to the appended claims rather than the foregoing embodiments, that citations are directed to the appended claims rather than the foregoing examples, and that falling within the scope of the claims All changes that come within the meaning and range of equivalents are therefore intended to be embraced herein.

All patents, patent applications and literature references cited in this specification are hereby incorporated by reference in their entirety. In case of inconsistency, the present disclosure, including definitions, will be persuasive.

Claims (9)

A coupled NO donor treprostinil derivative represented by the following formula I or a pharmaceutically acceptable salt thereof:

n is 0-6; R is -X-ONO ₂ , -OC(O)-X-ONO ₂ , -OX-ONO ₂ , or

Wherein X is straight chain or branched C ₁ -C ₁₀ alkyl, C _5-7 cycloalkyl or -C ₁ -C ₁₀ alkyl-aromatic ring-; wherein C ₁ -C ₁₀ alkyl, C _{5- 7} Cycloalkyl or aromatic ring can be substituted by one or more of the following substituents: halogen atom, hydroxyl, carboxyl, cyano or -(C ₁ -C ₁₀ alkyl)-ONO ₂ . The coupled NO donor treprostinil derivative or a pharmaceutically acceptable salt thereof according to claim 1, wherein said n is 0, 1, 2, 3 or 4. The coupled NO donor treprostinil derivative or a pharmaceutically acceptable salt thereof according to claim 1, wherein the compound comprises any one of the following structures:

A pharmaceutical composition, which contains a therapeutically effective amount of any one of claims 1-3 coupled NO donor treprostinil derivative or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. The pharmaceutical composition according to claim 4, wherein the carrier is a sustained-release agent, excipient, filler, binder, wetting agent, disintegrant, absorption promoter, adsorption carrier, surface Any one or a mixture of two or more active agents and lubricants. The pharmaceutical composition according to any one of claims 4-5, wherein the pharmaceutical composition is any one of external preparations, oral preparations and injection preparations. A kind of pharmaceutical composition according to claim 6, is characterized in that, described oral preparation is any one in granule, capsule and tablet. A coupled NO donor treprostinil derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, used as a procyclin analogue. A coupling NO donor treprostinil derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, used in the preparation of drugs for the treatment of pulmonary hypertension, respiratory distress syndrome, chronic arterial occlusive disease, etc. in the application.

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