Prodrug of treprostinil and its application

Abstract

The present invention provides a prodrug of treprostinil as shown in formula I and a pharmaceutical composition thereof, wherein the prodrug can specifically bind to prostaglandin receptors, relax vascular smooth muscle, and can be used to treat pulmonary hypertension, chronic arterial occlusive disease, idiopathic pulmonary fibrosis and other diseases. The prodrug also optimizes the pharmacokinetic characteristics of treprostinil and the irritation of the drug, reduces the dosage and frequency of administration, and improves the effectiveness and safety of the drug.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and in particular relates to a prodrug of treprostinil, a pharmaceutical composition thereof and uses thereof. Background Art

[0002] Pulmonary hypertension (including pulmonary arterial hypertension, PAH) is a type of disease characterized by increased pulmonary vascular resistance and right ventricular failure. After being diagnosed, patients have a short survival period and a high mortality rate. It is a malignant disease. Prostaglandins in the body activate adenylate cyclase by interacting with prostaglandin receptors on platelets or vascular smooth muscle, thereby increasing the concentration of intracellular cAMP and dilating blood vessels. Prostaglandins can counteract thromboxane A2 (TXA2). The synthesis of prostaglandins in PAH patients is reduced, and the concentration of TXA2 is increased, leading to vasoconstriction, endothelial cell proliferation and platelet activation (Tuder RM et al., Am J Respir Crit Care Med, 1999).

[0003] Treprostinil, as a prostaglandin analog, can specifically bind to prostaglandin receptors, relax vascular smooth muscle, reduce pulmonary artery pressure, and inhibit pulmonary artery vascular remodeling and in situ thrombosis. The drug can produce effects similar to prostacyclin (PGI2), including pulmonary vascular and circulatory vasodilation, inhibition of platelet aggregation and inhibition of smooth muscle cell proliferation (ClappLH, et al., Am J Respir Cell Mol Biol, 2002). It is also believed that the vasodilation and anti-cell proliferation effects of treprostinil are mediated by peroxisome proliferator-activated receptors (Li Y, et al., Am J Respir Cell Mol Biol, 2012). The injection preparation of the drug developed by United Therapeutics was approved by the FDA in 2002 for the treatment of pulmonary arterial hypertension PAH and is listed in the United States. In April 2021, the drug was further approved for the treatment of pulmonary arterial hypertension (PH-ILD) associated with interstitial lung disease. Compared with the shorter half-lives of epoprostenol and enoprostol, treprostinil has an elimination half-life of about 4 hours, which has a significant advantage. The main excretion route is urine (79%); the elimination is reduced by 80% in patients with liver dysfunction, while the exposure of patients with renal insufficiency after oral administration of the drug does not increase, and dialysis can reduce the exposure of the drug by 50% (Jenkins A, et al., Am J Respir Crit Care Med, 2011).

[0004] Treprostinil is the only prostacyclin drug currently used in the clinic to treat PAH that can be administered by multiple routes, subcutaneously / intravenously, orally, and by inhalation. It has a stable structure and a longer duration of action than prostacyclin. It takes effect quickly when administered by subcutaneous / intravenous routes and can exert biological effects within minutes. In clinical studies of PAH, subcutaneous, intravenous, and inhaled administration have shown good efficacy and safety. Among them, subcutaneous and intravenous preparations (treprostinil for injection, trade name: Remodulin) and inhalation solution (trade name: Tyvaso) sustained-release tablets (trade name: Orenitram) and injection (trade name: Remodulin) were approved for marketing by the FDA in 2002, 2009, 2013, and 2018, respectively.

[0005] Since long-term administration is required, injection and inhalation administration are very inconvenient. Oral administration is the best route of administration. However, the oral form has poor efficacy, systemic toxic side effects, and requires multiple administrations a day. Therefore, sustained-release tablets have been developed and marketed for clinical use. However, due to the defects of the compound itself, the current clinical use of oral preparations has not been good, and inhalation preparations are mainly used. For example, the latest summary report of the international multicenter clinical trial of inhaled treprostinil for the treatment of pulmonary hypertension was published in the New England Journal of Medicine in 2021. In order to improve patient compliance and reduce the toxic side effects of systemic administration, INSMED modified treprostinil to palmitic acid (treprostinilpalmitil) as a long-acting release prodrug inhaler (patents US09255064 and US09469600) for the treatment of pulmonary arterial hypertension, interstitial lung disease-associated pulmonary hypertension, and idiopathic fibrosis. The project has now entered the Phase II clinical trial stage.

[0006] Therefore, it is of positive clinical value to explore suitable prodrugs or dosage forms to further improve the in vivo metabolic characteristics of treprostinil, reduce the dosage and frequency of administration, and reduce the toxic side effects caused by direct exposure. Summary of the invention

[0007] The present invention aims at the problems existing in the prior art and provides a prodrug of treprostinil, which optimizes the pharmacokinetic characteristics of treprostinil, reduces the dosage and frequency of administration, and improves the effectiveness and safety of the drug.

[0008] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0009] In one aspect, the present invention provides a prodrug of treprostinil or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the prodrug structure is shown in the following formula I:

[0010]

[0011] in,

[0012] R 1 is hydrogen, alkyl, cycloalkyl, alkoxy or aryl;

[0013] R 2 is alkyl, cycloalkyl, alkoxy, alkylamino or hydroxyl;

[0014] n is an integer from 0 to 6.

[0015] In another aspect, the present invention provides a pharmaceutical composition comprising the above-mentioned prodrug of treprostinil or a pharmaceutically acceptable salt thereof.

[0016] In another aspect, the present invention provides a use of the above-mentioned prodrug of treprostinil or a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical composition as a procyclidine analog.

[0017] In another aspect, the present invention provides a use of the above-mentioned prodrug of treprostinil or its pharmaceutically acceptable salt or the above-mentioned pharmaceutical composition in the preparation of drugs for treating pulmonary hypertension, chronic arterial occlusive disease, idiopathic pulmonary fibrosis, etc.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] The series of treprostinil prodrugs provided by the present invention can specifically bind to prostaglandin receptors, relax vascular smooth muscles, and can be used to treat diseases such as pulmonary hypertension, chronic arterial occlusive disease, and idiopathic pulmonary fibrosis.

[0020] The prodrug of treprostinil provided by the present invention optimizes the pharmacokinetic characteristics of treprostinil and the irritation of the drug, reduces the dosage and frequency of administration, and improves the effectiveness and safety of the drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 These are the in vivo test results of Experimental Example 2 on rats with hypoxic pulmonary hypertension. DETAILED DESCRIPTION

[0022] The present invention is described in detail below.

[0023] In one aspect, the present invention provides a prodrug of treprostinil as shown in the following formula I:

[0024]

[0025] or a pharmaceutically acceptable salt or stereoisomer thereof;

[0026] in,

[0027] R 1is hydrogen, alkyl, cycloalkyl, alkoxy or aryl;

[0028] R 2 is alkyl, cycloalkyl, alkoxy, alkylamino or hydroxyl;

[0029] n is an integer of 0-6, for example, n can be 0, 1, 2, 3, 4, 5 or 6.

[0030] In some embodiments of the present invention, R 1 For hydrogen, C 1-20 Alkyl, C 3-20 Cycloalkyl, C 1-20 Alkoxy or C 7-20 Aryl; R 2 C 1-20 Alkyl, C 3-20 Cycloalkyl, C 1-20 Alkoxy, C 1-20 Alkylamine or hydroxyl.

[0031] In some embodiments of the present invention, R 1 is hydrogen or C 1-10 Alkyl; R 2 C 1-10 Alkyl, C 3-7 Cycloalkyl, C 1-6 Alkylamine, hydroxyl or C 1-6 Alkoxy.

[0032] In some embodiments of the present invention, R 1 is hydrogen, methyl or isopropyl; R 2 is methyl, ethyl, n-propyl, isopropyl, tert-butyl, cyclopropyl, cyclohexyl, hydroxy, methoxy, ethoxy, propoxy, methylamino, ethylamino, butylamino, 2-methylpropan-1-amine, n-hexyl, n-heptyl or n-octyl; n is 0, 1, 2 or 3.

[0033] In some embodiments of the present invention, the prodrug of treprostinil is selected from any of the following structures:

[0034]

[0035]

[0036] On the other hand, the present invention provides a pharmaceutical composition comprising the above-mentioned prodrug of treprostinil or a pharmaceutically acceptable salt or stereoisomer thereof.

[0037] In some embodiments of the present invention, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient.

[0038] The pharmaceutical excipients can be those widely used in the field of drug production. Excipients are mainly used to provide a safe, stable and functional pharmaceutical composition, and can also provide methods to dissolve the active ingredient at a desired rate after the subject receives the administration, or promote the effective absorption of the active ingredient after the subject receives the composition. The pharmaceutical excipients can be inert fillers, or provide a certain function, such as stabilizing the overall pH value of the composition or preventing the degradation of the active ingredient of the composition. The pharmaceutical excipients can include one or more of the following excipients: adhesives, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adhesive agents, glidants, wetting agents, gelling agents, absorption delay agents, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavoring agents and sweeteners.

[0039] In some embodiments of the present invention, the pharmaceutical composition is generally formulated into a dosage form suitable for oral administration, injection administration or inhalation administration to a patient.

[0040] In some embodiments of the invention, the dosage form is an oral dosage form, such as tablets, capsules, caplets, pills, lozenges, powders, syrups, elixirs, suspensions, solutions, emulsions, granules, cachets, and the like.

[0041] The pharmaceutical composition of the present invention can be prepared according to the disclosed content using any method known to those skilled in the art, such as conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding or lyophilizing processes.

[0042] In another aspect, the present invention provides a use of the above-mentioned prodrug of treprostinil or its pharmaceutically acceptable salt, stereoisomer or the above-mentioned pharmaceutical composition as a procyclidine analog.

[0043] On the other hand, the present invention provides a use of the above-mentioned prodrug of treprostinil or its pharmaceutically acceptable salt, stereoisomer or the above-mentioned pharmaceutical composition in the preparation of drugs for treating cardiovascular diseases, cerebrovascular diseases, respiratory diseases, etc., especially in drugs for pulmonary hypertension, chronic arterial occlusive disease, idiopathic pulmonary fibrosis, etc.

[0044] Definitions and general terms

[0045] The term "pharmaceutically acceptable salt" refers to organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art, as described in Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmacol Sci, 1977, 66, 1-19. Pharmaceutically acceptable salts formed by non-toxic acids include, but are not limited to, inorganic acid salts formed by reaction with amino groups, such as hydrochlorides, hydrobromides, phosphates, sulfates, perchlorates, etc.; and organic acid salts, such as acetates, oxalates, maleates, tartrates, citrates, succinates, malonates, etc., or other methods described in books and documents, such as ion exchange methods, to obtain these salts. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cyclopentylpropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, enanthate, caproate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. The salt obtained by appropriate base includes alkali metal, alkaline earth metal, ammonium salt. Alkali metal or alkaline earth metal salt includes sodium, lithium, potassium, calcium, magnesium, etc. Pharmaceutically acceptable salts further include appropriate, non-toxic ammonium, quaternary ammonium and amine cations that counter-form counterions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C1-8 sulfonates and aromatic sulfonates.

[0046] The term "stereoisomer" refers to compounds with the same chemical structure, but with different arrangements of atoms or groups in space. Stereoisomers include enantiomers, diastereomers, conformers (rotamers), geometric isomers, (cis / trans) isomers, atropisomers, and the like. "Enantiomers" refer to two isomers of a compound that are not superimposable but are mirror images of each other. "Diastereomers" refer to stereoisomers that have two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting points, boiling points, spectral properties, and reactivity. Diastereomeric mixtures can be separated by high-resolution analytical procedures such as electrophoresis and chromatography, such as HPLC.

[0047] The term "alkyl" or "alkyl group" refers to a saturated straight or branched monovalent hydrocarbon radical containing 1 to 20 carbon atoms. The alkyl group may be independently and optionally substituted with one or more substituents. Examples of alkyl or alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl isobutyl, sec-butyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, n-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, n-heptyl, n-octyl, etc.

[0048] The term "alkoxy" means an alkyl group attached to the rest of the molecule via an oxygen atom, wherein the alkyl group has a meaning as described herein. Examples of alkoxy include, but are not limited to, methoxy, ethoxy, 1-propoxy, 2-propoxy, 1-butoxy, 2-methyl-1-propoxy, 2-butoxy, 2-methyl-2-propoxy, 1-pentoxy, 2-pentoxy, 3-pentoxy, 2-methyl-2-butoxy, 3-methyl-2-butoxy, 3-methyl-1-butoxy, 2-methyl-1-butoxy, and the like.

[0049] The term "aryl" refers to a monocyclic, bicyclic or tricyclic carbon ring system containing 6-20 ring members, wherein at least one ring system is aromatic, wherein each ring system contains 3-7 ring members and has only one point of attachment to the rest of the molecule. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, anthracene, and the like.

[0050] The term "cycloalkyl" refers to a C3-C20 cycloaliphatic ring structure; examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.

[0051] The term "alkylamino" denotes a group -NHR or -NRR, wherein R represents an alkyl or cycloalkyl group as defined above.

[0052] The term "hydroxy" refers to -OH.

[0053] The technical scheme of the present invention will be clearly and completely described below in conjunction with specific embodiments. Obviously, the described embodiments are only a part of embodiments of the present invention, rather than all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work, all belong to the scope of protection of the present invention. In addition, it is worth noting that the raw materials involved in the present invention are all common commercial products unless otherwise specified.

[0054] Example 1

[0055]

[0056] Synthesis route:

[0057]

[0058] Synthesis of compound 1

[0059] Treprostinil (20 mg) protected by O-TES, potassium iodide (25 mg) and potassium carbonate (21 mg) were dissolved in 10 mL of acetonitrile, stirred at room temperature for 10 min, and then chloromethyl acetate (15 mg) was slowly added. The mixture was reacted at 80° C. for 8 h, and the reaction was stopped. The solvent was dried by spin drying, dichloromethane was added, and the mixture was washed twice with water and once with saturated brine. The organic phase was concentrated, purified by column chromatography, concentrated, dissolved in THF / water, and 2N HCl was added. The mixture was stirred at room temperature for reaction. The reaction was complete when detected by TLC. The solvent was dried by spin drying, and purified by HPLC to obtain Example 1 with a yield of 42%. 1 HNMR(300MHz,DMSO-d)δ7.13(t,J=7.5Hz,1H),7.02–6.96(m,1H),6.88(dd,J=7.3,2.0Hz,1H),5.35– 5.12(m,2H),4.75(s,2H),3.70(d,J=7.7Hz,2H),3.06(d,J=12.8Hz,2H),2.90–2.63(m,2H),2.60–2. 37(m,2H),2.12(s,3H),2.09–1.96(m,2H),1.78–1.67(m,2H),1.61(dd,J=12.4,1.9Hz,2H),1.59–1. 39(m,4H),1.39–1.27(m,4H),1.11(d,J=12.5Hz,1H),0.86(t,J=5.3Hz,3H).ESI-MSm / z:463.6[M+H] + .

[0060] Example 2

[0061]

[0062] Referring to the synthesis method of Example 1, Compound 2 can be prepared by replacing chloromethyl acetate with methyl chloropropionate. 1HNMR(300MHz,DMSO-d)δ7.11(t,J=7.5Hz,1H),7.00–6.85(m,2H),5.45–5.27(m,2H),4.75(s,2H),3.73(d,J=7.9 Hz,2H),3.01(d,J=11.8Hz,2H),2.87–2.65(m,2H),2.56(d,J=13.0Hz,1H),2.35(s,2H),2.08–1.92(m,3H),1.77–

[0063] 1.58(m,4H),1.58–1.37(m,4H),1.37–1.27(m,4H),1.17–1.03(m,4H),0.90(t,J=5.2Hz,3H).ESI-MS m / z:477.3[M+H] + .

[0064] Example 3

[0065]

[0066] Referring to the synthesis method of Example 1, Compound 3 can be prepared by replacing chloromethyl acetate with ethyl 1-chloroacetate. 1 HNMR(300MHz,DMSO-d)δ7.22(t,J=7.2Hz,1H),6.97(dd,J=7.5,2.0Hz,2H),5.82–5.77(m, 1H),4.68(d,J=1.8Hz,2H),3.77(d,J=4.9Hz,1H),3.63(d,J=4.9Hz,1H),3.06(d,J=12.8H z,1H),2.85–2.65(m,2H),2.56(d,J=13.0Hz,2H),2.14–1.93(m,5H),1.79–1.59(m,7H),1 .59–1.37(m,4H),1.37–1.26(m,4H),1.11(d,J=12.5Hz,1H),0.91(t,J=5.3Hz,3H).ESI-MS m / z:477.3[M+H] + .

[0067] Example 4

[0068]

[0069] Referring to the synthesis method of Example 1, Compound 4 can be prepared by replacing chloromethyl acetate with ethyl 1-chloropropionate. 1HNMR(300MHz,DMSO-d)δ7.07(t,J=7.7Hz,1H),6.99–6.89(m,2H),5.84–5.79(m,1H),4 .04(d,J=5.1Hz,1H),3.77(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.85–2.62(m,2H), 2.56(d,J=13.0Hz,1H),2.38(d,J=27.7Hz,3H),2.05–1.93(m,4H),1.80–1.57(m,7H), 1.56–1.39(m,4H),1.39–1.26(m,4H),1.23–1.03(m,4H),0.91(t,J=5.3Hz,3H).ESI-MS m / z:513.3[M+Na] + .

[0070] Example 5

[0071]

[0072] Referring to the synthesis method of Example 1, Compound 5 can be prepared by replacing chloromethyl acetate with chloromethyl isobutyrate. 1 HNMR(300MHz,DMSO-d)δ7.14(t,J=7.2Hz,1H),6.97–6.85(m,2H),4.92–4.61(m,4H),4.18(d,J=5.1Hz,1H),3 .77(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.85–2.66(m,2H),2.63–2.47(m,2H),2.08–1.90(m,4H),1.81–

[0073] 1.58(m,4H),1.58–1.38(m,4H),1.38–1.27(m,4H),1.22–1.08(m,7H),0.86(t,J=5.2Hz,3H).ESI-MS m / z:491.3[M+H] + .

[0074] Example 6

[0075]

[0076] Referring to the synthesis method of Example 1, Compound 6 can be prepared by replacing chloromethyl acetate with chloromethyl pivalate. 1HNMR(300MHz,DMSO-d)δ7.15(t,J=7.2Hz,1H),7.01(t,J=7.5Hz,1H),6.89(dd,J=7.3,2.0Hz,1H),5.25 –5.04(m,2H),4.86–4.73(m,2H),4.04(d,J=5.1Hz,1H),3.63(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2 .84–2.66(m,2H),2.56(d,J=13.0Hz,1H),2.07–1.93(m,2H),1.86–1.67(m,4H),1.61(dd,J=12.4,1.9Hz ,2H),1.58–1.37(m,4H),1.31(dd,J=3.8,0.8Hz,4H),1.13–1.07(m,10H),0.89(t,J=5.5Hz,3H).ESI-MS m / z:505.3[M+H] + .

[0077] Example 7

[0078]

[0079] Referring to the synthesis method of Example 1, Compound 7 can be prepared by replacing chloromethyl acetate with methyl chlorobutyrate. 1 HNMR(300MHz,DMSO-d)δ7.17(t,J=7.2Hz,1H),7.02–6.91(m,2H),5.29–5.02(m,2H),4.75–4.66(m, 2H),4.18(d,J=5.1Hz,1H),3.77(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.86–2.68(m,2H),2.56(d ,J=13.0Hz,1H),2.46–2.28(m,3H),2.08–1.92(m,4H),1.78–1.59(m,6H),1.58–1.48(m,2H),1.48– 1.38(m,2H),1.38–1.25(m,4H),1.11(d,J=12.5Hz,1H),1.04–0.84(m,6H).ESI-MSm / z:513.3[M+Na] + .

[0080] Example 8

[0081]

[0082] Referring to the synthesis method of Example 1, Compound 8 can be prepared by replacing chloromethyl acetate with ethyl 1-chlorobutyrate.1 HNMR(300MHz,DMSO-d)δ7.15(t,J=7.3Hz,1H),7.01–6.91(m,2H),5.81–5.74(m,1H),4.79–4 .59(m,2H),4.15(d,J=5.1Hz,1H),3.77(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.88–2.63( m,2H),2.56(d,J=13.0Hz,1H),2.36–2.20(m,3H),2.08–1.94(m,4H),1.82–1.57(m,9H),1.5 7–1.39(m,4H),1.31(dd,J=3.8,0.8Hz,4H),1.14(d,J=9.5Hz,1H),1.01–0.84(m,6H).ESI-MS m / z:503.3[M+H] + .

[0083] Example 9

[0084]

[0085] Referring to the synthesis method of Example 1, Compound 9 can be prepared by replacing chloromethyl acetate with 1-chloroethyl isobutyrate. 1 H NMR(300MHz,DMSO-d)δ7.13(t,J=7.2Hz,1H),7.03–6.92(m,2H),5.83–5.74(m,1H),4. 79–4.60(m,2H),4.19(d,J=5.2Hz,1H),3.63(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H), 2.84–2.67(m,2H),2.61–2.46(m,2H),2.12–1.94(m,4H),1.82–1.57(m,7H),1.57–1.3 7(m,4H),1.31(dd,J=3.8,0.8Hz,4H),1.25–1.06(m,7H),0.89(t,J=5.3Hz,3H).ESI-MS m / z:505.3[M+H] + .

[0086] Example 10

[0087]

[0088] Referring to the synthesis method of Example 1, compound 10 can be prepared by replacing chloromethyl acetate with 1-chloroethyl pivalate. 1HNMR(300MHz,DMSO-d)δ7.15(t,J=7.0Hz,1H),7.03–6.85(m,2H),5.86–5.74(m,1H),4. 78–4.54(m,2H),4.04(d,J=5.1Hz,1H),3.77(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2. 85–2.66(m,2H),2.56(d,J=13.0Hz,1H),2.05–1.93(m,4H),1.82–1.57(m,7H),1.57–1.3 6(m,4H),1.31(dd,J=3.8,0.8Hz,4H),1.13–1.03(m,10H),0.89(t,J=5.3Hz,3H).ESI-MS m / z:541.3[M+

[0089] Na] + .

[0090] Embodiment 11

[0091]

[0092] Referring to the synthesis method of Example 1, compound 11 can be prepared by replacing chloromethyl acetate with 1-chloroethyl cyclohexanecarboxylate. 1 HNMR(300MHz,DMSO-d)δ7.08(dd,J=7.8,7.2Hz,1H),7.02–6.84(m,2H),5.77–5.64(m,1H),4.80 –4.56(m,2H),4.15(d,J=5.1Hz,1H),3.77(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.83–2.65( m,2H),2.56(d,J=13.0Hz,1H),2.39(d,J=17.9Hz,2H),2.07(d,J=13.0Hz,2H),2.01–1.87(m,4H ),1.84–1.67(m,7H),1.67–1.26(m,16H),1.11(d,J=12.5Hz,1H),0.92(t,J=5.3Hz,3H).ESI-MS m / z:545.4[M+H] + .

[0093] Example 12

[0094]

[0095] Referring to the synthesis method of Example 1, Compound 12 can be prepared by replacing chloromethyl acetate with 1-chloro-2-methylpropyl isobutyrate.1 H NMR(300MHz,DMSO-d)δ7.15(t,J=7.5Hz,1H),7.04–6.86(m,2H),5.65–5.47(m,1H),4.68(d ,J=1.8Hz,2H),4.04(d,J=5.1Hz,1H),3.77(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.84– 2.66(m,2H),2.65–2.47(m,2H),2.19(d,J=29.5Hz,2H),2.09–1.90(m,4H),1.84–1.67(m,2 H),1.67–1.38(m,6H),1.36–1.19(m,10H),1.19–0.99(m,7H),0.94(t,J=5.3Hz,3H).ESI-MS m / z:533.3[M+H] + .

[0096] Example 13

[0097]

[0098] Synthetic route

[0099]

[0100] Synthesis of compound 13

[0101] Methylamine (2mmol) was dissolved in dichloromethane. At 0℃, N,N,N',N'-tetramethyl-1,8-naphthalenediamine (2.2mmol) was added. After stirring for 2min, chloromethyl chloroformate (2.2mmol) was added dropwise. The reaction was allowed to react at room temperature overnight. TLC detected that the reaction was complete. The mixture was washed with water twice, washed with saturated brine once, dried over anhydrous sodium sulfate, and the solvent was dried by spin drying. The above-prepared compound (0.2mmol) and O-TES-protected treprostinil (0.1mmol) and DIPEA (0.2mmol) were dissolved in dichloromethane. After stirring at room temperature for two hours, the reaction was complete by TLC detection. The mixture was washed with water twice, washed with saturated brine once, dried over anhydrous sodium sulfate, concentrated by column chromatography, dissolved in THF / water, added with 2N HCl, stirred at room temperature, the reaction was complete by TLC detection, the solvent was dried by spin drying, and purified by HPLC to obtain compound 13 with a yield of 51%. 1H NMR(300MHz,DMSO-d)δ7.13(t,J=7.5Hz,1H),7.04–6.87(m,2H),5.29(d,J=2.0Hz,2H),4.7 3–4.61(m,2H),4.18(d,J=5.1Hz,1H),3.63(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.87–2 .65(m,5H),2.56(d,J=13.0Hz,1H),2.07–1.93(m,4H),1.76–1.58(m,4H),1.58–1.37(m,4H ),1.36–1.25(m,4H),1.11(d,J=12.5Hz,1H),0.95(t,J=5.3Hz,3H).ESI-MSm / z:478.3[M+H] + .

[0102] Embodiment 14

[0103]

[0104] Referring to the synthesis method of Example 13, Compound 14 can be prepared. 1 H NMR(300MHz,DMSO-d)δ7.15(t,J=7.2Hz,1H),7.04–6.91(m,2H),5.30(d,J=2.0Hz,2H),4.73–4.66 (m,2H),4.04(d,J=5.1Hz,1H),3.77(d,J=4.9Hz,1H),3.26–3.13(m,2H),3.06(d,J=12.8Hz,1H),2 .87–2.65(m,2H),2.56(d,J=13.0Hz,1H),2.07–1.90(m,4H),1.78–1.66(m,2H),1.61(dd,J=12.4, 1.9Hz,2H),1.57–1.38(m,4H),1.38–1.26(m,4H),1.17–1.04(m,4H),0.96(t,J=5.3Hz,3H).ESI-MS m / z:492.3[M+H] + .

[0105] Embodiment 15

[0106]

[0107] Referring to the synthesis method of Example 13, Compound 15 can be prepared. 1H NMR(300MHz,DMSO-d)δ7.14(t,J=7.0Hz,1H),7.03–6.88(m,2H),5.30(d,J=1.1Hz,2H),4 .82–4.66(m,2H),4.15(d,J=5.1Hz,1H),3.63(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2 .85–2.65(m,2H),2.56(d,J=13.0Hz,1H),2.10–1.91(m,2H),1.84–1.69(m,2H),1.68–1. 37(m,8H),1.31(dd,J=3.8,0.8Hz,4H),1.11(d,J=12.5Hz,1H),1.04–0.89(m,6H).ESI-MS m / z:506.3[M+H] + .

[0108] Example 16

[0109]

[0110] Referring to the synthesis method of Example 13, Compound 16 can be prepared. 1 H NMR(300MHz,DMSO-d)δ7.14(t,J=7.2Hz,1H),7.00–6.80(m,2H),5.30(d,J=1.0Hz,2H),4.83– 4.71(m,2H),3.77(d,J=4.9Hz,1H),3.69–3.55(m,2H),3.31(d,J=12.3Hz,1H),3.06(d,J=12.8 Hz,1H),2.85–2.65(m,2H),2.56(d,J=13.0Hz,1H),2.09–1.90(m,4H),1.83–1.68(m,2H),1.6 7–1.37(m,6H),1.31(dd,J=3.8,0.8Hz,4H),1.11(d,J=12.5Hz,1H),1.00–0.78(m,9H).ESI-MS m / z:520.3[M+H] + .

[0111] Embodiment 17

[0112]

[0113] Synthetic route

[0114]

[0115] O-TES protected treprostinil (0.1 mmol), 1,2-ethylene glycol monoacetate (0.11 mmol), DCC (0.2 mmol) and DMAP (0.02 mmol) were dissolved in dichloromethane and reacted overnight. The reaction was complete when detected by TLC. The solvent was dried and concentrated by column chromatography, dissolved in THF / water, and 2N HCl was added. The reaction was stirred at room temperature. The reaction was complete when detected by TLC. The solvent was dried and purified by HPLC to obtain compound 17. 1 H NMR(300MHz,DMSO-d)δ7.09(t,J=7.3Hz,1H),7.05–6.91(m,2H),4.78(s,2H),4 .39–4.13(m,5H),3.77(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.84–2.65(m, 2H),2.56(d,J=13.0Hz,1H),2.09–1.90(m,7H),1.75–1.57(m,4H),1.57–1.37( m,4H),1.37–1.27(m,4H),1.11(d,J=12.5Hz,1H),0.94(t,J=5.3Hz,3H).ESI-MS m / z:477.3[M+H] + .

[0116] Embodiment 18

[0117]

[0118] Referring to the synthesis method of Example 17, Compound 18 can be prepared. 1 H NMR(300MHz,DMSO-d)δ7.10(t,J=7.4Hz,1H),7.01–6.89(m,2H),4.77(d,J=2.2Hz,2 H),4.27–4.11(m,5H),3.77(d,J=4.9Hz,1H),3.63(d,J=4.9Hz,1H),3.06(d,J=12.8 Hz,1H),2.84–2.65(m,2H),2.56(d,J=13.0Hz,1H),2.14(d,J=1.4Hz,3H),2.09–1.9 1(m,7H),1.78–1.25(m,12H),1.11(d,J=12.5Hz,1H),0.99(t,J=5.3Hz,3H).ESI-MS m / z:513.3[M+Na] + .

[0119] Embodiment 19

[0120]

[0121] Referring to the synthesis method of Example 17, Compound 19 can be prepared. 1 H NMR(300MHz, DMSO-d)δ7.12(t,J=7.5Hz,1H),7.05–6.90(m,2H),4.87–4.61(m,2H),4.21–4.11(m, 5H),3.77(d,J=4.9Hz,1H),3.63(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.84–2.66(m,2H),2.56 (d,J=13.0Hz,1H),2.05–1.95(m,7H),1.89(d,J=0.7Hz,4H),1.84–1.67(m,2H),1.67–1.39(m,6H) ,1.31(dd,J=3.8,0.8Hz,4H),1.14(d,J=9.4Hz,1H),0.89(t,J=5.8Hz,3H).ESI-MSm / z:505.3[M+H] + .

[0122] Embodiment 20

[0123]

[0124] Referring to the synthesis method of Example 17, Compound 20 can be prepared. 1 H NMR(300MHz,DMSO-d)δ7.14(t,J=7.2Hz,1H),7.04–6.88(m,2H),4.77(s,2H),4.38–4.14( m,5H),4.04(d,J=5.1Hz,1H),3.77(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.86–2.65(m ,2H),2.56(d,J=13.0Hz,1H),2.39(d,J=16.5Hz,3H),2.08–1.94(m,4H),1.79–1.57(m,4H ),1.57–1.37(m,4H),1.37–1.26(m,4H),1.22–1.06(m,4H),0.95(t,J=5.8Hz,3H).ESI-MS m / z:513.3[M+Na] + .

[0125] Embodiment 21

[0126]

[0127] Referring to the synthesis method of Example 17, Compound 21 can be prepared. 1H NMR(300MHz,DMSO-d)δ7.11(t,J=7.4Hz,1H),7.03–6.90(m,2H),4.79(s,2H),4.39–4 .14(m,5H),4.04(d,J=5.1Hz,1H),3.63(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.8 4–2.64(m,2H),2.63–2.48(m,2H),2.09–1.91(m,4H),1.85–1.67(m,2H),1.67–1.37( m,6H),1.31(dd,J=3.8,0.8Hz,4H),1.28–1.06(m,7H),0.97(t,J=5.4Hz,3H).ESI-MS m / z:505.3[M+H] + .

[0128] Embodiment 22

[0129]

[0130] Referring to the synthesis method of Example 17, Compound 22 can be prepared. 1 H NMR(300MHz,DMSO-d)δ7.15(t,J=7.2Hz,1H),7.04–6.92(m,2H),4.86(s,2H),4.28–4.10( m,5H),4.04(d,J=5.1Hz,1H),3.77(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.86–2.67(m ,2H),2.56(d,J=13.0Hz,1H),2.39(d,J=16.7Hz,3H),2.25–2.10(m,3H),2.09–1.90(m,4H ),1.83–1.69(m,2H),1.67–1.26(m,10H),1.18–1.03(m,4H),0.94(t,J=5.8Hz,3H).ESI-MS m / z:505.3[M+H] + .

[0131] Embodiment 23

[0132]

[0133] Referring to the synthesis method of Example 23, Compound 23 can be prepared. 1H NMR(300MHz,DMSO-d)δ7.07(dd,J=7.9,7.1Hz,1H),7.01–6.90(m,2H),5.03–4.86(m,2H) ),4.73(s,2H),4.29(d,J=4.9Hz,1H),4.07(d,J=2.5Hz,2H),3.69–3.49(m,3H),3.06(d ,J=12.8Hz,1H),2.86–2.67(m,2H),2.65–2.46(m,2H),2.23–2.02(m,4H),1.87(s,2H), 1.82–1.68(m,4H),1.66–1.38(m,6H),1.37–1.04(m,11H),0.94(t,J=5.8Hz,3H).ESI-MS m / z:563.4[M+H] + .

[0134] Embodiment 24

[0135]

[0136] Referring to the synthesis method of Example 13, Compound 24 can be prepared. 1 H NMR(300MHz,DMSO-d)δ7.15(t,J=7.5Hz,1H),6.99(t,J=7.2Hz,1H),6.90(d,J=7.3Hz, 1H),5.82(s,1H),4.80–4.52(m,2H),4.18(d,J=5.1Hz,1H),3.63(d,J=4.9Hz,1H),3.06 (d,J=12.8Hz,1H),2.85–2.62(m,5H),2.56(d,J=13.0Hz,1H),2.06–1.91(m,4H),1.80 –1.38(m,11H),1.38–1.25(m,4H),1.13(d,J=7.5Hz,1H),0.95(t,J=5.4Hz,3H).ESI-MS m / z:514.3[M+Na] + .

[0137] Embodiment 25

[0138]

[0139] Referring to the synthesis method of Example 17, Compound 25 can be prepared. 1H NMR(300MHz,DMSO-d)δ7.09(t,J=7.4Hz,1H),7.04–6.90(m,2H),4.77(s,2H),4.32(d ,J=12.4Hz,1H),4.27–4.12(m,4H),4.04(d,J=5.1Hz,1H),3.77(d,J=4.9Hz,1H),3.06 (d,J=12.8Hz,1H),2.88–2.65(m,5H),2.56(d,J=13.0Hz,1H),2.11–1.92(m,4H),1.80 –1.57(m,4H),1.57–1.27(m,8H),1.16(d,J=9.5Hz,1H),0.93(t,J=5.8Hz,3H).ESI-MS m / z:514.2[M+Na] + .

[0140] Embodiment 26

[0141]

[0142] Referring to the synthesis method of Example 17, Compound 26 can be prepared. 1 H NMR(300MHz,DMSO-d)δ7.07(t,J=7.2Hz,1H),7.04–6.91(m,2H),4.88–4.70(m,2H),4.30–4.1 2(m,5H),4.04(d,J=5.1Hz,1H),3.63(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.88–2.65(m, 5H),2.56(d,J=13.0Hz,1H),2.23–2.08(m,3H),2.06–1.90(m,4H),1.84–1.70(m,2H),1.67–1 .37(m,6H),1.31(dd,J=3.8,0.8Hz,4H),1.14(d,J=7.4Hz,1H),0.89(t,J=5.8Hz,3H).ESI-MS m / z:528.3[M+Na] + .

[0143] Embodiment 27

[0144]

[0145] Referring to the synthesis method of Example 17, Compound 27 can be prepared. 1H NMR(300MHz,DMSO-d)δ7.07(t,J=7.3Hz,1H),7.04–6.90(m,2H),4.87(s,2H),4.15(d,J =4.4Hz,5H),4.04(d,J=5.1Hz,1H),3.63(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.86–

[0146] 2.65(m,5H),2.56(d,J=13.0Hz,1H),2.04–1.96(m,4H),1.89(d,J=4.2Hz,4H),1.83–1.69(m,2H),1 .67–1.37(m,6H),1.31(dd,J=3.8,0.8Hz,4H),1.12(d,J=10.5Hz,1H),0.92(t,J=5.5Hz,3H).ESI-MS m / z:520.3[M+H] + .

[0147] Embodiment 28

[0148]

[0149] Referring to the synthesis method of Example 1, compound 28 can be prepared. 1 H NMR(300MHz,DMSO-d)δ7.11(t,J=7.2Hz,1H),7.04–6.76(m,2H),5.28–5.00(m,2H),4.74 (d,J=2.4Hz,2H),4.15(d,J=5.1Hz,1H),3.77(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2 .87–2.64(m,2H),2.56(d,J=13.0Hz,1H),2.39(d,J=18.8Hz,3H),2.09–1.92(m,4H),1.8 4–1.69(m,2H),1.69–1.22(m,18H),1.14(d,J=11.4Hz,1H),0.97(t,J=5.9Hz,6H).ESI-MS m / z:555.3[M+Na] + .

[0150] Embodiment 29

[0151]

[0152] Referring to the synthesis method of Example 1, compound 29 can be prepared. 1H NMR(300MHz,DMSO-d)δ7.17–7.05(m,1H),7.05–6.86(m,2H),5.31–4.97(m,2H),4.74 (d,J=2.4Hz,2H),4.15(d,J=5.1Hz,1H),3.63(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H) ,2.85–2.65(m,2H),2.56(d,J=13.0Hz,1H),2.47–2.28(m,3H),2.08–1.93(m,4H),1. 86–1.67(m,2H),1.66–1.22(m,20H),1.15(d,J=8.2Hz,1H),1.08–0.93(m,6H).ESI-MS m / z:569.3[M+Na] + .

[0153] Embodiment 30

[0154]

[0155] Referring to the synthesis method of Example 1, compound 30 can be prepared. 1 H NMR(300MHz,DMSO-d)δ7.08(dd,J=7.8,7.2Hz,1H),7.01–6.89(m,2H),5.28–5.00(m,2H),4.7 4(d,J=2.4Hz,2H),4.29(d,J=4.9Hz,1H),3.63(d,J=4.9Hz,1H),3.06(d,J=12.8Hz,1H),2.82– 2.65(m,2H),2.56(d,J=13.0Hz,1H),2.45–2.26(m,3H),2.07(d,J=13.0Hz,2H),1.84–1.64(m ,3H),1.64–1.34(m,8H),1.34–1.20(m,14H),1.17(d,J=5.2Hz,1H),1.04–0.94(m,6H).ESI-MS m / z:561.3[M+H] + .

[0156] Test Example 1: Pharmacokinetics of the compounds of the present application

[0157] 1. Experimental instruments and materials

[0158] High-speed refrigerated centrifuge, vortex oscillator (Vortex Genius3), high-speed centrifuge (Eppendorf5415D), disposable syringes, pipettes (Eppendorf), EDTA-K2 vacuum blood collection tubes, and normal saline.

[0159] The SD male rats used in the experiment were purchased from Yangzhou University. All rats in the oral group were fasted for 12 h before administration and had free access to water and food during the administration period.

[0160] (II) Experimental steps

[0161] The compound was dissolved in DMSO / solutol / water (10 / 10 / 80) to prepare a clear solution. The compound was administered orally at a dose of 25 mg / kg, and the compound was administered by tail vein at a dose of 5 mg / kg. At 2 min, 10 min, 30 min, 1 h, 2 h, 3 h, 5 h, 8 h, 12 h, 16 h, and 24 h after tail vein administration, 0.5 mL of blood was continuously collected from the venous plexus of the fundus into a heparin tube. At 5 min, 15 min, 30 min, 1 h, 2 h, 3 h, 5 h, 8 h, 12 h, 16 h, and 24 h after oral administration, 0.5 mL of blood was continuously collected from the venous plexus of the fundus into a heparin tube. After the sample was centrifuged at 8000 r and 4 ° C for 10 min, 0.15 mL of the upper plasma was collected and stored at -20 ° C, and then analyzed by LC-MS / MS. The data were analyzed by WinNolin non-compartmental model to obtain key pharmacokinetic parameters.

[0162] (III) Experimental results

[0163] Table 1 Pharmacokinetic parameters of the compounds in the examples

[0164]

[0165] From the pharmacokinetic data of the example compounds, it can be seen that the bioavailability of the example compounds of the present invention is significantly improved compared with the oral bioavailability of treprostinil of 17% (Chinese Journal of New Drugs 2014, 23, 2585-2589), half-life of 1h and peak time of 0.5h. The compounds of the present invention have excellent oral administration potential, and the peak time and half-life are also significantly improved.

[0166] Test Example 2: In vivo test in rats with hypoxic pulmonary hypertension

[0167] 1. Experimental instruments and materials

[0168] HX-200 animal ventilator. SD male rats used in the experiment were purchased from Yangzhou University, and physiological saline was used. All control groups were raised in normal environment, and the intervention group and model group were raised in a low-pressure hypoxic chamber (air pressure 50 kPa, oxygen concentration 10%).

[0169] (II) Experimental steps

[0170] Compounds 8, 13, 19 and 25 were dissolved in DMSO / solutol / water (10 / 10 / 80) to prepare clear solutions. The intervention group was gavaged with compounds 8, 13, 19 and 25 at a dose of 5 mg / kg starting from the second day of hypoxia. All rats were weighed weekly, and survival was recorded. Pulmonary artery pressure was measured four weeks later. Rats were anesthetized with chloral hydrate (100 g / L) (3 mL / kg), fixed in the supine position, tracheotomy, and assisted breathing with a small animal ventilator (frequency 60 times / min, tidal volume 5 mL, respiratory ratio 4:5). The third rib on the left side was freed, and a catheter with a tension transducer at one end was sent to the pulmonary artery, and the mean pulmonary artery pressure (mPAP) was recorded by the BL-420E biofunctional experimental system. Pleural effusion and ascites were examined and collected, and finally blood was drawn from the abdominal aorta to kill the rats.

[0171] (III) Experimental results

[0172] See attached Figure 1 Compared with the control group, the mPAP of the model group rats was significantly increased, and the mPAP of the intervention groups treated with compounds 8, 13, 19 and 25 was lower than that of the model group.

[0173] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A prodrug of treprostinil as shown in the following formula I or a pharmaceutically acceptable salt or stereoisomer thereof: in, R1 is hydrogen, methyl or isopropyl; R2 is methyl, ethyl, n-propyl, isopropyl, tert-butyl, cyclopropyl, cyclohexyl, hydroxy, methoxy, ethoxy, propoxy, methylamino, ethylamino, butylamino, 2-methylpropan-1-amine, n-hexyl, n-heptyl or n-octyl; n is 0, 1, 2 or 3.

2. The prodrug of treprostinil or its pharmaceutically acceptable salt or stereoisomer according to claim 1, characterized in that: The compound of formula I is selected from any one of the following structures:

3. A pharmaceutical composition comprising the prodrug of treprostinil according to any one of claims 1 to 2 or a pharmaceutically acceptable salt, stereoisomer thereof and a pharmaceutically acceptable excipient.

4. The pharmaceutical composition according to claim 3, characterized in that The pharmaceutical composition is in the form of oral administration, injection administration or inhalation administration.

5. The pharmaceutical composition according to claim 4, characterized in that: The pharmaceutical composition is in the form of an oral administration, and the dosage form includes tablets, capsules, caplets, pills, lozenges, powders, syrups, elixirs, suspensions, solutions, emulsions, granules or cachets.

6. Use of the prodrug of treprostinil according to any one of claims 1 to 2 or a pharmaceutically acceptable salt, stereoisomer thereof or the pharmaceutical composition according to any one of claims 3 to 5 as a procyclidine analog.

7. Use of the prodrug of treprostinil according to any one of claims 1 to 2 or its pharmaceutically acceptable salt, stereoisomer or the pharmaceutical composition according to any one of claims 3 to 5 in the preparation of a drug for treating pulmonary hypertension, chronic arterial occlusive disease and idiopathic pulmonary fibrosis.

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