Method for preventing or treating pulmonary arterial hypertension with 7-dehydrocholesterol or salt, oxide, metabolite or derivative thereof

Abstract

The present disclosure provides a method for preventing or treating pulmonary arterial hypertension in a subject in need thereof, the method comprising administering to the subject an effective amount of 7-dehydrocholesterol, a salt, oxide, metabolite, or derivative thereof.

Description

Technical Field

[0001] This disclosure relates to a method for preventing or treating pulmonary hypertension, particularly a method for preventing or treating pulmonary hypertension by using 7-dehydrocholesterol or its salts, oxides, metabolites or derivatives. Background Technology

[0002] Pulmonary arterial hypertension (PAH) is a devastating pulmonary vascular disease characterized by a persistently elevated mean arterial pressure in the pulmonary vessels (≥25 mmHg at rest) and a normal pulmonary capillary wedge pressure not exceeding 15 mmHg, accompanied by increased vascularity. This progressively leads to right heart failure and even death. 1,2 Pulmonary hypertension can be primary, hereditary, drug- or toxin-induced, or associated with other conditions, including connective tissue diseases, HIV infection, portal hypertension, congenital heart disease, and schistosomiasis. 3 However, the pathogenesis of pulmonary hypertension is still not fully understood.

[0003] Early pathological changes in pulmonary hypertension include vascular endothelial fibrosis, smooth muscle cell proliferation, and peripheral pulmonary artery obstruction. 4 Pleural lesions are a pathological change in advanced severe pulmonary hypertension, originating from the proliferation of pulmonary artery endothelial cells, smooth muscle cells, and circulating cells. 5,6 Related genetic studies have shown that bone morphogenetic protein receptor type 2 (BMPR2) is an important gene affecting more than 70% of patients with familial pulmonary arterial hypertension (FPAH) and 10 to 20% of patients with idiopathic pulmonary arterial hypertension (IPAH). 7-10 The new classification system has reclassified these patients as having hereditary pulmonary arterial hypertension (HPAH).

[0004] BMPR2 is a member of the superfamily of transforming growth factor-β (TGF-β) receptors. TGF-β is a multifunctional cytokine involved in cell growth, differentiation, apoptosis, angiogenesis, wound healing, neuroprotection, and immune regulation. Pathologies associated with TGF-β include immunosuppression, inflammation, arteriosclerosis, neurodegeneration, tissue fibrosis, and cancer development. Promoting or inhibiting TGF-β has become a target for many new drug development efforts.

[0005] 7-Dehydrocholesterol (7-DHC) is a TGF-β receptor inhibitor. As disclosed in U.S. Patent No. 8,946,201, a certain amount of oxidized 7-DHC can effectively inhibit the activity of TGF-β in the body of a subject, thereby treating and / or preventing skin conditions such as skin fibrosis, skin wounds, inflammation, and alopecia. PCT Patent Publication No. WO2009 / 138582 also discloses that compositions containing 7-DHC, its derivatives, or natural extracts of plants or animal microorganisms containing 7-DHC can be used as cosmetics or food additives. U.S. Patent No. 10,683,324 discloses that 7-DHC can be used to treat or prevent cancer, as well as to treat or prevent uncontrolled angiogenesis.

[0006] Traditional medications for treating pulmonary hypertension include calcium channel blockers, anticoagulants, diuretics, and cardiotonics. In recent years, several new drugs have been developed for the treatment of pulmonary hypertension, which can be classified into three categories based on their mechanisms of action: (1) endothelin receptor antagonists; (2) phosphodiesterase type V inhibitors; and (3) prostaglandin analogs. However, both traditional and new drugs can only slow the progression of the disease and temporarily improve the patient's clinical symptoms.

[0007] Therefore, there is still an urgent need to develop a method or drug that can effectively prevent or treat pulmonary hypertension. Summary of the Invention

[0008] This disclosure provides a method for using a pharmaceutical composition comprising a TGF-β receptor inhibitor and a pharmaceutically acceptable excipient for the prevention or treatment of pulmonary hypertension.

[0009] In one respect, in view of the foregoing, this disclosure provides a pharmaceutical composition for the prevention or treatment of pulmonary hypertension in a subject in need, comprising an effective amount of a TGF-β receptor inhibitor and a pharmaceutically acceptable excipient thereof.

[0010] In at least one embodiment of this disclosure, the TGF-β receptor inhibitor is 7-dehydrocholesterol, its salt, oxide, metabolite, or derivative.

[0011] In at least one embodiment of this disclosure, the 7-dehydrocholesterol, its salts, oxides, metabolites, or derivatives are administered to the subject at a dose range of 1 mg / kg to 50 mg / kg.

[0012] In some embodiments of this disclosure, the 7-dehydrocholesterol, its salts, oxides, metabolites, or derivatives are administered to the subject at a dose range of 10 mg / kg to 20 mg / kg.

[0013] In at least one embodiment of this disclosure, the 7-dehydrocholesterol, its salts, oxides, metabolites, or derivatives are a single active ingredient in the pharmaceutical composition. In other embodiments of this disclosure, the pharmaceutical composition further comprises other active ingredients, wherein the other active ingredients are selected from the group consisting of calcium channel blockers, anticoagulants, diuretics, cardiotonics, endothelin receptor antagonists, phosphodiesterase type V inhibitors, prostaglandin analogs, and any combination thereof.

[0014] On the other hand, in view of the foregoing, this disclosure provides a method for preventing or treating pulmonary hypertension in a subject in need, comprising administering to the subject a pharmaceutical composition comprising an effective amount of a TGF-β receptor inhibitor and a pharmaceutically acceptable excipient thereof.

[0015] In at least one embodiment of this disclosure, the TGF-β receptor inhibitor can reduce the proliferation of pulmonary artery endothelial cells, smooth muscle cells, or a combination thereof in the subject.

[0016] In at least one embodiment of this disclosure, the TGF-β receptor inhibitor is 7-dehydrocholesterol, its salt, oxide, metabolite, or derivative.

[0017] In at least one embodiment of this disclosure, the salt of 7-dehydrocholesterol includes acetate or benzoate of (3β)-7-dehydrocholesterol or a derivative thereof.

[0018] In at least one embodiment of this disclosure, the 7-dehydrocholesterol derivative comprises cholecalciferol (i.e., vitamin D3) or a compound of formula (I) below, wherein R1 is CR5 or N; R3 is selected from the group consisting of -O(CR5)nR6, -OC(-O)(CR5)nR6, -OC(=O)(CR5)nOR5 and -OC(=O)C(R5)=C(R5)2; R2 is selected from the group consisting of oxygen, sulfur, C(R4)2 and N(R4); and R4 is independently selected each time it appears from hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, aralkyl The group consisting of substituted aralkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, substituted heteroarylalkyl, OR5, and N(R5)2; R5 is independently selected each time it appears from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, aralkyl, substituted aralkyl, heteroaryl, substituted heteroaryl, heteroarylalkyl, and substituted heteroarylalkyl; R6 is selected from the group consisting of fluorine, chlorine, bromine, iodine, methanesulfonyl, toluenesulfonyl, -OSi(R5)3, -C(=O)OR5, and -C(=O)R5; the dashed line represents a single or double bond; and n is an integer from 1 to 10.

[0019]

[0020] Compound of formula (I)

[0021]

[0022] Cholecalciferol

[0023] In at least one embodiment of this disclosure, the compound of formula (I) is a compound of formula (Ia) or a salt or solvation thereof, wherein R1 to R3 are as defined above:

[0024]

[0025] In at least one embodiment of this disclosure, the compound of formula (I) is a compound of formula (Ib) or a salt or solvation thereof, wherein R1 to R3 are as defined above:

[0026]

[0027] In at least one embodiment of this disclosure, R1 of the compound of formula (I) is N.

[0028] In at least one embodiment of this disclosure, R2 is N(R4), and R4 is as defined above.

[0029] In at least one embodiment of this disclosure, the compound of formula (I) is a compound of formula (Ic) or a salt or solvation thereof, wherein R3 and R4 are as defined above:

[0030]

[0031] In at least one embodiment of this disclosure, the compound of formula (I) is a compound of formula (Id) or a salt or solvation thereof, wherein R3 and R4 are as defined above:

[0032]

[0033] In at least one embodiment of this disclosure, the compound of formula (I) is

[0034] In at least one embodiment of this disclosure, the subject is a mammal.

[0035] In at least one embodiment of this disclosure, the subject is a human.

[0036] In at least one embodiment of this disclosure, the compound of formula (I) is

[0037]

[0038] In at least one embodiment of this disclosure, the 7-dehydrocholesterol, its salts, oxides, metabolites, or derivatives are administered to the subject at a dose range of 1 mg / kg to 50 mg / kg.

[0039] In at least one embodiment of this disclosure, the 7-dehydrocholesterol, its salts, oxides, metabolites, or derivatives are administered to the subject at a dose range of 10 mg / kg to 20 mg / kg.

[0040] In at least one embodiment of this disclosure, the TGF-β receptor inhibitor may be administered to the subject in combination with other active ingredients to prevent or treat pulmonary hypertension.

[0041] In at least one embodiment of this disclosure, the 7-dehydrocholesterol, its salt, oxide, metabolite, or derivative is a single active ingredient in the pharmaceutical composition for the prevention or treatment of pulmonary hypertension.

[0042] In other embodiments of this disclosure, the other active ingredient is selected from the group consisting of calcium channel blockers, anticoagulants, diuretics, cardiotonics, endothelin receptor antagonists, phosphodiesterase type V inhibitors, prostaglandin analogs, and any combination thereof.

[0043] In other embodiments of this disclosure, the TGF-β receptor inhibitor reduces the proliferation of pulmonary artery endothelial cells, smooth muscle cells, or a combination thereof in the subject.

[0044] In other embodiments of this disclosure, the pharmaceutically acceptable excipient includes fillers, binders, preservatives, disintegrants, lubricants, suspending agents, wetting agents, solvents, surfactants, acids, flavoring agents, polyethylene glycol, alkyl glycol, sebacic acid, dimethyl sulfoxide, alcohols, or any combination thereof.

[0045] In other embodiments of this disclosure, the pharmaceutical composition is a formulation selected from the group consisting of lozenges, tablets, liquids, powders, granules, powders, pills, drops, capsules, ointments, creams, emulsions, gels, patches, injections, inhalers, sprays, and suppositories.

[0046] In other embodiments of this disclosure, the pharmaceutical composition is administered to the subject subcutaneously, intravenously, intradermally, intraperitoneally, orally, buccally, sublingually, intramuscularly, through the respiratory tract, or into the lungs.

[0047] In other embodiments of this disclosure, the pharmaceutical composition is administered to the subject at least once daily.

[0048] In other embodiments of this disclosure, the pharmaceutical composition is administered to the subject for at least one month.

[0049] In addition to the above, this disclosure also provides the use of a pharmaceutical composition comprising an effective amount of 7-dehydrocholesterol, its salts, oxides, metabolites, or derivatives in the preparation of a medicament for the prevention or treatment of pulmonary hypertension in a subject of need. Furthermore, this disclosure also provides the use of an effective amount of 7-dehydrocholesterol, its salts, oxides, metabolites, or derivatives in the preparation of a medicament for the prevention or treatment of pulmonary hypertension in a subject of need.

[0050] Furthermore, this disclosure also provides a pharmaceutical composition comprising an effective amount of 7-dehydrocholesterol, its salts, oxides, metabolites, or derivatives thereof, for the prevention or treatment of pulmonary hypertension in a subject of need. Attached Figure Description

[0051] This disclosure can be more fully understood by reading the following description of the embodiments and referring to the relevant drawings.

[0052] Figure 1The results of the MTT assay show the cell viability of 7-DHC (code 039) in smooth muscle cells (CON1) of normal mouse pulmonary artery, smooth muscle cells (rPASMC MCT4) of pulmonary artery of mouse with pulmonary hypertension (PAH), human pulmonary artery endothelial cells (hPAEC), and human pulmonary artery smooth muscle cells (hPASMC).

[0053] Figure 2 The study showed the inhibitory effect of 7-DHC (code 039) on the cell growth of normal mouse pulmonary artery smooth muscle cells (rPASMC CON1), pulmonary hypertension (PAH) mouse pulmonary artery smooth muscle cells (rPASMC MCT4), human pulmonary artery endothelial cells (hPAEC), and human pulmonary artery smooth muscle cells (hPASMC) under 10% FBS conditions in BrdU cell proliferation assays.

[0054] Figure 3 The study showed the inhibitory effect of BrdU cell proliferation assay and 7-DHC (code 039) on the cell growth of normal mouse pulmonary artery smooth muscle cells (rPASMC CON3), pulmonary hypertension (PAH) mouse pulmonary artery smooth muscle cells (rPASMC MCT2), human pulmonary artery endothelial cells (hPAEC), and human pulmonary artery smooth muscle cells (hPASMC) under the conditions of 0.1% FBS and 100 pM TGF-β.

[0055] Figure 4 This study depicts the drug administration timeline, animal survival status, and changes in body weight (BW) in an animal model of pulmonary hypertension. The rats were divided into a control group (CON), an MCT-induced pulmonary hypertension rat group (MCT), a rat group treated with 10 mg / kg 7-DHC (code 039 10 mg / kg), a rat group treated with 20 mg / kg 7-DHC (code 039 20 mg / kg), and a rat group treated with 50 mg / kg compound 042 (code 042 50 mg / kg). The slashes represent dead rats in different groups.

[0056] Figure 5 The study showed physiological parameters of rats in the pulmonary hypertension animal model after drug administration, including right ventricular pressure (RVP), mean blood pressure (MBP), heart rate (HR), and degree of right ventricular hypertrophy (RV / S+LV; right ventricle (RV) / septum (S)+left ventricle (LV)).

[0057] Figure 6Immunohistochemical staining images of pulmonary artery vascular tissue sections from rats in the pulmonary hypertension animal model, and bar charts showing the degree of pulmonary artery thickening in rats of different treatment groups. The different treatment groups include the control group (CON), the pulmonary hypertension rat group (MCT), the 10 mg / kg 7-DHC treatment group for pulmonary hypertension rats (represented as "039 10 mg"), the 20 mg / kg 7-DHC treatment group for pulmonary hypertension rats (represented as "039 20 mg"), and the 50 mg / kg compound 042 treatment group for pulmonary hypertension rats (represented as "042 50 mg").

[0058] Figure 7 This displays arterial gas analysis values ​​of test rats in an animal model of pulmonary hypertension after drug administration, including blood pH, oxygen pressure (PaO2), carbon dioxide pressure (PaCO2), and total carbon dioxide in the blood (TCO2, including CO2, H2CO3, and dissociated HCO3). - (ion), dissociated HCO3 - Ionic bicarbonate (HCO3) and base excess in the extracellular fluid compartment (BEecf).

[0059] Figure 8 This diagram depicts the drug administration timeline, animal survival status, and changes in body weight (BW) in an animal model of pulmonary hypertension. The rats were divided into a control group (CON), a pulmonary hypertension rat group (MCT), a rat group treated with 10 mg / kg 7-DHC (represented as "039 10 mg / kg"), a rat group treated with 20 mg / kg 7-DHC (represented as "039 20 mg / kg"), and a rat group treated with 30 mg / kg macitentan (represented as "macitentan 30 mg / kg"). The slashes represent dead rats in different groups.

[0060] Figure 9 This study showed physiological parameters of rats in an animal model of pulmonary hypertension after drug administration, including right ventricular pressure (RVP), mean blood pressure (mbp), heart rate (HR), and degree of right ventricular hypertrophy (RV / S+LV).

[0061] Figure 10Immunohistochemical staining images of pulmonary artery tissue sections from rats in the pulmonary hypertension animal model, and bar charts showing the degree of pulmonary artery thickening (statistical vessel wall thickness of vessels with diameters between 50-100 μM) in different drug administration groups. The different drug administration groups include the control group (CON), pulmonary hypertension rats (MCT), 10 mg / kg 7-DHC (represented as "039 10 mg / kg"), 20 mg / kg 7-DHC (represented as "039 20 mg / kg"), and 30 mg / kg macitentan group (represented as "macitentan 30 mg / kg").

[0062] Figure 11 This displays arterial gas analysis values ​​in rats after drug administration in an animal model of pulmonary hypertension, including carbon dioxide pressure (PaCO2), oxygen pressure (PaO2), total carbon dioxide (TCO2), oxygen saturation (sO2), extracellular fluid alkali excess (BEecf), and bicarbonate (HCO3). The normal range for sO2 is 93% to 100%. At a specific PaO2, the percentage of oxyhemoglobin to total hemoglobin in blood samples reflects the degree of O2 binding to hemoglobin in the blood, but its sensitivity to hypoxia is lower than its sensitivity to PaO2.

[0063] Figure 12A This shows the inhibition of Pai-1 luciferase activity by compound 039 at different concentrations (0, 0.5, 1, 10, 25, 50, and 75 μM) with and without TGF-β induction.

[0064] Figure 12B This shows the inhibition of TGF-β-induced Pai-1 luciferase activity (%) by compound 039 at different concentration ranges (0 to 80 μM).

[0065] Figure 13A This shows the inhibition of Pai-1 luciferase activity induced by TGF-β with or without compound 042 at different concentrations (0, 0.01, 0.1, 1, 5, 10 and 25 μM).

[0066] Figure 13B This shows the inhibition of TGF-β-induced Pai-1 luciferase activity (%) by compound 042 at different concentration ranges (0 to 30 μM).

[0067] Figure 14AThis shows the inhibition of Pai-1 luciferase activity induced by TGF-β with or without compound 050 at different concentrations (0, 1, 5, 10, 25, 50, and 100 μM).

[0068] Figure 14B This shows the inhibition of TGF-β-induced Pai-1 luciferase activity (%) by compound 050 at different concentration ranges (0 to 120 μM).

[0069] Figure 15A This shows the inhibition of Pai-1 luciferase activity induced by TGF-β with or without compound 053 at different concentrations (0, 50, 75, 100, 150 and 200 μM).

[0070] Figure 15B This shows the inhibition of TGF-β-induced Pai-1 luciferase activity (%) by compound 053 at different concentration ranges (0 to 200 μM).

[0071] Figure 16A This shows the inhibition of Pai-1 luciferase activity induced by TGF-β with or without compound 056 at different concentrations (0, 0.1, 1, 10, 50 and 100 μM).

[0072] Figure 16B This shows the inhibition of TGF-β-induced Pai-1 luciferase activity (%) by compound 056 at different concentration ranges (0 to 120 μM).

[0073] Figure 17A This shows the inhibition of Pai-1 luciferase activity induced by TGF-β with or without compound 057 at different concentrations (0, 0.01, 0.1, 1, 10 and 100 μM).

[0074] Figure 17B This shows the inhibition of TGF-β-induced Pai-1 luciferase activity (%) by compound 057 at different concentration ranges (0 to 120 μM).

[0075] Figure 18A This shows the inhibition of Pai-1 luciferase activity induced by TGF-β with or without compound 203 at different concentrations (0, 5, 10, 25, 50, and 75 μM).

[0076] Figure 18B This shows the inhibition of TGF-β-induced Pai-1 luciferase activity (%) by compound 203 at different concentration ranges (0 to 70 μM). Detailed Implementation

[0077] The following embodiments are used to describe the content of this disclosure. Based on the invention described in this specification, those skilled in the art will readily conceive of other advantages and effects of this disclosure. Furthermore, this disclosure can be implemented or applied in the manner described in the various embodiments. Modifications or changes can be made to these embodiments without departing from the spirit and scope of this disclosure to carry out the disclosure in different aspects and applications.

[0078] It should be noted that the singular forms “a,” “an,” and “the” used in this disclosure include their plural forms unless explicitly and unambiguously limited to a single reference. Furthermore, unless the context clearly indicates otherwise, the term “or” may be used interchangeably with the terms “and / or.”

[0079] As used herein, the terms “comprising,” “having,” “including,” or “containing,” etc., are used to refer to the components, methods, and elements thereof required by this disclosure, and are open to including elements not specifically specified, whether or not such element is necessary. Furthermore, the compositions of this disclosure can be used to implement the methods of this disclosure.

[0080] As used herein, the term "treating / treatment" refers to administering an effective amount of 7-DHC or its salts or derivatives to a subject in need to cure, alleviate, relieve, remedy, improve, or prevent a disease, its symptoms, or predisposition to it. This subject may be identified by a healthcare professional based on results from any appropriate diagnostic method.

[0081] This disclosure provides a method for treating pulmonary hypertension in a subject in need, the method comprising administering to the subject an effective amount of 7-DHC or a salt or derivative thereof.

[0082] As used herein, the term “effective dose” means a therapeutic dose sufficient to achieve the following objectives: prevention or treatment of the onset, recurrence, or initiation of pulmonary hypertension and one or more of its symptoms; enhancement or improvement of the preventive effect of other therapies; reduction of the severity and duration of symptoms; improvement of the symptoms of one or more of the symptoms; prevention of the progression of pulmonary hypertension; and / or enhancement or improvement of the therapeutic effect of other therapies.

[0083] The term "alkyl" refers to a straight-chain or branched saturated monovalent hydrocarbon chain having 1 to 12 carbon atoms. Preferably, straight-chain or branched alkyl groups have 1 to 6 carbon atoms. Examples include methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, isobutyl, pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl, octyl, 2,2,4-trimethylpentyl, nonyl, decyl, and their various branched isomers. Furthermore, if desired, the alkyl group may optionally and independently be substituted with 1 to 4 substituents, as described below.

[0084] The term "cycloalkyl" refers to a saturated monocyclic or bicyclic hydrocarbon ring having 3 to 12 carbon atoms. More preferably, it refers to a monocyclic saturated hydrocarbon group having 3 to 7 carbon atoms. Examples of such groups are monocyclic and bicyclic alkyl groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, etc. If necessary, these groups may be optionally and independently substituted with 1 to 4 substituents, as described below.

[0085] The term "alkenyl" refers to a straight-chain or branched monovalent hydrocarbon chain having 2 to 12 carbon atoms and containing at least one double bond. Preferred alkenyl groups are straight-chain or branched alkenyl groups having 1 to 6 carbon atoms. Examples include vinyl, 2-propenyl, 3-butenyl, 2-butenyl, 4-pentenyl, 3-pentenyl, 2-hexenyl, 3-hexenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 3-octenyl, 3-nonenyl, 4-decenyl, 3-undecenyl, 4-dodecenyl, 4,8,12-tetradecanetriene, etc. If necessary, the alkenyl group may optionally and independently be substituted with 1 to 4 substituents, as described below.

[0086] The term "alkynyl" refers to a straight-chain or branched monovalent hydrocarbon chain having at least one triple bond. Preferred alkynyl groups are straight-chain or branched alkynyl groups having 1 to 6 carbon atoms. Examples include 2-propynyl, 3-butynyl, 2-butynyl, 4-pentynyl, 3-pentynyl, 2-hexynyl, 3-hexynyl, 2-heptynyl, 3-heptynyl, 4-heptynyl, 3-octyynyl, 3-nonynyl, 4-decynyl, 3-undecanynyl, 4-dodecynyl, etc. If necessary, the alkynyl group may be selectively and independently substituted with 1 to 4 substituents, as described below.

[0087] The term "aryl" refers to a monocyclic or bicyclic monovalent aromatic hydrocarbon group having 6 to 10 carbon atoms. Examples include phenyl and naphthyl (including 1-naphthyl and 2-naphthyl). If necessary, these groups may be optionally and independently substituted with 1 to 4 substituents, as described below.

[0088] The term "aralkyl," whether used alone or as part of other groups, refers to an alkyl group as described above that has an aryl substituent. If desired, aralkyl groups may optionally and independently be substituted with 1 to 4 substituents, as described below.

[0089] The term "heteroaryl" refers to a monocyclic or bicyclic monovalent aromatic hydrocarbon group having 6 to 10 carbon atoms, wherein at least one carbon atom is substituted with at least one heteroatom such as N, O, or S. If desired, the heteroaryl group may optionally and independently be substituted with 1 to 4 substituents, as described below.

[0090] The term "heteroaryl" refers, whether used alone or as part of other groups, to an alkyl group as described above that has a heteroaryl substituent. If desired, heteroaryl groups may optionally and independently be substituted with 1 to 4 substituents, as described below.

[0091] The substituents of the above groups include, for example, halogen atoms (such as fluorine, chlorine, bromine, iodine), nitro, cyano, oxo, hydroxyl, mercapto, carboxyl, sulfonyl, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heterocyclic, alkoxy, and cycloalkoxy, but are not limited to these.

[0092] In some specific embodiments of this disclosure, the effective amount of 7-DHC or its salts and derivatives can be from 1 mg / kg to 50 mg / kg. In some embodiments, the lower limit of this dose can be 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, or 30 mg / kg, and the upper limit of this dose can be 50 mg / kg, 48 mg / kg, 45 mg / kg, 43 mg / kg, 40 mg / kg, 38 mg / kg, 35 mg / kg, 33 mg / kg, 30 mg / kg, 25 mg / kg, 24 mg / kg, 23 mg / kg, 22 mg / kg, 21 mg / kg, or 20 mg / kg. For example, the dosage of 7-DHC or its salts or derivatives may be from 1 mg / kg to 40 mg / kg, 5 mg / kg to 40 mg / kg, 5 mg / kg to 35 mg / kg, 10 mg / kg to 30 mg / kg, 10 mg / kg to 20 mg / kg, about 42 mg / kg, about 40 mg / kg, about 37 mg / kg, about 35 mg / kg, about 32 mg / kg, about 30 mg / kg, about 28 mg / kg, about 25 mg / kg, about 20 mg / kg, about 18 mg / kg, about 15 mg / kg, about 13 mg / kg, or about 10 mg / kg.

[0093] As used herein, when numbers or ranges are mentioned, it will be understood by one of ordinary skill in the art that they are intended to cover an appropriate and reasonable range in the particular field relating to this disclosure.

[0094] In some embodiments of this disclosure, 7-DHC or its salts or derivatives may be administered once every two days, once daily, twice daily, three times daily, or four times daily. In some embodiments of this disclosure, 7-DHC may be administered three times weekly.

[0095] In some embodiments of this disclosure, 7-DHC or its salts or derivatives may be administered to a subject via subcutaneous, intravenous, intradermal, intraperitoneal, oral, intramuscular, or intracranial routes.

[0096] In some embodiments of this disclosure, 7-DHC or its salts or derivatives may be administered to a subject for a sufficient period of time to prevent or treat pulmonary hypertension. In some embodiments of this disclosure, this sufficient period may vary depending on factors such as the subject's species, sex, weight or age, stage of disease, symptoms or severity, and route, time, or frequency of administration. In some embodiments of this disclosure, 7-DHC or its salts or derivatives are administered once daily for at least one month. For example, the administration period of 7-DHC or its salts or derivatives may last 1, 2, 3, 4, or 6 months, or 1, 2, 3, or 4 years, or even longer, provided that no side effects occur during treatment; no specific limitations are set in this disclosure. In one embodiment of this disclosure, the administration period may range from 1 month to 2 years. In other embodiments of this disclosure, the administration period may range from 4 weeks to 12 months. In still other embodiments of this disclosure, daily administration of 7-DHC or its salts or derivatives lasts for at least 2 months.

[0097] In at least one embodiment of this disclosure, 7-DHC or its salts or derivatives may be administered in an oral dosage form. In at least one embodiment of this disclosure, 7-DHC or its salts or derivatives administered to a subject may be included in a pharmaceutical composition. In at least one embodiment of this disclosure, the pharmaceutical composition comprises 7-DHC or its salts or derivatives and a pharmaceutically acceptable excipient. In at least one embodiment, the composition of this disclosure is formulated into a form suitable for oral administration, thereby allowing administration to a subject via an oral route. In other embodiments of this disclosure, the composition may be formulated as lozenges, tablets, liquids, powders, granules, powders, pills, drops, capsules, ointments, creams, emulsions, gels, patches, injections, inhalers, sprays, and suppositories. In some embodiments of this disclosure, pharmaceutically acceptable excipients include, but are not limited to, fillers, binders, preservatives, disintegrants, lubricants, suspending agents, wetting agents, solvents, surfactants, acids, flavoring agents, polyethylene glycol (PEG), alkylene glycols, sebacic acid, dimethyl sulfoxide, alcohols, or any combination thereof.

[0098] The pharmaceutical compositions disclosed herein may use 7-DHC or its salts or derivatives as a single active ingredient for the prevention or treatment of pulmonary hypertension. In other words, 7-DHC may be the sole active ingredient in the pharmaceutical compositions of this disclosure for the prevention or treatment of pulmonary hypertension. In this embodiment, this disclosure provides a safe and effective therapy for the prevention or treatment of pulmonary hypertension by using 7-DHC or its salts or derivatives as the active ingredient alone.

[0099] In other embodiments of this disclosure, the composition may be administered to a subject in combination with other active ingredients unless this would inhibit the effects of the disclosure. In some embodiments of this disclosure, 7-DHC or its salts or derivatives and other active ingredients may be administered to a subject in need as a single composition or as separate compositions.

[0100] In at least one embodiment, the administration of 7-DHC or its salts or derivatives in the methods provided in this disclosure can be combined with any suitable conventional therapy for pulmonary hypertension. In at least one embodiment of this disclosure, conventional therapy for pulmonary hypertension includes, but is not limited to, calcium channel blockers, anticoagulants, diuretics, cardiotonics, endothelin receptor antagonists, phosphodiesterase type V inhibitors, and prostaglandin analogs.

[0101] As described herein, the term "mammal" refers to all animals classified as mammals, including, but not limited to, humans, livestock and farm animals, as well as zoo, sporting or pet animals such as dogs, horses, cats, cattle, etc. Preferably, the mammal is human.

[0102] This disclosure has been illustrated through various embodiments. The following embodiments should not be considered as any limitation on the scope of this disclosure.

[0103] Example

[0104] Experimental methods and procedures

[0105] The principle of the MTT assay is based on MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazazole bromide], a yellow compound that accepts hydrogen ions and functions in mitochondria. It produces purple formazan crystals under the catalysis of succinate dehydrogenase (SDH). The amount of formazan crystals produced is directly proportional to the number of viable cells. The absorbance (OD) at 570 nm is measured after dissolving the formazan crystals in DMSO. This OD value represents mitochondrial activity, i.e., the number of viable cells; therefore, the MTT assay can be used as an indicator of cell viability (toxicity). After co-culturing the cells with the test drug in the culture medium, the MTT assay can be used to detect the drug's toxicity to the cells and determine the effective concentration range of the drug. The experimental procedure for the MTT assay includes adding 1.5 × 10⁻⁶... 4 Cells were seeded in 48-well plates and cultured overnight at 37°C in medium containing 10% FBS and different drug concentrations. The cell culture medium was then aspirated, washed with phosphate-buffered saline (PBS), and 10% MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) solution was added. The plates were then incubated at 37°C for 4 hours. Next, the MTT solution was aspirated, and 100 μl / well of DMSO was added to completely dissolve the purple MTT crystals. After thorough mixing, the absorbance was measured at 570 nm.

[0106] BrdU (5-bromo-2-deoxyuridine) cell proliferation assay

[0107] Use a 48-well plate, inoculate 1.5 × 10⁶ cells per well. 4 Cells were collected. After 24 hours, the medium was replaced with serum-free medium and synchronized for another 24 hours. A double-concentration compound was prepared in serum-free medium and the cells were pretreated for 1 hour. An equal volume of medium containing 20% ​​fetal bovine serum (FBS) was then added to dilute the compound, resulting in a final concentration of 10% FBS medium and a double-concentration compound. In the TGF-β-added experimental group, an equal volume of medium containing 10 ng / mL TGF-β was added, resulting in a final concentration of 5 ng / mL TGF-β and a double-concentration compound. In the TGF-β-added experimental group, the entire experiment was conducted in medium containing 0.1% serum. Approximately 6 hours later, BrdU reagent was added. Cells were collected the following day for enzyme-linked immunosorbent assay (ELISA) analysis. The compound treatment time was approximately 24 hours, while the BrdU treatment time was approximately 18 hours.

[0108] TGF-β-induced Pai-1 luciferase assay

[0109] The effects of different compounds on downstream genes induced by TGF-β, specifically plasminogen activator inhibitor 1 (PAI-1), were determined using the MLECs-clone32 cell line. MLECs-clone32 was used as the expression construct for stable transfection of the Mv1Lu cell line. This PAI-1 luciferase construct contained a truncated (PAI-1) promoter fused with a firefly luciferase reporter gene. 11,12 The activity induced by the truncated PAI-I promoter was quantified using a luciferase assay. The effect of each compound on TGF-β-induced PAI-I promoter activity was compared, with TGF-β-induced luciferase activity in untreated cells as 100%. The assay procedure for Pai-1 luciferase activity involved seeding 1.5 × 10⁶ cells per well in a 96-well plate (DMEM medium containing 10% FBS). 5 Cells were cultured overnight in an incubator. The next day, the medium was discarded, and cells in DMEM medium with or without the test compound were treated with recombinant TGF-β (50 pM) and incubated at 37°C for 4 hours. The medium was then aspirated, the cells were washed with PBS, and the activity of Pai-1 luciferase was assessed by a luciferase assay.

[0110] Animal experimental model of pulmonary hypertension

[0111] This disclosure utilizes the injection of monocrotaline to induce symptoms of pulmonary hypertension in rats, thereby establishing an animal experimental model of pulmonary hypertension suitable for screening and testing drugs.

[0112] The clinical diagnostic criteria for pulmonary hypertension (PH) are right heart catheterization, defined as a mean pulmonary arterial pressure (mPAP) greater than or equal to 20 mmHg. In addition to increased mPAP, PH is further diagnosed when pulmonary vascular resistance (PVR) is greater than 3 Wood units. This disclosure utilizes the Millar Model PCU200 pressure system to measure right ventricular pressure (RVP) and mean blood pressure (mbP) in rats with PH, while simultaneously monitoring heart rate (HR). After measuring hemodynamic data, arterial blood can be drawn from the carotid artery of pulmonary hypertension rats. Arterial gas analysis data can then be measured using an Abbott i-STAT pulse oximeter with an Abbott i-STAT Cartridge G3+. This includes pH, PaO2, PaCO2, HCO3, and base excess (BE) values ​​in a quantitative whole blood sample. Normal pH values ​​are between 7.35 and 7.45, indicating whether the experimental animal exhibits acidemia, alkalemia, acidosis, or alkalosis. Normal PaO2 values ​​are between 80 and 100 mmHg, reflecting the effectiveness of gas exchange (ventilation / perfusion). Normal PaCO2 values ​​are between 35 and 45 (e.g., 38 to 42) mmHg, evaluating the effectiveness of alveolar ventilation. Acids produced by food, tissue breakdown, inflammation, and hypoxia are primarily expelled through the lungs as volatile acidic CO2 (short-term) and through the kidneys as bicarbonate (HCO3). - (Long-term) neutral and non-volatile acids. Normal bicarbonate (HCO3) - The BE value is 22 to 26 meq / L. BE is the excess alkali value, with a normal range of -2 to +2. A negative value indicates a deficiency of alkali ions in the blood. An excessively high negative BE value (-) indicates metabolic acidosis, and an excessively high positive value (+) indicates metabolic alkalosis. At the end of the experiment, the rats were sacrificed, their hearts were removed, the left and right ventricles were separated, and their weight (RV) / (S)+(LV) was measured.

[0113] Example 1: Screening for TGF-β receptor inhibitors using a pulmonary hypertension cell model

[0114] Five test compounds were screened using the MTT assay and the BrdU (5-bromo-2-deoxyuridine) cell proliferation assay. Information on these test compounds is listed in Table 1. Tables 2 and 3 show the growth inhibition of the five test compounds on human pulmonary artery endothelial cells (hPAEC), human pulmonary artery smooth muscle cells (hPASMC), normal rat pulmonary artery smooth muscle cells (rPASMC, as a control group, denoted as "CON"), and rat pulmonary artery smooth muscle cells (denoted as "MCT") under the conditions of 10% FBS, 0.1% FBS, and 100 pM TGF-β, respectively. The results showed that treatment with 7-DHC (code 039) resulted in the most significant growth inhibition.

[0115] Table 1: Codes, names, and structures of seven compounds

[0116]

[0117] Table 2: Cell growth inhibition of six compounds in 10% FBS

[0118]

[0119] *EC50: Half-maximal effective concentration (concentration for 50% of maximal effect)

[0120] Table 3: Cell growth inhibition of five compounds in the presence of 100 pM TGF-β

[0121]

[0122] Table 4: IC50 values ​​of seven compounds inhibiting TGF-β-induced Pai-1 luciferase activity (%)

[0123]

[0124]

[0125] Cell viability assays revealed that cytotoxicity began to occur in all types of pulmonary artery smooth muscle cells (rPASMCCON1), (rPASMC MCT4), human pulmonary artery endothelial cells (hPAEC), and human pulmonary artery smooth muscle cells (hPASMC) when the concentration of 7-DHC (code 039) increased to 100 μM. Figure 1 ).

[0126] In BrdU cell proliferation assays, such as Figure 2As shown, under 10% FBS conditions, when the concentration of 7-DHC (code 039) was 10 μM, it had a significant inhibitory effect on both normal rat pulmonary artery smooth muscle cells (rPASMC CON1) and rat pulmonary artery smooth muscle cells (rPASMC MCT4), and cell growth inhibition was also observed on human pulmonary artery endothelial cells (hPAEC) and human pulmonary artery smooth muscle cells (hPASMC).

[0127] like Figure 3 As shown, under conditions of 0.1% FBS and 100 pM TGF-β, a concentration of 25 μM of 7-DHC (code 039) significantly inhibited the growth of both normal rat pulmonary artery smooth muscle cells (rPASMC CON3) and rat pulmonary artery smooth muscle cells (rPASMC CT2), with the inhibitory effect on rat pulmonary artery smooth muscle cells being particularly pronounced. Similarly, a concentration of 25 μM of 7-DHC (code 039) also inhibited the growth of human pulmonary artery endothelial cells (hPAEC) and human pulmonary artery smooth muscle cells (hPASMC).

[0128] Example 2: Physiological parameters, pulmonary artery thickening, and arterial gas analysis in animals with pulmonary hypertension after administration of 7-DHC.

[0129] A rat model of pulmonary hypertension was induced using monocrotaline (MCT). The TGF-β receptor inhibitors 7-DHC (code 039) and 042 were administered intraperitoneally (IP). Physiological parameters were collected, including right ventricular pressure, systemic arterial pressure, heart rate, and the degree of right ventricular hypertrophy. Pulmonary artery thickening and blood oxygenation were observed and analyzed through lung tissue histopathological sections. Specifically, 7-DHC (code 039) was administered at doses of 10 mg / kg or 20 mg / kg, while the 042 group received 50 mg / kg. Each administration group contained 3 rats (n=3), while the control group (CON) and the pulmonary hypertension rat group (MCT) each contained 6 rats (n=6).

[0130] like Figure 4 As shown, the rats in the 042 treatment group had significantly decreased body weight and activity level by the second week of treatment, and therefore died prematurely on day 21 after induction with limonene. On day 23 after induction with limonene, rats in the 20 mg / kg 7-DHC (039) treatment group had already died, so pulmonary artery pressure could not be obtained. Therefore, right ventricular pressure (RVP) was measured starting on day 24.

[0131] like Figure 5As shown, the mean right ventricular pressure (RVP) of pulmonary hypertension rats (MCT) was 28 mmHg, but administration of 7-DHC (code 039) (10 mg / kg or 20 mg / kg) reduced RVP. After sacrificing the rats, the hearts were removed, and the left and right ventricles were separated and weighed (RV / S+LV). The left-to-right ventricular ratio of normal rats was approximately 25%, while the mean right ventricular hypertrophy rate in MCT rats exceeded 50%. However, administration of 7-DHC (code 039) (10 mg / kg or 20 mg / kg) reduced right ventricular hypertrophy, and 7-DHC (code 039) did not reduce systemic blood pressure or heart rate. This indicates that 10 mg / kg of 7-DHC (code 039) has the effect of reducing right ventricular pressure and right ventricular hypertrophy. Furthermore, the right ventricular hypertrophy rate in the group treated with code 042 was slightly lower than that in MCT rats.

[0132] In addition, pulmonary artery tissue sections from rats in the treatment group (code 042) were observed, and the location of the pulmonary artery was confirmed by immunohistoscopy staining for α-smooth muscle actin (α-SM actin or α-SMA). The results are as follows: Figure 6 As shown, the mean pulmonary artery thickness (median wall thickness) in pulmonary hypertension rats (MCT) exceeded 55%, however, the 10 mg / kg 7-DHC (code 039) group showed a significant reduction in pulmonary artery thickness (p<0.001). This result demonstrates that 10 mg / kg 7-DHC (code 039) is effective in reducing pulmonary artery thickness in rats with pulmonary hypertension.

[0133] like Figure 7As shown, there were no significant differences in pH, PaO2, and PaCO2 among the control group (CON), the pulmonary hypertension rat group (MCT), the 10 mg / kg 7-DHC (code 039) group (represented as "code 039-10"), and the 20 mg / kg 7-DHC (code 039) group (represented as "code 039-20"). This may be because the prolonged surgery time affected the function of pulmonary oxygen exchange. Interestingly, the normal value for alkali excess is generally between -2 and +2, but the BE value in the MCT group was -5.5, indicating metabolic acidosis. This may be related to tissue hypoxia caused by tissue inflammation or tissue edema. The BE value in the 10 mg / kg 7-DHC (code 039) group was -2, and the BE value in the 20 mg / kg 7-DHC (code 039) group was 0. It is evident that 7-DHC (code 039) can improve tissue inflammation or edema, allowing tissues to regain oxygen and thus ending hypoxia. As a result, the lactic acid produced by hypoxia is reduced, and metabolic acidosis is also improved.

[0134] Example 3: Comparison of physiological parameters, pulmonary artery thickening, and arterial gas analysis values ​​in rats with pulmonary hypertension after administration of 7-DHC or macitentan.

[0135] Macitentan is a novel dual endothelin receptor antagonist (ERA) currently used to treat pulmonary arterial hypertension. It has high affinity and can occupy endothelin (ET) receptors on the smooth muscle of human pulmonary arteries for a long time, thereby preventing endothelin-1 (ET-1) from binding to its receptors (ETA and ETB).

[0136] The rat model of pulmonary hypertension was induced by lily alkaloids. The TGF-β receptor inhibitor 7-DHC (code 039) was administered intraperitoneally (IP) and macitentan (dissolved in methylcellulose) was administered orally. The 7-DHC (code 039) group was divided into two groups according to the dosage: 10 mg / kg or 20 mg / kg. The macitentan group was administered 30 mg / kg.

[0137] Rats in the macitentan group did not show significant impact on body weight and activity level during the second week of administration in the pulmonary hypertension animal model. However, on day 23 after induction with limonene, rats in the macitentan group died, making it impossible to obtain pulmonary artery pressure data for this group subsequently. In contrast, rats in the 20 mg / kg 7-DHC (code 039) group showed a significant decrease in body weight, but their activity level was not affected, and their survival rate was 100%. Figure 8 ).

[0138] On day 25 after induction with lily alkaloid, right ventricular pressure (RVP) was measured in each group of rats. Figure 9 As shown, the mean pulmonary artery pressure (RVP) of pulmonary hypertension rats (MCT) was 28.97 mmHg. The right ventricular pressure decreased to 17.61 mmHg and 19.96 mmHg in the 10 mg / kg and 20 mg / kg groups of 7-DHC (code 039), respectively. After sacrificing the rats, the hearts were removed, and the left and right ventricles were separated and weighed (right ventricle / middle septum + left ventricle; RV / S+LV). The left-to-right ventricular ratio in normal rats is approximately 26.7%, while the average right ventricular hypertrophy rate in pulmonary hypertension rats (MCT) is approximately 71.64%. In contrast, the average right ventricular hypertrophy rates in the 10 mg / kg and 20 mg / kg groups treated with 7-DHC (code 039) were 53.96% and 62.97%, respectively, both significantly reducing right ventricular hypertrophy. Furthermore, 7-DHC (code 039) did not significantly reduce the average blood pressure and heart rate. Therefore, it can be concluded that the administration of 7-DHC (code 039) has the effect of reducing right ventricular pressure.

[0139] Pulmonary artery vascular tissue sections from rats in each treatment group were observed, and the location of the pulmonary artery was confirmed by immunohistochemical staining for α-smooth muscle actin. Figure 10 As shown, the results indicated that the average pulmonary arterial thickness in MCT rats exceeded 65%, but the 10 mg / kg and 20 mg / kg groups treated with 7-DHC (code 039) significantly reduced the pulmonary artery thickening (###p<0.001). Similarly, since macitentan is an endothelin-1 receptor antagonist, a reduction in vascular thickening was also observed in the macitentan group, but its survival rate was significantly worse than that of the 7-DHC (code 039) group. Therefore, the TGF-β receptor inhibitor 7-DHC can inhibit pulmonary vascular cell proliferation and remodeling by reducing TGF-β-related signaling.

[0140] Figure 11The results showed no significant differences in PaO2 and PaCO2 among the control group (CON), the pulmonary hypertension rat group (MCT), the 10 mg / kg 7-DHC (code 039) group, and the 20 mg / kg 7-DHC (code 039) group. This may be because the operation time was long, thus affecting the function of pulmonary blood oxygen exchange. The normal range for excess alkali (BE) is -2 to +2. In the MCT group, the BE value was -6, indicating metabolic acidosis, which may be related to tissue hypoxia caused by tissue inflammation or edema. The BE value of the 10 mg / kg 7-DHC (code 039) group was -3, and the BE value of the 20 mg / kg 7-DHC (code 039) group was -1.4. This shows that 7-DHC (code 039) can improve tissue inflammation or edema, allowing tissues to regain oxygen and thus stopping hypoxia. The lactic acid produced by hypoxia is reduced, and metabolic acidosis is thus improved. This is consistent with the results of Example 2.

[0141] Figures 12 to 18 show the inhibition of different compounds at different concentration ranges, including compounds 7-DHC (code 039), begain (code 042), styraxin (code 050), macitentan (code 053), tanshinone (code 056), melatonin (code 057), and MeTC7 (code 203) on Pai-1 luciferase activity induced or not induced by TGF-β, as well as the inhibition of TGF-β-induced Pai-1 luciferase activity (%) by different compounds at different concentration ranges.

[0142] The inhibition results of TGF-β-induced Pai-1 luciferase activity (%) revealed that among the tested compounds 7-DHC, becaine, styraxine, macitentan, tanshinone, melatonin, or MeTC7, only 7-DHC and its derivative MeTC7 significantly inhibited TGF-β-induced Pai-1 luciferase activity at concentrations increased to 10 μM. Other compounds failed to show the same effect at similar concentrations. Specifically, for example, such as... Figure 12A and 12B As shown, 7-DHC concentrations exceeding 10 μM significantly inhibited TGF-β-induced Pai-1 luciferase activity, and the inhibitory effect increased with increasing concentration. For example, according to... Figure 13A and 13B When the concentration of the compound begain exceeds 10 μM, a significant inhibitory effect on TGF-β-induced Pai-1 luciferase activity is observed, and the inhibitory effect increases with concentration. For example, as... Figure 18A and 18BAs shown, the concentration of compound MeTC7 exceeding 10 μM significantly inhibited the activity of TGF-β-induced Pai-1 luciferase, and the inhibitory effect increased with increasing concentration.

[0143] Furthermore, as shown in Table 4, compounds 7-DHC, becaine, styraxine, macitentan, tanshinone, melatonin, and MeTC7 caused a 50% (IC50) inhibition of TGF-β-induced Pai-1 luciferase activity. 50 The values ​​were calculated using the IC50 values ​​of three compounds: 7-DHC, begain, and MeTC7. 50 The concentrations were lower than those of macitentan, tanshinone, styrax, and melatonin, and these compounds did not show any inhibitory effect. Clearly, compounds 7-DHC, begain, and MeTC7 were more effective than the other four compounds in inhibiting TGF-β-induced Pai-1 luciferase activity.

[0144] While the foregoing has described various features and advantages of this disclosure, and detailed the structure and features of the invention, it is merely illustrative for the purposes of this invention. Variations may be made on the details, particularly in the shape, size, and arrangement of components, within the scope of the invention principles which are formed by referring to the terms recited in the claims in their broad general definitions.

[0145] References:

[0146] 1. McGoon M., Gutterman D., Steen V., Barst R., McCrory DC, Fortin TA, et al., "Screening, early detection, and diagnosis of pulmonary arterial hypertension: ACCP evidence-based clinical practice guidelines." CHESTJournal. 2004; 126 (suppl.): 14S-34S.

[0147] 2.GalièN.,Humbert M.,Vachiery J.L.,et al.“2015ESC / ERS Guidelines forthe diagnosis and treatment of pulmonary hypertension:the Joint Task Forcefor the Diagnosis and Treatment of Pulmonary Hypertension of the EuropeanSociety of Cardiology(ESC)and the European Respiratory Society(ERS)”;endorsedby:Association for European Paediatric and Congenital Cardiology(AEPC),International Society for Heart and Lung Transplantation(ISHLT).Eur.HeartJ.2016;37:67-119。

[0148] 3.Simonneau G.,Gatzoulis M.A.,Adatia I.,Celermajer D.,Denton C.,Ghofrani A.,et al.,“Updated clinicalclassification of pulmonaryhypertension.”Journal of theAmerican College of Cardiology.2013;62(25):D34-D41。

[0149] 4.Humbert M.,Morrell N.W.,Archer S.L.,Stenmark K.R.,MacLean M.R.,LangI.M.,et al.“Cellular and molecularpathobiology of pulmonary arterialhypertension.”Journal ofthe American College of Cardiology.2004;43(12s1):S13-S24。

[0150] 5.Cool C.D.,Stewart J.S.,Werahera P.,Miller G.J.,Williams R.L.,Voelkel N.F.,et al.“Three-dimensionalreconstruction of pulmonary arteries inplexiform pulmonaryhypertension using cell-specific markers:evidence foradynamic and heterogeneous process of pulmonary endothelialcell growth.”TheAmerican Journal of Pathology.1999;155(2):411-419。

[0151] 6.Jonigk D.,Golpon H.,Bockmeyer C.L.,Maegel L.,HoeperM.M.,GottliebJ.,et al.“Plexiform lesions in pulmonaryarterial hypertension:composition,architecture,andmicroenvironment.”The American Journal of Pathology.2011;179(1):167-179。

[0152] 7.Aldred M.A.,Vijayakrishnan J.,James V.,Soubrier F.,Gomez-SanchezM.A.,Martensson G.,et al.“BMPR2 generearrangements account for a significantproportion ofmutations in familial and idiopathic pulmonaryarterialhypertension.”Human Mutation.2006;27(2):212-213。

[0153] 8.Cogan J.D.,Vnencak-Jones C.L.,Phillips J.A.,LaneK.B.,Wheeler L.A.,Robbins I.M.,et al.“Gross BMPR2 generearrangements constitute a new cause forprimary pulmonaryhypertension.”Genetics in Medicine.2005;7(3):169-174。

[0154] 9.Cogan J.D.,Pauciulo M.W.,Batchman A.P.,Prince M.A.,Robbins I.M.,Hedges L.K.,et al.“High frequency of BMPR2exonic deletions / duplications infamilial pulmonary arterialhypertension.”American Journal of Respiratory andCriticalCare Medicine.2006;174(5):590-598。

[0155] 10.Thomson J.R.,Machado R.D.,Pauciulo M.W.,MorganN.V.,Humbert M.,Elliott G.C.,et al.“Sporadic primarypulmonary hypertension is associated withgermline mutationsof the gene encoding BMPR-II,a receptor member of the TGF-βfamily.”Journal of Medical Genetics.2000;37(10):741-745。

[0156] 11.An assay for transforming growth factor-beta usingcellstransfected with a plasminogen activator inhibitor-1promoter-luciferaseconstruct.Anal Biochem 1994,216:276–284。

[0157] 12.Quantification of active and total transforming growthfactor-βlevels in serum and solid organ tissues.BMCResearch Notes 2012,5:636。

Claims

1. A method for preventing or treating pulmonary hypertension in a subject in need, comprising administering to the subject a pharmaceutical composition comprising an effective amount of a TGF-β receptor inhibitor and a pharmaceutically acceptable excipient thereof.

2. The method as described in claim 1, wherein, The TGF-β receptor inhibitor is 7-dehydrocholesterol, its salt, oxide, metabolite, or derivative.

3. The method as described in claim 2, wherein, The salts of the 7-dehydrocholesterol include acetates or benzoates of (3β)-7-dehydrocholesterol or its derivatives.

4. The method of claim 2, wherein, The 7-dehydrocholesterol derivatives include cholecalciferol or compounds of formula (I): Wherein, R1 is CR5 or N; R3 is selected from the group consisting of -O(CR5)nR6, -OC(-O)(CR5)nR6, -OC(=O)(CR5)nOR5, and -OC(=O)C(R5)=C(R5)2; R2 is selected from the group consisting of oxygen, sulfur, C(R4)2, and N(R4); R4 is independently selected each time it appears from hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, aralkyl, substituted aralkyl, heteroaryl, substituted heteroaryl, heteroaryl, substituted The group consisting of heteroaryl, OR5, and N(R5)2; R5 is independently selected each time it appears from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, aryl, substituted aryl, aralkyl, substituted aralkyl, heteroaryl, substituted heteroaryl, heteroaryl, and substituted heteroaryl; R6 is selected from the group consisting of fluorine, chlorine, bromine, iodine, methanesulfonyl, toluenesulfonyl, -OSi(R5)3, -C(=O)OR5, and -C(=O)R5; the dashed line indicates a single or double bond; and n is an integer from 1 to 10.

5. The method of claim 4, wherein, The compound of formula (I) is a compound of formula (Ia) or a salt or solvation thereof: R1 to R3 are as defined in claim 4.

6. The method of claim 5, wherein, The compound of formula (I) is a compound of formula (Ib) or a salt or solvation thereof: R1 to R3 are as defined in claim 4.

7. The method of claim 4, wherein, R1 is N.

8. The method of claim 7, wherein, R2 is N(R4), and R4 is as defined in claim 4.

9. The method of claim 8, wherein, The compound of formula (I) is a compound of formula (Ic) or a salt or solvation thereof: R3 and R4 are as defined in claim 4.

10. The method of claim 9, wherein, The compound of formula (I) is a compound of formula (Id) or a salt or solvation thereof: R3 and R4 are as defined in claim 4.

11. The method of claim 4, wherein, The compound of formula (I) is 12. The method of claim 4, wherein, The subjects were mammals.

13. The method of claim 4, wherein, The compound of formula (I) is:

14. The method of claim 2, wherein, The 7-dehydrocholesterol, its salts, oxides, metabolites, or derivatives were administered to the subject at a dose range of 1 mg / kg to 50 mg / kg.

15. The method of claim 1, wherein, The 7-dehydrocholesterol, its salts, oxides, metabolites, or derivatives are the single active ingredient in the pharmaceutical composition for the prevention or treatment of pulmonary hypertension.

16. The method of claim 1, wherein, The pharmaceutical composition further comprises other active ingredients, wherein the other active ingredients are selected from the group consisting of calcium channel blockers, anticoagulants, diuretics, cardiotonics, endothelin receptor antagonists, phosphodiesterase type V inhibitors, prostaglandin analogs, and any combination thereof.

17. The method of claim 1, wherein, The TGF-β receptor inhibitor reduces the proliferation of pulmonary artery endothelial cells, smooth muscle cells, or a combination thereof in the subject.

18. The method of claim 1, wherein, The pharmaceutical composition is administered to the subject subcutaneously, intravenously, intradermally, intraperitoneally, orally, buccally, sublingually, intramuscularly, through the respiratory tract, or into the lungs.

19. The method of claim 1, wherein, The pharmaceutical composition is administered to the subject at least once a day.

20. The method of claim 1, wherein, The pharmaceutical composition was administered to the subject for at least one month.

Images