阅读说明：本技术 马兜铃烷型倍半萜类化合物在制备预防和/或治疗心脑血管疾病药物中的用途 (Application of aristolochiane type sesquiterpene compound in preparation of medicine for preventing and/or treating cardiovascular and cerebrovascular diseases ) 是由 王启隆 吴红华 高秀梅 王跃飞 刘二伟 房景梅 李冉 于 2021-08-11 设计创作，主要内容包括：本申请提供了马兜铃烷型倍半萜类化合物在制备预防和/或治疗心脑血管疾病药物中的用途,其中,所述马兜铃烷型倍半萜类化合物选自马兜铃酮和甘松香酮H中的至少一种。马兜铃烷型倍半萜类化合物能够舒张胸主动脉、肠系膜动脉和冠状动脉,舒张血管,增加血管内皮细胞中eNOS磷酸化,保护血管内皮细胞,降低血压,从而能够用于预防和/或治疗心脑血管疾病,进而用于制备预防和/或治疗心脑血管疾病的药物。本申请还提供了包含马兜铃烷型倍半萜类化合物的药物组合物及其在制备预防和/或治疗心脑血管疾病的药物中的用途,其中,所述马兜铃烷型倍半萜类化合物选自马兜铃酮和甘松香酮H中的至少一种。(The application provides an application of aristolochiane type sesquiterpene compounds in preparation of medicines for preventing and/or treating cardiovascular and cerebrovascular diseases, wherein the aristolochiane type sesquiterpene compounds are selected from at least one of aristolocone and nardosolone H. The aristolochiane type sesquiterpenoids can relax thoracic aorta, mesenteric artery and coronary artery, relax blood vessel, increase eNOS phosphorylation in vascular endothelial cells, protect vascular endothelial cells and reduce blood pressure, so that the aristolochiane type sesquiterpenoids can be used for preventing and/or treating cardiovascular and cerebrovascular diseases, and further can be used for preparing medicines for preventing and/or treating the cardiovascular and cerebrovascular diseases. The application also provides a pharmaceutical composition containing the aristolochiane type sesquiterpene compound and application thereof in preparing a medicament for preventing and/or treating cardiovascular and cerebrovascular diseases, wherein the aristolochiane type sesquiterpene compound is selected from at least one of aristolocone and nardosolone H.)

1. Use of aristolochiane type sesquiterpenes in preparation of drugs for preventing and/or treating cardiovascular and cerebrovascular diseases, wherein the aristolochiane type sesquiterpenes are selected from at least one of aristolocone and nardosolone H.

2. The use of claim 1, wherein the cardiovascular and cerebrovascular disease comprises at least one of atherosclerosis, coronary heart disease, myocardial infarction, hypertension, hypertensive heart disease, diabetic vascular complications, restenosis following angioplasty, stroke, pulmonary hypertension, pulmonary heart disease, vasculitis, vascular headache, microangiopathy.

3. A pharmaceutical composition comprising an aristolochiane-type sesquiterpene compound selected from at least one of aristolocone and narcolepsyn H.

4. The pharmaceutical composition according to claim 3, wherein the aristolochiane-type sesquiterpene compound is provided in the form of a monomer or in the form of a plant extract containing the same.

5. The pharmaceutical composition as claimed in claim 4, wherein the plant extract is at least one selected from the group consisting of nardostachys chinensis extract, aristolochia debilis extract, cyperus rotundus extract, acorus calamus extract and acorus gramineus soland extract.

6. The pharmaceutical composition of claim 3, wherein the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

7. The pharmaceutical composition according to claim 6, wherein the pharmaceutically acceptable carrier or excipient is selected from at least one of a solvent, a diluent, a disintegrant, a precipitation inhibitor, a surfactant, a glidant, a binder, a lubricant, a dispersant, a suspending agent, an isotonicity agent, a thickener, an emulsifier, a preservative, a stabilizer, a hydrating agent, an emulsification accelerator, a buffer, an absorbent, a colorant, a flavoring agent, a sweetener, an ion exchanger, a mold release agent, a coating agent, a flavoring agent, or an antioxidant.

8. Use of the pharmaceutical composition according to any one of claims 3-7 in the preparation of a medicament for the prevention and/or treatment of cardiovascular and cerebrovascular diseases.

9. The use of claim 8, wherein the cardiovascular and cerebrovascular disease comprises at least one of atherosclerosis, coronary heart disease, myocardial infarction, hypertension, hypertensive heart disease, diabetic vascular complications, restenosis following angioplasty, stroke, pulmonary hypertension, pulmonary heart disease, vasculitis, vascular headache, microangiopathy.

Technical Field

The application relates to the technical field of medicines, in particular to application of aristolochiane type sesquiterpenes in preparation of medicines for preventing and/or treating cardiovascular and cerebrovascular diseases.

Background

With the development of science and technology and medicine, cardiovascular and cerebrovascular diseases are continuously researched and treated with considerable results, but a large number of patients still die every year because of the failure of effective treatment. The occurrence of cardiovascular and cerebrovascular diseases is related to vascular conditions, vascular endothelial cells are important regulatory sites for maintaining vascular homeostasis, and vascular dysfunction can cause abnormal contraction of blood vessels, resulting in a series of cardiovascular and cerebrovascular diseases. Therefore, the medicine with the functions of relaxing blood vessels and protecting vascular endothelial cells is searched for, is used for preventing and treating cardiovascular and cerebrovascular diseases, and has important social significance and economic significance.

Disclosure of Invention

The present inventors have found, through intensive studies, that aristolochiane-type sesquiterpene compounds have a vasodilating effect and are capable of protecting vascular endothelial cells, and thus can be used for preventing and/or treating cardiovascular and cerebrovascular diseases, and have completed the present application based on this finding.

The first aspect of the application provides the use of aristolochiane type sesquiterpenes in the preparation of medicines for preventing and/or treating cardiovascular and cerebrovascular diseases, wherein the aristolochiane type sesquiterpenes are selected from at least one of aristolocone and nardostachyne H.

A second aspect of the present application provides a pharmaceutical composition comprising an aristolochiane-type sesquiterpene compound selected from at least one of aristolocone and narcolepsyn H.

A third aspect of the present application provides a use of the pharmaceutical composition of the second aspect of the present application in the preparation of a medicament for preventing and/or treating cardiovascular and cerebrovascular diseases.

The aristolochiane type sesquiterpenoids can relax thoracic aorta, mesenteric artery and coronary artery, relax blood vessel, increase eNOS phosphorylation in vascular endothelial cells, protect vascular endothelial cells and reduce blood pressure, so that the aristolochiane type sesquiterpenoids can be used for preventing and/or treating cardiovascular and cerebrovascular diseases, and further can be used for preparing medicines for preventing and/or treating the cardiovascular and cerebrovascular diseases. Furthermore, the medicine composition containing the aristolochiane type sesquiterpenoids can also be used for preparing medicines for preventing and/or treating cardiovascular and cerebrovascular diseases.

Drawings

In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present application, and it is also obvious for a person skilled in the art to obtain other embodiments according to the drawings.

FIG. 1 shows the effect of different concentrations of narcolesterone H on the diastolic rate of the thoracic aorta, mesenteric artery and coronary artery.

FIG. 2 shows the effect of different concentrations of aristolocone on thoracic aorta, mesenteric artery and coronary artery diastolic rates.

FIG. 3 shows the effect of aristolocone on the p-eNOS content in vascular endothelial cells.

Figure 4 shows the effect of aristolocone on blood pressure.

Detailed Description

The technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are only a part of the embodiments of the present application, and not all of the embodiments. All other embodiments that can be derived by one of ordinary skill in the art from the description herein are intended to be within the scope of the present disclosure.

The first aspect of the application provides the use of aristolochiane type sesquiterpenes in the preparation of medicines for preventing and/or treating cardiovascular and cerebrovascular diseases, wherein the aristolochiane type sesquiterpenes are selected from at least one of aristolocone and nardostachyne H.

In the application, the constitutional formula of aristolocone ((-) -aristolone, CAS number: 25274-27-5) is shown as formula I, and the constitutional formula of nardostachynone H (kanshone H, CAS number: 1445952-33-9) is shown as formula II.

The inventor finds in research that the aristolochiane type sesquiterpenoids can relax thoracic aorta, mesenteric artery and coronary artery, relax blood vessel, increase eNOS phosphorylation of vascular endothelial cells, protect vascular endothelial cells and reduce blood pressure, so that the aristolochiane type sesquiterpenoids can be used for preventing and/or treating cardiovascular and cerebrovascular diseases, and can be further used for preparing medicines for preventing and/or treating the cardiovascular and cerebrovascular diseases.

In the present application, the term "treatment" has its ordinary meaning and herein may refer to the treatment of a mammalian subject (preferably a human) already suffering from a cardiovascular or cerebrovascular disease with a medicament of the present application in order to produce a therapeutic, curative, palliative, etc. effect on said disease. Similarly, as used herein, the term "prevention" has its ordinary meaning and may refer herein to the treatment of a mammalian subject who may or is at risk of suffering from a cardiovascular or cerebrovascular disease with an agent of the present application in an effort to prevent, arrest, abrogate, etc. the disease.

In some embodiments of the first aspect of the present application, the cardiovascular and cerebrovascular diseases include, but are not limited to, at least one of atherosclerosis, coronary heart disease, myocardial infarction, hypertension, hypertensive heart disease, diabetic vascular complications, restenosis following angioplasty, stroke, pulmonary hypertension, pulmonary heart disease, vasculitis, vascular headache, microangiopathy.

A second aspect of the present application provides a pharmaceutical composition comprising an aristolochiane-type sesquiterpene compound selected from at least one of aristolocone and narcolepsyn H.

In some embodiments of the second aspect of the present application, the aristolochiane-type sesquiterpene compound is provided as a monomer or as a plant extract comprising the same; wherein the plant extract is at least one selected from Nardostachys chinensis extract, fructus Aristolochiae extract, rhizoma Cyperi extract, rhizoma Acori Calami extract and rhizoma Acori Graminei extract. In the present application, the extraction manner of the extract is not particularly limited as long as the purpose of the present application can be achieved, and the person skilled in the art can obtain the plant extract containing the aristolochiane-type sesquiterpene compound by any conventional manner, and exemplarily, the plant extract containing the aristolochiane-type sesquiterpene compound can be obtained by performing heating reflux extraction with 70-80% ethanol solution.

In some embodiments of the second aspect of the present application, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

In some embodiments of the second aspect of the present application, the pharmaceutically acceptable carrier or excipient is selected from at least one of a solvent, diluent, disintegrant, precipitation inhibitor, surfactant, glidant, binder, lubricant, dispersant, suspending agent, isotonic agent, thickener, emulsifier, preservative, stabilizer, hydrating agent, emulsification accelerator, buffer, absorbent, colorant, flavoring agent, sweetener, ion exchanger, mold release agent, coating agent, flavoring agent, or antioxidant.

Herein, "pharmaceutically acceptable" means having no substantial toxic effect when used in the usual dosage amounts, and thus being approved by the government or equivalent international organization or approved for use in animals, more particularly in humans, or registered in the pharmacopoeia.

The "pharmaceutically acceptable carrier or excipient" useful in the pharmaceutical compositions of the present application may be any conventional carrier in the art of pharmaceutical formulation, and the selection of a particular carrier will depend on the mode of administration or the type and state of the disease used to treat a particular patient. The preparation of suitable pharmaceutical compositions for a particular mode of administration is well within the knowledge of those skilled in the pharmaceutical art.

As used herein, the term "pharmaceutical composition" has its ordinary meaning. In addition, the "pharmaceutical composition" of the present application may also be present or provided in the form of a health product, an essential oil, a functional food, a food additive, or the like. The pharmaceutical compositions of the present application can be prepared by conventional techniques in the pharmaceutical field, particularly in the formulation field, by obtaining the active ingredients of the raw materials of the pharmaceutical compositions of the present application by extraction, separation and purification means commonly used in pharmaceutical manufacturing, optionally mixing with one or more pharmaceutically acceptable carriers or excipients, and then forming the desired dosage form. The pharmaceutical composition according to the present application is a pharmaceutical preparation which can be suitably used for oral administration, a pharmaceutical preparation (e.g., solution) suitable for parenteral injection (e.g., intravenous injection, subcutaneous injection), a pharmaceutical preparation (e.g., ointment, patch or cream) suitable for surface administration, or a pharmaceutical preparation (e.g., suppository) suitable for rectal administration, and the like. Dosage forms for oral administration may include, for example, tablets, pills, hard or soft capsules, solutions, suspensions, emulsions, syrups, powders, fine granules, pellets, elixirs and the like, without limitation. In addition to the active ingredient, these preparations may contain diluents (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and glycine), lubricants (e.g., silica, talc, stearic acid or its magnesium salt, calcium salt, and polyethylene glycol). Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, and polyvinylpyrrolidone. If necessary, it may further contain pharmaceutically acceptable additives such as disintegrating agents (e.g., starch, agar, alginic acid or sodium salt thereof), absorbents, coloring agents, flavoring agents, sweetening agents, and the like. Tablets may be prepared according to conventional mixing, granulating or coating methods.

A third aspect of the present application provides a use of the pharmaceutical composition of the second aspect of the present application in the preparation of a medicament for preventing and/or treating cardiovascular and cerebrovascular diseases.

In some embodiments of the third aspect of the present application, the cardiovascular and cerebrovascular diseases include, but are not limited to, at least one of atherosclerosis, coronary heart disease, myocardial infarction, hypertension, hypertensive heart disease, diabetic vascular complications, restenosis following angioplasty, stroke, pulmonary hypertension, pulmonary heart disease, vasculitis, vascular headache, microangiopathy.

Terms used herein have their ordinary meanings as known to those skilled in the art, if not explicitly stated or defined.

Materials: 9, 11-dideoxy-11 α,9 α -methyleneepoxyprostaglandin F2 α (11-Didexy-11 α,9 α -epoxyethane prostaglandin F2 α, u46619), acetylcholine (Ach), Dimethylsulfoxide (DMSO): sigma corporation; calcium channel agonists a 23187: MCE corporation; potassium chloride (KCl)): tianjin, Guang Compound science and technology development Co., Ltd; human Umbilical Vein Endothelial Cells (HUVEC), endothelial cell culture medium: ALLCELLS Biotechnology, Inc.; phosphorylated nitric oxide synthase 3(p-eNOS, Ser 117): abcam corporation; nitric oxide synthase 3(eNOS), antibody to glyceraldehyde-3-phosphate dehydrogenase (GAPDH), goat anti-rabbit ii antibody (Proteintech): CST corporation, USA; preparing a kit, a TBST solution, an electrophoresis solution and a membrane transferring solution by SDS-PAGE gel: kang is a century corporation; 5% BSA blocking solution: beijing Solaibao Biometrics, Inc.; medical tissue glue: beram medical spanish limited; deoxycorticosterone acetate (DOCA): TCI Corp; food-grade olive oil: osmunda pulchella. The instrument comprises the following steps: multichannel isolated vessel tonometry system 630 AM: DMT company; PowerLab biological signal acquisition and analysis system: ADInstructions Inc.; blood pressure remote sensing monitoring system: millar Instruments Inc.; body type microscope: LEICA corporation; biosafety cabinet and CO₂An incubator: likang biomedical science and technology Consortium Co., Ltd; a multifunctional microplate reader: TECAN corporation; gel imaging system: japan GE corporation; electrophoresis apparatus and transfer membrane appearance: Bio-Rad.

The experimental materials and methods used in the following examples are, unless otherwise specified, conventional materials and methods.

Example 1 Effect of Aristolochiane-type sesquiterpenes on thoracic aorta, mesenteric artery and coronary artery relaxation

Selecting 8-10 week-old SPF male C57BL/6 mice with weight of 18-25 g; male SD rats at 8 weeks of age, body weight 200-. Anaesthetizing, taking the thoracic aorta, mesenteric artery and coronary artery of SD rat of C57BL/6 mouse, removing fat and connective tissue around blood vessel, cutting into 3-4mm long blood vessel ring, placing the blood vessel ring in Krebs-Hennseleit (K-H) solution (composition of K-H solution: NaCl 118mmol/L, KCl 4.7.7 mmol/L, NaHCO)₃25mmol/L、KH₂PO₄1.2mmol/L、MgSO₄ 1.2mmol/L、CaCl₂1.3mmol/L, D-glucose 10mmol/L) at 37 ℃ and with continuous introduction of 95% O₂And 5% CO₂Mixing the gases, and under microscope, passing the blood vessel ring through two parallel steel needlesThe pressure sensor is connected, the other one is connected with the tension adjusting knob, no action force is given to the blood vessel at the moment, the blood vessel adapts for 15min under the unstressed state, and the instrument is zeroed; rotating a tension adjusting knob, adjusting the initial tension of the blood vessel to 5mN, replacing K-H liquid, replacing the K-H liquid once every 15min, if the tension is reduced, adjusting to 5mN again, and balancing the blood vessel for 60 min; adding 20nmol/L u46619 for stimulating vasoconstriction, and sequentially adding 10% of the final concentration after the maximum constriction value^-9、10^-8、10^-7、10^-6、10^-5measuring the contractility of the acetylcholine after the acetylcholine is added by adopting a multichannel in vitro vascular tension measuring system, and calculating the diastolic rate according to the following formula: the diastole rate is (1- (the contractility after adding acetylcholine-the initial tension)/u 46619 maximum contractility) multiplied by 100%, the maximum diastole rate of the blood vessel reaches 60-80%, which indicates that the blood vessel endothelium is intact, so that the next experiment can be carried out. After the vascular endothelial detection is completed, u46619 is added into a bath tank in which thoracic aorta, mesenteric artery and coronary artery are incubated respectively to stimulate vasoconstriction, after the constriction is stable, nardosinone H (taking DMSO as a solvent) with final concentration of 10, 25, 50, 75 and 100 mu mol/L or aristolocone (taking DMSO as a solvent) with final concentration of 10, 50, 100, 150 and 200 mu mol/L is added in sequence, meanwhile, equal volume of DMSO is added in sequence in a control group, the administration intervals are all 5min, the relaxation rates are recorded respectively, and a relaxation curve is drawn to obtain the influence results of the nardosinone H on the thoracic aorta, mesenteric artery and coronary artery relaxation rates as shown in figure 1, and the influence results of the aristolocone on the thoracic aorta, mesenteric artery and coronary artery relaxation rates are shown in figure 2.

As shown in fig. 1, galbanolone H significantly increased the diastolic rate of thoracic aorta in C57BL/6 mice (n 13, P <0.01, compared to the control group), significantly increased the diastolic rate of mesenteric artery in C57BL/6 mice (n 4, P <0.01, compared to the control group), and significantly increased the diastolic rate of coronary artery in SD rats (n 4, P <0.01, compared to the control group), indicating that galbanolone H significantly dilated the thoracic aorta, mesenteric artery, and coronary artery, and has a vasodilating effect. As can be seen from fig. 2, aristolocone significantly increased the diastolic rate of thoracic aorta in C57BL/6 mice (n 13, P <0.01, compared to the control group), significantly increased the diastolic rate of mesenteric artery in C57BL/6 mice (n 4, P <0.01, compared to the control group), and significantly increased the diastolic rate of coronary artery in SD rats (n 4, P <0.01, compared to the control group), indicating that aristolocone significantly dilated thoracic aorta, mesenteric artery, and coronary artery, and has a vasodilating effect.

Example 2 Effect of Aristolochiane-type sesquiterpenes on p-eNOS

The inventor finds in research that vascular endothelial cells can release vasodilation factors such as Nitric Oxide (NO) and the like to relax blood vessels, and the aristolocidine type sesquiterpenes of the application can stimulate the vascular endothelial cells to release NO by stimulating eNOS phosphorylation in the vascular endothelial cells, so as to relax blood vessels.

At 37 ℃ and 5% O₂And 95% CO₂Human Umbilical Vein Endothelial Cells (HUVEC) were cultured under the conditions, inoculated into 6-well plates, added with aristolocones (DMSO as a solvent) at concentrations of 10, 50, 100, 150, and 200. mu. mol/L, respectively, while DMSO was added to a control group, A23187 (DMSO as a solvent) was added at a concentration of 1. mu. mol/L to a positive administration group, and administration time was 15min, and the cells were harvested, extracted for protein, and the change in p-eNOS content of HUVEC cells after administration was detected by Western Blot, as shown in FIG. 3. As can be seen from fig. 3, different concentrations of aristolocone can significantly increase the P-eNOS content (n-4, P)<0.05, compared to the control group), indicating that aristolocone can increase eNOS phosphorylation of vascular endothelial cells, thereby protecting vascular endothelial cells and dilating blood vessels.

Example 3 Effect of Aristolochiane-type sesquiterpene Compounds on blood pressure

Selecting 6 male SD rats with the age of 8-10 weeks, anesthetizing the SD rats with isoflurane with the weight of 200-; injecting deoxycorticosterone acetate (DOCA)50mg/kg subcutaneously for one week; the drinking water was changed to 0.9% NaCl and 0.2% KCl for 4 weeks to induce hypertension in rats. Blood pressure in the abdominal aorta is continuously measured by implant telemetry. The aristolocone takes olive oil as a solvent, an injection solvent with the concentration of 100mg/ml is prepared, 1ml/kg is injected into the abdominal cavity, 1ml/kg of the olive oil is injected into the abdominal cavity of a control group, the abdominal aorta blood pressure is recorded, the difference value between the blood pressure before administration and the blood pressure before administration of the aristolocone and the blood pressure before administration of the olive oil is calculated according to the blood pressure value after administration, and according to the graph 4, the difference value between the systolic pressure, the mean arterial pressure and the diastolic pressure before administration and after administration of the aristolocone is negative, so that the aristolocone is shown to reduce the systolic pressure, the mean arterial pressure and the diastolic pressure and has significant difference (n is 3, P is less than 0.05 compared with the control group), and the result shows that the aristolocone has the function of reducing the blood pressure.

In conclusion, the aristolochiane type sesquiterpene compounds nardostachyne H and aristolochiane can relax thoracic aorta, mesenteric artery and coronary artery and blood vessel, and the aristolochiane can increase eNOS phosphorylation in vascular endothelial cells, protect the vascular endothelial cells and reduce blood pressure, so that the aristolochiane type sesquiterpene compounds can be used for preventing and/or treating cardiovascular and cerebrovascular diseases, and can be used for preparing medicines for preventing and/or treating the cardiovascular and cerebrovascular diseases. Furthermore, the medicine composition containing the aristolochiane type sesquiterpenoids can also be used for preparing medicines for preventing and/or treating cardiovascular and cerebrovascular diseases.

The above description is only for the preferred embodiment of the present application and is not intended to limit the scope of the present application. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application are included in the protection scope of the present application.