Application of tormentic acid in preparation of products for preventing and treating hypertension

Abstract

The invention discloses application of tormentic acid in preparation of products for preventing and treating hypertension. It is found for the first time that tormentic acid or pharmaceutically acceptable salt thereof can be applied to preparation of the medicine for preventing or treating hypertension or heart and vasculopathy caused by hypertension, the treatment effect is good, safety is achieved, and a new treatment approach is provided for treating hypertension or hypertension-related diseases.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to use of potentilla acid in preparing products for preventing and treating hypertension. Background Art

[0002] Hypertension is one of the major chronic diseases in the world today. The increasing morbidity and mortality of hypertension have attracted great attention from the medical community at home and abroad. Hypertension not only has a high incidence, but is also a major global public health problem and the leading risk factor for death. A study on atherosclerosis risk based on a population community conducted abroad showed that up to 25% of cardiovascular events (including coronary artery revascularization for coronary heart disease, stroke, and heart failure) are ultimately attributed to hypertension. It can be seen that the prevention and control of hypertension requires the search for safer and more effective prevention and treatment strategies, and it is urgent to find antihypertensive drugs with good therapeutic effects.

[0003] Potentilla acid is widely found in plants of Boraginaceae, Lamiaceae and Rosaceae. Studies in recent years have found that potentilla acid has pharmacological activities such as antioxidant, anti-inflammatory, analgesic, antibacterial, anti-tumor, lipid-lowering and liver-protecting, anti-atherosclerotic, platelet aggregation inhibition, abnormal vascular smooth muscle proliferation inhibition and blood sugar reduction. It can inhibit the activity of glycogen phosphorylase in vitro and improve blood sugar. For example, CN106581006 discloses that hawthorn acid and potentilla acid can be used to treat Parkinson's disease; CN111297820 discloses enteric-coated tablets made with thymosin and potentilla acid as active ingredients, and it was found that potentilla acid can not only effectively reduce the absorption fluctuation problem of thymosin enteric-coated preparations, but also promote the immunoregulatory effect of thymosin; TW104112036 discloses that potentilla acid, an extract of loquat leaf cells, can be used to lower blood sugar and / or reduce fatty liver; CN114053282A discloses the use of potentilla acid and anisodamine capsules in the preparation of drugs for treating diabetes; in addition, there are reports that potentilla acid improves prostate cancer and has a protective effect on Helicobacter pylori-induced GES-1 cell damage.

[0004] However, there are no reports on the effects of potentilla acid on cardiovascular diseases such as hypertension. Summary of the invention

[0005] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide a use of potentianic acid in preparing a product for preventing and treating hypertension.

[0006] To achieve the above objectives and other related objectives, the present invention is achieved through the following technical solutions.

[0007] The first aspect of the present invention protects the use of potentianic acid or a pharmaceutically acceptable salt thereof in the preparation of a product for preventing or treating hypertension.

[0008] In certain embodiments, the potentianic acid or a pharmaceutically acceptable salt thereof is used alone or in combination with other drugs.

[0009] In certain embodiments, the efficacy of the product comprises reducing one or more of systolic blood pressure, diastolic blood pressure, and mean arterial pressure.

[0010] In certain embodiments, the potentilla acid or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient constitute a pharmaceutical composition.

[0011] In certain specific embodiments, the pharmaceutically acceptable excipient is selected from one or more of a carrier, a diluent, a binder, a lubricant and a wetting agent.

[0012] In certain specific embodiments, the dosage form of the pharmaceutical composition is selected from one or more of solutions, suspensions, emulsions, injections, sprays, aerosols, tablets, capsules, powders and granules.

[0013] In certain specific embodiments, the pharmaceutical composition is an oral preparation.

[0014] The second aspect of the present invention is to protect the use of potentilla acid or its pharmaceutically acceptable salt in the preparation of a product for improving at least one of the following symptoms 1) to 4) caused by hypertension:

[0015] 1) Decreased vascular function;

[0016] 2) Thickening of blood vessel walls;

[0017] 3) Reduced heart function;

[0018] 4) Cardiomyocyte hypertrophy.

[0019] In certain embodiments, at least one of the following items A1)-A4) is included:

[0020] A1) Decreased vascular function refers to an increase in vascular resistance index or pulsatility index;

[0021] A2) Decreased cardiac function refers to decreased cardiac ejection fraction or stroke volume;

[0022] A3) Vascular thickening refers to the thickening of the blood vessel walls;

[0023] A4) Cardiomyocyte hypertrophy is an increase in the number of cells that causes the cardiomyocytes to become misaligned, smaller, or smaller.

[0024] The third aspect of the present invention protects a drug for preventing or treating hypertension, the drug comprising potentilla acid, and the drug has at least one of the following uses B1) to B6):

[0025] B1) Reduce high blood pressure;

[0026] B2) Improve decreased vascular function;

[0027] B3) Improve the thickness of blood vessel walls;

[0028] B4) Improve decreased heart function;

[0029] B5) Improve cardiomyocyte hypertrophy.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] The present invention discovers for the first time that cinquefoic acid or a pharmaceutically acceptable salt thereof can be used to prevent or treat hypertension or heart and vascular diseases caused by hypertension, and the treatment effect is good and safe, providing a new treatment approach for treating hypertension or hypertension-related diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1A A graph showing the effect of potentianic acid on the systolic blood pressure of mice after AngII modeling in the present application.

[0033] Figure 1B A graph showing the effect of potentianic acid on the diastolic blood pressure of mice after AngII modeling in the present application.

[0034] Figure 1C The graph shows the effect of potentianic acid on the mean arterial pressure of mice after AngII modeling in the present application.

[0035] Figure 1D The figure shows the effect of cinnamic acid on the body weight of mice after AngII modeling in the present application.

[0036] Figure 2A Ultrasound graph showing the effect of cinnamic acid on the abdominal aorta resistance index and pulsatility index of mice after AngII modeling in the present application.

[0037] Figure 2B The statistical analysis graph shows the effect of potentinic acid on the abdominal aorta pulsatility index of mice after AngII modeling in the present application.

[0038] Figure 2C The statistical analysis graph shows the effect of potentinic acid on the abdominal aorta resistance index of mice after AngII modeling in the present application.

[0039] Figure 2D Ultrasound images showing the effect of potentinic acid on the vascular wall thickness of the abdominal aorta of mice after AngII modeling in the present application.

[0040] Figure 2EThe statistical analysis graph shows the effect of potentilla acid on the vascular wall thickness of the abdominal aorta of mice after AngII modeling in the present application.

[0041] Figure 2F A diagram showing the effect of potentinic acid on the pathological morphology of the abdominal aorta in mice after AngII modeling in the present application.

[0042] Figure 3A Ultrasound diagram showing the cardiac function of mice after AngII modeling with potentinic acid in the present application (arrow: left ventricular end-diastolic / systolic diameter).

[0043] Figure 3B The diagram shows the analysis of the effect of potentianic acid on the cardiac function of mice after AngII modeling in the present application.

[0044] Figure 3C The analysis diagram shows the effect of cinnamic acid on the cardiac stroke volume function of mice after AngII modeling in the present application.

[0045] Figure 3D A diagram showing the effect of potentinic acid on cardiac pathological morphology in mice after AngII modeling in the present application. DETAILED DESCRIPTION

[0046] In the arterial system, the frictional force generated in the direction of blood flow when blood flows through the surface of the blood vessel is called wall shear stress (WSS); the circumferential stretching generated when blood pressure acts perpendicularly on the blood vessel wall and the force that changes periodically with blood pressure is called annular tension; both are the main forces that promote and maintain vascular remodeling.

[0047] AngⅡ increases intravascular WSS and annular tension by constricting blood vessels, putting systemic resistance vessels in a high stress state, increasing peripheral vascular resistance and pulsatility index, leading to reduced vascular elasticity; increased blood pressure causes changes in blood rheology patterns and increased blood turbulence. Hypertension can lead to left ventricular hypertrophy, high atrial pressure, and other conditions, reducing cardiac ejection fraction and cardiac stroke volume.

[0048] The first aspect of the present invention provides the use of cinnamic acid or a pharmaceutically acceptable salt thereof in preparing a product for preventing or treating hypertension. The cinnamic acid or a pharmaceutically acceptable salt thereof of the present invention can reduce the systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MAP) of the subject, and is safe to the subject and has no toxic side effects.

[0049] Clinical research and practice guidelines generally define hypertension as systolic blood pressure (SBP) ≥140 mm Hg and / or diastolic blood pressure ≥90 mm Hg. Normal blood pressure is defined as SBP less than 120 mm Hg and diastolic blood pressure less than 80 mm Hg.

[0050] Potentilla acid (2A, 19A-dihydroxyursolic acid, TA) is also a widely distributed natural triterpenoid compound, which exists in plants such as Potentilla, loquat leaves, willow herb, quince, root of the sedge, and root of golden cherry. Potentilla acid is a white powdery substance with a molecular formula of C 30 H 48 O5, molecular weight is 489, CAS number is 13850-16-3, and its structural formula is as follows:

[0051]

[0052] In addition, the above-mentioned potentilla acid can also be artificially synthesized potentilla acid. No matter it is naturally extracted or artificially synthesized, it can be used to prevent or treat hypertension.

[0053] In certain embodiments, the pharmaceutically acceptable salt refers to a salt prepared from a pharmaceutically acceptable non-toxic acid or base, wherein the acid or base includes an inorganic acid, an inorganic base, an organic acid, and an organic base. The inorganic acid is selected from hydrochloric acid, hydrobromic acid, phosphoric acid, hydroiodic acid, or sulfuric acid. The inorganic base is selected from calcium, magnesium, lithium, sodium, zinc, aluminum, or potassium. The organic acid is selected from formic acid, glycolic acid, propionic acid, acetic acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, maleic acid, glutamic acid, benzoic acid, stearic acid, alginic acid, benzenesulfonic acid, glucuronic acid, pamoic acid, or galacturonic acid. The organic base is selected from diethanolamine, choline, procaine, lysine, or 1,2-ethylenediamine.

[0054] In certain embodiments, the potentinic acid or its pharmaceutically acceptable salt is used alone or in combination with other drugs, that is, the potentinic acid or its pharmaceutically acceptable salt can be used as the only active ingredient or one of the active ingredients of the product.

[0055] In certain embodiments, the potentilla acid or its pharmaceutically acceptable salt and a pharmaceutically acceptable excipient constitute a pharmaceutical composition. The pharmaceutically acceptable means that when the drug is properly administered to an animal or a human, they will not produce adverse, allergic or other adverse reactions. The pharmaceutically acceptable excipient should be compatible with the potentilla acid, that is, it can be mixed with it without significantly reducing the effect of the potentilla acid under normal circumstances.

[0056] In certain specific embodiments, the pharmaceutically acceptable excipient is selected from one or more of a carrier, a diluent, a binder, a lubricant and a wetting agent.

[0057] In certain more specific embodiments, specific examples of some substances that can serve as pharmaceutically acceptable carriers, diluents, binders, lubricants and wetting agents are sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium methylcellulose, ethylcellulose and methylcellulose; tragacanth powder; malt; gelatin; talc; solid lubricants, such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils, such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and cocoa butter; polyols, such as propylene glycol, glycerol, sorbitol, mannitol and polyethylene glycol; alginic acid; emulsifiers, such as Tween; wetting agents, such as sodium lauryl sulfate; colorants; flavoring agents; tableting, stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline solution; and phosphate buffer, etc. These substances are used as needed to help the stability of the formulation or to help improve the activity or its biological effectiveness or to produce an acceptable taste or smell in the case of oral administration.

[0058] In certain embodiments, the dosage form of the pharmaceutical composition is selected from one or more of a solution, a suspension, an emulsion, an injection, a spray, an aerosol, a tablet, a capsule, a powder and a granule. The drugs in the above-mentioned various dosage forms can be prepared according to conventional methods in the pharmaceutical field. Preferably, it is one or more of a solution, a suspension, an emulsion, an injection, a spray, an aerosol, a tablet, a capsule, a powder and a granule.

[0059] In certain embodiments, the pharmaceutical composition is suitable for oral administration, and the dosage form suitable for oral administration includes solid dosage forms and liquid preparations. The solid dosage forms include capsules, tablets, pills, powders and granules. In these solid dosage forms, cinnamic acid or its pharmaceutically acceptable salt is obtained according to the prior art with at least one conventional inert excipient, filler or solubilizer, adhesive, humectant, disintegrant, solubilizer, adsorbent, lubricant. In capsules, tablets and pills, the dosage form may also include a buffer. Solid dosage forms such as tablets, sugar pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compound in this composition can be released in a delayed manner in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into a microcapsule form with one or more of the above-mentioned excipients. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers, and emulsifier adjuvants.

[0060] In certain embodiments, the pharmaceutical composition is suitable for enteral or parenteral administration, such as by injection, spraying, nasal drops, eye drops, penetration, absorption, physical or chemical mediated methods such as muscle, intradermal, subcutaneous, intravenous, mucosal tissue; or mixed or wrapped with other substances and then introduced into the body. Preferably, it is administered enterally. Therefore, preferably, the pharmaceutical composition also includes an antioxidant, a buffer, an antibacterial agent, and a solute that makes the preparation isotonic with the subject's blood, as well as an aqueous and non-aqueous sterile suspension, which may contain a suspending agent, a solubilizer, a thickener, a stabilizer and a preservative.

[0061] In certain embodiments, the preparation of the drug is preferably in unit dosage form. In this form, the preparation is subdivided into unit doses containing appropriate amounts of active ingredients. The unit dosage form can be a solution, injection, injection, capsule, tablet or any dosage form; in addition, the unit dosage form can also be a packaged preparation, such as a solution, injection, tablet, capsule and powder packaged in a vial or ampoule. The amount of the active ingredient in the unit dosage preparation can be changed or adjusted from 0.1 mg to 1000 mg, depending on the specific application and efficacy of the active ingredient.

[0062] In certain embodiments, the drug may also be used in combination with other treatments, including surgery, radiotherapy, chemotherapy, and targeted therapy.

[0063] Another aspect of the present invention provides the use of cinnamic acid or a pharmaceutically acceptable salt thereof in the preparation of a product for improving at least one of the following symptoms A1) to A4) caused by hypertension:

[0064] A1) Decreased vascular function refers to a decrease in vascular resistance index or pulsatility index;

[0065] A2) Decreased cardiac function refers to decreased cardiac ejection fraction or stroke volume;

[0066] A3) Vascular thickening refers to the thickening of the blood vessel walls;

[0067] A4) Cardiomyocyte hypertrophy refers to an increase in the number of cells that causes the cardiomyocytes to become misaligned, smaller, or smaller.

[0068] In certain embodiments, the reduced vascular function refers to a reduction in the vascular resistance index or pulsatility index, the improved reduced vascular function refers to an increase in the vascular resistance index or pulsatility index, and the increased vascular pulse wave propagation velocity refers to bringing the vascular resistance index or pulsatility index of the subject close to or reaching the velocity of healthy blood vessels. For example, based on healthy blood vessels, the product can restore the resistance index or pulsatility index of the subject's blood vessels to at least 70%, 80%, 90% or 100% of that of healthy blood vessels. The abdominal aorta resistance index and the abdominal aorta pulsatility index are ultrasonic indicators for evaluating vascular elasticity and function, vascular elasticity and functional ultrasonic indicators. Vascular elasticity refers to the ability of blood vessels to expand and contract normally, which is essential for maintaining normal blood pressure and blood flow. A decrease in vascular elasticity may lead to increased blood pressure, changes in hemodynamics, and an increased risk of cardiovascular disease. The present invention finds that potentilla acid or a pharmaceutically acceptable salt thereof can increase the abdominal aorta resistance index and the abdominal aorta pulsatility index, that is, increase the elasticity of blood vessels and reduce blood flow resistance.

[0069] In certain embodiments, the thickening of the vascular wall refers to the thickening of the vascular wall of the abdominal aorta, and the improvement of vascular thickening refers to reducing the thickness of the vascular wall, and reducing the thickness of the vascular wall refers to making the vascular wall of the subject approach or reach the thickness of a healthy blood vessel. For example, based on healthy blood vessels, the product can make the thickness of the vascular wall of the subject's blood vessel not exceed 130%, 120%, 110% or 100% of the thickness of healthy blood vessels. Hypertension is a common chronic disease, and the vascular wall may gradually thicken due to the impact of high-pressure blood flow for a long time, which may cause the thickening of the vascular wall; and the thickening of the vascular wall may cause the stenosis of the vascular lumen, increase blood flow resistance, and further aggravate the symptoms of hypertension. The present invention finds that potentilla acid or a pharmaceutically acceptable salt thereof can alleviate or inhibit the thickening of the vascular wall, that is, reduce the thickness of the vascular wall and relieve the pressure on the vascular wall.

[0070] In certain embodiments, the reduced cardiac function refers to a reduced cardiac ejection fraction or a reduced stroke volume, the improved reduced cardiac function refers to an increase in cardiac ejection fraction or stroke volume, and the increased cardiac ejection fraction or stroke volume refers to bringing the cardiac ejection fraction and stroke volume of the subject close to or reaching the fraction of a healthy heart. For example, based on a healthy heart, the product can restore the cardiac ejection fraction and stroke volume of the subject's heart to at least 70%, 80%, 90% or 100% of that of a healthy heart. The cardiac ejection fraction is an important indicator for measuring the heart's ability to pump blood, which indicates the percentage of the amount of blood discharged by the ventricle to the end-stage volume of the ventricle when the heart contracts each time. When the ejection fraction decreases, it means that the efficiency of the heart's pumping decreases, which may cause the heart to be unable to effectively transport blood to the whole body, thereby causing symptoms such as heart failure. Due to long-term high blood pressure, the heart of hypertensive patients needs more power to promote blood flow, which may lead to increased heart load, gradual thickening of the heart muscle, and decreased elasticity of the heart wall, which may eventually cause a decrease in the heart's pumping function, manifested as a decrease in ejection fraction or a decrease in stroke volume. The present invention finds that cinquefoil acid or its pharmaceutically acceptable salt can improve cardiac ejection fraction or stroke volume, that is, the heart's ability to pump blood, and transport blood to the whole body.

[0071] In certain embodiments, the myocardial cell hypertrophy refers to the disordered arrangement of myocardial cells, the enlargement of myocardial cells or the increase in the number of enlarged cells, the improvement of myocardial cell hypertrophy refers to the neat arrangement of myocardial cells, the restoration of myocardial cells to health or the reduction of the number of enlarged myocardial cells, and the improvement of myocardial cell hypertrophy refers to the approach or reaching of the healthy size of the myocardial cells of the object of action. For example, based on healthy myocardial cells, the product can make the size of the myocardial cells of the object of action no more than 130%, 120%, 110% or 100% of healthy myocardial cells. Under normal circumstances, myocardial cells are arranged in a certain order to form myocardial fibers, and neat arrangement helps the heart to contract and relax effectively. Hypertension or other heart diseases may cause disorder in the arrangement of myocardial cells, which may be caused by abnormal proliferation, fibrosis or necrosis of myocardial cells. Untidy arrangement of myocardial cells may affect the coordination of the contraction of the heart, resulting in a decrease in the efficiency of the heart's pumping. The present invention finds that cinquefoil acid or its pharmaceutically acceptable salt cinquefoil acid or its pharmaceutically acceptable salt can improve the arrangement of myocardial cells to make them neat and enhance the heart's pumping function.

[0072] In certain embodiments, the symptoms of reduced vascular function, thickened vascular walls, reduced cardiac function or myocardial cell hypertrophy are caused by hypertension, and are specifically related to angiotensin II.

[0073] In certain embodiments, the potentilla acid or its pharmaceutically acceptable salt is the sole active ingredient or one of the active ingredients of the product. Preferably, the potentilla acid, its pharmaceutically acceptable salt and pharmaceutically acceptable excipients form a pharmaceutical composition, more preferably the pharmaceutically acceptable excipient is selected from one or more of a carrier, a diluent, a binder, a lubricant and a wetting agent; more preferably the pharmaceutical composition is one or more of a solution, a suspension, an emulsion, an injection, a spray, an aerosol, a tablet, a capsule, a powder and a granule.

[0074] Based on the application of potentinic acid described above, the present invention further provides a drug for preventing or treating hypertension, the drug comprising potentinic acid, and the drug has at least one of the following uses B1) to B6):

[0075] B1) Reduce high blood pressure;

[0076] B2) Improve decreased vascular function;

[0077] B3) Improve the thickness of blood vessel walls;

[0078] B4) Improve decreased heart function;

[0079] B5) Improve cardiomyocyte hypertrophy.

[0080] In certain embodiments, the drug for preventing and treating hypertension includes pharmaceutically acceptable excipients, and more importantly, an effective dose of potentilla acid.

[0081] In certain specific embodiments, the effective dose of the potentinic acid refers to a therapeutically effective amount, which refers to an amount of potentinic acid sufficient to show benefit or clinical significance to an individual. It will be understood by those skilled in the art that the actual amount or dosage administered and the administration schedule will depend on the nature and severity of the disease being treated, the age and general condition of the subject being treated, and the mode of administration, etc.

[0082] In certain specific embodiments, the pharmaceutically acceptable excipient refers to any excipient known to those skilled in the art that is suitable for a specific mode of administration. For example, in a specific embodiment, the pharmaceutically acceptable excipient may include one or more pharmaceutically acceptable carriers, solvents, excipients, buffers, stabilizers, adjuvants, and prodrugs compatible with cinquefoil acid.

[0083] In certain specific embodiments, the pharmaceutically acceptable excipients should be non-toxic, do not interfere with or impair the effectiveness of cinquefoil acid. In addition, the pharmaceutically acceptable excipients can be flexibly selected according to the dosage form of the hypertension treatment drugs in the above-mentioned embodiments. As in a specific embodiment, the carrier includes but is not limited to sugars such as lactose, glucose and sucrose, starches such as corn starch and potato starch, cellulose and its derivatives such as sodium shuttle methyl cellulose, ethyl cellulose and cellulose acetate, Xihuang Pu gum powder, malt, gelatin, talc, Cremophor, Solutol, etc. At least one. Excipients include but are not limited to cocoa butter, suppository wax, oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil, glycols such as propylene glycol, esters such as ethyl oleate and ethyl dodecanoate, agar, etc. At least one. Buffers include but are not limited to magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethanol and phosphate buffer solution, etc. Of course, the pharmaceutically acceptable excipients may also include lubricants such as sodium lauryl sulfate and magnesium stearate, colorants, release agents, coating agents, sweeteners, flavoring agents and fragrances according to the dosage form or administration method of the drug, or according to the judgment of those skilled in the art, preservatives and antioxidants may also be present in the pharmaceutical excipients.

[0084] In certain embodiments, the drug for preventing and treating hypertension may be administered to animals, including humans.

[0085] In certain embodiments, the administration mode of the drug for preventing and treating hypertension includes topical, parenteral, intravenous, intraarterial, subcutaneous, intramuscular, intracranial, intraorbital, ophthalmic, intraventricular, intracapsular, intraspinal, intracisternal, intraperitoneal, intranasal, by aerosol, by suppository, or by oral delivery. Moreover, the above-mentioned drug for treating hypertension can be administered alone or in combination with other compositions if necessary.

[0086] In certain embodiments, according to the administration mode of the drug for preventing or treating hypertension, the drug-acceptable excipients can be flexibly selected and combined with an effective dose of cinquefoil acid to prepare different dosage forms such as creams, gels, lotions, solutions, pills, tablets, capsules, powders, pastes, aerosols, etc.

[0087] In certain embodiments, the effective dose of the potentinic acid or its pharmaceutically acceptable salt administered to the subject is 0.1-10 mg / kg body weight / time per day, for 20-30 consecutive days. Specifically, the effective dose is 0.1-10 mg / kg body weight / time per day, for 28 consecutive days.

[0088] The present invention also provides a method for preparing the above-mentioned drug for preventing or treating hypertension, the preparation method comprising treating the above-mentioned drug-acceptable excipients and the effective dose of cinquefoil acid according to a method that pharmacists can understand and a process that can be accepted in the preparation. Through this process treatment, a drug for preventing or treating hypertension in a specific administration mode is prepared. Therefore, the preparation method of the present invention can flexibly select drug-acceptable excipients according to the requirements of the dosage form, and the cinquefoil acid of the active drug ingredient can be treated with drug-acceptable excipients according to a method that pharmacists can understand and a process that can be accepted in the preparation to form a drug, and its preparation process is stable, which effectively ensures the activity stability of the drug and ensures the production cost.

[0089] The present invention also provides a method for preventing or treating hypertension, which comprises administering an effective amount of cinnamic acid or a pharmaceutically acceptable salt thereof as described above to a subject in need. The method may also be in vitro or non-therapeutic, wherein the subject is a mammal, such as but not limited to humans, primates, livestock, pets, laboratory test animals, or captured wild animals. The subject is preferably a primate. The subject is most preferably a human. The subject may be a patient with hypertension or an individual who expects to prevent hypertension. The cinnamic acid or pharmaceutical composition may be administered to the subject before, during, or after receiving treatment for hypertension.

[0090] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0091] Before further describing the specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the examples of the present invention are for describing the specific embodiments rather than for limiting the scope of protection of the present invention; in the present specification and claims, unless otherwise expressly stated herein, the singular forms "a", "an" and "the" include plural forms.

[0092] When the embodiments give numerical ranges, it should be understood that, unless otherwise specified in the present invention, both endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those generally understood by those skilled in the art. In addition to the specific methods, equipment, and materials used in the embodiments, according to the grasp of the prior art by those skilled in the art and the record of the present invention, any methods, equipment, and materials of the prior art similar or equivalent to the methods, equipment, and materials described in the embodiments of the present invention can also be used to realize the present invention.

[0093] In the examples of this application, the material information used is as follows:

[0094] Experimental animals: SPF-grade C57BL / 6 male mice were purchased from Slack Laboratory Animal Co., Ltd., with the experimental animal use license number: SYXK(Min)2020-0002.

[0095] Experimental drugs and main reagents: osmotic micropump (Alzet, 2004D); angiotensin II (AngII; Abcam, ab120183); potentilla acid (Shanghai Yuanye Biotechnology Co., Ltd., B30371); eosin staining solution (Beijing Solebold Technology Co., Ltd., g1100); valsartan (Sigma, PHR315); hematoxylin staining solution (Beijing Solebold Technology Co., Ltd., g1140); isoflurane (Shenzhen Ruiwode Life Science Co., Ltd., 970-00026-00); paraformaldehyde (Fuzhou Feijing Biotechnology Co., Ltd. LA0427), anhydrous ethanol (Xilong Science Co., Ltd., 1280340101602), xylene (Xilong Science Co., Ltd., 1430030101600) and other chemical reagents.

[0096] The main instruments of the experiment are: pipette (Raining Company, USA); electronic balance scale (Ohaus Instrument Co., Ltd., Shanghai); non-invasive rat tail sphygmomanometer (Kent Co., Ltd., USA); small animal ultrasound imaging system Vevo2100 (Fujifilm Investment Co., Ltd.); inhalation small animal anesthesia machine (Shenzhen Ruiwode Life Science Technology Co., Ltd.); pathological slicer (Leica Company, Germany); paraffin embedding machine (Hubei Xiaogan Yaguang Medical Electronic Technology Co., Ltd.).

[0097] Example 1

[0098] 1. Materials

[0099] 1.1 Experimental Animals

[0100] Eighteen healthy male C57BL / 6 mice (weight 19±1g) were housed in the SPF-level laboratory of the Experimental Animal Center of Fujian University of Chinese Medicine, with adequate diet and water, 12-h light / dark cycle, temperature of 23±1°C, and humidity controlled at 50%, all within the acceptable range for experimental animals. The mice were used in the experiment after 5-7 days of adaptive feeding.

[0101] All animal experiments were performed in compliance with the regulations of Laboratory Animal Ethics and the Animal Care Association of Fujian University of Traditional Chinese Medicine, and the Laboratory Animal Ethics Review Form has been approved with the approval number 2W2024005.

[0102] 2. Experimental methods

[0103] 2.1 Drug preparation

[0104] Weigh an appropriate amount of tormentic acid powder, dissolve it in a corresponding volume of distilled water at a dose of 10 mg / kg / d according to the average weight of mice, place it in an ultrasound machine to maintain low-temperature ultrasound for about 1 hour, and store it in a 4°C refrigerator for use to obtain a tormentic acid solution.

[0105] Weigh an appropriate amount of AngII powder and dissolve it in high-pressure water to prepare a mother solution with a concentration of 1 mM. After preparation, store it in a -80°C refrigerator for later use to obtain an AngII solution.

[0106] Weigh an appropriate amount of Valsartan powder and dissolve it in high-pressure water. Dissolve it in a corresponding volume of distilled water at a dose of 10 mg / kg / d according to the weight of the mouse. After preparation, store it in a refrigerator at 4°C for future use to obtain a valsartan solution.

[0107] 2.2 Animal grouping and model construction

[0108] Animal grouping: Twelve male C57BL / 6 mice were randomly divided into 4 groups according to basal blood pressure, with 3 mice in each group:

[0109] Control group: An osmotic mini-pump containing normal saline was implanted, and distilled water was used for intragastric gavage later.

[0110] AngII group: An osmotic mini-pump containing AngII was implanted, and distilled water was used for intragastric administration later.

[0111] AngII+Tormentic acid (10 mg / kg / d) group: An osmotic mini-pump containing AngII was implanted, and later the potentilla acid solution was gavaged.

[0112] AngII group + Valsartan (10 mg / kg / day): An osmotic mini pump containing AngII was implanted, and Valsartan was administered intragastricly later.

[0113] Osmotic mini pump preparation:

[0114] 1) One day before surgery, the osmotic micropumps were taken out in the clean bench, and 200 μL of AngII solution (2.952 mg / mL) was injected into the osmotic micropumps of the AngII group and the AngII+Tormentic acid (10 mg / kg / d) group, respectively, to obtain pre-set osmotic micropumps containing AngII.

[0115] The osmotic minipumps in the Control group were injected with 200 μL of saline solution to obtain osmotic minipumps pre-loaded with saline.

[0116] 2) All groups of osmotic mini-pumps were immersed in physiological saline in a 37°C cell culture incubator overnight and used for pump implantation surgery the next day.

[0117] Model construction and processing:

[0118] On the day of surgery, the operating table was routinely disinfected, the mice were anesthetized with isoflurane, placed in a prone position, the hair on the back of the head and neck was removed, the skin was disinfected with iodine, the skin was cut with scissors, the subcutaneous tissue was bluntly separated, and the mice in the AngII group and the AngII+Tormenticacid (10 mg / kg / d) group were subcutaneously implanted with a pre-placed osmotic mini-pump containing AngII; at the same time, the mice in the Control group were subcutaneously implanted with a pre-placed osmotic mini-pump containing saline. After the corresponding osmotic mini-pumps were implanted in each group, the skin was sutured and disinfected.

[0119] Starting from the second day after pump implantation, mice in the AngII+Tormentic acid (10 mg / kg / d) group were given 10 mg / kg / d of tormentic acid solution by gavage, and mice in the Control and AngII groups were given an equal volume of distilled water by gavage, once a day, for a total of 4 weeks.

[0120] 2.3 Blood pressure measurement

[0121] During the experiment, the CODA TM The mean arterial pressure (MAP), systolic blood pressure (SBP) and diastolic blood pressure (DBP) of each group of mice were measured by non-invasive precision blood pressure meter once a week, and the data were statistically analyzed. The weight changes of each mouse were recorded every week. The specific operation is as follows:

[0122] (1) Assemble the mouse heating plate, sensor, computer, etc.

[0123] (2) Place the mouse in a restrainer and place it steadily on a mouse heating plate. Turn on the mouse heating plate to maintain the mouse's body temperature and smooth blood flow in the tail artery.

[0124] (3) Place the occlusion suit and volume pressure sensor on the rat’s tail;

[0125] (4) Open Coda 4.1 software and set the software conditions to 15 cycles per cycle. Before measurement, place the mouse on a heating plate and let it rest for 10 minutes. Pay attention to the measurement period and keep the surrounding environment in a quiet state. After the measurement, the volume pressure sensor transmits the value of each measurement to the computer;

[0126] (5) The systolic blood pressure, diastolic blood pressure, and mean arterial pressure of each mouse were recorded each week, and the body weight was measured. The mean and standard deviation were calculated.

[0127] The calculation formula for mean arterial pressure is: mean arterial pressure = (systolic pressure + (2×diastolic pressure)) ÷ 3.

[0128] 2.4. Abdominal aorta resistance index, pulsatility index and thickness measurement

[0129] (1) Echocardiography: After the experiment, the abdominal aorta vascular function of each group of mice was detected using Vevo 2100 small animal ultrasound instrument.

[0130] (2) Abdominal hair removal and animal anesthesia: The abdominal hair of the mice was removed with a depilatory cream, and each mouse was anesthetized with 1.5% isoflurane inhalation and fixed in a supine position on a 37°C constant temperature heating plate.

[0131] (3) Obtaining abdominal aortic cross-section: Coupling agent was applied to the front chest and abdomen of the mouse to expose the abdominal aorta and obtain the abdominal aortic cross-section. The probe frequency was 30 MHz.

[0132] (4) Analysis and calculation of ultrasound results: After the operation, Vevo Strain Software (VevoLAB 1.7.1) was used to calculate the abdominal aortic resistance index (RI) and pulsatility index (PI) of each group of mice, and the average value was calculated by taking three times each. At the same time, the vascular thickness was analyzed by taking three times each and the average value was calculated.

[0133] 2.5. Echocardiography to detect cardiac function

[0134] The steps of abdominal hair removal and mouse anesthesia are the same as described in 2.4. Then, the ultrasound probe (frequency is 400MHz) is placed vertically on the left chest of the mouse, and the probe is adjusted so that its notch faces the head of the mouse. In B-Mode mode, slowly adjust the probe until a clear heart image is obtained. Then switch to M-Mode mode to collect the cardiac cycle diagram of the mouse. At least three cardiac cycle data are measured for each mouse. After completing the ultrasound scan, the average value is taken for subsequent statistical analysis. The cardiac function indicators measured are left ventricular ejection fraction (LVEF) and stroke volume (SV).

[0135] The main function of the heart is to pump blood. The amount of blood ejected by one ventricle in one heartbeat is called stroke volume, or stroke volume for short.

[0136] 2.6 HE staining

[0137] (1) Fixed

[0138] Mouse abdominal aorta and heart tissue samples were immersed in 4% paraformaldehyde for 48 h, placed in tissue embedding boxes and labeled with numbers.

[0139] (2) Dehydration and wax dipping

[0140] The heart tissue samples were immersed in the following reagents for dehydration and wax in turn: 70% anhydrous ethanol (60 min) → 80% anhydrous ethanol (60 min) → 90% anhydrous ethanol (60 min) → 95% anhydrous ethanol (60 min) → 100% anhydrous ethanol I (20 min) → 100% anhydrous ethanol II (20 min) → 100% anhydrous ethanol III (50 min) → xylene I (5 min) → xylene II (20 min) → paraffin I (10 min) → paraffin II (20 min) → paraffin III (30 min).

[0141] (3) Embedding

[0142] Preheat the embedding mold and wax block, and place the tissue upright in the center of the bottom of the embedding mold. Gently place the embedding box on top of the mold and fill it with paraffin from the top. Transfer the mold to a cooling table, wait for the paraffin to solidify, and place the wax block in a -20℃ refrigerator for 30 minutes. Carefully separate the wax block from the mold and store it at room temperature.

[0143] (4) Slicing

[0144] Fix the pre-cooled tissue wax block on the slicer, and set the rough trimming thickness to 10μm initially until the tissue structure is completely visible. Adjust the slice thickness to 4μm for fine trimming. Flatten the complete tissue slices in 39℃ warm water, gently remove them and place them on the adhesive slides one by one. Bake the slices at 60℃ for 1h, bake them for 3h, and store the slices at room temperature for subsequent experiments.

[0145] (5) Dewaxing

[0146] Dewax by immersing in reagents in the following order: xylene I (25 min) → xylene II (25 min) → xylene III (15 min) → 100% anhydrous ethanol I (5 min) → 100% anhydrous ethanol II (5 min) → 95% anhydrous ethanol (5 min) → 80% anhydrous ethanol (5 min) → 70% anhydrous ethanol (5 min) → ultrapure water I (5 min) → ultrapure water II (5 min).

[0147] (6) Hematoxylin staining

[0148] Immerse the heart slide in the hematoxylin solution for 30 seconds and then wash it with pure water.

[0149] (7) Eosin staining

[0150] The above slides were immersed in eosin solution for staining for 15 seconds, and then washed with pure water.

[0151] (8) Sealing

[0152] After staining, blow dry the sections immediately and add an appropriate amount of neutral gum to seal the sections.

[0153] (9) Microscope observation

[0154] Using an intelligent tissue section analysis system, the entire sample was photographed at a 12.5× field of view for each sample, and 6 areas were randomly selected at a 400× field of view to observe the pathological changes of each group of samples.

[0155] 2.7 Statistical Analysis

[0156] Data statistical analysis was performed using IBM SPSS Statistics (version 29.0.1.0) software.

[0157] The Shapiro-Wilk test was used to test the normality of the data. The data are expressed as mean ± standard deviation. When comparing the differences among the three groups, the data were normally distributed, and the ANOVA one-way analysis of variance was used for comparison (when the data variance was equal, LSD was used for post hoc test; otherwise, Games-Howell was used for post hoc test); when the data did not conform to the normal distribution, the nonparametric Kruskal-Wallis test was used to compare the differences between the groups. P < 0.05 was considered statistically significant.

[0158] 3. Experimental results

[0159] 3.1 Effects of Potentilla Acid on Blood Pressure and Body Weight in Mice

[0160] The results of the effect of potentilla acid on blood pressure are shown in Figure 1A , 1B and 1C.

[0161] like Figure 1A As shown in the figure, compared with the Control group, the SBP of the AngII group mice after AngII intervention increased significantly (*P<0.05), indicating that the model was successfully established. Compared with the AngII group, the SBP of the AngII+Tormentic acid group mice after potentinic acid intervention decreased significantly (#P<0.05). The results of potentinic acid intervention were similar to those of valsartan, and the effect of lowering SBP was even better than that of valsartan. This shows that potentinic acid can significantly inhibit the increase of SBP.

[0162] like Figure 1B As shown in the figure, compared with the Control group, the DBP of the AngII group mice after AngII intervention increased significantly (*P<0.05). Compared with the AngII group, the DBP of the AngII+Tormentic acid group mice after potentianic acid intervention decreased significantly (#P<0.05). The results of potentianic acid intervention were similar to those of valsartan, and the effect of lowering DBP was even better than that of valsartan. This shows that potentianic acid can significantly inhibit the increase of DBP.

[0163] like Figure 1C As shown in the figure, compared with the Control group, the MAP of the AngII group mice increased significantly after AngII intervention (*P<0.05). Compared with the AngII group, the MAP of the AngII+Tormentic acid group mice decreased significantly after potentinic acid intervention (#P<0.05). The result of MAP reduction after potentinic acid intervention was similar to that of valsartan. This shows that potentinic acid can significantly inhibit the increase of MAP.

[0164] At the same time, body weight measurements showed that potentilla acid intervention had no significant effect on the body weight of mice ( Figure 1D ).

[0165] 3.2 Effects of Potentilla acid on the resistance index, pulsatility index and thickness of the abdominal aorta in mice

[0166] Mice were first induced with AngII and then treated with potentilla acid for 4 weeks. The resistance index, pulsatility index and vascular thickness of each group of mice were detected and analyzed by small animal ultrasound. The results are shown in Figure 2A -E. From left to right, in the bar graph, the first bar represents the Control group; the second bar represents the AngII group; the third bar represents the AngII+Tormentic acid group; and the fourth bar represents the AngII+Valsartan group.

[0167] from Figure 2A and Figure 2B It can be seen that compared with the Control group, the abdominal aortic pulsatility index of mice in the AngII group was significantly increased, while the abdominal aortic pulsatility index of mice in the AngII+Tormentic acid group after potentianic acid intervention was significantly decreased (P<0.05); there was no significant difference in the abdominal aortic pulsatility index between the AngII+Tormentic acid group and the AngII+Valsartan group.

[0168] from Figure 2A and Figure 2C It can be seen that compared with the Control group, the abdominal aortic resistance index of mice in the AngII group was significantly increased, while the abdominal aortic resistance index of mice in the AngII+Tormentic acid group after potentianic acid intervention was significantly decreased (P<0.05); there was no significant difference in the abdominal aortic resistance index between the AngII+Tormentic acid group and the AngII+Valsartan group.

[0169] from Figure 2D and 2E It can be seen that compared with the Control group, the thickness of the abdominal aorta of mice in the AngII group was significantly thickened (P<0.05), while the thickness of the abdominal aorta of mice in the AngII+Tormentic acid group after potentianic acid intervention was significantly decreased (P<0.05); there was no significant difference in the thickness of the abdominal aorta between the AngII+Tormentic acid group and the AngII+Valsartan group.

[0170] In summary, AngII will lead to increased resistance of blood flow in blood vessels and increased pulsation under the action of resistance, while the administration of potentiostatic acid can improve the decrease of abdominal aortic resistance index and abdominal aortic pulsation index caused by hypertension, increase abdominal aortic resistance index and abdominal aortic pulsation index, and thus make blood vessels elastic.

[0171] 3.3. Effect of Potentilla acid on the pathological morphology of abdominal aorta in mice

[0172] HE staining was used to observe the changes of abdominal aorta morphology in mice induced by potentinic acid. Figure 2F .

[0173] from Figure 2F It can be seen that compared with the Control, the abdominal aorta wall of the AngII group mice was thickened, while the thickening of the abdominal aorta wall of the AngII+Tormentic acid group mice after intervention with potentinic acid was alleviated, and there was no significant difference between the AngII+Valsartan group, the AngII+Tormentic acid group and the AngII+Valsartan group after intervention with valsartan. This shows that potentinic acid can reduce the thickness of the abdominal aorta wall and can be used to improve the phenomenon of abdominal aorta wall thickening caused by hypertension.

[0174] 3.4 Effects of Potentilla acid on cardiac function in mice

[0175] Changes in cardiac function were assessed by small animal ultrasound. Figure 3A , 3B and 3C.

[0176] from Figure 3A , 3B According to the results of 3C, compared with the Control group, the left ventricular ejection fraction (LVEF) and stroke volume (SV) of the mice in the AngII group were significantly reduced (P<0.05); compared with the AngII group, the left ventricular ejection fraction and stroke volume of the mice in the AngII+Tormentic acid group after intervention with potentianic acid and the AngII+Valsartan group after intervention with valsartan were significantly increased (P<0.05), and there was no significant difference between the AngII+Tormentic acid group and the AngII+Valsartan group. This indicates that potentianic acid can improve ejection fraction and stroke volume, and can be used to improve the phenomenon of reduced ejection fraction and stroke volume caused by hypertension.

[0177] 3.5. Effects of Potentilla acid on cardiac pathology in mice

[0178] HE staining was used to evaluate the effect of potentilla acid on the pathological morphology of cardiac tissue in AngII model mice. Figure 3D The arrows point to the direction where the myocardial cells are disordered.

[0179] from Figure 3D It can be seen that the nuclei of the cardiomyocytes of the mice in the Control group were clear, the cytoplasm was evenly stained, and they were arranged tightly and neatly, without degeneration and necrosis; compared with the Control group, the cardiac tissue morphology of the mice in the AngII group showed obvious changes: the neatness of the arrangement of the cardiomyocytes was significantly reduced; while the cardiomyocytes of the mice in the AngII+Tormentic acid group after intervention with potentinic acid and the AngII+Valsartan group after intervention with valsartan were arranged neatly; there was no significant difference between the AngII+Tormentic acid group and the AngII+Valsartan group. This shows that potentinic acid can be used to significantly improve the pathological morphology of cardiac tissue caused by hypertension.

[0180] In summary, Potentilla acid significantly lowers blood pressure, improves vascular function, and relieves abdominal aorta thickening and pathological changes. It also has the effect of improving reduced heart function, myocardial cell hypertrophy and vascular wall thickening caused by hypertension.

[0181] The above examples are for the purpose of illustrating the embodiments disclosed by the present invention and are not to be construed as limitations of the present invention. In addition, the various modifications listed herein and the variations of methods and compositions in the invention are obvious to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been specifically described in conjunction with various specific preferred embodiments of the present invention, it should be understood that the present invention should not be limited to these specific embodiments. In fact, various modifications obvious to those skilled in the art as described above to obtain the invention should be included within the scope of the present invention.

Claims

1. Use of potentilla acid or its pharmaceutically acceptable salt in preparing a product for preventing or treating hypertension.

2. The use according to claim 1, characterized in that The potentilla acid or its pharmaceutically acceptable salt is used alone or in combination with other drugs.

3. The use according to claim 1 or 2, characterized in that The efficacy of the product includes reducing one or more of systolic blood pressure, diastolic blood pressure and mean arterial pressure.

4. The use according to claim 3, characterized in that The potentilla acid or its pharmaceutically acceptable salt and pharmaceutically acceptable excipients form a pharmaceutical composition.

5. The use according to claim 4, characterized in that The pharmaceutically acceptable excipient is selected from one or more of a carrier, a diluent, a binder, a lubricant and a wetting agent.

6. The use according to claim 4, characterized in that The dosage form of the pharmaceutical composition is selected from one or more of solutions, suspensions, emulsions, injections, sprays, aerosols, tablets, capsules, powders and granules.

7. The use according to claim 4, characterized in that The pharmaceutical composition is an oral preparation.

8. Use of potentilla acid or a pharmaceutically acceptable salt thereof in the preparation of a product for improving at least one of the following symptoms 1) to 4) caused by hypertension: 1) Decreased vascular function; 2) Thickening of blood vessel walls; 3) Reduced heart function; 4) Cardiomyocyte hypertrophy.

9. The use according to claim 8, characterized in that Include at least one of the following items A1)-A4): A1) Decreased vascular function refers to an increase in vascular resistance index or pulsatility index; A2) Decreased cardiac function refers to decreased cardiac ejection fraction or stroke volume; A3) Vascular thickening refers to the thickening of the blood vessel walls; A4) Cardiomyocyte hypertrophy is an increase in the number of cells that causes the cardiomyocytes to become misaligned, smaller, or smaller.

10. A drug for preventing or treating hypertension, characterized in that: The drug comprises potentilla acid, and the drug has at least one of the following uses B1) to B6): B1) Reduce high blood pressure; B2) Improve decreased vascular function; B3) Improve the thickness of blood vessel walls; B4) Improve decreased heart function; B5) Improve cardiomyocyte hypertrophy.

Images