Application of yohimbine hydrochloride and related products

Abstract

The invention relates to the technical field of medicines, in particular to application of yohimbine hydrochloride and a related product. The invention provides application of yohimbine hydrochloride or pharmaceutically acceptable salt thereof in preparation of products for preventing or treating hypertension, and further provides application of yohimbine hydrochloride or pharmaceutically acceptable salt thereof in preparation of products for preventing or treating vasculopathy caused by hypertension and / or heart disease caused by hypertension. It is found for the first time that yohimbine hydrochloride 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 belongs to the technical field of medicines, and in particular relates to uses of yohimbine hydrochloride and related products. Background Art

[0002] Hypertension is a common chronic disease in clinical practice and the most important risk factor for cardiovascular and cerebrovascular events. The situation of hypertension prevention and control is extremely severe, so finding a safe and effective prevention and treatment strategy is a crucial proposition.

[0003] Yohimbine hydrochloride is mainly derived from the bark of the African yohimbine tree (Pausinystalia johimbe). Recent studies have shown that yohimbine has a variety of pharmacological activities, including improving sexual function, promoting fat metabolism, improving athletic performance, anti-depression and anti-anxiety. These effects are mainly achieved by antagonizing α2-adrenaline receptors, thereby enhancing the release of norepinephrine and promoting blood flow and metabolism. In addition, yohimbine can also increase the activity of the central nervous system and improve psychological state by regulating the neurotransmitter system. However, there are no reports on the effects of yohimbine hydrochloride on cardiovascular diseases such as hypertension. Summary of the invention

[0004] In view of the above-mentioned deficiencies in the prior art, the present invention provides a use of yohimbine hydrochloride and related products.

[0005] The first aspect of the present invention provides the use of yohimbine hydrochloride and related products or the use of its pharmaceutically acceptable salt in the preparation of products for preventing or treating hypertension.

[0006] The second aspect of the present invention provides the use of yohimbine hydrochloride or a pharmaceutically acceptable salt thereof in the preparation of a product for preventing or treating vascular lesions caused by hypertension and / or cardiac lesions caused by hypertension.

[0007] In some embodiments of the present invention, the vascular lesions caused by hypertension and / or the heart lesions caused by hypertension have at least one of the following symptoms:

[0008] 1) Decreased vascular function;

[0009] 2) Thickening of blood vessel walls;

[0010] 3) Reduced heart function.

[0011] In some embodiments of the present invention, the reduced vascular function refers to an increase in the propagation velocity of vascular pulse waves.

[0012] In some embodiments of the present invention, the reduced cardiac function refers to a reduced cardiac ejection fraction or a reduced left ventricular fractional shortening.

[0013] The yohimbine hydrochloride in the present invention is used in its pharmaceutically acceptable salt alone or in combination with other drugs.

[0014] In some embodiments of the present invention, the yohimbine hydrochloride or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient, or a combination thereof, constitute a pharmaceutical composition.

[0015] In some embodiments of the present invention, the pharmaceutically acceptable excipient is selected from one or more of a diluent, a binder, a lubricant and a wetting agent.

[0016] In some embodiments of the present invention, the pharmaceutical composition is one or more of a solution, an injection, a spray, a nasal drop, an aerosol, a powder spray, a tablet, a capsule and a granule.

[0017] The third aspect of the present invention provides a product comprising yohimbine hydrochloride or a pharmaceutically acceptable salt thereof, wherein the product has at least one of the following functions:

[0018] 1) Lower blood pressure;

[0019] 2) Alleviate decreased vascular function;

[0020] 3) Relieve blood vessel wall thickening;

[0021] 4) Alleviate decreased heart function.

[0022] In some embodiments of the present invention, the products include medicines, health products and foods.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] 1. The present invention first discovered that yohimbine hydrochloride can be used in the preparation of drugs for treating hypertension or heart and vascular diseases caused by hypertension, with good and safe therapeutic effects, and provides a new treatment approach for treating hypertension or hypertension-related diseases.

[0025] 2. The present invention proves through experiments that yohimbine hydrochloride can significantly reduce the blood pressure of AngII model mice, improve vascular function, and relieve abdominal aorta thickening and pathological changes. It also has the effect of improving the pathological morphological changes of the heart caused by hypertension and reducing the function of the renal artery and mesenteric artery. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1A A graph showing the effect of yohimbine hydrochloride on systolic blood pressure in mice after AngII modeling in this application.

[0027] Figure 1B A graph showing the effect of yohimbine hydrochloride on diastolic blood pressure in mice after AngII modeling in this application.

[0028] Figure 1C A graph showing the effect of yohimbine hydrochloride on the mean arterial pressure of mice after AngII modeling in this application.

[0029] Figure 1D A graph showing the effect of yohimbine hydrochloride on the body weight of mice after AngII modeling in this application.

[0030] Figure 2A Ultrasound graph showing the effect of yohimbine hydrochloride on the abdominal aorta resistance index and pulsatility index of mice after AngII modeling in the present application.

[0031] Figure 2B The statistical analysis graph showing the effect of yohimbine hydrochloride on the abdominal aorta resistance index of mice after AngII modeling in the present application.

[0032] Figure 2C The statistical analysis graph showing the effect of Yohimbine Hydrochloride on the abdominal aorta pulsatility index of mice after AngII modeling in the present application.

[0033] Figure 2D Ultrasound images showing the effect of yohimbine hydrochloride on the vascular wall thickness of the abdominal aorta of mice after AngII modeling in the present application.

[0034] Figure 2E The statistical analysis graph showing the effect of yohimbine hydrochloride on the vascular wall thickness of the abdominal aorta of mice after AngII modeling in the present application.

[0035] Figure 2F A diagram showing the effect of yohimbine hydrochloride on the pathological morphology of the abdominal aorta of mice after AngII modeling in this application.

[0036] Figure 3A Ultrasound images showing the effect of Yohimbine hydrochloride on the cardiac function of mice after AngII modeling in the present application (arrow: left ventricular end-diastolic / systolic diameter).

[0037] Figure 3B The statistical analysis graph shows the effect of Yohimbine Hydrochloride on the left ventricular ejection fraction of the mouse heart after AngII modeling in the present application.

[0038] Figure 3C The statistical analysis graph showing the effect of Yohimbine Hydrochloride on the short axis shortening rate of the left ventricle in the mouse heart after AngII modeling in the present application.

[0039] Figure 3D A diagram showing the effect of yohimbine hydrochloride on the pathological morphology of cardiac tissue in mice after AngII modeling in this application.

[0040] Figure 4A Ultrasound diagram showing the effect of yohimbine hydrochloride on the renal artery resistance index and pulsatility index of mice after AngII modeling in the present application.

[0041] Figure 4B The statistical analysis graph showing the effect of Yohimbine Hydrochloride on the renal artery resistance index of mice after AngII modeling in the present application.

[0042] Figure 4C The statistical analysis graph showing the effect of Yohimbine Hydrochloride on the renal artery pulsatility index of mice after AngII modeling in the present application. DETAILED DESCRIPTION

[0043] The inventors of the present invention accidentally discovered that the compound yohimbine hydrochloride has an antihypertensive effect and can improve heart disease and vascular disease caused by hypertension during the process of screening a large number of compounds for their effects on hypertension. On this basis, the present invention was completed.

[0044] The first aspect of the present invention provides use of yohimbine hydrochloride or a pharmaceutically acceptable salt thereof in preparing a product for preventing or treating hypertension.

[0045] In some specific embodiments of the first aspect of the present invention, the product for preventing or treating hypertension is a drug for preventing or treating hypertension. The yohimbine hydrochloride 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.

[0046] Yohimbine hydrochloride is an α-2 adrenergic receptor inhibitor with a molecular formula of C 21 H 26 N 2 O 3 HCL, CAS No. 65-19-0, has the following structural formula:

[0047]

[0048] The "pharmaceutically acceptable salt" refers to an acid addition salt or a base addition salt. All compounds of the present invention that exist in the form of free base or free acid can be converted into their pharmaceutically acceptable salts by treatment with appropriate inorganic or organic bases or acids according to methods known to those skilled in the art. Salts of compounds of the present invention can be converted into their free bases or acids by standard techniques.

[0049] The pharmaceutically acceptable salts of Yohimbine hydrochloride of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable nontoxic acid addition salts are salts with an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods such as ion exchange used in the art. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, citrate, dodecylsulfate, ethanesulfonate, formate, fumarate, gluconate heptanoate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Salts derived from suitable bases include alkali metal salts, alkaline earth metal salts, ammonium salts. Representative alkali metal salts or alkaline earth metal salts include sodium salts, lithium salts, potassium salts, calcium salts, magnesium salts, etc. Where appropriate, other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, sulfonates and aryl sulfonates. Other pharmaceutically acceptable salts include salts formed by quaternization of amines, which are carried out using suitable electrophilic reagents (e.g., alkyl halides) to form quaternized alkylated amino salts.

[0050] The product of the present invention necessarily contains yohimbine hydrochloride or a pharmaceutically acceptable salt thereof, and uses yohimbine hydrochloride or a pharmaceutically acceptable salt thereof as an effective ingredient for preventing or treating hypertension.

[0051] In the product of the present invention, the effective ingredient for preventing or treating hypertension may be only yohimbine hydrochloride, or may contain other chemicals that can play a similar role.

[0052] The product of the present invention may be a single-component substance or a multi-component substance.

[0053] The second aspect of the present invention provides the use of yohimbine hydrochloride or a pharmaceutically acceptable salt thereof in the preparation of a product for preventing or treating vascular lesions caused by hypertension and / or cardiac lesions caused by hypertension.

[0054] In some embodiments of the second aspect of the present invention, the vascular lesions caused by hypertension and / or the cardiac lesions caused by hypertension have at least one of the following symptoms:

[0055] 1) Decreased vascular function;

[0056] 2) Thickening of blood vessel walls;

[0057] 3) Reduced heart function.

[0058] In the second aspect of the present invention, the reduced vascular function refers to an increase in the vascular pulse wave velocity (PWV). Pulse wave velocity is an important indicator for evaluating the degree of vascular sclerosis. The higher the PWV, the stiffer the blood vessels and the worse the elasticity, resulting in increased blood flow resistance. Specifically, the vascular function includes the status of the abdominal aorta, renal arteries and mesenteric arteries. Reduced abdominal aortic function will affect the blood supply to the abdominal cavity and lower limbs, renal artery sclerosis will affect renal function, leading to problems such as hypertension and renal failure, while mesenteric artery sclerosis will affect the intestinal blood supply, resulting in impaired digestion and absorption, causing symptoms such as abdominal pain and indigestion.

[0059] In the second aspect of the present invention, the thickening of the vascular wall refers specifically to the thickening of the vascular wall of the abdominal aorta. The abdominal aorta is an important large blood vessel that supplies blood to the abdominal cavity and lower limbs, and its vascular wall thickening is usually caused by pathological changes such as atherosclerosis. The thickening of the vascular wall not only leads to a decrease in vascular elasticity, but also increases vascular resistance, restricting the normal flow of blood. This situation may lead to insufficient blood supply to the abdominal organs and lower limbs, causing a series of clinical symptoms, such as abdominal pain, lower limb weakness, edema, etc.

[0060] In the second aspect of the present invention, the reduced cardiac function refers to a reduction in the left ventricular ejection fraction (LVEF) or the left ventricular shortening fraction (LVSF). These parameters are important indicators for evaluating the heart's pumping function. A reduction in the left ventricular ejection fraction means that the amount of blood pumped out by the heart during each contraction is reduced, which directly reflects a decrease in the heart's overall pumping efficiency. A reduction in the left ventricular shortening fraction means that the left ventricle fails to shorten effectively during contraction, resulting in a reduction in the amount of blood discharged by the heart. This reduction in cardiac function may lead to clinical symptoms such as heart failure, fatigue, and dyspnea.

[0061] In the present invention, the yohimbine hydrochloride or a pharmaceutically acceptable salt thereof is used alone or in combination with other drugs.

[0062] In some embodiments of the present invention, the yohimbine hydrochloride or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient, or a combination thereof, constitute a pharmaceutical composition.

[0063] In some embodiments of the present invention, the pharmaceutically acceptable excipient is selected from one or more of a diluent, a binder, a lubricant and a wetting agent.

[0064] "Pharmaceutically acceptable" refers to molecular entities and compositions that do not produce adverse, allergic or other untoward reactions when appropriately administered to an animal or a human.

[0065] "Pharmaceutically acceptable carriers or excipients" should be compatible with yohimbine hydrochloride, that is, they can be mixed with it without significantly reducing the effect of the pharmaceutical composition under normal circumstances. Specific examples of some substances that can be used as pharmaceutically acceptable carriers or excipients are sugars, such as lactose, glucose and sucrose; binders, specifically starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium methylcellulose, ethyl cellulose and methyl cellulose; tragacanth powder; malt; gelatin; lubricants, specifically 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; flavorings; tablets, stabilizers; antioxidants; preservatives; diluents, such as pyrogen-free water; isotonic saline solution; and phosphate buffer, etc. These materials are used as needed to aid in the stability of the formulation or to help increase the activity or its bioavailability or to produce an acceptable taste or flavor in the case of oral administration.

[0066] In the present invention, unless otherwise specified, the dosage form of the pharmaceutical composition is not particularly limited. In some embodiments of the present invention, the pharmaceutical composition is one or more of a solution, an injection, a spray, a nasal drop, an aerosol, a powder spray, a tablet, a capsule and a granule, and can be prepared by a conventional method. Preferably, it is a solution.

[0067] The dosage form of the pharmaceutical composition should be matched with the mode of administration. The pharmaceutical composition of the present invention can be introduced into the body such as muscle, intradermal, subcutaneous, intravenous, mucosal tissue by injection, spraying, nasal drops, eye drops, penetration, absorption, physical or chemical mediation; or introduced into the body after being mixed or wrapped with other substances. Preferably, it is introduced into the body subcutaneously. The pharmaceutical composition can also be used in combination with other treatment methods, including surgery, radiotherapy, chemotherapy, and targeted therapy.

[0068] The third aspect of the present invention provides a product, comprising yohimbine hydrochloride or a pharmaceutically acceptable salt thereof, and the product has at least one of the following functions:

[0069] 1) Lower blood pressure;

[0070] 2) Alleviate decreased vascular function;

[0071] 3) Relieve blood vessel wall thickening;

[0072] 4) Alleviate decreased heart function.

[0073] In some embodiments of the third aspect of the present invention, the lowering of blood pressure is achieved by dilating blood vessels, reducing vascular resistance, and improving blood flow, thereby effectively lowering arterial blood pressure.

[0074] In some embodiments of the third aspect of the present invention, the alleviation of reduced vascular function refers to improving the elasticity and function of blood vessels and reducing the vascular pulse wave velocity (PWV) by promoting the health of vascular endothelial cells and enhancing blood circulation, thereby improving the overall health of blood vessels. For example, based on healthy blood vessels, the product can restore the vascular pulse wave velocity of the target blood vessels to at least 70%, 80%, 90% or 100% of that of healthy blood vessels.

[0075] In some embodiments of the third aspect of the present invention, the relieving of blood vessel thickening refers to inhibiting the formation and development of atherosclerotic plaques, promoting the recovery and regeneration of the blood vessel wall, thereby reducing the thickness of the blood vessel wall and restoring the normal function of the blood vessel. For example, based on a healthy blood vessel, the product can make the thickness of the blood vessel wall of the target blood vessel not exceed 130%, 120%, 110% or 100% of the thickness of a healthy blood vessel.

[0076] In some embodiments of the third aspect of the present invention, the relief of reduced cardiac function refers to the restoration of normal cardiac function by enhancing myocardial contractility and improving myocardial energy metabolism, improving the heart's pumping efficiency and capacity, thereby helping patients alleviate heart failure and other heart disease symptoms, which can be demonstrated by improving the cardiac ejection fraction or the left ventricular shortening fraction. Improving the cardiac ejection fraction or the left ventricular shortening fraction refers to bringing the cardiac ejection fraction and the left ventricular shortening fraction 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 the left ventricular shortening fraction of the subject's heart to at least 70%, 80%, 90% or 100% of that of a healthy heart.

[0077] In the third aspect of the present invention, the reduced vascular function, thickened vascular wall and reduced cardiac function are caused by hypertension, and are specifically related to angiotensin II.

[0078] In some embodiments of the present invention, the products include medicines, health products and foods.

[0079] When the drug of the present invention is used to treat hypertension in a subject, an effective dose of the product needs to be administered to the subject. Using this method, the hypertension or the decreased vascular function, thickening of the vascular wall, decreased cardiac function or disordered arrangement of myocardial cells caused by hypertension are inhibited, reduced or alleviated. The subject is an organism for prevention and / or treatment, including mammals. The mammal is preferably a rodent, an artiodactyl, a perissodactyl, a lagomorph, a primate, etc. The primate is preferably a monkey, an ape or a human.

[0080] In the third aspect of the present invention, the effective dose of yohimbine hydrochloride or a pharmaceutically acceptable salt thereof administered to a subject is 0.1-10 mg / kg body weight / time per day, for continuous administration for 20-30 days. Specifically, the effective dose is 0.1-10 mg / kg body weight / time per day, for continuous administration for 28 days.

[0081] Treatment as used herein refers to slowing, interrupting, preventing, controlling, stopping, alleviating, reversing the progression or severity of a sign, symptom, disorder, condition, or disease after the disease has begun to develop, but does not necessarily involve the complete elimination of all disease-related signs, symptoms, conditions, or disorders.

[0082] Prevention as described in the present invention refers to all actions to suppress symptoms or delay the onset of a specific state of tension by administering the product described in the present invention.

[0083] 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 specific embodiments rather than for limiting the scope of protection of the present invention.

[0084] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meanings as those commonly understood by those skilled in the art. In addition to the specific methods, equipment, and materials used in the embodiments, any methods, equipment, and materials of the prior art that are similar or equivalent to the methods, equipment, and materials in the embodiments of the present invention may be used to implement the present invention, based on the prior art mastery of those skilled in the art and the description of the present invention.

[0085] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt conventional techniques in the field of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related fields.

[0086] In the examples of this application, the material information used is as follows:

[0087] 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.

[0088] Experimental drugs and main reagents: osmotic micropump (Alzet, 2004D); angiotensin II (AngII; Abcam, ab120183); yohimbine hydrochloride (Shanghai Yuanye Biotechnology Co., Ltd., T94263); eosin staining solution (Beijing Solebold Technology Co., Ltd., g1100); 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.

[0089] 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.).

[0090] Example 1: Animal model construction and treatment

[0091] 1.1 Experimental animals

[0092] Twenty-four healthy male C57BL / 6 mice (weight 19±1 g) were purchased from Slake Experimental Animal Co., Ltd. (license number: SYXK(Min)2020-0002). All animals were housed in the SPF-level laboratory of the Experimental Animal Center of Fujian University of Traditional Chinese Medicine, with adequate diet and water, 12h light / dark cycle, temperature 23±1°C, and humidity controlled at 50%, all within the acceptable range for experimental animals. Mice were used for experiments after 5-7 days of adaptive feeding. All animal experiments were conducted in compliance with the regulations of the Laboratory Animal Ethics and the Animal Management Association of Fujian University of Traditional Chinese Medicine, and the Laboratory Animal Ethics Review Form has been approved with the approval number 2W2024005.

[0093] 1.2 Drug preparation

[0094] Weigh an appropriate amount of yohimbine hydrochloride 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 ultrasonic instrument after preparation to maintain low-temperature ultrasound for about 1 hour, and store it in a 4°C refrigerator for use to obtain a yohimbine hydrochloride solution.

[0095] Weigh an appropriate amount of AngII powder and dissolve it in high-pressure water to prepare a mother solution concentration of 1 mM. After preparation, store it in a -80°C refrigerator for later use to obtain an AngII solution.

[0096] 1.3 Animal grouping and model construction

[0097] Animal grouping: 24 male C57BL / 6 mice were randomly divided into 4 groups according to basal blood pressure: Control group, AngII group, AngII+Yohimbine (10 mg / kg / d) group, and AngII+Valsartan (10 mg / kg / d) group, with 6 mice in each group.

[0098] Preparation of osmotic micropumps: The day before the operation, the osmotic micropumps were taken out in the clean bench, and 200 μL of AngII solution was injected into the osmotic micropumps of the AngII group, AngII+Yohimbine (10 mg / kg / d) group, and AngII+Valsartan (10 mg / kg / d) group, respectively, to obtain preset osmotic micropumps containing AngII.

[0099] The osmotic minipumps in the Control group were injected with 200 μL of saline solution to obtain osmotic minipumps containing saline.

[0100] The osmotic mini-pumps of all groups were immersed in physiological saline in a 37°C cell culture incubator overnight and used for pump implantation surgery the next day.

[0101] Model construction and treatment: 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 cut with scissors after disinfection with iodine, and the subcutaneous tissue was bluntly separated. The mice in the AngII group and the AngII+Yohimbine (10 mg / kg / d) group were subcutaneously implanted with a pre-placed osmotic mini-pump containing AngII, and 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.

[0102] Starting from the second day after pump implantation, mice in the AngII+Yohimbine (10 mg / kg / d) group were gavaged with 10 mg / kg / d of yohimbine hydrochloride solution, and mice in the AngII+Valsartan (10 mg / kg / d) group were gavaged with 10 mg / kg / d of valsartan solution. The control group and the AngII group were gavaged with an equal volume of distilled water, once a day, for a total of 4 weeks of intervention.

[0103] 1.4 Blood pressure measurement

[0104] In this experiment, the changes in mouse blood pressure were monitored by non-invasive tail arterial blood pressure detection. The American Kent non-invasive tail arterial blood pressure detector was used to monitor the systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MAP) of the tail artery of the mouse. When measuring the blood pressure of the mouse, it must be ensured that it is in a quiet state. The non-invasive blood pressure meter with tail cannula is installed. Check whether the instrument has abnormal conditions such as leakage. Fix the mouse in the restraint, put the occlusion sleeve 1 cm from the root of the tail, and then place the volume pressure sensor (VPR) behind the occlusion sleeve. When the sensor can no longer move toward the tail, it can be stopped. Put the mouse on the heating plate and select the appropriate heating gear according to the room temperature to heat the tail. Cover the mouse with a black towel to ensure that the environment where the mouse is located is dark and quiet. After the mouse is calm for 10 minutes in the dark state, start the measurement. Each mouse is measured for at least 10 cycles, each cycle is 30 seconds, and the mouse SBP, DBP and MAP are recorded and the average value is taken for statistics. 1.5 Measurement of abdominal aorta pulse wave velocity and thickness

[0105] Vevo 2100 ultra-high resolution small animal ultrasound instrument was used to detect vascular function. Before the test, the chest and abdominal hair of the mice were removed with depilatory cream, and care was taken not to burn the surface skin of the mice. The mice were anesthetized by inhalation of isoflurane (500 ml / min) and fixed in a supine position on a 37°C constant temperature heating plate. Coupling agent was applied to the front chest and abdomen of the mice, and the long axis section of the left ventricle was found using an ultrasound probe (probe frequency of 30 MHz). The probe was moved down parallel to the midline of the abdomen, and the abdominal aorta was visible at this time, and the aortic section image was obtained. After the operation, Vevo Strain Software (Vevo LAB 1.7.1) was used to calculate the pulse wave velocity (PWV) and vascular wall thickness values ​​of each group of mice according to the software manual. The pulse wave velocity was calculated according to the following formula: PWV (m / s) = vessel length / (distal-proximal time difference), and each mouse was calculated 3 times and the average value was taken.

[0106] 1.6 Echocardiography to detect cardiac function

[0107] One day before the examination, the left chest of the mouse was depilated with depilatory cream, and the mouse was anesthetized with 1.5% isoflurane in 95% oxygen and 5% carbon dioxide, and fixed in a supine position on a 37°C constant temperature heating plate. A high-resolution small animal ultrasound system (Vevo2100, visualsonic) was used to perform non-invasive cardiac function measurement of the mouse, and the probe frequency was 400MHz. The probe was placed 45 degrees to the left of the midline of the sternum to display the long axis of the left ventricle, and rotated 90 degrees clockwise to display the short axis of the left ventricle. M-mode ultrasound was used at the level of the papillary muscle to record the movement of the left ventricle, and the left ventricular short axis shortening rate (LVFS), left ventricular ejection fraction (LVEF) and heart rate were measured to evaluate the cardiac function of the mouse.

[0108] 1.7 HE staining

[0109] (1) After the abdominal aorta was fixed in 4% paraformaldehyde for 24 hours, it was taken out and placed in an embedding box. The vessel number was clearly marked and the embedding box was then immersed in 70% ethanol for 24 hours.

[0110] (2) Dehydration and wax immersion: The vascular tissue was dehydrated and waxed in the following order: 80% ethanol (30 min) → 90% ethanol (30 min) → 95% ethanol (30 min) → 100% ethanol I (10 min) → 100% ethanol II (10 min) → 100% ethanol III (30 min) → xylene I (5 min) → xylene II (10-15 min) → paraffin I (10 min) → paraffin II (10 min) → paraffin III (25 min).

[0111] (3) Embedding: After removing the wax from the abdominal aorta, place the tissue upright in the center of the bottom of the mold, gently place the embedding box, add paraffin from above, and then transfer it to the cold table. After the wax block solidifies, pry it out of the mold and store it at room temperature for subsequent experiments.

[0112] (4) Sectioning: Precool the wax block in a refrigerator and fix it on a microtome. Roughly trim the block to a thickness of 10 μm. After exposing the tissue, fine trim the block to a thickness of 4 μm and then section the block. After unfolding and scooping the block, place it in a 37°C oven.

[0113] (5) Place the slide in a 60°C oven for 30 to 60 min and then dewax in the following order: xylene I (15 min) → xylene II (15 min) → 100% ethanol I (5 min) → 100% ethanol II (5 min) → 95% ethanol (5 min) → 80% ethanol (5 min) → 70% ethanol (5 min) → ultrapure water (5 min).

[0114] (6) Place the blood vessel slide in the hematoxylin stain solution for 1 min and then rinse with pure water;

[0115] (7) Place the blood vessel slide in hydrochloric acid ethanol for differentiation for 1 second, wash with pure water, and place in pure water for blueing for 10 minutes;

[0116] (8) Place the blood vessel slide in eosin stain for 20 seconds and then rinse with pure water;

[0117] (9) The stained slides were air-dried and gently sealed with neutral resin. After drying, the slides were observed and images were collected under a 400× field of view using an optical microscope to analyze the pathological morphological changes of the blood vessels.

[0118] 1.8 Statistical analysis

[0119] SPSS 22.0 software was used for statistical analysis. The Shapiro-Wilk test method was first used to analyze whether the data conformed to the normal distribution, and then the variance was tested to see whether it was homogeneous. If the data conformed to the normal distribution and the variance was homogeneous, the t test was used for two groups and the one-way ANOVA was used for three groups; if the data did not conform to the normal distribution or the variance was unequal, the Kruskal-Wallis test was used, and P < 0.05 was considered to be statistically significant.

[0120] Example 2: Analysis results

[0121] 2.1 Effects of Yohimbine Hydrochloride on Blood Pressure and Body Weight in Mice

[0122] Figure 1A , 1B and 1C demonstrate the effects of yohimbine hydrochloride on blood pressure. Figure 1A The data showed that the systolic blood pressure (SBP) of mice significantly increased after AngII intervention compared with the Control group, P<0.05. Compared with the AngII group, the SBP of the AngII+Yohimbine group was significantly reduced after Yohimbine hydrochloride intervention. This indicates that Yohimbine hydrochloride intervention can significantly inhibit the increase of SBP in the AngII group.

[0123] exist Figure 1B In the experiment, it was found that the diastolic blood pressure (DBP) of mice after AngII intervention was significantly increased compared with the Control group, P<0.05. In the AngII group, the DBP level of the AngII+Yohimbine group that received Yohimbine hydrochloride intervention was significantly decreased, confirming the significant inhibitory effect of Yohimbine hydrochloride on the increase of DBP in the AngII group.

[0124] Figure 1CThe results showed that compared with the Control group, the mean arterial pressure (MAP) of mice after AngII intervention was also significantly increased, P < 0.05. In contrast, the MAP of the AngII + Yohimbine group after Yohimbine hydrochloride intervention was significantly reduced, indicating that Yohimbine hydrochloride can significantly inhibit the increase of MAP in the AngII group.

[0125] Finally, according to Figure 1D The results showed that the intervention of yohimbine hydrochloride did not show a significant effect on the body weight of mice.

[0126] 2.2 Effects of Yohimbine Hydrochloride on the Resistance Index, Pulsatility Index and Thickness of the Abdominal Aorta in Mice

[0127] After AngII induction, mice were treated with yohimbine hydrochloride for four weeks. Using small animal ultrasound technology, this application detected and analyzed the resistance index, pulsatility index and vascular thickness of each group of mice. The relevant results are shown in Figure 2A -E.

[0128] according to Figure 2A and Figure 2B The results showed that compared with the Control group, the abdominal aorta pulsatility index of mice in the AngII group was significantly increased, while after the intervention of yohimbine hydrochloride, the index was significantly decreased. The statistical results showed that P<0.05.

[0129] Figure 2A and Figure 2C The results showed that the abdominal aorta resistance index of mice in the AngII group was significantly higher than that in the Control group, but after intervention with yohimbine hydrochloride, the index decreased significantly (P<0.05).

[0130] Figure 2D and Figure 2E The data showed that the thickness of the abdominal aorta of mice in the AngII group was significantly increased compared with that in the Control group, P<0.05, while after intervention with yohimbine hydrochloride, the thickness of the abdominal aorta was significantly decreased, P<0.05.

[0131] 2.3 Study on the pathological morphology of abdominal aorta in mice treated with yohimbine hydrochloride

[0132] The present application uses HE staining technology to observe the effect of yohimbine hydrochloride on the morphology of the abdominal aorta of mice. Figure 2F The results showed that compared with the Control group, the abdominal aorta wall of the mice in the AngII group was significantly thickened, and after the intervention of yohimbine hydrochloride, the thickening of the abdominal aorta wall of the mice was significantly improved.

[0133] 2.4 Effect of Yohimbine Hydrochloride on Cardiac Function in Mice

[0134] Small animal ultrasound was used to assess changes in cardiac function. Figure 3A , 3B and 3C results showed that compared with those of the Control group, the left ventricular ejection fraction (LVEF) and left ventricular fraction shortening (LVFS) of the mice in the AngII group were significantly decreased, P<0.05; compared with the AngII group, after the intervention of yohimbine hydrochloride, the left ventricular ejection fraction and cardiac output were significantly increased, P<0.05.

[0135] 2.5 Study on the pathological morphology of heart disease in mice treated with yohimbine hydrochloride

[0136] HE staining was used to evaluate the effect of yohimbine hydrochloride on the pathological morphology of AngII-induced mouse heart tissue. Figure 3D As shown in the figure, the myocardial cell nuclei of the mice in the Control group were clear, the cytoplasm was evenly stained and arranged neatly, and no degeneration or necrosis was observed; in contrast, the cardiac tissue morphology of the mice in the AngII group was significantly changed: the neat arrangement of myocardial cells was significantly reduced. After the intervention of yohimbine hydrochloride, the pathological morphology of the myocardial tissue of the model mice was significantly improved.

[0137] 2.6 Study on the effect of yohimbine hydrochloride on renal artery function in mice

[0138] Changes in renal artery function were assessed by small animal ultrasound.

[0139] like Figure 4A , 4B As shown in Figure 4C, compared with those in the Control group, the right renal artery pulsatility index (RRA PI) and renal artery resistance index (RRA RI) of the mice in the Ang II group were significantly increased, P < 0.05; compared with the Ang II group, the renal artery pulsatility index and renal artery resistance index were significantly decreased after yohimbine hydrochloride intervention, P < 0.05.

[0140] In summary, yohimbine hydrochloride can significantly reduce the blood pressure of AngII model mice, improve vascular function, relieve abdominal aortic thickening and related pathological changes, and also has the effect of improving the decline in heart function and vascular wall thickening caused by hypertension. In summary, yohimbine hydrochloride can significantly reduce the blood pressure of AngII model mice, improve vascular function, relieve abdominal aortic thickening and pathological changes, and also has the effect of improving the pathological morphological changes of the heart caused by hypertension and reducing the function of the renal artery and mesenteric artery.

[0141] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. Use of yohimbine hydrochloride or a pharmaceutically acceptable salt thereof in preparing a product for preventing or treating hypertension.

2. Use of yohimbine hydrochloride or a pharmaceutically acceptable salt thereof in the preparation of a product for preventing or treating vascular lesions caused by hypertension and / or cardiac lesions caused by hypertension.

3. The use according to claim 2, characterized in that The vascular lesions caused by hypertension and / or the heart lesions caused by hypertension have at least one of the following symptoms: 1) Decreased vascular function; 2) Thickening of blood vessel walls; 3) Reduced heart function.

4. The use according to claim 3, characterized in that Includes at least one of the following features: a) The reduced vascular function refers to an increase in the propagation velocity of vascular pulse waves; b) The reduced cardiac function refers to a reduced cardiac ejection fraction or a reduced left ventricular fractional shortening.

5. The use according to any one of claims 1 to 4, characterized in that: The yohimbine hydrochloride or a pharmaceutically acceptable salt thereof is used alone or in combination with other drugs.

6. The use according to claim 5, characterized in that The yohimbine hydrochloride or its pharmaceutically acceptable salt and a pharmaceutically acceptable carrier or auxiliary material, or a combination thereof, form a pharmaceutical composition.

7. The use according to claim 6, characterized in that The pharmaceutically acceptable excipient is selected from one or more of a diluent, a binder, a lubricant and a wetting agent.

8. The use according to claim 7, characterized in that The pharmaceutical composition is one or more of a solution, an injection, a spray, a nasal drop, an aerosol, a powder spray, a tablet, a capsule and a granule.

9. A product, characterized in that The product comprises yohimbine hydrochloride or a pharmaceutically acceptable salt thereof, and the product has at least one of the following functions: 1) Lower blood pressure; 2) Alleviate decreased vascular function; 3) Relieve blood vessel wall thickening; 4) Alleviate decreased heart function.

10. The product according to claim 9, characterized in that The products include medicines, health products and food.

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