Application of L-pyroglutamic acid and related products

Abstract

The invention relates to the technical field of medicines, in particular to application of L-pyroglutamic acid and related products. It is found for the first time that L-pyroglutamic acid 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 present invention relates to the field of medical technology, and in particular to a use of L-pyroglutamic acid and related products. Background Art

[0002] Hypertension is one of the most common chronic diseases and an important risk factor for cardiovascular disease. The Global Burden of Disease study shows that cardiovascular disease has the highest mortality rate and seriously endangers human health. At present, the incidence of hypertension is increasing year by year. It is estimated that by 2025, the number of people with hypertension in the world will reach 1.56 billion. In my country, with the change of people's lifestyle and the advancement of the aging process of the population, 16.5% of the annual deaths are caused by hypertension. By 2030, it is estimated that the annual death toll will reach 23.5 million. According to research reports, nearly 9.4 million people died of hypertension worldwide, accounting for 17.8% of total deaths. The number of deaths due to hypertension in my country accounted for 24.6% of all deaths, up to 2.043 million (including 1.154 million men and 889,000 women), and the direct medical expenses reached 36.6 billion yuan. The extremely high prevalence, large number of patients, and extremely low control rate make hypertension a major chronic disease in my country, which seriously endangers human health and also brings a huge burden to society. Therefore, it is of great significance to find safer and more effective prevention and treatment strategies.

[0003] L-pyroglutamic acid (Pyroglutamic acid) is widely present in many organisms. In recent years, studies have found that L-pyroglutamic acid has pharmacological activities such as anti-oxidation, anti-inflammatory, promoting nerve growth, regulating blood sugar and blood lipids, and anti-tumor. L-pyroglutamic acid can support energy metabolism by participating in the tricarboxylic acid cycle. At the same time, as a coenzyme or cofactor of certain important enzymes, it plays a key role in maintaining cell function. In addition, it also shows certain antioxidant properties, which helps to remove free radicals in the body and protect cells from oxidative stress damage. However, there is currently a lack of sufficient scientific evidence on its specific effects 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 L-pyroglutamic acid and related products.

[0005] The first aspect of the present invention provides use of L-pyroglutamic acid or a pharmaceutically acceptable salt thereof in preparing a product for preventing or treating hypertension.

[0006] The second aspect of the present invention provides the use of L-pyroglutamic acid 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] 4) Myocardial cells are disordered.

[0012] In some embodiments of the present invention, the reduced vascular function refers to an increase in the propagation velocity of vascular pulse waves.

[0013] 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.

[0014] The L-pyroglutamic acid or its pharmaceutically acceptable salt in the present invention is used alone or in combination with other drugs.

[0015] In some embodiments of the present invention, the L-pyroglutamic acid or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient, or a combination thereof, constitute a pharmaceutical composition.

[0016] 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.

[0017] 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.

[0018] The third aspect of the present invention provides a product, comprising L-pyroglutamic acid or a pharmaceutically acceptable salt thereof, wherein the product has at least one of the following functions:

[0019] 1) Lower blood pressure;

[0020] 2) Alleviate decreased vascular function;

[0021] 3) Relieve blood vessel wall thickening;

[0022] 4) Alleviate decreased heart function;

[0023] 5) Relieve the disorder of myocardial cell arrangement.

[0024] In some embodiments of the present invention, the products include medicines, health products and foods.

[0025] Compared with the prior art, the present invention has the following beneficial effects:

[0026] 1. The present invention first discovered that L-pyroglutamic acid 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, providing a new treatment approach for treating hypertension or hypertension-related diseases.

[0027] 2. The present invention has proved through experiments that L-pyroglutamate 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 improving the function of the renal artery and mesenteric artery. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1A The figure shows the effect of L-pyroglutamate on the systolic blood pressure of mice after AngII modeling in the present invention. The data are presented as mean ± standard deviation, and one-way analysis of variance and turkey test are used; * indicates that P < 0.05 is statistically significant compared with the Control group, and # indicates that P < 0.05 is statistically significant compared with the Ang II group.

[0029] Figure 1B The figure shows the effect of L-pyroglutamate on the diastolic blood pressure of mice after AngII modeling. * indicates that P<0.05 is statistically significant compared with the Control group, and # indicates that P<0.05 is statistically significant compared with the Ang II group.

[0030] Figure 1C The figure shows the effect of L-pyroglutamate on the mean arterial pressure of mice after AngII modeling. * indicates that P<0.05 is statistically significant compared with the Control group, and # indicates that P<0.05 is statistically significant compared with the Ang II group.

[0031] Figure 1D The figure shows the effect of L-pyroglutamate on the body weight of mice after AngII modeling in the present invention.

[0032] Figure 2A Ultrasound graph showing the effect of L-pyroglutamate on the resistance index and pulsatility index of the abdominal aorta of mice after AngII modeling.

[0033] Figure 2B Statistical analysis diagram showing the effect of L-pyroglutamate on the abdominal aorta pulsatility index of mice after AngII modeling. * indicates that P<0.05 is statistically significant compared with the Control group, and # indicates that P<0.05 is statistically significant compared with the Ang II group.

[0034] Figure 2CStatistical analysis diagram showing the effect of L-pyroglutamate on the abdominal aorta resistance index of mice after AngII modeling. * indicates that P<0.05 is statistically significant compared with the Control group, and # indicates that P<0.05 is statistically significant compared with the Ang II group.

[0035] Figure 2D Ultrasound images showing the effect of L-pyroglutamate on the vascular wall thickness of the abdominal aorta in mice after AngII modeling in the present invention.

[0036] Figure 2E Statistical analysis diagram showing the effect of L-pyroglutamate on the thickness of the abdominal aorta wall of mice after AngII modeling. * indicates that P<0.05 is statistically significant compared with the Control group, and # indicates that P<0.05 is statistically significant compared with the Ang II group.

[0037] Figure 2F The figure shows the effect of L-pyroglutamate on the pathological morphology of abdominal aorta in mice after AngII modeling in the present invention, wherein the scale bar is 50 μm.

[0038] Figure 3 A diagram showing the effect of L-pyroglutamate on the pathological morphology of heart in mice after AngII modeling in the present invention.

[0039] Figure 4A Ultrasound diagram showing the effect of L-pyroglutamate on renal function in mice after AngII modeling in the present invention.

[0040] Figure 4B The analysis diagram showing the effect of L-pyroglutamate on the renal pulsatility index (RRA PI) of mice after Ang II modeling in the present invention. * indicates that P<0.05 is statistically significant compared with the Control group, and # indicates that P<0.05 is statistically significant compared with the Ang II group.

[0041] Figure 4C The analysis diagram showing the effect of L-pyroglutamate on the renal resistance index (RRA RI) of mice after Ang II modeling in the present invention. * indicates that P<0.05 is statistically significant compared with the Control group, and # indicates that P<0.05 is statistically significant compared with the Ang II group.

[0042] Figure 5A Ultrasound images showing the effect of L-pyroglutamate on the function of mesenteric arteries in mice after AngII modeling in the present invention.

[0043] Figure 5BThe analysis diagram showing the effect of L-pyroglutamate on the mesenteric artery pulsatility index (SMAPI) of mice after AngII modeling in the present invention. * indicates that P<0.05 is statistically significant compared with the Control group, and # indicates that P<0.05 is statistically significant compared with the Ang II group.

[0044] Figure 5C The analysis diagram showing the effect of L-pyroglutamate on the mesenteric artery resistance index (SMARI) of mice after AngII modeling in the present invention. * indicates that P<0.05 is statistically significant compared with the Control group, and # indicates that P<0.05 is statistically significant compared with the Ang II group. DETAILED DESCRIPTION

[0045] The inventors of the present invention accidentally discovered that the compound L-pyroglutamic acid has antihypertensive effects and can improve heart and vascular lesions 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.

[0046] The first aspect of the present invention provides use of L-pyroglutamic acid or a pharmaceutically acceptable salt thereof in preparing a product for preventing or treating hypertension.

[0047] 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 L-pyroglutamic 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.

[0048] The molecular formula of L-pyroglutamic acid (also known as 5-oxoproline or 5-OPA) is C5H7NO3, CAS No. 98-79-3, and its structural formula is as follows:

[0049]

[0050] 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.

[0051] The pharmaceutically acceptable salts of L-pyroglutamic acid 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 an inorganic acid such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with an organic acid 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.

[0052] The product of the present invention necessarily contains L-pyroglutamic acid or a pharmaceutically acceptable salt thereof, and uses L-pyroglutamic acid or a pharmaceutically acceptable salt thereof as an effective ingredient for preventing or treating hypertension.

[0053] In the product of the present invention, the effective ingredient for preventing or treating hypertension may be only L-pyroglutamic acid, or may contain other chemicals that can play a similar role.

[0054] The product of the present invention may be a single-component substance or a multi-component substance.

[0055] The second aspect of the present invention provides the use of L-pyroglutamic acid 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.

[0056] 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:

[0057] 1) Decreased vascular function;

[0058] 2) Thickening of blood vessel walls;

[0059] 3) Reduced heart function;

[0060] 4) Myocardial cells are disordered.

[0061] In the second aspect of the present invention, the reduced vascular function refers to an increase in the vascular pulse wave propagation velocity. Specifically, the vascular function refers to the abdominal aorta, renal artery and mesenteric artery.

[0062] In the second aspect of the present invention, the vascular wall thickening refers to the vascular wall thickening of the abdominal aorta.

[0063] In the second aspect of the present invention, the reduced cardiac function refers to a reduced cardiac ejection fraction or a reduced left ventricular fractional shortening.

[0064] In the second aspect of the present invention, the disordered arrangement of cardiomyocytes refers to abnormal orientation and disordered arrangement of some cardiomyocytes, and abnormal changes in the cell nuclei.

[0065] In the present invention, the L-pyroglutamic acid or its pharmaceutically acceptable salt is used alone or in combination with other drugs.

[0066] In some embodiments of the present invention, the L-pyroglutamic acid or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient, or a combination thereof, constitute a pharmaceutical composition.

[0067] 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.

[0068] "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.

[0069] "Pharmaceutically acceptable carriers or excipients" should be compatible with L-pyroglutamic acid, 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.

[0070] 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.

[0071] 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.

[0072] The third aspect of the present invention provides a product, comprising L-pyroglutamic acid or a pharmaceutically acceptable salt thereof, wherein the product has at least one of the following functions:

[0073] 1) Lower blood pressure;

[0074] 2) Alleviate decreased vascular function;

[0075] 3) Relieve blood vessel wall thickening;

[0076] 4) Alleviate decreased heart function;

[0077] 5) Relieve the disorder of myocardial cell arrangement.

[0078] In some embodiments of the third aspect of the present invention, the mitigation of reduced vascular function refers to reducing the vascular pulse wave propagation velocity, and the reduction of the vascular pulse wave propagation velocity refers to making the vascular pulse wave propagation velocity 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 vascular pulse propagation velocity of the subject's blood vessels to at least 70%, 80%, 90% or 100% of that of healthy blood vessels.

[0079] In some embodiments of the third aspect of the present invention, the relieving of blood vessel thickening refers to reducing the thickness of the blood vessel wall, and reducing the thickness of the blood vessel wall refers to making the blood vessel wall of the subject approach or reach the thickness of a healthy blood vessel. For example, based on a healthy blood vessel, the product can make the thickness of the blood vessel wall of the subject blood vessel not exceed 130%, 120%, 110% or 100% of the thickness of a healthy blood vessel.

[0080] In some embodiments of the third aspect of the present invention, the alleviation of decreased cardiac function refers to improving cardiac ejection fraction or left ventricular shortening fraction, and the improvement of cardiac ejection fraction or left ventricular shortening fraction refers to making the cardiac ejection fraction and 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 left ventricular shortening fraction of the subject's heart to at least 70%, 80%, 90% or 100% of that of a healthy heart.

[0081] In some embodiments of the third aspect of the present invention, the relief of disordered myocardial cell arrangement refers to making myocardial cells arranged neatly, restoring myocardial cells to health, or reducing the number of myocardial cells with abnormal nuclei. For example, based on a healthy heart, the product can restore the orderly arrangement of myocardial cells in the heart of the subject to at least 70%, 80%, 90% or 100% of that of a healthy heart.

[0082] In the third aspect of the present invention, the symptoms of reduced vascular function, thickened vascular wall, reduced cardiac function or disordered myocardial cell arrangement are caused by hypertension, and are specifically related to angiotensin II.

[0083] In some embodiments of the present invention, the products include medicines, health products and foods.

[0084] 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.

[0085] In the third aspect of the present invention, the effective dose of the trilobatin 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.

[0086] 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.

[0087] 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.

[0088] 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.

[0089] 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.

[0090] 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.

[0091] The following examples all used IBM SPSS Statistics (version 29.0.1.0) software for data statistical analysis.

[0092] The Shapiro-Wilk test was used to test the normality of the data. Indicates. When comparing differences between three or more groups, ANOVA was used to compare the data under the premise of normal distribution (when the data variances were equal, LSD was used for post hoc test; otherwise, Games-Howell was used for post hoc test); when the data did not conform to normal distribution, the non-parametric Kruskal-Wallis test was used to compare the differences between groups. P < 0.05 indicated a significant difference and was considered statistically significant.

[0093] Example 1: Animal model construction and treatment

[0094] 1.1 Experimental animals

[0095] 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.

[0096] 1.2 Drug preparation

[0097] Weigh an appropriate amount of L-pyroglutamic 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 an L-pyroglutamic acid solution.

[0098] Weigh an appropriate amount of angiotensin 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.

[0099] Meanwhile, an appropriate amount of valsartan powder was weighed and dissolved in a corresponding volume of distilled water at a dose of 10 mg / kg / d according to the weight of the mice. After preparation, the mixture was stored in a refrigerator at 4° C. for later use to obtain a valsartan solution.

[0100] 1.3 Animal grouping and model construction

[0101] Animal grouping: 24 male C57BL / 6 mice were randomly divided into 4 groups according to basal blood pressure: Control group, AngII group, AngII+Pyroglutamic acid (10 mg / kg / d) group, and AngII+Valsartan (7.5 mg / kg / d) group, with 6 mice in each group.

[0102] Preparation of osmotic micropumps: The 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, AngII + Pyroglutamic acid (10 mg / kg / d) group, and AngII + Valsartan (7.5 mg / kg / d) group, respectively, to obtain pre-set osmotic micropumps containing AngII.

[0103] The osmotic minipumps in the Control group were injected with 200 μL of saline solution to obtain osmotic minipumps containing saline.

[0104] The osmotic mini-pumps of all groups were immersed in physiological saline in a 37°C cell culture incubator for 24 hours and used for pump implantation surgery the next day.

[0105] Model construction and treatment: On the day of surgery, the operating table was routinely disinfected, and the mice were anesthetized with isoflurane, placed in a prone position, and the hair on the back of the head and neck was removed. The skin was cut with scissors and the subcutaneous tissue was bluntly separated after disinfection with iodine. The mice in the AngII group, AngII+Pyroglutamic acid (10 mg / kg / d) group, and AngII+Valsartan (7.5 mg / kg / d) group were subcutaneously implanted with pre-placed osmotic micropumps containing AngII, and the mice in the Control group were subcutaneously implanted with pre-placed osmotic micropumps containing saline. After the corresponding osmotic micropumps were implanted in each group, the skin was sutured and disinfected.

[0106] Starting from the second day after pump implantation, mice in the AngII+Pyroglutamic acid group were given intragastric administration of 10 mg / kg / d L-pyroglutamic acid solution, and mice in the AngII+Valsartan group were given intragastric administration of 7.5 mg / kg / d valsartan solution. The Control group and the AngII group were given intragastric administration of an equal volume of distilled water once a day for a total of 4 weeks.

[0107] Example 2: Effect of L-pyroglutamate on blood pressure and body weight in mice

[0108] 2.1 Blood pressure and weight measurement

[0109] During the experiment, the CODA TMThe non-invasive precision blood pressure meter measures the mean arterial pressure (MAP), systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the experimental mice once a week, and performs statistical analysis on the data. The weight changes of each mouse are also recorded weekly. The specific operation is as follows:

[0110] (1) Check whether there are any abnormalities in the equipment and assemble the mouse heating plate, sensor and computer completely;

[0111] (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.

[0112] (3) Ensure that the mouse is in a quiet state and put the occlusion suit and volume pressure sensor on the mouse tail;

[0113] (4) Open Coda 4.1 software and set the blood pressure test to 15 cycles per cycle. The first 5 cycles are adaptive tests and the last 10 cycles are formal tests. Wait for the mouse to calm down for 10 minutes before starting the measurement. Pay attention to the measurement period and keep the surrounding environment in a quiet state. After the measurement is completed, the volume pressure sensor transmits the value of each measurement to the computer;

[0114] (5) Record the systolic blood pressure, diastolic blood pressure, and mean arterial pressure of each animal each week and weigh the body weight each week, and take the mean and standard deviation.

[0115] (6) After the measurement, weigh the mice and place them back in their cages.

[0116] 2.2 Analysis results

[0117] Effects of L-pyroglutamate on blood pressure Figure 1A -C.

[0118] like Figure 1A As shown in the figure, compared with the Control group, the SBP of mice after AngII intervention was significantly increased (P<0.05). Compared with the AngII group, after L-pyroglutamic acid intervention, the SBP of the AngII+Pyroglutamic acid group was significantly reduced (P<0.05). The results showed that L-pyroglutamic acid intervention could significantly inhibit the increase of SBP in the AngII group, and was equivalent to the SBP after valsartan intervention.

[0119] like Figure 1BAs shown in the figure, compared with the Control group, the DBP of mice after AngII intervention was significantly increased (P<0.05). Compared with the AngII group, the DBP of the AngII+Pyroglutamic acid group was significantly reduced after L-pyroglutamic acid intervention (P<0.05). The results showed that L-pyroglutamic acid intervention could significantly inhibit the increase of DBP in the AngII group, and was equivalent to the DBP after valsartan intervention.

[0120] like Figure 1C As shown in the figure, compared with the Control group, the MAP of mice after AngII intervention was significantly increased (P<0.05). Compared with the AngII group, the MAP of the AngII+Pyroglutamic acid group was significantly decreased after L-pyroglutamic acid intervention (P<0.05). The results showed that L-pyroglutamic acid intervention could significantly inhibit the increase of MAP in the AngII group, and was equivalent to the MAP after valsartan intervention.

[0121] Figure 1A -C showed that L-pyroglutamate could reduce systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MAP) of mice, and the effect was significant.

[0122] like Figure 1D As shown, L-pyroglutamate intervention had no significant effect on the body weight of mice.

[0123] Example 3: Effect of L-pyroglutamate on abdominal aorta function and vascular wall thickness in mice

[0124] 3.1 Ultrasonic measurement of abdominal aorta pulsatility index, resistance index and vascular wall thickness in mice

[0125] (1) Ultrasound detection time: After the L-pyroglutamate intervention period, the abdominal aorta vascular function of each group of mice was detected using Vevo 2100 small animal ultrasound instrument.

[0126] (2) Abdominal hair removal and animal anesthesia: The abdominal hair of the animal was removed with a depilatory cream, and the mouse was anesthetized by inhalation of 1.5% isoflurane. The mouse was placed in a supine position on a 37° C. constant temperature heating plate.

[0127] (3) Obtaining the abdominal aorta section: Coupling agent was applied to the mouse (including the anterior chest, abdomen, etc.), and an ultrasound probe (the probe frequency was 30 MHz) was used to find the long axis section of the left ventricle. The probe was moved downward parallel to the midline of the abdomen to completely expose the abdominal aorta and obtain an abdominal aorta section image.

[0128] (4) Analysis and calculation of ultrasound results: Vevo Strain Software (Vevo LAB 1.7.1) was used to calculate the abdominal aorta resistance index, pulsatility index and vascular wall thickness of each group of animals according to the software manual, and the average value was calculated three times.

[0129] Calculated according to the following formula:

[0130] Pulsatility index: PI = (peak velocity during systole - average velocity during diastole) / average velocity;

[0131] Resistance index: RI = (peak systolic velocity - end diastolic velocity) / peak systolic velocity.

[0132] 3.2 Analysis results

[0133] The resistance index, pulsatility index and vascular wall thickness of each group of mice were detected and analyzed by small animal ultrasound technology. Figure 2A -E as shown.

[0134] like Figure 2A and Figure 2B As shown, compared with the Control group, the abdominal aortic resistance index of mice in the AngII group was significantly increased; compared with the AngII group, after L-pyroglutamate intervention, the abdominal aortic resistance index was significantly decreased, P<0.05, and was comparable to this index after valsartan intervention.

[0135] like Figure 2A and Figure 2C As shown, compared with the Control group, the abdominal aorta pulsatility index of the mice in the AngII group was significantly increased; compared with the AngII group, after L-pyroglutamate intervention, the abdominal aorta pulsatility index was significantly decreased, P<0.05, and was comparable to that after valsartan intervention.

[0136] like Figure 2D and Figure 2E As shown, compared with the Control group, the thickness of the abdominal aorta of mice in the AngII group was significantly increased, P<0.05; compared with the AngII group, the thickness of the abdominal aorta of mice after L-pyroglutamate intervention was significantly decreased, P<0.05, and the effect was comparable to the thickness of the abdominal aorta after valsartan intervention.

[0137] Depend on Figure 2A -E showed that L-pyroglutamate could reduce the abdominal aortic resistance index, aortic pulsatility index and abdominal aorta thickness of mice.

[0138] Example 4: Effects of L-pyroglutamate on pathological morphology of aorta and heart

[0139] 4.1 HE staining to detect the pathological morphology of aorta and heart

[0140] (1)Fixed:

[0141] Mouse abdominal aorta and heart tissue samples were immersed in 4% paraformaldehyde for fixation for 48 h, taken out, trimmed, placed in an embedding box, and placed in 70% ethanol.

[0142] (2) Alcohol gradient dehydration and wax immersion:

[0143] The heart tissue samples were immersed in the following reagents for dehydration: 70% anhydrous ethanol for 60 min → 80% anhydrous ethanol for 60 min → 90% anhydrous ethanol for 60 min → 95% anhydrous ethanol for 60 min → 100% anhydrous ethanol I for 20 min → 100% anhydrous ethanol II for 20 min → 100% anhydrous ethanol III for 50 min → xylene I for 5 min → xylene II for 20 min. The dehydrated tissues were immersed in wax in the following order: paraffin I for 10 min → paraffin II for 20 min → paraffin III for 30 min.

[0144] The abdominal aorta tissue samples were immersed in the following reagents for dehydration: 70% anhydrous ethanol for 30 min → 80% anhydrous ethanol for 30 min → 90% anhydrous ethanol for 30 min → 95% anhydrous ethanol for 30 min → 100% anhydrous ethanol I for 10 min → 100% anhydrous ethanol II for 10 min → 100% anhydrous ethanol III for 30 min → xylene I for 5 min → xylene II for 15 min. The dehydrated tissues were immersed in wax in the following order: paraffin I for 10 min → paraffin II for 10 min → paraffin III for 25 min.

[0145] (4) Embedding:

[0146] First inject a little liquid paraffin into the bottom of the embedding box, then place the abdominal aorta vertically into the embedding box, and then fill it with liquid paraffin to prevent the formation of bubbles. After the paraffin solidifies, pry the wax block out of the mold and trim it.

[0147] (5) Slicing:

[0148] Place the wax block in a -20℃ refrigerator for precooling and then fix it on the slicer. First, use 10μm thickness for rough trimming. When the tissue sample is cut, change to 0.4μm thickness for fine trimming. Use a brush to pick up the complete slice and spread it in 41℃ water. Use a glass slide to gently pick it up and stick it flat on the carrier. Bake the slice at 60℃ for 2h, dry it for 3h, and store the slice at room temperature for subsequent experiments.

[0149] (6) Dewaxing:

[0150] Immerse the heart tissue in the reagents for dewaxing in the following order: xylene I 25 min→xylene II 20 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.

[0151] The abdominal aorta tissue was immersed in reagents for dewaxing in the following order: xylene I 20 min→xylene II 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.

[0152] (7) Hematoxylin staining:

[0153] The slides were immersed in hematoxylin solution for 1 min and then washed with pure water.

[0154] (8) Eosin staining:

[0155] The above slides were immersed in eosin solution for staining for 3 seconds, and then washed with pure water.

[0156] (9) Sealing:

[0157] After staining, blow dry the slices immediately, gently add an appropriate amount of neutral gum, take a cover slip and stick it on the slide along one side, then squeeze it hard. There should be no bubbles around the tissue.

[0158] (10) Microscope observation:

[0159] Using the intelligent tissue section analysis system, the stained tissue sections of each sample were imaged at 400× magnification, and 6 fields of view were randomly selected to take pictures and save them to observe the pathological changes of each group of samples.

[0160] 4.2 Analysis results

[0161] HE staining was used to observe the effect of L-pyroglutamate on the morphology of abdominal aorta in mice.

[0162] like Figure 2F As shown, compared with the Control, the abdominal aorta wall of the mice in the AngII group was thickened; compared with the AngII group, the thickening of the abdominal aorta wall of the mice was alleviated after L-pyroglutamate intervention, and the effect was comparable to that after valsartan intervention.

[0163] HE staining was used to evaluate the effect of L-pyroglutamate on the pathological morphology of cardiac tissue in AngII model mice.

[0164] like Figure 3 As shown, 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 AngII group showed obvious changes: the neatness of the arrangement of the cardiomyocytes was significantly decreased; compared with the AngII group, the pathological morphology of the myocardial tissue of the mice after modeling with L-pyroglutamate intervention was significantly improved, and the effect was comparable to that after valsartan intervention.

[0165] Example 5: Effect of L-pyroglutamate on right renal artery function in mice

[0166] 5.1 Ultrasonic measurement of mouse abdominal aorta pulsatility index and resistance index

[0167] The steps of hair removal and animal anesthesia are the same as 3.1. After the mouse enters the anesthetized state, the full picture of the two renal arteries on both sides of the abdominal aorta is displayed. The sampling volume is placed within 1 / 2 of the length of one end of the starting point of the right renal artery to obtain the right renal artery section picture. The angle is <60 degrees to read 3-5 frequency spectrums of various blood flow parameters, and the average value of each index is taken to calculate and analyze the changes in renal artery blood flow. The calculation formula of pulsatility index and resistance index is the same as 3.1.

[0168] 5.2 Analysis results

[0169] Changes in renal artery function were assessed by small animal ultrasound.

[0170] like Figure 4A , 4B As shown in Figure 4C, compared with those of the Control group, the right renal artery pulsatility index (RRAPI) and renal artery resistance index (RRARI) of the mice in the AngII group were significantly increased, P<0.05; compared with the AngII group, after L-pyroglutamate intervention, the renal artery pulsatility index and renal artery resistance index were significantly decreased, P<0.05, which were comparable to those after valsartan intervention.

[0171] Example 6: Effect of L-pyroglutamate on mesenteric artery function in mice

[0172] 6.1 Ultrasonic measurement of mouse mesenteric artery pulsatility index and resistance index

[0173] The steps of hair removal and animal anesthesia are the same as 3.1. The probe is placed in the middle of the abdomen along the longitudinal section to display the abdominal aorta, and the mesenteric artery is gradually found downward. The blood flow spectrum is displayed with pulsed Doppler. After adjusting the angle of blood flow and sound beam, the cross-sectional image of the mesenteric artery is obtained. The average value of each index is taken to calculate and analyze the changes in mesenteric artery blood flow. The calculation formula of pulsatility index and resistance index is the same as 3.1.

[0174] 6.2 Analysis results

[0175] Changes in renal artery function were assessed by small animal ultrasound.

[0176] like Figure 5A , 5B As shown in Figure 5C, compared with those of the Control group, the mesenteric artery pulsatility index (SMAPI) and mesenteric artery resistance index (SMARI) of the mice in the AngII group were significantly increased, P<0.05; compared with the AngII group, after L-pyroglutamate intervention, the mesenteric artery pulsatility index and mesenteric artery resistance index were significantly decreased, P<0.05, which were comparable to those after valsartan intervention.

[0177] In summary, L-pyroglutamate can significantly reduce blood pressure, improve vascular function, and alleviate abdominal aortic thickening and pathological changes in AngII model mice. It also has the effect of improving cardiac pathological morphological changes caused by hypertension and reducing the function of renal arteries and mesenteric arteries.

[0178] 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 L-pyroglutamic acid or a pharmaceutically acceptable salt thereof in preparing a product for preventing or treating hypertension.

2. Use of L-pyroglutamic acid 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) Myocardial cells are disordered.

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 L-pyroglutamic acid 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 L-pyroglutamic acid or its pharmaceutically acceptable salt and 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 L-pyroglutamic acid 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; 5) Relieve the disorder of myocardial cell arrangement.

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

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