阅读说明：本技术 乙氧基血根碱的用途及相关产品 (Use of ethoxy sanguinarine and related products ) 是由 彭军 沈阿灵 褚剑锋 程瑛 李加鹏 张秀丽 魏丽慧 林珊 徐南辉 于 2021-01-05 设计创作，主要内容包括：本发明属于生物医药研究领域,具体涉及乙氧基血根碱在制备具有以下一项或多项功能的产品中的用途：高血压治疗；改善血管功能；改善血管病理改变；缓解心脏功能降低；改善心脏病理改变。本发明首次发现乙氧基血根碱可以应用在制备治疗高血压或高血压引起的血管、心脏病变的药物中,治疗效果良好。为治疗高血压或高血压相关疾病提供了新的治疗途径。(The invention belongs to the field of biomedical research, and particularly relates to an application of ethoxy sanguinarine in preparing a product with one or more functions as follows: treatment of hypertension; improving the function of blood vessels; improving vascular pathological changes; relieving decreased cardiac function; improving the pathological change of the heart. The invention discovers for the first time that the ethoxy sanguinarine can be applied to the preparation of the medicine for treating hypertension or vascular and cardiac pathological changes caused by hypertension, and has good treatment effect. Provides a new treatment way for treating hypertension or hypertension-related diseases.)

1. Use of ethoxysanguinarine in the manufacture of a product having one or more of the following functions:

(1) treatment of hypertension;

(2) improving the function of blood vessels;

(3) improving vascular pathological changes;

(4) relieving decreased cardiac function;

(5) improving the pathological change of the heart.

2. Use according to claim 1, wherein the ethoxysanguinarine is the sole active ingredient or one of the active ingredients of the product.

3. The use of claim 1, wherein the vascular function, vascular pathology, reduction in cardiac function, or cardiac pathology is due to hypertension.

4. The use of claim 1, wherein said ameliorating a pathological change in the heart comprises ameliorating cardiomyocyte hypertrophy.

5. A medicament, comprising ethoxysanguinarine; the medicament has one or more of the following functions:

(1) treatment of hypertension;

(2) improving the function of blood vessels;

(3) improving vascular pathological changes;

(4) relieving decreased cardiac function;

(5) improving the pathological change of the heart.

6. The medicament for treating hypertension according to claim 5, wherein the ethoxysanguinarine is the sole active ingredient or one of the active ingredients of the medicament.

7. The medicament for treating hypertension according to claim 5, wherein the reduction in cardiac function or the pathological changes in the heart are caused by hypertension.

8. The medicament for treating hypertension according to claim 5, further comprising a pharmaceutically acceptable carrier or adjuvant.

9. The medicament for treating hypertension according to claim 5, which is an injectable medicament or an oral medicament.

10. Use of ethoxylated sanguinarine in the manufacture of a product for reducing thickening of blood vessel thickness.

Technical Field

The invention belongs to the field of biomedical research, and particularly relates to application of ethoxy sanguinarine and a related product.

Background

Ethoxy sanguinarine (ethoxyanguinarine) is a class of phenylisoquinoline alkaloids, found primarily in the papaveraceae and rutaceae families. To date, it has been found to be present in many natural herbal ingredients with good biological activity, such as antibacterial, antifungal, anti-inflammatory, antioxidant and antitumor properties. Ethoxy sanguinarine is reported to have good antiviral activity by down-regulating the cytopathic effect induced by porcine reproductive and respiratory syndrome virus; in addition, reports further prove that 0.5mg/kg of ethoxy sanguinarine can effectively inhibit CIP2A protein expression of colorectal cancer in mice, inhibit activity of colorectal cancer cells and induce apoptosis; research shows that in lung cancer cells, the ethoxysanguinarine with the concentration of 1.5uM can also inhibit the expression of CIP2A, thereby inhibiting the proliferation of the lung cancer cells and simultaneously enhancing the apoptosis effect of the lung cancer cells. Another study showed that sanguinarine at 3uM can act as a potent TMEM16A (transmembrane member) inhibitor, effectively inhibiting TMEM16A-Ca in lung cancer cells²⁺Activated Cl^-A channel, thereby inhibiting proliferation and invasion of cells and inducing apoptosis of lung cancer cells; researches prove that the ethoxy sanguinarine plays an important role in intestinal flora and has different influences on the functions of different intestinal flora; in a breast cancer model mouse, the ethoxy sanguinarine can be used as a novel AMPK kinase activator, can inhibit the growth of breast cancer of the mouse, and induces the breast cancer cells to generate autophagy, so that the ethoxy sanguinarine is expected to become a new target point for treating the breast cancer. However, the influence and mechanism of ethoxy sanguinarine on cardiovascular diseases such as hypertension are not reported yet.

Disclosure of Invention

In order to overcome the problems in the prior art, the invention aims to provide the application of the ethoxy sanguinarine and related products.

In order to achieve the above objects and other related objects, the present invention adopts the following technical solutions:

in a first aspect the present invention provides the use of ethoxylated sanguinarine in the manufacture of a product having one or more of the following functions:

(1) treatment of hypertension;

(2) improving the function of blood vessels;

(3) improving vascular pathological changes;

(4) relieving decreased cardiac function;

(5) improving the pathological change of the heart.

In a second aspect, the invention provides a medicament comprising ethoxysanguinarine; the medicament has one or more of the following functions:

(1) treatment of hypertension;

(2) improving the function of blood vessels;

(3) improving vascular pathological changes;

(4) relieving decreased cardiac function;

(5) improving the pathological change of the heart.

In a third aspect, the invention provides the use of ethoxylated sanguinarine in the manufacture of a product for reducing thickening in blood vessel thickness.

Compared with the prior art, the invention has the following beneficial effects:

the invention discovers for the first time that the ethoxy sanguinarine can be applied to the preparation of the medicine for treating hypertension or heart diseases caused by hypertension, and the treatment effect is good. Provides a new treatment way for treating hypertension or hypertension-related diseases.

Drawings

FIG. 1A Effect of ethoxysanguinarine on AngII-stimulated blood pressure (systolic blood pressure) in mice.

FIG. 1B Effect of ethoxysanguinarine on AngII-stimulated blood pressure (diastolic blood pressure) in mice.

FIG. 1C Effect of ethoxysanguinarine on AngII-stimulated blood pressure in mice (mean arterial pressure).

FIG. 1D Effect of ethoxysanguinarine on AngII-stimulated mouse body weight.

FIG. 2A is an ultrasound image of the effect of ethoxysanguinarine on the propagation velocity of pulse waves in the abdominal aorta of AngII-stimulated mice.

FIG. 2B is a bar graph of the effect of ethoxysanguinarine on the speed of propagation of pulse waves in the abdominal aorta of AngII-stimulated mice.

FIG. 2C histogram analysis of the effect of ethoxysanguinarine on AngII stimulated mouse abdominal aorta thickness.

FIG. 3 Effect of ethoxysanguinarine on AngII stimulation of pathological morphology of mouse aorta.

FIG. 4A sonogram of the effect of ethoxysanguinarine on AngII stimulated mouse cardiac function (arrow: left ventricular end diastolic/systolic inside diameter).

FIG. 4B is an analysis of the effect of ethoxysanguinarine on AngII stimulation of mouse cardiac function (EF%).

FIG. 4C analysis of the effect of ethoxysanguinarine on AngII-stimulated mouse cardiac function (FS%).

Figure 5 effect of ethoxysanguinarine on ang ii stimulated heart pathology in mice.

Detailed Description

Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention. Test methods in which specific conditions are not specified in the following examples are generally carried out under conventional conditions or under conditions recommended by the respective manufacturers.

When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.

Unless otherwise indicated, the experimental methods, detection methods, and preparation methods disclosed herein all employ techniques conventional in the art of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related arts.

One embodiment of the present invention provides the use of ethoxylated sanguinarine in the preparation of a product having one or more of the following functions:

(1) treatment of hypertension;

(2) improving the function of blood vessels;

(3) improving vascular pathological changes;

(4) relieving decreased cardiac function;

(5) improving the pathological change of the heart.

The ethoxy sanguinarine is the only effective component or one of the effective components of the medicine.

In one embodiment, the vascular function, the change in vascular pathology, the reduction in cardiac function, or the change in cardiac pathology is due to hypertension.

Optionally, the improving the vascular function comprises reducing the propagation speed of the vascular pulse wave.

Optionally, the ameliorating the vascular pathological change comprises alleviating vascular thickening.

Optionally, the alleviating reduced cardiac function comprises increasing cardiac ejection fraction and left ventricular shortening fraction.

Optionally, the ameliorating the pathological change of the heart comprises ameliorating cardiac myocyte hypertrophy.

One embodiment of the present invention provides a medicament comprising ethoxysanguinarine; the medicament has one or more of the following functions:

(1) treatment of hypertension;

(2) improving the function of blood vessels;

(3) improving vascular pathological changes;

(4) relieving decreased cardiac function;

(5) improving the pathological change of the heart.

The ethoxy sanguinarine is the only effective component or one of the effective components of the medicine.

In one embodiment, the vascular function, the change in vascular pathology, the reduction in cardiac function, or the change in cardiac pathology is due to hypertension.

Optionally, the improving the vascular function comprises reducing the propagation speed of the vascular pulse wave.

Optionally, the ameliorating the vascular pathological change comprises alleviating vascular thickening.

Optionally, the alleviating reduced cardiac function comprises increasing cardiac ejection fraction and left ventricular shortening fraction.

Optionally, the ameliorating the pathological change of the heart comprises ameliorating cardiac myocyte hypertrophy.

Further, the medicine also comprises a pharmaceutically acceptable carrier or auxiliary material.

By "pharmaceutically acceptable" is meant that the molecular entities and compositions do not produce adverse, allergic, or other untoward reactions when properly administered to an animal or human.

The "pharmaceutically acceptable carrier or adjuvant" should be compatible with the active ingredient, i.e., capable of being blended therewith without substantially diminishing the effectiveness of the drug under ordinary circumstances. Specific examples of some substances that can serve as pharmaceutically acceptable carriers or adjuvants are sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium methylcellulose, ethylcellulose and methylcellulose; powdered gum tragacanth; malt; gelatin; talc; solid lubricants, such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and cocoa butter; polyhydric alcohols such as propylene glycol, glycerin, sorbitol, mannitol, and polyethylene glycol; alginic acid; emulsifiers, such as Tween; wetting agents, such as sodium lauryl sulfate; a colorant; a flavoring agent; tabletting agents, stabilizers; an antioxidant; a preservative; pyrogen-free water; isotonic saline solution; and phosphate buffer, and the like. These materials are used as needed to aid in the stability of the formulation or to aid in the enhancement of the activity or its bioavailability or to produce an acceptable mouthfeel or odor upon oral administration.

The medicament can be an injection medicament or an oral medicament.

In the present invention, unless otherwise specified, the pharmaceutical dosage form is not particularly limited, and may be prepared into injection, oral liquid, tablet, capsule, dripping pill, spray, etc., and may be prepared by a conventional method. The choice of the pharmaceutical dosage form should be matched to the mode of administration.

Further, where the medicament is for treating hypertension in a subject, it is desirable to administer an effective amount of the medicament to the subject. With this method, the hypertension is suppressed.

The subject may be a mammal. The mammal is preferably a rodent, artiodactyla, perissodactyla, lagomorpha, primate, or the like. The primate is preferably a monkey, ape or human.

One embodiment of the invention provides the use of ethoxylated sanguinarine in the preparation of a product for alleviating thickening of blood vessel thickness.

Example 1

1 Material

1.1 Experimental animals

SPF grade C57BL/6 male mice, weighing 25 + -5 g, were used in this experiment. The experimental animals were purchased from shanghai slaike experimental animals limited, and the experimental animals used license numbers: SCKY 2017-. Animal experiments were performed according to the "guidelines for experimental animal treatment" published by the national department of science and technology in 2006. The experimental animals are all raised in the specific nonpathogenic experimental animal center of Fujian Chinese medicinal university, an SPF laboratory is used for freely and movably taking food and drinking water, the indoor ventilation and illumination are sufficient, the room temperature is controlled to be (23 +/-1) DEG C, the relative humidity is about 50-60%, the illumination/dark cycle is 12h, the raising environment is kept quiet and is not disturbed, and the mice are used for experiments after being adaptively fed for 5-7 days.

1.2 Experimental drugs and Primary reagents

Angiotensin II (Abcam, ab 120183); osmotic minipumps (Alzet, 2004D); ethoxysanguinarine (Shanghai leaf Biotech limited, B21511); eosin staining solution (beijing solibao science and technology ltd., g 1100); hematoxylin staining solution (beijing solibao science and technology ltd., g 1140); isoflurane (Shenzhen, Riwold Life technologies, Ltd., 970-; chemical reagents such as paraformaldehyde (LA 0427, feijing, inc.), absolute ethanol (1280340101602, west longa science co., ltd.), and xylene (1430030101600, west longa science co., ltd.).

1.3 Main Instrument of experiment

Pipettors (rying corporation, usa); electronic balance scales (Shanghai Aohaus instruments Co., Ltd.); a non-invasive rat tail sphygmomanometer (Kent corporation, usa); small animal ultrasound imaging system Vevo2100 (fuji film invest ltd); inhalation type small animal anesthesia machine (Shenzhen Riwode Life technologies, Ltd.); pathological microtomes (lycra, germany); paraffin embedding machine (Hubei filial tract sub-optical medical electronic technology Co., Ltd.).

2 method of experiment

2.1 pharmaceutical formulation

Dissolving ethoxy sanguinarine (Ethoxysanguiarine) in normal saline at dosage of 200 μ l/per day at administration concentration of 1 mg/kg/day, placing in ultrasonicator, treating at low temperature for 30min until completely dissolved, and storing in refrigerator at-20 deg.C.

2.2 animal grouping and model construction

Male C57BL/6 mice, 8-10 weeks old, were randomized into three groups according to basal blood pressure: control group (n ═ 5), AngII + ethoxysguinare group (AngII + Eth; n ═ 5). One day before operation, the osmotic mini-pumps were taken out in a super clean bench, 200. mu.l of AngII solution (500 ng/kg/min) was injected into the osmotic mini-pumps of AngII group and AngII + Eth group, and 200. mu.l of physiological saline solution was injected into the osmotic mini-pumps of Control group, and all the osmotic mini-pumps were immersed in physiological saline in a cell culture chamber at 37 ℃ overnight. On the day of operation, an operating table is used for carrying out conventional disinfection, mice are anesthetized by isoflurane, the mice are in prone position, the skin of the head, the neck and the back is depilated, the skin is disinfected by iodophor, the skin is cut by scissors, subcutaneous tissues are separated in a blunt way, mice in an AngII group and an AngII + Eth group are subcutaneously implanted with a preset osmotic micropump containing AngII, the mice in a Control group are subcutaneously implanted with a preset osmotic micropump containing normal saline, and the skin is sutured and disinfected. The pump is buried for the next day, 200. mu.l/body of normal saline is intraperitoneally injected into the Control group and the AngII group, and 200. mu.l/body of ethoxysanguinarine solution (1 mg/kg/day) is intraperitoneally injected into the AngII + Eth group, 1 time per day, for 4 weeks.

2.3 blood pressure measurement

In the experiment, a noninvasive rat tail artery sphygmomanometer is adopted to monitor the blood pressure change of each group of mice. Keeping the experimental animal in a calm state, then loading the experimental animal into a restraint device, sleeving a blocking sleeve at a proper position (proper elasticity) of a rat tail, then sleeving a volume pressure sensor (VPR), ensuring that one part of the rat tail is exposed out of the sensor, placing the animal restraint device on a heating plate, selecting a proper heating gear according to indoor temperature to heat the rat tail, covering the head of the animal with a black towel and ensuring the quiet of the experimental environment, standing the animal in the dark state for 10min, starting to detect the blood pressure, circularly inflating and deflating the blocking sleeve and the VPR sensor for 15 times, setting computer software, measuring for 15 times, and taking the average value. The tail systolic arterial pressure (SBP), diastolic pressure (DBP) and Mean Arterial Pressure (MAP) of the small animals were measured by the instrument.

2.4 measurement of conduction velocity and thickness of Abdominal aortic pulse wave

Pulse wave velocity and vessel thickness measurements of the abdominal aorta were performed using a Vevo2100 animal ultrasound machine (VisualSonics, Toronto, Ontario, Canada). The mice are fixed on a physiological information monitoring table in a supine position under 2 percent isoflurane inhalation anesthesia to maintain the anesthesia concentration at 1.5 to 2 percent. Depilatory cream is used for belly depilation, and proper amount of coupling agent is smeared on belly and physiological information monitor. The probe is placed in the middle of the abdomen in parallel, B-mode ultrasound is used for recording abdominal aorta, the probe is perpendicular to the middle of the abdomen, M-mode ultrasound is used for recording conduction conditions of a proximal end and a distal end, the length of an abdominal aorta blood vessel and the delay time of a tip are measured to calculate the pulse propagation speed of the mouse, and meanwhile, the blood vessel thickness of an ultrasound result is analyzed.

2.5 echocardiography for cardiac function

After the mouse is anesthetized by isoflurane, an MS400C probe is adopted to collect the long-axis section beside the sternum of the mouse, B-type sampling is carried out, the image is stored, the image is amplified on the long-axis section, M-type sampling is carried out, and the image is stored. Each mouse is continuously scanned for 3 cardiac cycles, and the heart function index is calculated according to the measurement result.

2.6 HE staining

The abdominal aorta and heart tissues of the mouse are taken and placed in 4% paraformaldehyde for fixation for 24 hours, then the gradient ethanol is adopted for dehydration treatment, the tissue blocks are placed in xylene for transparency, and paraffin embedding is carried out on the tissues after wax dipping. The tissue was cut into 4 μm slices, placed in an oven at 37 ℃ overnight, rehydrated with xylene I, II clear and graded ethanol, and the sections were stained in hematoxylin solution for 1min and eosin solution for 10s, respectively. After air drying, the samples were mounted on neutral gum and the pathological morphological changes of the tissues were observed by microscopy.

2.8 statistical analysis

Firstly, analyzing whether the data accord with normal distribution or not by adopting a Shapiro-Wilk test method, and carrying out the homogeneity test of the variance on the data of the normal distribution. For data that fit normal distribution and are homogeneous in variance, ANOVA multiple comparisons were used. For data with non-normal distributions or non-uniform variances, the Kruskal-Wallis test was used. P <0.05 was considered statistically different. All data analyses were performed in SPSS 26.0.

3 results of the experiment

3.1 Effect of Ethoxysanguinarine on blood pressure in AngII-stimulated mice

In the research, a C57BL/6 hypertension mouse model is constructed by implanting an osmotic pump subcutaneously, and the intervention is given by ethoxy sanguinarine. Blood pressure monitoring revealed that mice in the AngII group had significantly elevated systolic (SBP; FIG. 1A), diastolic (DBP; FIG. 1B) and mean arterial (MAP; FIG. 1C) pressures compared to the Control group, and significantly decreased in all three after ethoxysanguinarine intervention (FIG. 1A-FIG. 1C;. P <0.05 vs. Control group, # P <0.05 vs. AngII group). Meanwhile, body weight measurements found that ethoxysanguinarine had no significant effect on mice (fig. 1D).

3.2 Effect of Ethoxysanguinarine on the speed of propagation and thickness of pulse waves in the Abdominal aorta of AngII-stimulated mice

After the mice are induced by AngII and intervened by ethoxy sanguinarine for 4 weeks, the pulse wave propagation speed and the blood vessel thickness of each group of mice are detected and analyzed by adopting the ultrasonic of the mice. As shown in FIGS. 2A-2B, the pulse wave propagation speed of abdominal aorta of mice in AngII group is significantly higher than that of Control group, and the pulse wave propagation speed of mice in AngII + Eth group is significantly reduced (P <0.05, compared with Control; P <0.05, compared with AngII group), and is similar to that of Control group; the abdominal aorta thickness was significantly higher in the ANGII group than in the Control group, and significantly decreased in the ANGII + Eth group (. about.P <0.05, vs. Control;. about.P <0.05, vs. ANGII) and similar to the Control group (FIG. 2C).

3.3 Effect of Ethoxysanguinarine on AngII stimulation of pathological morphology of mouse aorta

The study further used hematoxylin-eosin (HE) staining to observe the changes of ethoxysanguinarine to the morphology of mouse abdominal aorta. As shown in fig. 3, the abdominal aorta of the AngII group mice had some thickening compared to Control, while the ethoxysanguinarine intervention alleviated the thickening of vessel thickness induced by AngII.

3.4 Effect of Ethoxysanguinarine on AngII stimulation of cardiac function in mice

The study assessed cardiac functional changes by small animal ultrasound. The cardiac function index Ejection Fraction (EF) (FIGS. 4A and 4B) and left ventricular shortening Fraction (FS) (FIGS. 4A and 4C) of mice in the AngII group were significantly decreased compared to the Control group, while the dry prognosis of ethoxysanguinarine, both significantly increased (. P <0.05, compare to Control;. P <0.05, compare to AngII group), was close to the Control group.

3.5 Effect of Ethoxysanguinarine on AngII stimulation of the pathologic morphology of the mouse Heart

This study observed changes in the pathomorphic structure of cardiac tissue by HE staining. The results are shown in fig. 5, and compared with the Control group, the angium cardiomyocytes in the AngII group showed hypertrophic expression, while the ethoxysanguinarine stem prognosis, the cell hypertrophy was significantly relieved.

And (4) conclusion:

the ethoxy sanguinarine can remarkably reduce the blood pressure, the blood vessel function, the thickness and the pathological changes of an AngII model mouse, and simultaneously has the function of improving the reduction of the heart function and the pathological changes caused by hypertension.