Use of asd in the preparation of medicaments for the relief / treatment of hypertension and complications

Abstract

The application discloses a new use of alectra saponin D, that is, application in preparation of a medicine for relieving and treating hypertension and complications, can significantly reduce blood pressure, reduce hypertension vascular remodeling and target organ damage, and provides a target for developing a new medicine for relieving and / or treating hypertension and complications thereof. The molecular formula of the alectra saponin D is as follows:

Description

Technical Field

[0001] This invention relates to the technical field of medicine, and more particularly to the application of ASD in the preparation of drugs for relieving / treating hypertension and its complications. Background Technology

[0002] Hypertension, a prevalent and serious chronic disease, is experiencing a rising prevalence and mortality rate, drawing significant attention from the medical community. Currently, the five major classes of first-line drugs widely used domestically and internationally for treating hypertension include ACE inhibitors, ARBs, calcium channel blockers, beta-blockers, and diuretics. However, the treatment of hypertension faces numerous challenges, such as refractory hypertension and drug resistance. Therefore, it remains necessary to identify new drug targets to more safely and effectively prevent and treat hypertension, further refine clinical hypertension treatment protocols, and improve hypertension control rates.

[0003] Figure 1 This is a schematic diagram of the molecular formula of Akebia saponin D (ASD). ASD is a triterpenoid saponin compound and the main active ingredient of the traditional Chinese medicine Dipsacus asper. Dipsacus asper is the dried root of the plant Dipsacus asper, belonging to the Dipsacaceae family, and is a traditional Chinese medicine. The *Shennong Bencao Jing* (Shennong's Classic of Materia Medica) records that it has the effects of tonifying the liver and kidneys, strengthening tendons and bones, healing fractures, and regulating meridians. Current research on Akebia saponin D is mostly based on its estrogen-like antioxidant effects in the prevention and treatment of Alzheimer's disease; some studies have also shown its clinical significance in fatty liver disease. It is used to prevent and treat menopausal syndrome and osteoporosis in women.

[0004] Currently, there are no reports of the use of Dipsacus asperoides and Akebia trifoliata saponin D in the preparation of drugs to relieve and / or treat hypertension and its complications. Summary of the Invention

[0005] To overcome the shortcomings of the prior art, the technical problem to be solved by the present invention is to provide an application of ASD in the preparation of drugs for relieving / treating hypertension and its complications, which can significantly reduce blood pressure, reduce vascular remodeling in hypertension and target organ damage.

[0006] The technical solution of this invention is: the application of this ASD in the preparation of drugs for relieving / treating hypertension and its complications. The molecular formula of ASD is as follows:

[0007]

[0008] This invention provides a new use for Akebia saponin D, namely, its application in the preparation of drugs to relieve / treat hypertension and its complications. It can significantly reduce blood pressure, alleviate vascular remodeling and target organ damage in hypertension, and provide a target for the development of new drugs to relieve and / or treat hypertension and its complications.

[0009] Preferably, the hypertension and its complications include: primary hypertension, secondary hypertension, hypertensive heart disease, hypertensive heart failure, hypertensive encephalopathy, hypertensive retinopathy, and hypertensive nephropathy.

[0010] Preferably, the active ingredient of the drug is akebia saponin D, 25-100 mg / kg, once daily for 28 consecutive days.

[0011] Preferably, the ASD reduces systolic and diastolic blood pressure.

[0012] Preferably, the ASD reduces the thickness of the aortic media and the degree of aortic fibrosis.

[0013] Preferably, the ASD reduces the degree of myocardial cell damage and fibrosis.

[0014] Preferably, the ASD reduces the degree of damage to the glomeruli and renal tubules.

[0015] Preferably, the drug further comprises pharmaceutically acceptable components and / or carriers for ASD.

[0016] Preferably, the pharmaceutically acceptable carrier for ASD includes any one or more combinations of diluents, excipients, disintegrants, fillers, binders, lubricants, flavoring agents, surfactants, and stabilizers.

[0017] Preferably, the dosage form of the drug is any one of injection, syringe, tablet, granule, pill, capsule, microcapsule / microsphere formulation, spray, or targeted formulation. Attached Figure Description

[0018] Figure 1 A schematic diagram of the molecular formula of Akebia saponin D is shown.

[0019] Figure 2 This graph shows the results of akebia saponin D reducing systolic blood pressure, mean blood pressure, and diastolic blood pressure in a rat model of hypertension. * and ** indicate p < 0.05, and *** and **** indicate p < 0.01, indicating statistically significant differences.

[0020] Figure 3 The results of hematoxylin-eosin (HE) staining of the rat thoracic aorta are shown.

[0021] Figure 4 The staining results of elastic fibers and collagen fibers in the rat thoracic aorta are shown.

[0022] Figure 5 The results of hematoxylin-eosin (HE) staining of rat heart are shown.

[0023] Figure 6The results of hematoxylin-eosin (HE) staining of rat kidneys are shown. Detailed Implementation

[0024] This invention proposes a novel role and application of akebia saponin D in alleviating / treating hypertension and its complications. Akebia saponin D can significantly reduce blood pressure and alleviate vascular remodeling and target organ damage in hypertensive patients. Using SD rats as experimental subjects, this invention administered akebia saponin D (25 mg / kg / d, 50 mg / kg / d, and 100 mg / kg / d, with physiological saline as the solvent, via gavage) and established a rat hypertension model by adding N-nitro-L-arginine methyl ester (L-NAME) (40 mg / kg / d) to the drinking water. The protective effects of akebia saponin D on blood pressure, vascular remodeling, and target organ damage in the rat hypertension model were observed.

[0025] The embodiments of the present invention will be described in detail below.

[0026] Laboratory animals and their care: Male SD rats, 8 weeks old, were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. and housed in the Animal Center (SPF grade) of the Clinical Medical Research Institute of China-Japan Friendship Hospital. The housing conditions were: temperature between 22-24℃, humidity between 40-70%, light and dark alternation lighting time of 12h, and free access to water and food.

[0027] Example 1: Akebia saponin D reduced systolic blood pressure, diastolic blood pressure, and mean blood pressure in rats.

[0028] 1. Rat model of hypertension

[0029] (1) Rats were acclimatized to the environment for one week in an SPF-grade standardized animal center.

[0030] (2) Add 40 mg / kg / d of N-nitro-L-arginine methyl ester (L-NAME) solution (solvent is physiological saline) to 200 mL of drinking water, feed rats, change the water daily for 28 days, and thus construct a rat hypertension model.

[0031] 2. Administration method of Akebia saponin D

[0032] Starting on day one of administration of N-nitro-L-arginine methyl ester (L-NAME), Akebia saponin D was administered by gavage at doses of 25 mg / kg, 50 mg / kg, and 100 mg / kg, once daily for 28 consecutive days, in physiological saline. The control group received an equal amount of placebo (physiological saline) at the same time.

[0033] 3. Measurement of blood pressure in the tail artery of rats

[0034] Blood pressure (systolic, diastolic, and mean blood pressure) in the tail arteries of rats was measured using a rat intelligent non-invasive blood pressure monitor (BP-2010A, Beijing Ruanlong Biotechnology Co., Ltd.) before the hypertension model and at weeks 1, 2, 3, and 4 of the experiment. Each measurement was performed for 15 cycles (the first 5 cycles were adaptive measurements and their data were not included), and the average value was taken as the rat's blood pressure (systolic, diastolic, and mean blood pressure). In addition, rats underwent blood pressure measurement adaptation training 3-5 times before the experiment.

[0035] The results of systolic blood pressure, diastolic blood pressure and mean blood pressure in rats are as follows: Figure 2 As shown, Akebia saponin D can reduce systolic and diastolic blood pressure in a rat hypertension model in a dose-dependent manner.

[0036] Example 2: Akebia saponin D reduces vascular remodeling caused by hypertension.

[0037] 1. Measurement of thoracic aortic media thickness

[0038] (1) Preparation of paraffin specimens and sections

[0039] 1) Four weeks after the hypertension model was established, the rats were euthanized, and the intact thoracic aorta was dissected and fixed in 4% paraformaldehyde solution. After 72 hours, the aortic tissue was removed from the 4% paraformaldehyde solution, trimmed in a fume hood, and the trimmed aortic tissue and corresponding labels were placed in a dehydration box.

[0040] 2) Dehydration and wax impregnation: Place the dehydration box into a dehydrator for dehydration and wax impregnation, then hydrate it in a descending gradient of ethanol. Xylene I 20 min; Xylene II 20 min; 100% ethanol I 2 min; 100% ethanol II 2 min; 95% ethanol 2 min; 90% ethanol 2 min; 80% ethanol 2 min; 75% ethanol 2 min, then rinse three times with distilled water for 3 min each time.

[0041] 3) Tissue embedding: First, inject molten paraffin into the embedding frame. While the paraffin is still wet, remove the aortic tissue from the dehydration box, place it in the embedding frame with the transverse section of the vascular ring facing down, and attach the corresponding label. Place it horizontally on a -20℃ freezing stage to cool. After the paraffin has solidified, remove the paraffin block from the embedding frame.

[0042] 4) Tissue sectioning: Trim and fix the paraffin block onto the paraffin microtome, sectioning to a thickness of 4-5 μm. Rotate the microtome handle continuously to section, allowing the sections to float in warm water and flatten them. Use a glass slide to retrieve the sections, centering them on the slide, and bake them in a 60℃ oven for later use.

[0043] (2) Hematoxylin-eosin (HE) staining of the thoracic aorta

[0044] Using the paraffin sections described above, perform the following operations in sequence:

[0045] 1) Reagent (A) Hematoxylin staining solution: Immerse for 6 minutes, then wash away excess dye with distilled water.

[0046] 2) Reagent (B) Differentiation solution: Soak for 30 seconds, rinse off the surface color with tap water, soak twice for 3 minutes each time, and adjust the water flow to the minimum flow rate for continuous rinsing.

[0047] 3) Reagent (C) Eosin staining solution: Immerse for 1 min 30s-1 min 45s, then pour off the excess staining solution without rinsing.

[0048] 4) Dehydration and clearing: The sections were subjected to 75% ethanol for 2 seconds; 80% ethanol for 2 seconds; 90% ethanol for 2 seconds; 95% ethanol for 2 seconds; 100% ethanol I for 2 seconds; 100% ethanol II for 1 minute; xylene I for 3 minutes; xylene II for 3 minutes.

[0049] 5) Seal with neutral resin. Place in a fume hood overnight.

[0050] 6) After the slide has dried, take a picture and record it using an optical microscope.

[0051] The results of aortic media thickness are as follows: Figure 3 As shown, in a rat model of hypertension, Akebia saponin D significantly reduced the thickness of the aortic media, indicating that Akebia saponin D significantly reduced vascular remodeling in hypertension.

[0052] 2. Thoracic aortic fibrosis measurement

[0053] (1) Staining of elastic fibers and collagen fibers in the thoracic aorta

[0054] 1) Dewax the above paraffin sections to 75% ethanol.

[0055] 2) Wash the slices in 70% ethanol for 2 minutes.

[0056] 3) Immerse the sections in Victoria Blue solution for staining, and incubate overnight at 37°C.

[0057] 4) Directly immerse in 95% ethanol and control the color separation under a microscope for several seconds.

[0058] 5) Wash away excess dye with anhydrous ethanol.

[0059] 6) Rinse with running water for 2 minutes.

[0060] 7) Add Sirius red saturated bitter acid solution for 1 hour.

[0061] 8) Rinse with tap water for 10 minutes.

[0062] 9) Stepwise ethanol dehydration: 50% → 60% → 70% → 80% → 90% → 100%.

[0063] 10) Xylene is transparent.

[0064] 11) Seal with neutral resin.

[0065] 12) After staining, the sections were photographed and recorded using an optical microscope.

[0066] Results of aortic fibrosis degree as follows Figure 4 As shown, in a rat model of hypertension, Akebia saponin D significantly reduced the degree of aortic fibrosis, indicating that Akebia saponin D significantly reduced vascular remodeling in hypertension.

[0067] Example 3: Akebia saponin D reduces cardiac damage caused by hypertension.

[0068] 1. Cardiac Injury Assessment

[0069] (1) Hematoxylin-eosin (HE) staining of the heart

[0070] Four weeks after the hypertension model was established, rats were euthanized, and their hearts were collected and fixed in 4% paraformaldehyde solution. After 72 hours, the hearts were removed from the 4% paraformaldehyde solution, and the tissue was trimmed in a fume hood. Subsequent procedures were the same as described above for preparing paraffin-embedded specimens and sections. Staining procedures and observations were also performed as described above.

[0071] Results of the degree of heart damage as follows Figure 5 As shown, in a rat model of hypertension, Akebia saponin D significantly reduced myocardial cell damage and fibrosis, indicating that Akebia saponin D significantly reduced cardiac damage caused by hypertension.

[0072] Example 4: Akebia saponin D reduces kidney damage caused by hypertension.

[0073] 1. Kidney Injury Assessment

[0074] (1) Kidney hematoxylin-eosin (HE) staining

[0075] Four weeks after the hypertension model was established, rats were euthanized, and their kidneys were collected and fixed in 4% paraformaldehyde solution. After 72 hours, the kidneys were removed from the 4% paraformaldehyde solution, and the tissue was trimmed in a fume hood. Subsequent procedures were the same as described above for preparing paraffin-embedded specimens and sections. Staining procedures and observations were also performed as described above.

[0076] Results of kidney damage degree as follows Figure 6 As shown, in a rat model of hypertension, Akebia saponin D significantly reduced the degree of glomerular and tubular damage, indicating that Akebia saponin D significantly reduced kidney damage caused by hypertension.

[0077] The above results indicate that in a rat model of hypertension, Akebia saponin D significantly reduced aortic media thickness, aortic fibrosis, and cardiac and renal damage, suggesting that Akebia saponin D significantly reduced vascular remodeling and target organ damage caused by hypertension.

[0078] The above results indicate that, in a rat model of hypertension, akebia saponin D significantly reduces blood pressure and alleviates hypertensive vascular remodeling and target organ damage. Akebia saponin D can be used as an active ingredient in the preparation of drugs to alleviate and / or treat hypertension and its related complications.

[0079] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any simple modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of the present invention shall still fall within the protection scope of the present invention.

Claims

1. Application of ASD in the preparation of drugs to relieve / treat hypertension and its complications. The molecular formula of ASD is as follows: ； The hypertension and its complications include: Secondary hypertension, hypertensive nephropathy.

2. The application of ASD according to claim 1 in the preparation of drugs for relieving / treating hypertension and its complications, characterized in that: The active ingredient of the drug is ASD, 25-100 mg / kg, once daily for 28 consecutive days.

3. The application of ASD according to claim 2 in the preparation of drugs for relieving / treating hypertension and its complications, characterized in that: The ASD reduces systolic and diastolic blood pressure.

4. The application of ASD according to claim 2 in the preparation of drugs for relieving / treating hypertension and its complications, characterized in that: The ASD reduces the thickness of the aortic media and the degree of aortic fibrosis.

5. The application of ASD according to claim 2 in the preparation of drugs for relieving / treating hypertension and its complications, characterized in that: The ASD reduces the degree of damage to the glomeruli and renal tubules.

6. The application of ASD according to claim 2 in the preparation of drugs for relieving / treating hypertension and its complications, characterized in that: The drug also contains pharmaceutically acceptable components and / or carriers for ASD.

7. The application of ASD according to claim 6 in the preparation of drugs for relieving / treating hypertension and its complications, characterized in that: The pharmaceutically acceptable carriers for ASD include any one or more combinations of diluents, excipients, disintegrants, fillers, binders, lubricants, flavoring agents, surfactants, and stabilizers.

8. The application of ASD according to claim 7 in the preparation of drugs for relieving / treating hypertension and its complications, characterized in that: The dosage form of the drug is any one of the following: injection, tablet, granule, pill, capsule, microcapsule / microsphere formulation, spray, or targeted formulation.

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