Application of radix linderae-sourced sesquiterpenoids derivative in medicine preparation

Abstract

The invention discloses application of a radix linderae-derived sesquiterpenoid derivative isolindelactone in preparation of a medicine, the medicine is used for treating or preventing atherosclerosis diseases, and particularly, the medicine inhibits release of inflammatory factors and activation of NLRP3 inflammasomes, so that lipid plaque deposition is reduced, and the atherosclerosis pathological process is relieved.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology. Specifically, the present invention relates to the use of the sesquiterpenoid derivative isolinderalactone (I) derived from Lindera aggregata in the preparation of a medicament for treating atherosclerosis. Additionally, the present invention also relates to the use of isolinderalactone in the preparation of an NLRP3 inhibitor with anti-inflammatory activity.

[0002] Background Art

[0003] Atherosclerosis (AS), as a highly threatening cardiovascular disease globally, imposes a heavy burden on human health. In recent years, its incidence has also shown an increasing trend year by year. The pathogenesis of AS is a complex multi-factorial process, involving aspects such as dyslipidemia, hypertension, smoking, diabetes, obesity, genetic factors, and endothelial dysfunction. AS is typically characterized by lipid deposition under vascular endothelial cells to form lipid streaks, which then develop into fibrous plaques with a fibrous cap and a lipid core, accompanied by an inflammatory response throughout, leading to thickening and remodeling of the vascular wall, weakened vascular elasticity, narrowed lumen, and some unstable plaques are prone to rupture and trigger thrombosis, which may cause vascular occlusion and serious cardiovascular events.

[0004] The sesquiterpenoid derivative isolinderalactone (ISO) derived from Lindera aggregata is a sesquiterpenoid lactone compound extracted and isolated from plants such as Lindera aggregata, and has a variety of biological activities, showing certain effects in anti-inflammatory, antibacterial, antitumor, etc. Research on the mechanism of action of isolinderalactone in relieving acute inflammatory diseases found that isolinderalactone mainly treats diseases by inhibiting inflammatory signaling pathways. However, the role of isolinderalactone in atherosclerosis has not been reported. Summary of the Invention

[0005] The technical problem to be solved by the present invention is to provide a new use of the sesquiterpenoid derivative isolinderalactone derived from Lindera aggregata in the treatment or prevention of atherosclerosis.

[0006] The present invention provides an application of a sesquiterpenoid derivative derived from Lindera aggregata in the preparation of a medicament, and the medicament is used for treating or preventing atherosclerosis and its complications.

[0007] In the present invention, we screened the activities of various sesquiterpenoid derivatives in Lindera aggregata, including isolinderalactone, linderalactone, linderane, linderene, etc. The results showed that isolinderalactone had the most significant effect on atherosclerosis and its complications.

[0008] Preferably, we explored the therapeutic effect of isolinderalactone on atherosclerosis. We used high-fat feeding of APOE - / -An atherosclerotic model was established in mice, and isolinderalactone (10 mg / kg / every other day) was intragastrically administered to the mice for 8 weeks. The results showed that isolinderalactone significantly alleviated the deposition of atherosclerotic lipid plaques (see Example 1 for details). At the same time, oral administration of isolinderalactone did not affect the body weight and lipid levels of atherosclerotic mice, indicating that its effect was not achieved by affecting blood lipid levels (see Example 3 for details).

[0009] As a further preference, the lindera - derived sesquiterpenoid derivative isolinderalactone is used to reduce collagen deposition in the atherosclerotic model (see Example 2 for details).

[0010] As a preference, the lindera - derived sesquiterpenoid derivative isolinderalactone plays a role in treating or preventing atherosclerosis by inhibiting the release of inflammatory factors and the activation of NLRP3 inflammasome (see Examples 4 and 5 for details).

[0011] In specific applications, the drug is a preparation formulated from the lindera - derived sesquiterpenoid derivative isolinderalactone and pharmaceutically acceptable adjuvants. Among them, powders, tablets, pills, capsules and oral liquids for oral administration are preferred. The pharmaceutical composition of the present invention can be various preparations formulated by combining isolinderalactone and pharmaceutically acceptable adjuvants, preferably solid preparations and liquid preparations. The preparations of the present invention can be in unit - dose forms, such as tablets, pills, capsules (including sustained - release or delayed - release forms), powders, suspensions, granules, tinctures, syrups, emulsions, suspensions, injections, etc., as well as various sustained - release dosage forms, so as to be suitable for various administration routes, such as oral, parenteral injection, mucosal, intramuscular, intravenous, subcutaneous, intraocular, intradermal or transdermal administration forms (among which powders, tablets, pills, capsules (including sustained - release or delayed - release forms) and oral liquids for oral administration are preferred).

[0012] The beneficial effects achieved by the present invention are as follows: It is found that the lindera - derived sesquiterpenoid derivative isolinderalactone can effectively treat atherosclerosis.

[0013] For the convenience of understanding, the present invention will be described in detail below through specific drawings and examples. It should be particularly noted that the specific examples and drawings are only for illustration and do not constitute a limitation on the scope of the present invention. Obviously, those of ordinary skill in the art can make various modifications and changes to the present invention within the scope of the present invention according to the description herein, and these modifications and changes are also included in the scope of the present invention. In addition, the present invention cites published literatures, and the full text of these literatures is also incorporated into the present invention for reference as if their full text has been repeated in the specification of the present invention. Brief Description of the Drawings

[0014] Figure 1 : Inhibitory effect of isolinderalactone on lipid plaque formation in the aorta of atherosclerotic mice.

[0015] Figure 2 : Mitigative effect of isolinderalactone on collagen deposition in atherosclerotic mice.

[0016] Figure 3 : Isolinderalactone does not affect the lipid metabolism level of atherosclerotic mice.

[0017] Figure 4 : Inhibitory effect of isolinderalactone on the inflammatory response of atherosclerotic mice.

[0018] Figure 5 : Inhibitory effect of isolinderalactone on the activation of NLRP3 inflammasome in atherosclerotic mice. Detailed implementation mode

[0019] The present invention is further illustrated in the following embodiments. These embodiments are for illustrative purposes only and are not used to limit the scope of the present invention.

[0020] Example 1 Inhibitory effect of isolinderalactone on lipid plaque formation in the aorta of atherosclerotic mice.

[0021] Male APOE - / - mice were purchased from Jiangsu GICC Biotech Co., Ltd. The mice were fed with standard rodent food and water and housed in an animal room with a constant 12 - 12h day - night rhythm. The animals spent at least one week for environmental adaptation growth before the experiment. The protocols involving animal use were approved by the Animal Policy and Welfare Committee of Hangzhou Normal University (Approval document: HSD - 20250407 - 01).

[0022] Male APOE - / - mice at 8 - 10 weeks old were randomly divided into 3 groups, with 12 mice in each group, namely: low - fat control group (LFD group): APOE - / - mice were fed with low - fat diet for 16 weeks and gavaged with 1% CMC - Na solvent as a control; HFD model group (HFD group): APOE - / - mice were fed with high - fat diet for 16 weeks and gavaged with 1% CMC - Na solvent as a control; ISO treatment group (HFD + ISO group, 10mg / kg): APOE - / -At 16 weeks of age, mice were administered the drug starting from week 8. The drug ISO was dissolved in 1% CMC-Na and administered by gavage every other day. During this period, the body weights of mice in each group were measured and recorded weekly. After 8 weeks of drug administration, the mice were sacrificed, and mouse serum was collected. After the whole aorta of the mouse was isolated and fixed in 4% paraformaldehyde, the adipose tissue and connective tissue attached to the adventitia of the blood vessel were carefully dissected under a stereomicroscope. The whole aorta was dissected from the aortic arch to the iliac artery and stained with Oil Red O. The heart and proximal aortic root were collected and embedded in OCT and frozen at -20°C. Frozen sections of the aortic root, 5 μm thick, were obtained from each mouse for Oil Red O staining. The experimental data are shown in Figure 1 . It was found that isolinderal significantly alleviated the formation of lipid plaques in the aorta of atherosclerotic mice.

[0023] Among them, Figure 1 Figure A is a representative diagram of Oil Red O staining of the whole aorta. The results showed that compared with the LFD group, the lipid plaque deposition in the HFD group was significantly increased, while after treatment with isolinderal, the lipid deposition was significantly reduced. The quantitative statistical results of the positive plaque lesion area are shown in Figure 1 Figure B. In addition, Figure 1 The results of Oil Red O staining of the aortic root in Figure C showed that isolinderal alleviated lipid formation in the atherosclerotic mouse model of the HFD group after treatment. This indicates that isolinderal alleviated the formation of lipid plaques and pathological lesions in the aorta of atherosclerotic mice.

[0024] Example 2: The alleviating effect of isolinderal on collagen deposition in atherosclerotic mice.

[0025] Grouping and experiments were carried out according to Example 1. At 16 weeks, the mice were sacrificed to collect whole blood, and the serum was collected by centrifugation. The heart and proximal aortic root were collected and embedded in OCT and frozen at -20°C. Frozen sections of the aortic root, 5 μm thick, were obtained from each mouse for Masson and Sirius red staining. The results of Masson and Sirius red staining of the proximal aortic root showed ( Figure 2 Figure A - D) that isolinderal had a certain alleviating effect on collagen deposition lesions during atherosclerosis. This indicates the alleviating effect of isolinderal on collagen deposition in atherosclerotic mice.

[0026] Example 3: Isolinderal does not affect the lipid metabolism level of atherosclerotic mice.

[0027] Grouping and experiments were carried out according to Example 1. The body weights were measured and recorded weekly. At 16 weeks, the mice were sacrificed to collect whole blood, and the serum was collected by centrifugation. The contents of triglyceride (TG), total cholesterol (T-CHO), and low-density lipoprotein cholesterol (LDL-C) in the serum were detected using a lipid metabolism kit. The results showed that compared with the LFD group, the body weight of the HFD group was significantly increased (Figure 3 A), and the lipid metabolism level also increased significantly ( Figure 3 B-D). Compared with the HFD group, there was no difference in body weight change in the isolinderalactone treatment group, and there was also no difference in the contents of triglyceride (TG), total cholesterol (T-CHO) and low-density lipoprotein cholesterol (LDL-C) ( Figure 3 A-D). This indicates that isolinderalactone does not affect the lipid metabolism level of atherosclerotic mice.

[0028] Example 4 Inhibitory effect of isolinderalactone on the inflammatory response of atherosclerotic mice. Grouping and conducting experiments according to Example 1, sacrificing the mice at the 16th week to collect whole blood, centrifuging to separate and collect serum, freezing the aorta at -80 °C, taking part of the tissue to extract total mRNA using Trizol reagent, and detecting the gene level changes by RT-qPCR experiment. Detecting the levels of inflammatory factors in the serum using an ELISA kit, and the results showed that compared with the LFD group, the levels of inflammatory factors such as IL-1β, TNF-α, and IL-6 in the HFD group increased significantly, while after treatment with isolinderalactone, the levels of inflammatory factors such as IL-1β, TNF-α, and IL-6 were significantly inhibited ( Figure 4 A-C). Subsequently, detecting the mRNA levels of related inflammatory factors in the tissue, such as Il1b ( Figure 4 D), Tnf ( Figure 4 E) and Il6 levels ( Figure 4 F), compared with the HFD group, the isolinderalactone treatment group significantly inhibited the expression of these inflammatory genes. This indicates the inhibitory effect of isolinderalactone on the inflammatory response of atherosclerotic mice.

[0029] Example 5 Inhibitory effect of isolinderalactone on the activation of NLRP3 inflammasome in atherosclerotic mice.

[0030] Grouping and conducting experiments according to Example 1, after 16 weeks of modeling, sacrificing the mice in each group, taking the aorta tissue and freezing it at -80 °C. After homogenizing the aorta tissue, extracting total protein using RIPA reagent (containing 1% phosphatase inhibitor and 1% protease inhibitor), and analyzing the protein content changes by Western blot. Additionally, taking part of the tissue to extract total mRNA using Trizol reagent, and detecting the gene level changes by RT-qPCR experiment. As Figure 5As shown in Figure 5 B, compared with the LFD group, the caspase-1 activity in the HFD group was significantly enhanced, while the caspase-1 activity was significantly inhibited after treatment with isolinderalactone. Subsequently, the content of IL-1β in tissue samples was detected by ELISA Figure 5 C. Compared with the LFD group, the level of IL-1β in the HFD group was significantly enhanced, while the level of IL-1β was significantly inhibited after treatment with isolinderalactone. When detecting the tissue mRNA levels, such as Nlrp3 Figure 5 D and Gsdmd levels Figure 5 E, compared with the HFD group, the isolinderalactone treatment group significantly inhibited the expression of these two genes. This indicates the inhibitory effect of isolinderalactone on the activation of NLRP3 inflammasome in atherosclerotic mice.

Claims

1. Use of a sesquiterpene derivative derived from Lindera aggregata (Sims) Kosterm. in the preparation of a drug, characterized in that, The sesquiterpene derivative is isolinderalactone; The active ingredient in the drug includes isolinderalactone; The drug is used for treating or preventing atherosclerosis or diseases related to atherosclerosis.

2. Use of the sesquiterpene derivatives derived from Lindera aggregata (Sims) Kosterm. according to claim 1 in the preparation of a medicament, characterized in that, The diseases related to atherosclerosis are lesions such as lipid plaque deposition in the aorta.

3. Use of the sesquiterpenoid derivatives derived from Lindera aggregata (Sims) Kosterm. according to claim 2 in the preparation of a medicament, characterized in that, The isolinderalactone is used to inhibit collagen deposition in the heart and proximal aortic root, and has the effect of alleviating pathological symptoms.

4. Use of the sesquiterpenoid derivatives derived from Lindera aggregata (Sims) Kosterm. according to any one of claims 1 to 3 in the preparation of a medicament, characterized in that, The atherosclerosis is atherosclerosis caused by high fat.

5. Use of the sesquiterpenoid derivatives derived from Lindera aggregata (Sims) Kosterm. according to any one of claims 1 to 3 in the preparation of a medicament, characterized in that, The drug is a preparation formulated from isolinderalactone and pharmaceutically acceptable excipients.

6. Use of the sesquiterpenoid derivatives derived from Lindera aggregata (Sims) Kosterm. according to claim 5 in the preparation of a medicament, characterized in that, The drug includes powders, tablets, pills, capsules and oral liquids for oral administration.

7. Use of the sesquiterpenoid derivatives derived from Lindera aggregata (Sims) Kosterm. according to any one of claims 1 to 3 in the preparation of a medicament, characterized in that, The isolinderalactone does not affect the lipid metabolism level during the process of atherosclerosis.

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