Use of cinnamoylflavan alkaloids and phenylpropanoid-substituted ester catechins in the preparation of drugs for treating or preventing aging and lipid lowering

Abstract

This invention discloses the application of cinnamyl flavanone alkaloids and phenylpropanol-substituted ester-type catechins in the preparation of drugs for treating or preventing aging and lowering lipids. In vivo experiments have confirmed that the cinnamyl flavanone alkaloids and phenylpropanol-substituted ester-type catechins of this invention, without reproductive toxicity to a *C. elegans* model, can prolong the average lifespan of nematodes, improve their motility, enhance their stress resistance, reduce endogenous ROS levels and lipofuscin content, and decrease the body fat content of nematodes. Therefore, the cinnamyl flavanone alkaloids and phenylpropanol-substituted ester-type catechins of this invention can be used in the preparation of drugs for treating or preventing aging and lowering lipids.

Description

Technical Field

[0001] This invention relates to the field of pharmaceuticals, specifically to the use of cinnamyl flavanone alkaloids and phenylpropanol-substituted ester-type catechins in the preparation of drugs for treating or preventing aging and lowering lipids. Background Technology

[0002] Population aging and declining birth rates are inevitable consequences of human societal development at a certain stage. Currently, China's aging population is rapidly increasing, projected to reach 450 million elderly by 2050. With declining birth rates and the arrival of an aging population, human aging and age-related high-risk diseases have become widespread social concerns. Natural active ingredients in plants have become a hot topic in anti-aging research due to their high efficiency, safety, and low cost. Therefore, studying the anti-aging functions of tea and identifying compounds with anti-aging activity and their health mechanisms can enable it to play a greater role in protecting human health.

[0003] *Caenorhabditis elegans* (Caenorhabditis elegans for short) is an invertebrate worm about 1 mm in length, primarily fed on *Escherichia coli* OP50 in laboratory cultures. Their average lifespan is about two to three weeks. Through self-fertilization, a nematode can lay about 300 eggs in its lifetime. Eggs from hermaphroditic nematodes undergo four larval stages (L1-L4) after hatching. Under normal circumstances, the eggs develop into adults and begin oviposition within 48 hours of hatching, a process that takes about four days. Genetic screening yields a large number of homozygous mutants, and the nematode is easy to culture and avoids ethical issues. Most importantly, the high homology between nematodes and human genes, as well as the extensive metabolic similarities, make them ideal models for biological research. Currently, the demand for extended lifespan and improved quality of life among Chinese residents is constantly increasing. Based on our previous research, we have discovered that a catechin compound can extend the lifespan of model organisms and reduce the body fat content of nematodes. These catechin compounds have multiple functions, including extending human lifespan and reducing lipid levels.

[0004] Based on this, the present invention proposes the application of cinnamyl flavanone alkaloids and phenylpropanol-substituted ester-type catechins in the preparation of drugs for treating or preventing aging and lowering lipids, thereby prolonging human lifespan and reducing the body fat content. Summary of the Invention

[0005] The purpose of this invention is to address the problem of significant adverse reactions in existing drugs for treating or preventing aging and lowering lipids, and to provide the application of cinnamyl flavan alkaloids and phenylpropanol-substituted ester-type catechins in the preparation of drugs for treating or preventing aging and lowering lipids.

[0006] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is as follows:

[0007] This invention discloses a cinnamyl flavane alkaloid having the structural formulas shown in (I) and (II) below:

[0008]

[0009] The present invention also discloses the application of the above-mentioned cinnamyl flavan alkaloids in the preparation of drugs for treating or preventing aging.

[0010] The present invention also discloses the application of the above-mentioned cinnamyl flavan alkaloids in the preparation of lipid-lowering drugs.

[0011] Furthermore, the concentration of the cinnamyl flavan alkaloid in the preparation of the drug for treating or preventing aging is one of 50 μM, 100 μM, 200 μM, and 400 μM.

[0012] Furthermore, the concentration of the cinnamyl flavan alkaloid in the preparation of the lipid-lowering drug is one of 50 μM, 100 μM, 200 μM, and 400 μM.

[0013] Furthermore, the cinnamyl flavan alkaloid is one of CF1-4.

[0014] The present invention also discloses a phenylpropanoid-substituted ester-type catechin having the following structural formulas (III), (IV), (V), (VI) and (VII):

[0015]

[0016]

[0017] The present invention also discloses the use of the above-described ester-type catechin in the preparation of drugs for treating or preventing aging.

[0018] The present invention also discloses the application of the above-described ester-type catechin in the preparation of lipid-lowering drugs.

[0019] Furthermore, the concentration of the ester-type catechin in the preparation of drugs for treating or preventing aging is one of 50 μM, 100 μM, 200 μM, and 400 μM.

[0020] Furthermore, the concentration of the ester-type catechin in the preparation of the lipid-lowering drug is one of 50 μM, 100 μM, 200 μM, and 400 μM.

[0021] Furthermore, the phenylpropanoid-substituted ester-type catechin is one of Zijuanins A-D, PSEC650, and PSEC620.

[0022] The beneficial effects of this invention are as follows:

[0023] The cinnamylflavonoid alkaloids (CF1-4) and phenylpropanol-substituted ester-type catechins (Zijuanins A-D, PSEC650, PSEC620) of this invention can prolong the average lifespan of nematodes, improve their motility, enhance their stress resistance, reduce endogenous ROS levels and lipofuscin content, and decrease the body fat content of nematodes, without exhibiting reproductive toxicity in a *C. elegans* model. Therefore, the cinnamylflavonoid alkaloids (CF1, CF2, CF3, CF4) and phenylpropanol-substituted ester-type catechins (Zijuanins AD, PSEC650, PSEC620) of this invention can be used to prepare drugs for treating or preventing aging and lowering lipids. Attached Figure Description

[0024] Figure 1 Figure showing the effect of cinnamyl flavanine alkaloids on the lifespan of nematodes;

[0025] Figure 2 The effect of phenylpropanol-substituted ester-type catechins on the lifespan of nematodes.

[0026] Figure 3 Plot of relative lipid content in nematodes treated with 100 μM Zijuanin A and CF1. Detailed Implementation

[0027] The present invention will be further described below with reference to the accompanying drawings and specific embodiments.

[0028] In view of the problems existing in the prior art, the present invention aims to invent a catechin compound with the effects of prolonging life and lowering lipids, and its application.

[0029] The first objective of this invention is achieved by selecting the catechin compounds with life-extending and lipid-lowering effects from: cinnamyl flavan alkaloids (CF1-4) or ester-type catechins with phenylpropanoid substitution (Zijuanins A-D, PSEC 650, PSEC 620).

[0030] Example 1

[0031] Application of cinnamyl flavanine alkaloids in the preparation of drugs for treating or preventing aging and lowering lipids

[0032] This cinnamyl flavan alkaloid has the following structural formulas (I) and (II):

[0033]

[0034] The concentration of this cinnamyl flavan alkaloid in the preparation of drugs for treating or preventing aging is one of 50 μM, 100 μM, 200 μM, or 400 μM; and in the preparation of drugs for treating lipid-lowering diseases, the concentration is one of 50 μM, 100 μM, 200 μM, or 400 μM.

[0035] Example 2

[0036] Application of phenylpropanol-substituted ester-type catechins in the preparation of drugs for treating or preventing aging and lowering lipids

[0037] The phenylpropanol-substituted ester catechin has the following structural formulas as shown in (III), (IV), (V), (VI), and (VII):

[0038]

[0039] The phenylpropanol-substituted ester-type catechin is used in the preparation of drugs for treating or preventing aging at a concentration of 50 μM, 100 μM, 200 μM, or 400 μM; and in the preparation of drugs for treating lipid-lowering diseases at a concentration of 50 μM, 100 μM, 200 μM, or 400 μM.

[0040] Test case

[0041] 1 Experimental Methods

[0042] 1.1 Preparation of nematode culture medium:

[0043] Weigh out 3g of sodium chloride, 2.5g of peptone, and 17g of agar, add deionized water to a final volume of 1L, autoclave, cool to 60℃, and then pour into plates.

[0044] 1.2 Life test:

[0045] Synchronized L1-stage nematodes were inoculated onto NGM plates containing E. coil OP50 and cultured to the L4 stage. The nematodes were then transferred to media containing different concentrations (0, 50, 100, 200, 400, 1000 μM) of the compound. The nematodes were transferred to new treatment plates every other day. The number of surviving individuals was counted daily, and mortality was recorded until 100% mortality was achieved. Each group had three replicates, with 50 nematodes per replicate, and statistical analysis was performed using Graphad Prism 6. Metformin and resveratrol served as positive controls.

[0046] 1.3 Determination of relative lipid content:

[0047] After culturing, the nematodes were washed off the culture medium, collected, centrifuged, and the supernatant was removed. In the dark, 800 μL of 3 mg / mL Oil Red O staining solution was added to the centrifuge tube, and the tube was stored overnight at 20°C in the dark with shaking. The mixture was then centrifuged again, the supernatant was removed, and the tube was washed with phosphate buffer. This process was repeated several times. In the dark, 10 μL of the nematode suspension was transferred to a glass slide and covered with a coverslip. The lipid content of the nematodes was measured by fluorescence intensity, and the total fluorescence of the nematode sample was quantified using ImageJ image analysis software.

[0048] 2 Experimental Results

[0049] The effects of four compounds, cinnamyl flavanine alkaloids CF1-4 (50-400 μM), on the lifespan of nematodes were investigated. The results showed that the mean lifespans of the CF1 treatment groups were 27.8 days (50 μM), 28.4 days (100 μM), 33.4 days (200 μM), and 23.4 days (400 μM), respectively; the mean lifespans of the CF2 treatment groups were 23.3 ± 2.2 days (50 μM), 28.7 days (100 μM), 32.9 days (200 μM), and 27.7 days (400 μM), respectively; and the mean lifespans of the CF3 treatment groups were 24.8 days (50 μM), 26.5 days (100 μM), and 33.0 days (200 μM), respectively. The average lifespan of the nematodes was 25.7 days (400 μM) in the CF4 treatment group, 21.4 days (50 μM), 25.2 days (100 μM), 31.1 days (200 μM), and 24.2 days (400 μM) in the CF4 treatment group, respectively. All cinnamyl flavanone alkaloids at the four concentrations significantly prolonged the lifespan of nematodes, with the 200 μM concentration showing the best effect. Compared with the control, the average lifespan of nematodes in the treatment groups increased by 72.9% (CF1), 70.3% (CF2), 71.0% (CF3), and 61.1% (CF4), respectively.

[0050] Phenylpropionic acid esters are nucleophilic substitution products formed by catechins and active phenolic acids, and are widely found in tea and other plants. Due to their structural diversity, we selected six compounds (50, 100, and 200 μM) – Zijuanins AD, PSEC650, and PSEC620 – for testing. The results showed that the average lifetimes of the Zijuanin A treatment group were 26.0 days (50 μM), 29.8 days (100 μM), and 29.0 days (200 μM), respectively; the average lifetimes of the Zijuanin B treatment group were 21.3 days (50 μM), 29.7 days (100 μM), and 26.1 days (200 μM), respectively; and the average lifetimes of the Zijuanin C treatment group were 24.9 days (50 μM), 28.7 days (100 μM), and 27.8 days (200 μM), respectively. The average lifespans of the D treatment group were 24.4 days (50 μM), 27.7 days (100 μM), and 26.3 days (200 μM), respectively; the average lifespans of the PSEC650 treatment group were 23.4 days (50 μM), 29.3 days (100 μM), and 25.5 days (200 μM), respectively; and the average lifespans of the PSEC620 treatment group were 24.6 days (50 μM), 29.4 days (100 μM), and 24.4 days (200 μM), respectively. All compounds at all three concentrations significantly prolonged the lifespan of nematodes, with the 100 μM concentration showing the best effect. In particular, Zijuanin A increased the average lifespan of nematodes in the Zijuanin A treatment group by 54.2% compared to the control, demonstrating superior efficacy compared to the positive control drug metformin.

[0051] The higher the degree of aging, the easier it is for body fat to accumulate. Therefore, based on the above experimental results, we further evaluated the lipid-lowering activity of catechins on nematodes. The results are shown in Table 1: The body fat content of nematodes treated with catechins was significantly reduced. Compared with the control group, Zijuanin A and CF1 decreased by 15.4% and 23.7%, respectively.

[0052] Table 1. Effects of catechin compounds on the lifespan of nematodes.

[0053]

[0054]

[0055] Based on the above embodiments, it can be seen that the cinnamyl flavan alkaloids and phenylpropanol-substituted ester-type catechins in this invention have a certain effect on wild-type nematode models. In specific applications, the cinnamyl flavan alkaloids and phenylpropanol-substituted ester-type catechins can be used to prepare drugs for treating or preventing aging and lowering lipids.

[0056] The specification and drawings of this invention are intended to be illustrative rather than restrictive. Based on this invention, those skilled in the art can make substitutions and modifications to some of the technical features without creative effort, and all such modifications are within the scope of protection of this invention.

Claims

1. The application of a cinnamyl flavanone alkaloid in the preparation of lipid-lowering drugs, wherein: The cinnamyl flavan alkaloids described herein have the following structural formulas (I) and (II): Furthermore, the concentration of the cinnamyl flavan alkaloid is one of 50 μM, 100 μM, 200 μM, or 400 μM.

2. The application of an ester-type catechin in the preparation of lipid-lowering drugs, wherein: The phenylpropanol-substituted ester catechin has the following structural formulas as shown in (III), (IV), (V), (VI), and (VII): Furthermore, the concentration of this ester-type catechin is one of 50μM, 100μM, 200μM, or 400μM.

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