A composition for reducing blood lipid and cholesterol, its preparation method and uses

Abstract

The present invention relates to the field of pharmaceutical technology, and specifically relates to a composition for reducing blood lipid and cholesterol, its preparation method and uses. The composition comprises the following components in parts by weight: 200 - 1000 parts by weight of phytosterol or its analogues, 50 - 500 parts by weight of red yeast rice extract, and 10 - 500 parts by weight of lycopene. In the above composition, by compounding phytosterol or its analogues, red yeast rice extract and lycopene, it is found that phytosterol or its analogues, red yeast rice extract and lycopene have a synergistic effect in reducing blood lipid and cholesterol, and can be used for preparing products for reducing blood lipid and cholesterol, and have uses in preparing products for preventing, alleviating, adjuvantly treating or treating hyperlipidemia and / or hypercholesterolemia.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and particularly to a composition for reducing blood lipid and cholesterol, its preparation method and uses. Background Art

[0002] In recent years, the blood lipid levels of the population have gradually increased, and the prevalence of dyslipidemia has increased significantly. The overall prevalence of adult dyslipidemia is as high as 40.40%. At the same time, the prevalence of hypercholesterolemia in children and adolescents has also increased significantly, and the total prevalence of dyslipidemia is 23.5%. This indicates that the burden of adult dyslipidemia and related diseases will continue to increase in the future. Dyslipidemia is an important risk factor for cardiovascular diseases. Currently, cardiovascular diseases have gradually become the disease group with the largest number of deaths in the world population, especially in developing countries. Hyperlipidemia, as one of the "three highs", namely the increase in serum cholesterol and triglycerides, has a significant correlation with cardiovascular diseases such as hypertension and atherosclerosis. Therefore, reducing blood lipid and cholesterol has become an urgent problem for patients with the "three highs". Summary of the Invention

[0003] Therefore, the technical problem to be solved by the present invention is to provide a composition for reducing blood lipid and cholesterol, its preparation method and uses.

[0004] For this purpose, the present invention provides the following technical solutions:

[0005] A composition for reducing blood lipid and cholesterol, comprising the following components in parts by weight: 200 - 1000 parts by weight of phytosterol or its analog, 50 - 500 parts by weight of red yeast rice extract, and 10 - 500 parts by weight of lycopene.

[0006] Optionally, it comprises the following components in parts by weight: 400 - 800 parts by weight of phytosterol or its analog, 150 - 300 parts by weight of red yeast rice extract, and 50 - 200 parts by weight of lycopene.

[0007] Optionally, it comprises the following components in parts by weight: 754 parts by weight of phytosterol or its analog, 114.9 parts by weight of red yeast rice extract, and 99.5 parts by weight of lycopene; or

[0008] 682 parts by weight of phytosterol or its analog, 280 parts by weight of red yeast rice extract, and 99 parts by weight of lycopene.

[0009] A preparation method of the composition for reducing blood lipid and cholesterol as described above, comprising: weighing raw materials according to the formula and mixing them.

[0010] A preparation, using the composition for reducing blood lipid and cholesterol as the active ingredient.

[0011] Optionally, it further comprises excipients or carriers permitted by the preparation;

[0012] Optionally, the form of the preparation includes liquid preparation and solid preparation;

[0013] Optionally, the form of the preparation includes injection, tablet, capsule, powder, granule or ointment.

[0014] The lipid-lowering and cholesterol-lowering composition or the preparation has the following uses:

[0015] (1) Use in the preparation of lipid-lowering and / or cholesterol-lowering products;

[0016] (2) Use in the preparation of products for preventing, alleviating, adjuvantly treating or treating hyperlipidemia and / or hypercholesterolemia;

[0017] (3) Use in the preparation of products for reducing triglyceride;

[0018] (4) Use in the preparation of products for reducing the expression level of hmgcra gene;

[0019] (5) Use in the preparation of weight loss products.

[0020] Optionally, the product includes food, food additive or medicine.

[0021] Optionally, the food includes functional food, health care product or ordinary food.

[0022] The technical solution of the present invention has the following advantages:

[0023] 1. A lipid-lowering and cholesterol-lowering composition provided by the present invention includes the following components in parts by weight: 200-1000 parts by weight of phytosterol or its analog, 50-500 parts by weight of red yeast rice extract, and 10-500 parts by weight of lycopene; in the above composition, by compounding phytosterol or its analog, red yeast rice extract and lycopene, it is found that phytosterol or its analog, red yeast rice extract and lycopene have a synergistic effect on reducing blood lipid and cholesterol, and can be used for preparing lipid-lowering and cholesterol-lowering products, and have uses in the preparation of products for preventing, alleviating, adjuvantly treating or treating hyperlipidemia and / or hypercholesterolemia. Detailed implementation manners

[0024] The following embodiments are provided to better further understand the present invention, and are not limited to the best implementation manner, and do not constitute a limitation to the content and protection scope of the present invention. Any product identical or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with other prior art features falls within the protection scope of the present invention.

[0025] For those embodiments in which specific experimental procedures or conditions are not specified, the operations or conditions of the conventional experimental procedures described in the literature in this field can be followed. For reagents or instruments whose manufacturers are not specified, they are all conventional reagent products that can be obtained through commercial purchase.

[0026] The plant sterol or its analog is a commercially available product, and its product forms include but are not limited to microcapsule powder, paste or oil of the plant sterol or its analog, and the total sterol content is more than 50%. It can be purchased from any manufacturer. In the following embodiments, the plant sterol ester is selected as the plant sterol or its analog, and the plant sterol ester is purchased from BASF Corporation.

[0027] The lycopene is a commercially available product, and its product forms include but are not limited to lycopene microcapsule powder, lycopene paste, lycopene oil or lycopene crystalline powder, and the lycopene content is more than 5%. It can be purchased from any manufacturer. In the following embodiments, the lycopene oil is purchased from Chenguang Biotech Group Co., Ltd.

[0028] The red yeast rice extract is a commercially available product, and its product forms include but are not limited to red yeast rice powder, red yeast rice or red yeast rice extract solution, and the lovastatin content is ≥1%. It can be purchased from any manufacturer. In the following embodiments and experimental examples, the red yeast rice extract is the red yeast rice powder purchased from Hangzhou Shuangma Biotech Co., Ltd.

[0029] Embodiment

[0030] The formulations of the lipid-lowering and cholesterol-lowering compositions of Embodiments 1-7 are as shown in the following table.

[0031] Table 1. Formulations of the lipid-lowering and cholesterol-lowering compositions

[0032]

[0033] The preparation method of the lipid-lowering and cholesterol-lowering compositions of Embodiments 1-7 includes: weighing the raw materials according to the formulation in Table 1 above and mixing them evenly to obtain the composition.

[0034] Experimental Example

[0035] 1. Test materials

[0036] 1.1. Sample preparation information

[0037] Samples: The lipid-lowering and cholesterol-lowering compositions of Embodiments 1-7, separate plant sterol esters, separate red yeast rice extracts or separate lycopene oils are formulated into 100 mg / mL stock solutions with DMSO (analytical grade) and stored at -20°C.

[0038] Positive control: Atorvastatin Calcium Tablets (hereinafter referred to as atorvastatin calcium), white tablets, batch number 202105219C, Lepu Medical, stored in a cool place protected from light. Prepared into a stock solution of 11.6 mg / mL with DMSO and stored in aliquots at -20°C.

[0039] 1.2. Experimental animals

[0040] Zebrafish were all raised in fish culture water at 28°C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, with a conductivity of 450 - 550 μS / cm; pH of 6.5 - 8.5; hardness of 50 - 100 mg / L CaCO3). The license number for the use of experimental animals was: SYXK(Zhe)2022 - 0004. The feeding management met the requirements of international AAALAC accreditation (accreditation number: 001458).

[0041] The zebrafish were a melanin allele mutant semi - transparent Albino strain of zebrafish, and were bred by natural paired mating. Zebrafish at 5 days post - fertilization (5 dpf) were used for the determination of the maximum test concentration (MTC) for lipid - lowering and cholesterol - lowering effects and their efficacy evaluation.

[0042] 1.3. Instruments, consumables and reagents

[0043] Dissecting microscope (SZX7, OLYMPUS, Japan); Microinjector (IM300, Narishige, Japan); Needle puller (PC - 10, Narishige, Japan); CCD camera (VertA1, Shanghai Tusen Vision Technology Co., Ltd., China); Motorized focusing continuous - zoom fluorescence microscope (AZ100, Nikon, Japan); Precision electronic balance (CP214, OHAUS, USA); 6 - well plate (Nest Biotech, China); Ordinary PCR amplifier (T100, BIO - RAD, Singapore); Fluorescent quantitative PCR instrument (CFXConnect, BIO - RAD, Singapore); High - speed refrigerated centrifuge (Heraeus Fresco17, ThermoFisher, Germany); Ultraviolet - visible spectrophotometer (Nanodrop2000, Thermo, Austria); Microplate mini - centrifuge (BE - 6100, Qilinbeier Instrument Manufacturing Co., Ltd., Haimen, China); PCR 96 - well plate (product number MQ50801S×5, Mona Biotechnology Co., Ltd., China); Optical adhesive sealing film B (MSB1001, Bio - rad, USA).

[0044] Dimethyl sulfoxide (DMSO, batch number BCCD8942, Sigma, Switzerland); methylcellulose (batch number B2006074, Shanghai Aladdin Biochemical Technology Co., Ltd., China); cholesterlyl BODIPY TM 542 / 563C11 (cholesterol fluorescence probe, batch number 2291600, invitrogen, USA); Oil Red O (batch number SHBM5455, Sigma, USA); 1,2 - propylene glycol (batch number 20210817, Sinopharm Chemical Reagent Co., Ltd., China); pure egg yolk powder (batch number 20200809, Zhejiang Aige Biotechnology Co., Ltd., China); D-(+)-glucose (batch number 20201105, Sinopharm Chemical Reagent Co., Ltd., China); absolute ethanol (batch number 20210107, Sinopharm Chemical Reagent Co., Ltd., China); FastQuant RT Kit (With gDNase) kit (product number KR106, TIANGEN, China); RNA-Quick Purification Kit (RNA rapid extraction kit) (product number RN001, YiShan Biotech, China); PowerUp TM SYBR TM Green Master Mix (product number A25742, Thermo Fisher Scientific (China) Co., Ltd., China).

[0045] 2. Detection methods

[0046] 2.1. MTC determination

[0047] Randomly select 5dpf melanin allele mutant translucent Albino strain zebrafish into beakers, with 30 tails in each beaker (experimental group). Atorvastatin calcium and samples (final concentrations are shown in Table 2) were administered by water solution respectively. At the same time, a normal control group and a model control group were set up, and the volume of each beaker was 25 mL. Except for the normal control group, the remaining experimental groups were all administered a high-fat diet by water solution (the high-fat diet had a final concentration of glucose of 3 wt% and a final concentration of egg yolk powder of 1.5 mg / mL) to establish a zebrafish hyperlipidemia model. The samples and the high-fat diet were co-treated for 15 h (co-treated for 7.5 h every day, and the remaining time of each day was raised in fish-raising water at 28°C). During the sample treatment period, the number of dead zebrafish in each experimental group was counted every day and removed in time. After treatment at 28°C for 48 h, the minimum toxic concentration (MTC) of atorvastatin calcium and samples on the model zebrafish was measured.

[0048] 2.2. Evaluation of triglyceride-lowering efficacy

[0049] Randomly select 5 dpf melanin allele mutant translucent Albino strain zebrafish into beakers, with 30 fish in each beaker. Atorvastatin calcium and the sample (final concentrations are shown in Table 3) were dissolved in water and administered respectively. At the same time, a normal control group and a model control group were set up, and the volume of each beaker was 25 mL. Except for the normal control group, the other experimental groups were administered a high-fat diet (the high-fat diet was glucose with a final concentration of 3 wt% and egg yolk powder with a final concentration of 1.5 mg / mL) dissolved in water to establish a zebrafish hyperlipidemia model. The sample and the high-fat diet were co-treated for 15 h (co-treated for 7.5 h every day, and the remaining time of each day was raised in fish-raising water at 28 °C). After treatment at 28 °C for 48 h, Oil Red O was given for whole-body fat staining. After the staining was completed, 10 zebrafish were randomly selected from each experimental group and photographed under a dissection microscope. The data were analyzed and collected using Image-J advanced image processing software. The staining intensity of the tail blood vessels of zebrafish was analyzed, and the triglyceride-lowering effects of atorvastatin calcium and the sample were evaluated based on the statistical analysis results of this index. The statistical treatment results were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0050] 2.3. Evaluation of cholesterol-lowering efficacy

[0051] Randomly select 5 dpf melanin allele mutant translucent Albino strain zebrafish into beakers, with 30 fish in each beaker. Atorvastatin calcium and the sample (final concentrations are shown in Table 4) were dissolved in water and administered respectively. At the same time, a normal control group and a model control group were set up, and the volume of each beaker was 25 mL. Except for the normal control group, the other experimental groups were administered a high-fat diet (the high-fat diet was glucose with a final concentration of 3 wt% and egg yolk powder with a final concentration of 1.5 mg / mL) dissolved in water to establish a zebrafish hyperlipidemia model. The sample and the high-fat diet were co-treated for 15 h (co-treated for 7.5 h every day, and the remaining time of each day was raised in fish-raising water at 28 °C). After treatment at 28 °C for 32 h, a cholesterol fluorescent probe (final concentration 1 mg / mL) was given. After continuing the treatment for 16 h, 10 zebrafish were randomly selected from each experimental group and photographed under a fluorescence microscope. The data were analyzed and collected using NIS-Elements D3.20 advanced image processing software. The cholesterol fluorescence intensity of the tail blood vessels of zebrafish was analyzed, and the cholesterol-lowering effect of the sample was evaluated based on the statistical analysis results of this index. The statistical treatment results were expressed as mean ± SE. SPSS 26.0 software was used for statistical analysis, and p < 0.05 indicated that the difference was statistically significant.

[0052] 2.4. Effects on the relative expression level of the hmgcra gene

[0053] Randomly select 5dpf melanin allele mutant translucent Albino strain zebrafish into beakers, with 30 tails in each beaker. Atorvastatin calcium and the sample (final concentration is shown in Table 7) were dissolved in water respectively. At the same time, a normal control group and a model control group were set up, and the volume of each beaker was 25 mL. Except for the normal control group, the other experimental groups were given a high-fat diet dissolved in water (the high-fat diet was 3 wt% glucose and 1.5 mg / mL egg yolk powder) to establish a zebrafish hyperlipidemia model. The sample and the high-fat diet were co-treated for 15 h (co-treated for 7.5 h every day). After treatment at 28 °C for 48 h, the total RNA of zebrafish in each group was extracted using a rapid RNA extraction kit, and the concentration and purity of the total RNA were measured using an ultraviolet-visible spectrophotometer. Take 2.00 μg of the total RNA of the zebrafish sample, and operate according to the instructions of the cDNA first-strand synthesis kit to synthesize 20.0 μL of cDNA. The expressions of β-actin and hmgcra genes were detected by q-PCR (the primer sequences are shown in Table 6). Use β-actin as the internal reference for gene expression, and calculate the relative RNA expression level of the hmgcra gene. The results of statistical processing were expressed as mean±SE. Statistical analysis was performed using SPSS 26.0 software, and p<0.05 indicated that the difference was statistically significant.

[0054] 3. Detection results

[0055] 3.1. MTC

[0056] Under the experimental conditions of this experiment, the MTC of atorvastatin calcium for model zebrafish was 11.6 μg / mL. See the following table for details.

[0057] Table 2. MTC experimental results of the sample (n = 30)

[0058]

[0059] 3.2. Evaluation of triglyceride-lowering efficacy

[0060] The results are shown in the following table.

[0061] Table 3. Evaluation of triglyceride-lowering efficacy

[0062]

[0063] Note: Compared with the model control group, *P < 0.05, **P < 0.01, ***P < 0.001; different letters indicate significant differences (P < 0.05).

[0064] As can be seen from the above table, compared with the model control group, the positive control group, the lipid-lowering and cholesterol-lowering compositions of Examples 1-7 of the present invention, as well as individual phytosterol esters, individual monascus extracts or individual lycopene oils, can all significantly reduce triglycerides (P<0.05). There are significant differences in the reduction of triglycerides between the lipid-lowering and cholesterol-lowering compositions of Examples 1-7 and individual phytosterol esters, individual monascus extracts or individual lycopene oils, indicating that the lipid-lowering and cholesterol-lowering compositions of the present invention can synergistically reduce triglycerides.

[0065] 3.3. Evaluation of cholesterol-lowering efficacy

[0066] The results are shown in the following table.

[0067] Table 4. Evaluation of cholesterol-lowering efficacy

[0068]

[0069] Note: Compared with the model control group, *P<0.05, **P<0.01, ***P<0.001;

[0070] As can be seen from the above table, compared with the model control group, the positive control group, the lipid-lowering and cholesterol-lowering compositions of Examples 1-7 of the present invention can significantly reduce cholesterol (P<0.05). There are significant differences in the reduction of cholesterol between the lipid-lowering and cholesterol-lowering compositions of Examples 1-7 and individual phytosterol esters, individual monascus extracts or individual lycopene oils, indicating that the lipid-lowering and cholesterol-lowering compositions of the present invention can synergistically reduce cholesterol.

[0071] 3.4. Effect on the relative expression level of hmgcra gene

[0072] At the end of the experiment, total RNA of zebrafish was extracted, and the concentration of RNA and the A 260 / A 280 ratio (Table 5) were measured using an ultraviolet-visible spectrophotometer. The A 260 / A 280 ratios were all between 1.8 and 2.2, indicating that the quality of the total RNA extracted from zebrafish was good and could be used for subsequent q-PCR experiments. The primer sequences are shown in Table 6.

[0073] Table 5. Concentration of total RNA and A 260 / A 280 ratio (n = 30)

[0074]

[0075] Table 6. Primer sequence information

[0076]

[0077] The results are shown in the following table.

[0078] Table 7, Relative expression levels of hmgcra gene

[0079]

[0080]

[0081] Note: Compared with the model control group, *P < 0.05, **P < 0.01, ***P < 0.001. Different letters indicate significant differences (P < 0.05).

[0082] As can be seen from the above table, compared with the model control group, the positive control group, the lipid-lowering and cholesterol-lowering compositions of Examples 1-7 of the present invention, as well as individual phytosterol esters, individual red yeast rice extracts or individual lycopene oils, can significantly reduce the relative expression level of the hmgcra gene (P < 0.05). The lipid-lowering and cholesterol-lowering compositions of Examples 1-7 are more significant than individual phytosterol esters, individual red yeast rice extracts or individual lycopene oils in reducing the relative expression level of the hmgcra gene, indicating that the lipid-lowering and cholesterol-lowering compositions of the present invention can synergistically reduce the relative expression level of the hmgcra gene.

[0083] Obviously, the above examples are only for clear illustration and not a limitation of the implementation manners. For those of ordinary skill in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the implementation manners here. And the obvious changes or modifications derived therefrom are still within the protection scope of the present invention.

Claims

1. A lipid-lowering composition, characterized in that, It consists of the following components in parts by weight: 754 parts by weight of phytosterol esters, 114.9 parts by weight of red yeast rice extract, and 99.5 parts by weight of lycopene; Or, 800 parts by weight of phytosterol esters, 150 parts by weight of red yeast rice extract, and 50 parts by weight of lycopene; Or, 1000 parts by weight of phytosterol esters, 500 parts by weight of red yeast rice extract, and 10 parts by weight of lycopene; Or, 200 parts by weight of phytosterol esters, 300 parts by weight of red yeast rice extract, and 500 parts by weight of lycopene; Or, 682 parts by weight of phytosterol esters, 280 parts by weight of red yeast rice extract, and 99 parts by weight of lycopene.

2. A preparation method of the lipid-lowering composition according to claim 1, characterized in that, Weigh the raw materials according to the formula and mix them.

3. A preparation, characterized in that, Using the lipid-lowering composition described in claim 1 as the active ingredient.

4. The preparation according to claim 3, characterized in that, It also includes excipients or carriers permitted in the preparation.

5. The preparation according to claim 4, characterized in that, The forms of the preparation include liquid preparations and solid preparations.

6. The preparation according to claim 5, characterized in that, The forms of the preparation include injections, tablets, capsules, powders, granules or ointments.

7. The lipid-lowering composition according to claim 1 or the preparation according to any one of claims 3-6 has the following uses: (1) Use in the preparation of lipid-lowering drugs; (2) Use in the preparation of weight-loss drugs.

8. The use according to claim 7, characterized in that, The uses in the preparation of lipid-lowering drugs include: (1) The use in the preparation of drugs for reducing cholesterol; (2) The use in the preparation of drugs for preventing, alleviating, adjuvantly treating or treating hyperlipidemia and / or hypercholesterolemia; (3) The use in the preparation of drugs for reducing triglycerides; (4) The use in the preparation of drugs for reducing the expression level of the hmgcr gene.

9. The lipid-lowering composition according to claim 1 or the preparation according to any one of claims 3-6 has the following uses: (1) Use in the preparation of food or food additives for assisting lipid-lowering; (2) Use in the preparation of food or food additives for weight loss.

10. The use according to claim 9, characterized in that, The uses in the preparation of food or food additives for adjuvantly reducing blood lipids include: (1) The use in the preparation of food or food additives for adjuvantly reducing cholesterol; (2) The use in the preparation of food or food additives for adjuvantly reducing triglycerides.

11. The use according to claim 9 or 10, characterized in that, The food includes functional foods, health products or ordinary foods.

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