Application of pentadecanoic acid in preparation of product for treating and / or preventing atherosclerosis

Abstract

The invention relates to the technical field of biological medicine, and discloses application of pentadecanoic acid in preparation of a product for treating and / or preventing atherosclerosis, and the molecular formula of the pentadecanoic acid is CH3 (CH2) 13COOH. Experiments show that pentadecanoic acid can significantly reduce the lipid plaque area of an atherosclerosis model mouse, the content of low-density lipoprotein cholesterol in blood and inflammatory response, has good safety and can be used for atherosclerosis treatment.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and in particular to the application of pentadecanoic acid in the preparation of products for treating and / or preventing atherosclerosis. Background Art

[0002] Atherosclerosis is a chronic arterial disease, mainly characterized by the formation of atherosclerotic plaques under the intima of the arterial wall, resulting in thickening of the vascular wall, reduced elasticity, and stenosis of the lumen. Related research has found that with the intensification of unhealthy lifestyles, atherosclerosis shows a trend of becoming younger. Therefore, it is urgent to strengthen the prevention and treatment of atherosclerosis.

[0003] In related technologies, atherosclerosis is mainly prevented by statin drugs through lipid-lowering, but the effect is not ideal. Pentadecanoic acid (hereinafter referred to as PA) is an odd-chain long-chain saturated fatty acid, and human fatty acids are synthesized by successively adding two-carbon compounds on the basis of two-carbon compounds, so humans do not have the ability to synthesize odd-chain long-chain saturated fatty acids. Related research has shown that pentadecanoic acid can promote the uptake of glucose by adipocytes and skeletal muscle cells through the AMPK pathway, and does not affect the fasting blood glucose level of type 2 diabetic mice, and can be used for the adjuvant treatment of non-alcoholic fatty liver disease, but there is no report on its application in the treatment of atherosclerosis.

[0004] Based on this, the present invention has developed a drug capable of intervening in atherosclerosis, which can reduce the lipid plaque area, the content of low-density lipoprotein cholesterol in the blood, and the inflammatory response in atherosclerotic model mice, and has good safety. Summary of the Invention

[0005] The present invention aims to solve at least one of the technical problems existing in the prior art. For this purpose, the present invention provides the application of pentadecanoic acid in the preparation of products for treating and / or preventing atherosclerosis. Through experiments, the present invention has found that pentadecanoic acid can reduce the lipid plaque area, the content of low-density lipoprotein cholesterol in the blood, and the inflammatory response in atherosclerotic model mice, and has good safety and can be used for the treatment of atherosclerosis.

[0006] In the first aspect of the present invention, there is provided the application of pentadecanoic acid or a pharmaceutically acceptable salt thereof in the preparation of products for treating and / or preventing atherosclerosis.

[0007] In some embodiments of the present invention, the molecular formula of the pentadecanoic acid is CH3(CH2) 13 COOH.

[0008] In some embodiments of the present invention, the pharmaceutically acceptable salts refer to salts of the free acids of the compounds referred to herein, which are non-toxic, biocompatible or otherwise biologically suitable for administration to a subject. See generally, S.M. Berge et al., "Pharmaceutical Salts", J. Pharm. Sci., 1977, 66, 1-19. Preferred pharmaceutically acceptable salts are those that are pharmacologically effective and suitable for contact with the tissues of a subject without excessive toxicity, irritation or allergic reaction.

[0009] In some embodiments of the present invention, the pharmaceutically acceptable salts include sodium salts, potassium salts or calcium salts.

[0010] In some embodiments of the present invention, the product for treating and / or preventing atherosclerosis has at least one of the following effects: A) Reducing arterial lipid plaques; B) Lowering triglyceride, total cholesterol or low-density lipoprotein cholesterol levels; C) Reducing inflammation.

[0011] In some embodiments of the present invention, the product includes a drug.

[0012] In some embodiments of the present invention, the drug exists in the form of an oral preparation, an injection or a topical administration preparation.

[0013] In some embodiments of the present invention, the oral preparation includes tablets, capsules, pills, powders, granules, syrups or solutions.

[0014] In some embodiments of the present invention, the injection includes an injection dosage form or a freeze-dried powder injection dosage form for injection.

[0015] In some embodiments of the present invention, the topical administration preparation includes creams, ointments, sprays, aerosols, gels, cataplasms or patches.

[0016] In a second aspect of the present invention, there is provided a drug for treating atherosclerosis, the active ingredient of which comprises the pentadecanoic acid or its pharmaceutically acceptable salt described in the first aspect.

[0017] In some embodiments of the present invention, the drug further comprises a pharmaceutically acceptable excipient.

[0018] In some embodiments of the present invention, the pharmaceutically acceptable excipient includes at least one of a diluent, an excipient, a filler, a binder, a disintegrant, an absorption enhancer, a surfactant, an adsorption carrier, a lubricant, a sweetening agent and a flavoring agent.

[0019] In some embodiments of the present invention, the excipient includes water.

[0020] In some embodiments of the present invention, the filler includes at least one of starch and sucrose.

[0021] In some embodiments of the present invention, the binder includes at least one of cellulose derivatives, alginates, gelatin, and polyvinylpyrrolidone.

[0022] In some embodiments of the present invention, the wetting agent includes glycerol.

[0023] In some embodiments of the present invention, the disintegrant includes at least one of agar, calcium carbonate, and sodium bicarbonate.

[0024] In some embodiments of the present invention, the absorption enhancer includes quaternary ammonium compounds.

[0025] In some embodiments of the present invention, the surfactant includes cetyl alcohol.

[0026] In some embodiments of the present invention, the adsorption carrier includes at least one of kaolin and saponite.

[0027] In some embodiments of the present invention, the lubricant includes at least one of talc, calcium stearate, magnesium stearate, and polyethylene glycol.

[0028] In some embodiments of the present invention, the drug further comprises a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is generally recognized for this purpose and serves as an inactive ingredient of the medicament. Compilations of pharmaceutically acceptable carriers can be found in reference books such as Handbook of Pharmaceutical Excipients (2nd Edition, edited by A. Wade and P. J. Weller; published by the American Pharmaceutical Association, Washington and The Pharmaceutical Press, London, 1994), etc.

[0029] In some embodiments of the present invention, when the pentadecanoic acid or a pharmaceutically acceptable salt thereof in the present invention is used for treating atherosclerosis, it can be used alone or in the form of a pharmaceutical composition containing the pentadecanoic acid or a pharmaceutically acceptable salt thereof, that is, the drug further includes a co-administered drug.

[0030] In some embodiments of the present invention, the drug is administered via the gastrointestinal tract or / and parenteral administration routes.

[0031] In some embodiments of the present invention, the parenteral administration route is selected from injection administration, respiratory administration, cutaneous administration, mucosal administration or cavity administration.

[0032] In some embodiments of the present invention, the pharmaceutical dosage forms for parenteral administration are selected from injections, sprays, aerosols, patches, etc.

[0033] In some embodiments of the present invention, the pharmaceutical dosage forms for enteral administration are selected from tablets, capsules, powders, granules, pills, solutions, emulsions or syrups, etc.

[0034] In some embodiments of the present invention, the drug exists in the form of an oral preparation, an injection or a topical administration preparation.

[0035] In some embodiments of the present invention, the oral preparations include tablets, capsules, pills, powders, granules, syrups or solutions; the injections include injection dosage forms or freeze-dried powder for injection dosage forms; the topical administration preparations include creams, ointments, sprays, aerosols, gels, cataplasms or patches.

[0036] In some embodiments of the present invention, the carriers that can be selected when preparing oral preparations can be conventional pharmaceutical excipients such as starch, dextrin or cyclodextrin and various chemically modified cyclodextrins, sucrose, stearates, etc. When preparing freeze-dried powder for injection, it can be prepared by methods such as sterile spray drying, low-temperature vacuum drying, freeze drying, etc. The subsequent preparation processes and equipment for each preparation all belong to the conventional technologies in the pharmaceutical field, and the present invention does not limit this.

[0037] The present invention has at least the following beneficial effects: Through experiments, the present invention finds that pentadecanoic acid can significantly reduce the lipid plaque area, the content of low-density lipoprotein cholesterol in the blood, and the inflammatory response in atherosclerotic model mice, and has good safety, and can be used for the treatment of atherosclerosis.

[0038] Other features and advantages of the present invention will be described in the subsequent specification. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] The following further describes the present invention with reference to the drawings and embodiments, wherein: Figure 1 It is a schematic diagram for constructing atherosclerotic model mice of the present invention.

[0040] Figure 2 It is the oil red staining result of the whole aorta sample of the present invention, wherein A is the staining result of a single representative sample, and B is the statistical result of multiple samples.

[0041] Figure 3These are the oil red staining results of the aortic root frozen sections of the present invention. Among them, A shows the staining results of a single representative sample, with a scale bar of 200 μm, and B shows the statistical results of multiple samples.

[0042] Figure 4 These are the blood lipid analysis results of mice in each treatment group of the present invention. Among them, A shows the statistical results of triglycerides in mouse blood samples, B shows the statistical results of total cholesterol, C shows the statistical results of high-density lipoprotein cholesterol, and D shows the statistical results of low-density lipoprotein cholesterol.

[0043] Figure 5 These are the flow cytometry analysis results of monocytes (CD11B+, Ly6C+) of the present invention in CD45+ cells. Among them, A shows the results of a single representative sample, and B shows the statistical results of multiple samples.

[0044] Figure 6 These are the flow cytometry analysis results of macrophages (CD11B+, F4 / 80+) of the present invention in CD45+ cells. Among them, A shows the results of a single representative sample, and B shows the statistical results of multiple samples.

[0045] Figure 7 These are the CCK8 toxicity test results of PA of the present invention on HepG2 cells. Detailed implementation manners

[0046] The following will clearly and completely describe the concept of the present invention and the technical effects produced in combination with the embodiments, so as to fully understand the purpose, features and effects of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, other embodiments obtained by those skilled in the art without creative efforts shall fall within the scope of protection of the present invention.

[0047] The terms "preferably", "more preferably", etc. in the present invention refer to the embodiments of the present invention that can provide certain beneficial effects in certain cases. However, in the same or other cases, other embodiments may also be preferred. In addition, the description of one or more preferred embodiments does not imply that other embodiments are not available, nor is it intended to exclude other embodiments from the scope of the present invention.

[0048] When a numerical range is disclosed herein, the above range is considered continuous and includes the minimum and maximum values of the range, as well as each value between such minimum and maximum values. Further, when the range refers to integers, it includes each integer between the minimum and maximum values of the range. In addition, when multiple ranges are provided to describe features or characteristics, the ranges can be combined. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all sub-ranges subsumed therein.

[0049] In the description of the present invention, the reference term "and / or" includes all and any combinations of one or more of the related listed items.

[0050] In the description of the present invention, the description of reference terms such as "one embodiment", "some embodiments", "schematic embodiments", "examples", "specific examples", or "some examples", etc. means that the specific features, structures, materials, or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic expressions of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in a suitable manner in any one or more embodiments or examples.

[0051] In the embodiment of the present invention, the CAS number of pentadecanoic acid (PA) is 1002 - 84 - 2.

[0052] For those not specifying specific conditions in the examples, they are carried out according to conventional conditions or the conditions recommended by the manufacturer. For reagents or instruments without indicating the manufacturer, they are all conventional products that can be obtained by commercial purchase.

[0053] Example 1: Effects on lipid plaques caused by atherosclerosis This example detected the effects of pentadecanoic acid on lipid plaques caused by atherosclerosis, specifically including the following content.

[0054] 1. Experimental method (1) Construction of an atherosclerotic mouse model: The construction method of the atherosclerotic mouse model used in this experiment is as Figure 1 shown, and the specific method is as follows: 6 - week - old ApoE - deficient mice (C57BL / 6 mice, purchased from Jiangsu Jicui Yakang Biotechnology Co., Ltd.) were used. The control group was fed a normal diet (Chow diet, hereinafter referred to as CHD) throughout the experiment, while the treatment group started to be fed a high - fat diet (High Fat Diet, HFD) from 7 weeks of age and continued to be fed until 20 weeks of age to construct an atherosclerotic mouse model.

[0055] (2) Administration method: The control group of model mice was gavaged with 100 μL of corn oil (as a solvent control) at a frequency of 3 times a week until 20 weeks of age; The model mice in the HFD treatment group were gavaged with 100 μL of corn oil containing pentadecanoic acid (PA). The dosage of pentadecanoic acid was 25 mg / kg of mouse body weight. The mice started gavage at a frequency of 3 times a week from 8 weeks of age until 20 weeks of age; The model mice in the positive treatment group were intragastrically administered with the lipid-lowering drug atorvastatin calcium (ATO) at a dosage of 25 mg / kg of mouse body weight. The intragastric administration frequency started from 8 weeks of age, three times a week until 20 weeks of age. (3)Oil red staining treatment: At 20 weeks of age, the mice in each group were euthanized and the hearts and aortas were collected. Among them, the heart and whole aorta samples were separated and fixed in 4% PFA (paraformaldehyde), and then the excess perivascular adipose tissue was removed and dehydrated in 20% sucrose solution for 24 hours. The heart was embedded in OCT embedding medium to prepare frozen sections of the aortic root. Then, the frozen sections of the aortic root and the whole aorta were stained with oil red O to detect lipid plaques. The plaque area was quantitatively analyzed by ImageJ software.

[0056] 2. Experimental results The oil red staining results of the whole aorta samples are as Figure 2 shown. The positive area of oil red O staining in the whole aorta of mice fed with normal diet (CHD) was about 1.5%, which was the experimental baseline. In the treatment group of mice fed with high-fat diet (HFD) + corn oil solvent control (Corn oil), the corresponding area was close to 4%, indicating that the atherosclerotic model was successfully constructed. In the mice intragastrically administered with PA, the corresponding area decreased to less than 2%, indicating that PA has the effect of preventing and alleviating atherosclerosis.

[0057] The oil red staining results of the frozen sections of the aortic root are as Figure 3 shown. Among them, A is the result of a single representative sample, and B is the statistical result of the samples, further showing that PA has the effect on the lipid plaque area.

[0058] Example 2: Effect on cholesterol content caused by atherosclerosis Low-density lipoprotein cholesterol (LDL-C) in the blood is one of the possible driving factors of atherosclerosis. To confirm the LDL-C level in mice, the blood lipids of mice were analyzed in this example. The specific experimental methods are as follows: Refer to Example 1 above to construct an atherosclerotic mouse model and perform drug administration treatment. At 20 weeks of age, the mice in each group were euthanized and blood samples were collected for triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C).

[0059] The detection results are as Figure 4 shown, indicating that PA treatment can significantly reduce the contents of triglyceride, total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol in the blood samples.

[0060] Example 3: Effect on inflammation caused by atherosclerosis The inflammatory level of the body is one of the possible driving factors of atherosclerosis. To measure the inflammatory level of mice, flow cytometry analysis was performed on the peripheral blood cells of mice in this example. The specific method is as follows: Refer to Example 1 above to construct an atherosclerotic mouse model and perform drug administration treatment. When the mice reach 20 weeks of age, collect peripheral blood samples for detection. Specifically, in this example, monocytes were defined by CD45+, CD11B+, and Ly6C+, and macrophages were defined by CD45+, CD11B+, and F4 / 80+ to measure the levels of monocytes and macrophages to reflect the inflammatory level of mice.

[0061] The detection method is as follows: Take peripheral blood cells and incubate them with 1× red blood cell lysis buffer to lyse and remove red blood cells. The remaining nucleated peripheral blood cells were washed once with PBS and then incubated on ice for 20 minutes with an antibody solution prepared in PBS containing 0.2% bovine serum albumin. Flow cytometry analysis was performed using an Agilent Novocyte D2060R flow cytometer. Monocytes were stained with FITC-labeled anti-CD45, PE-labeled anti-CD11b, and PE-Cy7-labeled anti-Ly6C, while macrophages were stained with FITC-labeled anti-CD45, PE-labeled anti-CD11b, and PE-Cy5-labeled anti-F4 / 80. The obtained flow cytometry data were then analyzed using FlowJo software.

[0062] The results of flow cytometry analysis of monocytes (CD11B+, Ly6C+) in CD45+ cells are as Figure 5 shown, and the results of flow cytometry analysis of macrophages (CD11B+, F4 / 80+) in CD45+ cells are as Figure 6 shown, showing that the relative abundances of monocytes and macrophages in the peripheral blood of mice in the PA treatment group were significantly lower than those in its solvent control (Corn oil), indicating that the inflammatory level in the bodies of mice in the PA treatment group was lower. It is speculated that PA may also prevent atherosclerosis by reducing the body's inflammatory level.

[0063] Example 4: Cytotoxicity test To test the toxicity of PA, a CCK-8 (Cell Counting Kit-8) toxicity experiment was carried out in this example to evaluate the toxicity of PA to cells. The specific method is as follows: HepG2 cells were seeded at an appropriate density (1×10 4Cells (at a density of [number of cells per well]) were seeded in 96-well plates, with 100 μL of culture medium in each well. The plates were incubated in a 37°C, 5% CO₂ incubator for 24 hours to allow the cells to adhere and reach the desired growth state. Different concentrations of the test substance solutions were added to the 96-well plates, setting several concentration gradients, with 3 replicates in each group. At the same time, a blank control group (containing only the culture medium) and a negative control group (containing only cells and the culture medium) were set up. After the cell treatment was completed, 10 μL of CCK8 reagent was added to each well, gently mixed, and then incubated in a 37°C, 5% CO₂ incubator for 12 hours. After the incubation, the optical density (OD) values of each well were measured at a wavelength of 450 nm using a microplate reader, and the data were recorded and the cell viability of each experimental group was calculated.

[0064] The detection results are as Figure 7 shown, indicating that PA has no toxicity at 100 μM, and its IC50 is about 1.77 mM. Compared with ATO, PA has relatively lower toxicity and is safer.

[0065] In summary, the present invention provides the application of pentadecanoic acid in the preparation of products for treating and / or preventing atherosclerosis. Specifically, the present invention discovers that pentadecanoic acid can reduce the lipid plaque area, the content of low-density lipoprotein cholesterol in the blood, and the inflammatory response in atherosclerotic model mice, and has good safety, and can be used for the treatment of atherosclerosis.

[0066] The above has described the embodiments of the present invention in detail, but the present invention is not limited to the above embodiments. Within the knowledge scope of those of ordinary skill in the art to which the present invention pertains, various changes can be made without departing from the purpose of the present invention. In addition, without conflict, the embodiments of the present invention and the features in the embodiments can be combined with each other.

Claims

1. Use of pentadecanoic acid or a pharmaceutically acceptable salt thereof in preparing a product for treating and / or preventing atherosclerosis.

2. The use according to claim 1, characterized in that: The molecular formula of the pentadecanoic acid is CH3(CH2) 13 COOH.

3. The use according to claim 1, characterized in that: The pharmaceutically acceptable salt includes at least one of a sodium salt, a potassium salt and a calcium salt.

4. The use according to claim 1, characterized in that: The product for treating and / or preventing atherosclerosis has at least one of the following effects: A) Reduce arterial lipid plaque; B) lowering of triglycerides, total cholesterol or LDL cholesterol levels; C) reduce inflammation.

5. The use according to claim 1, characterized in that: The products include pharmaceuticals.

6. The use according to claim 5, characterized in that: The drug is in the form of oral preparation, injection or local administration preparation.

7. The use according to claim 6, characterized in that: The oral preparations include tablets, capsules, pills, powders, granules, syrups or solutions; The injection includes an injection liquid dosage form or a lyophilized powder injection dosage form; The topical administration preparations include creams, ointments, sprays, aerosols, gels, cataplasms or patches.

8. A drug for treating atherosclerosis, characterized in that: The active ingredient comprises the pentadecanoic acid or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 7.

9. The requirement according to claim 8, characterized in that: The drug further comprises pharmaceutically acceptable excipients.

10. The requirement according to claim 8 or 9, characterized in that: The pharmaceutically acceptable excipients include at least one of diluents, excipients, fillers, binders, disintegrants, absorption promoters, surfactants, adsorption carriers, lubricants, sweeteners and flavoring agents.

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