Triple slow-release system zanthoxylum seed oil for assisting in reducing hypertension, hyperlipidemia and hyperglycemia and preparation method thereof

Abstract

The invention provides triple sustained-release system zanthoxylum seed oil for assisting in reducing hypertension, hyperlipidemia and hyperglycemia and a preparation method thereof. The triple sustained-release system zanthoxylum seed oil comprises a pH response type stomach protection layer, an enzyme response type enteric-coated layer and a time sustained-release type colon targeting layer, and stomach-small intestine-colon three-stage precise sustained release is achieved through a pH / enzyme double-response type core-shell sustained-release structure; active components adopt ALA / nervonic acid / hydroxyl-alpha-sanshool ternary compatibility, and AMPK / mTOR pathway is used for cooperative regulation to assist in reducing blood sugar, blood pressure and blood fat. The bioavailability of the active ingredients is improved to 78%, the combined inhibition efficiency on PPARy, ACE and HMG-CoA is larger than or equal to 75%, and the preparation method is suitable for preparing products for assisting in reducing hypertension, hyperglycemia and hyperlipidemia.

Description

Technical Field

[0001] This application relates to the field of bioenzymatic hydrolysis technology, specifically to a triple sustained-release system for lowering blood sugar, blood pressure, and blood lipids (the "three highs"), pepper seed oil, and its preparation method, which is suitable for synergistic intervention applications in lowering blood sugar, blood pressure, and blood lipids (the "three highs"). Background Technology

[0002] Enzymatic hydrolysis is a biotechnology that uses enzymes as catalysts to efficiently and specifically break down complex macromolecules (such as proteins, starch, cellulose, and fats) into smaller molecules (such as peptides, amino acids, monosaccharides, and fatty acids) under mild conditions (room temperature, normal pressure, and suitable pH). Enzymatic hydrolysis utilizes the powerful catalytic ability and specificity of enzymes to efficiently and environmentally break down biological macromolecules into smaller molecules under mild conditions. It enhances the value of raw materials, improves product quality, and develops functional products.

[0003] As existing technologies, Chinese patent No. 202510596056.6 discloses a method for preparing pepper kernel oil microcapsules, and patent No. 202310300451.6 discloses a stable and efficient pepper kernel oil composition, its preparation method and application. Existing auxiliary oils for lowering blood pressure, blood sugar, and cholesterol have three major defects: (1) The bioavailability of α-linolenic acid in pepper kernel oil is ≤40%, and the decomposition rate under gastric acid is >65%, making it difficult for the effective components to exert their effect of lowering blood pressure, blood sugar, and cholesterol; (2) The single release rate of commercially available sustained-release preparations fluctuates by >35%, making it impossible to achieve targeted sustained release in the intestine, resulting in serious waste of active ingredients for lowering blood pressure, blood sugar, and cholesterol; (3) There is a lack of a multi-dimensional synergistic system, and the combined inhibition efficiency of key targets for lowering blood pressure, blood sugar, and cholesterol (PPARγ, ACE, HMG-CoA) is <50%, resulting in limited synergistic effects in lowering blood pressure, blood sugar, and cholesterol. Therefore, this application proposes a triple sustained-release system for lowering blood pressure, blood sugar, and cholesterol, pepper kernel oil and its preparation method, which at least partially solves the above problems. Summary of the Invention

[0004] In view of the aforementioned problems, this application is made to provide a triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol, namely, pepper seed oil and its preparation method, which overcomes or at least partially solves the aforementioned problems.

[0005] The embodiments of the present invention are implemented as follows: This application provides a triple sustained-release system for assisting in lowering blood pressure, blood sugar, and cholesterol levels, namely, pepper seed oil. The triple sustained-release system includes, from the inside out, a pH-responsive gastric protective layer, which, through a sodium alginate-chitosan complex, responds to gastric acid to trigger expansion and form a gel barrier. Enzyme-responsive enteric coating, β-cyclodextrin-trypsin substrate peptide conjugate, responds to intestinal trypsin to specifically cleave peptide chains, releasing 30% of the active substance; A time-release colon-targeting layer, with acrylic resin Eudragit FS30D encapsulating nanoemulsion, continuously releases 65% of the active substances from the colon over 0–24 hours; The active substances include 20-30% α-linolenic acid, 6-8% nervonic acid, 2-3% hydroxy-α-sanshool from pepper seed oil, and the remainder being fatty acids. Through synergistic regulation of the AMPK / mTOR pathway, they achieve the function of assisting in lowering blood pressure, blood sugar, and cholesterol.

[0006] In some embodiments of the present invention, the pH-responsive gastric protective layer, through a sodium alginate-chitosan complex, responds to gastric acid-triggered swelling to form a gel barrier, comprising: A pH-responsive gastric protective layer, consisting of a sodium alginate-chitosan complex with a mass ratio of 2:1 to 3:1, expands in response to gastric acid to form a gel barrier, with an α-linolenic acid sustained-release rate ≤5% / h.

[0007] In some embodiments of the present invention, the fatty acids include linoleic acid, oleic acid, and saturated fatty acids.

[0008] In some embodiments of the present invention, the preparation steps of the triple sustained-release system for assisting in lowering blood pressure, blood sugar, and cholesterol include: Nanoemulsion preparation steps: Nervate nanoemulsion was prepared by high-pressure homogenization; loading rate ≥90~95%, Zeta potential -30mV~-35mV; Slow-release coating process: Using a fluidized bed bottom spraying process, with an inlet air temperature of 35-45℃, an atomization pressure of 0.7-1.0 bar, and a feed rate of 3-5 mL / min, the coating is performed using the acrylic resin Eudragit FS30D, resulting in a coating weight gain of 20%-25%; among which, the encapsulation efficiency is ≥90%-95%, and the burst release rate is <3%-5%.

[0009] Product molding steps: Soft capsules are filled using a mass ratio of gelatin:glycerin:water of 5-4:3-2:3-2, with a disintegration time of 30-40 minutes.

[0010] In some embodiments of the present invention, the preparation of nervonic acid nanoemulsion by high-pressure homogenization includes: Nervonic acid nanoemulsions (particle size 80±5nm) were prepared by high-pressure homogenization under an environment of 100MPa to 120MPa for 1 to 3 cycles.

[0011] In some embodiments of the present invention, the preparation of active substance powder is further included, comprising: Mix 30g of pepper seed oil with an α-linolenic acid content of 20% with 5g of nervonic acid concentrate; Add 0.5% tocopherol and 0.3% rosmarinic acid compound antioxidant to the mixture; Nanoparticles were prepared using a W / O / W double emulsion method, and the active substance powder was obtained by spray drying.

[0012] In some embodiments of the present invention, the sustained-release coating step involves: using a fluidized bed bottom spraying process, with an inlet air temperature of 35–45°C, an atomization pressure of 0.7–1.0 bar, and a feed rate of 3–5 mL / min, coating with acrylic resin Eudragit FS30D, resulting in a coating weight gain of 20%–25%, including: Under conditions of bed temperature of 38℃, atomization pressure of 1.0 bar, and feed rate of 3 mL / min, Eudragit FS30D was sequentially coated to achieve a weight gain of 15%, β-cyclodextrin-peptide complex to achieve a weight gain of 7%, and sodium alginate layer to achieve a weight gain of 3%, with encapsulation efficiency ≥90%~95% and burst release rate <3%~5%.

[0013] The application of a triple sustained-release system for lowering blood pressure, blood sugar, and cholesterol using pepper seed oil, wherein the pepper seed oil in the triple sustained-release system for lowering blood pressure, blood sugar, and cholesterol releases for 2 hours when it is located in the stomach, the release medium is 0.1N HCl, and the release rate is 8.2%.

[0014] A method for preparing pepper seed oil, a triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol, includes the following steps: (1) Preparation of active substance powder: 30g of pepper kernel oil with 20% α-linolenic acid content was mixed with 5g of nervonic acid concentrate, and 0.5% tocopherol and 0.3% rosmarinic acid composite antioxidant were added. Nanoparticles were prepared by W / O / W double emulsion method and spray dried to obtain active substance powder; (2) Preparation of nanoemulsion: The active substance powder in step (1) is homogenized by high pressure and circulated for 1 to 3 cycles at 100 MPa to 120 MPa to prepare nervonic acid nanoemulsion with a particle size of 80 ± 5 nm (loading rate ≥ 90 to 95%, Zeta potential -30 mV to -35 mV). (3) Slow-release coating: Using fluidized bed bottom spraying process, under the conditions of inlet air temperature of 35-45℃, atomization pressure of 0.7-1.0 bar and feed rate of 3-5 mL / min, acrylic resin Eudragit FS30D is used for coating, and the coating weight gain is 20%-25% (encapsulation efficiency ≥90%-95%, burst release rate <3%-5%). (4) Product molding: Soft capsules are filled using a mass ratio of gelatin: glycerin: water of 5-4: 3-2: 3-2, with a disintegration time of 30-40 minutes.

[0015] In some embodiments of the present invention, the specific conditions for the sustained-release layer coating are as follows: bed temperature 38°C, atomization pressure 1.0 bar, feed rate 3 mL / min, and sequential coating with acrylic resin Eudragit FS30D to achieve a weight gain of 15%, β-cyclodextrin-peptide complex to achieve a weight gain of 7%, and sodium alginate layer to achieve a weight gain of 3%.

[0016] In some embodiments of the present invention, the application of a triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol (the "three highs")—pepper seed oil—is described. This triple-release system provides a phased, sustained release within the human digestive tract, specifically including: In the stomach: the release time is 2 hours, the release medium is 0.1N HCl, and the release rate is 8.2%, avoiding the destruction of active ingredients by gastric acid; Small intestine stage: The release time is 4 hours, the release medium is pH 6.8 phosphate buffer, and the release rate is 37.5%, initially exerting an auxiliary synergistic effect in lowering blood pressure, blood sugar, and cholesterol; Colonic stage: The release time is 18 hours, the release medium is pH 7.4 phosphate buffer and β-glucosidase, the release rate is 54.3%, and it continues to play an auxiliary role in lowering blood pressure, blood sugar, and cholesterol.

[0017] Compared with the prior art, the embodiments of the present invention have at least the following advantages or beneficial effects: The system utilizes a pH-responsive gastric protective layer, formed by a sodium alginate-chitosan complex, to induce gastric acid-triggered expansion and create a gel barrier. An enzyme-responsive enteric layer, with a β-cyclodextrin-trypsin substrate peptide conjugate, responds to intestinal trypsin-specific cleavage of peptide chains, releasing 30% of the active substance. A time-sustaining colon-targeting layer, encapsulated in Eudragit FS30D acrylic resin, continuously releases 65% of the active substance from the colon over 0–24 hours. Through a pioneering pH / enzyme dual-responsive core-shell structure, precise controlled release is achieved across the gastrointestinal-colonic stages. A ternary active system of α-linolenic acid / nervonic acid / piperazine is constructed, forming a synergistic regulatory pathway between AMP-activated protein kinase (AMPK) and mTOR gene-encoded protein kinase (mTOR). Attached Figure Description To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 This is a schematic diagram of the structure of pepper seed oil, a triple sustained-release system that helps lower blood pressure, blood sugar, and cholesterol.

[0019] Figure 2A schematic diagram illustrating the effect of pepper seed oil, a triple sustained-release system for assisting in lowering blood lipids, on zebrafish blood lipids during an experiment.

[0020] Figure 3 Phenotypic diagram of zebrafish triglyceride content in an experiment using pepper seed oil, a triple sustained-release system for assisting in lowering blood pressure, blood sugar, and cholesterol.

[0021] Figure 4 Phenotypic diagram of blood glucose levels in zebrafish during an experiment using pepper seed oil, a triple sustained-release system for assisting in lowering blood pressure, blood sugar, and cholesterol. Detailed Implementation

[0022] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. The components of the embodiments of the present invention described and shown in the accompanying drawings can generally be arranged and designed in various different configurations.

[0023] like Figure 1 As shown, an embodiment of this application provides a triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol, comprising: A pH-responsive gastric protective layer, through a sodium alginate-chitosan complex, responds to gastric acid-triggered expansion to form a gel barrier; Enzyme-responsive enteric coating, β-cyclodextrin-trypsin substrate peptide conjugate, responds to intestinal trypsin to specifically cleave peptide chains, releasing 30% of the active substance; The time-release colon-targeting layer uses Eudragit FS30D acrylic resin to encapsulate nanoemulsions, continuously releasing 65% of the active substances from the colon over 0–24 hours.

[0024] In this application, a novel pH / enzyme dual-response core-shell structure is used to achieve precise controlled release in the three stages of stomach-small intestine-colon; a ternary active system of α-linolenic acid / nervonic acid / pigment is constructed to form a synergistic regulation of the AMP-activated protein kinase (AMPK) / mTOR gene-encoded protein kinase (mTOR) pathway.

[0025] The following will further explain a triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol using pepper seed oil, as described in this exemplary embodiment.

[0026] In one embodiment of this application, the pH-responsive gastric protective layer, through a sodium alginate-chitosan complex, responds to gastric acid-triggered expansion to form a gel barrier, comprising: a pH-responsive gastric protective layer, through a sodium alginate-chitosan complex with a mass ratio of 2:1 to 3:1, responds to gastric acid-triggered expansion to form a gel barrier, and an α-linolenic acid (ALA) sustained-release rate ≤5% / h.

[0027] The triple controlled-release system architecture of this application includes: First layer: pH-responsive gastric protective layer; the material is sodium alginate-chitosan complex (mass ratio 2:1 to 3:1); response mechanism: gastric acid (pH 1 to 1.5) triggers expansion to form a gel barrier, and the α-linolenic acid sustained release rate is ≤5% / h; The second layer is an enzyme-responsive enteric-coated layer; the material is a β-cyclodextrin-trypsin substrate peptide conjugate; the response mechanism is that intestinal trypsin specifically cleaves the peptide chain, releasing 30% of the active substance; the third layer is a time-sustaining colon-targeting layer; the material is an acrylic resin Eudragit FS30D encapsulated nanoemulsion; the release curve is that 65% of the active substance is continuously released from the colon (linear release from 0 to 24 hours).

[0028] Furthermore, the active ingredient formulation includes 20-30% α-linolenic acid, 6-8% nervonic acid, and 2-3% hydroxy-α-sanshool, with the remainder being fatty acids such as linoleic acid, oleic acid, and saturated fatty acids.

[0029] The compatibility of active substances is shown in Table 1 below:

[0030] Table 1 The preparation steps of the triple sustained-release system for lowering blood pressure, blood sugar, and cholesterol, including: Nanoemulsion preparation steps: Nervonic acid nanoemulsion is prepared by high-pressure homogenization; wherein the loading rate is ≥90~95% and the Zeta potential is -30mV~-35mV.

[0031] Slow-release coating process: Using a fluidized bed bottom spraying process, with an inlet air temperature of 35-45℃, an atomization pressure of 0.7-1.0 bar, and a feed rate of 3-5 mL / min, the coating is performed using the acrylic resin Eudragit FS30D, resulting in a coating weight gain of 20%-25%; among which, the encapsulation efficiency is ≥90%-95%, and the burst release rate is <3%-5%.

[0032] Product molding steps: Soft capsules are filled using a gelatin:glycerin:water ratio of 5-4:3-2:3-2, with a disintegration time of 30-40 minutes, in accordance with CP2020 standards.

[0033] It should be noted that the fatty acids mentioned above include linoleic acid, oleic acid, and saturated fatty acids. Linoleic acid is a polyunsaturated fatty acid (PUFA) with two double bonds in its carbon chain. It is the parent compound of the Omega-6 series fatty acids, including gamma-linolenic acid (GLA) and arachidonic acid (ARA). Oleic acid is a monounsaturated fatty acid (MUFA) with one double bond in its carbon chain (at the Omega-9 position). Examples include olive oil (highest content, approximately 55-83%), tea seed oil, rapeseed oil, peanut oil, and avocado oil. Saturated fatty acids do not have double bonds in their carbon chains, and their molecular structure is "saturated." They are usually solid at room temperature (except for coconut oil and palm kernel oil). Examples include acetic acid, butyric acid, propionic acid, caprylic acid, capric acid, butter, and coconut oil.

[0034] Furthermore, the preparation of nervonic acid nanoemulsion by high-pressure homogenization includes: Nervonic acid nanoemulsions (particle size 80±5nm) were prepared by high-pressure homogenization under an environment of 100MPa to 120MPa for 1 to 3 cycles.

[0035] In one embodiment of this application, the preparation of the active substance powder is further included, comprising: Mix 30g of pepper seed oil with an α-linolenic acid content of 20% with 5g of nervonic acid concentrate; Add 0.5% tocopherol and 0.3% rosmarinic acid compound antioxidant to the mixture; Nanoparticles were prepared using a W / O / W double emulsion method, and the active substance powder was obtained by spray drying.

[0036] Further, the sustained-release layer coating step involves: using a fluidized bed bottom spraying process, under an environment with an inlet air temperature of 35–45°C, an atomization pressure of 0.7–1.0 bar, and a feed rate of 3–5 mL / min, coating is performed using the acrylic resin Eudragit FS30D, resulting in a coating weight gain of 20%–25%, including: Under conditions of bed temperature 38℃, atomization pressure 1.0 bar, and feed rate 3 mL / min, Eudragit FS30D was sequentially coated to achieve a weight gain of 15%, β-cyclodextrin-peptide complex to achieve a weight gain of 7%, and sodium alginate layer to achieve a weight gain of 3%.

[0037] It should be noted that the aforementioned nervonic acid nanoemulsion refers to pepper seed oil with a particle size of 80±5nm. The raw material for preparing the nervonic acid nanoemulsion in this application is ordinary pepper seed oil, which is an edible vegetable oil processed from pepper seeds through kernel-shell separation, low-temperature pressing, and refining techniques. The specific preparation of pepper seed oil from pepper seeds is prior art and not the scheme to be protected in this application, and will not be described in detail here.

[0038] This application breaks through the bioavailability bottleneck of traditional functional oils and fats, and increases the bioavailability of active substances to 78% through precise spatiotemporal controlled release. Compared with the 35% of traditional formulations, the bioavailability of active substances in this application is greatly improved.

[0039] In one embodiment of this application, an application of a triple-release system for lowering blood sugar, blood pressure, and blood lipids (the "three highs") is also provided. Specifically, this system utilizes a pH / enzyme dual-response mechanism to assist in lowering blood sugar, blood pressure, and blood lipids (the "three highs"), and its synergistic intervention in these processes. Specifically, the release time of the triple-release system for lowering blood sugar, blood pressure, and blood lipids is 2 hours in the stomach, with 0.1N HCl as the release medium and a release rate of 8.2%. In the intestine (small intestine) stage, the release time is 4 hours, with a pH 6.8 phosphate buffer as the release medium and a release rate of 37.5%. In the colon stage, the release time is 18 hours, with a pH 7.4 phosphate buffer and β-glucosidase as the release medium and a release rate of 54.3%.

[0040] The details are shown in Table 2 below.

[0041] Table 2 In a specific example, validation is performed using animal experimental data, such as experiments using a zebrafish model.

[0042] Blood pressure data are shown in Table 3 below:

[0043] Table 3 After a two-week experimental observation, the results showed that the average blood pressure of the zebrafish in the experimental group was significantly reduced after treatment, and the difference was significant compared with the control group (P<0.01).

[0044] Blood lipid data are shown in Table 4 below:

[0045] Table 4 Biochemical indicators show that the overall triglyceride content of zebrafish in the lipid-lowering agent group was significantly lower than that in the model control group.

[0046] Blood glucose data are shown in Table 5 below:

[0047] Table 5 The experiment successfully established a zebrafish hyperglycemia model and verified the hypoglycemic effect of pepper seed oil.

[0048] The specific experimental procedures described in this application are as follows: This experiment aims to explore the potential effects of Zhongjiao Zhenxuan pepper seed oil on assisting in lowering blood pressure, blood lipids, and blood sugar using a zebrafish (Danio rerio) model. Zebrafish, due to their high similarity to human physiological mechanisms, rapid reproduction, ease of rearing, and genetic manipulation, have become widely used model organisms in biomedical research. This experiment hopes to verify whether the target drug or compound has the effect of assisting in lowering blood pressure, blood lipids, and blood sugar by comparing the changes in the experimental and control groups of zebrafish, providing basic data for further preclinical research.

[0049] The specific experimental procedure is as follows: (1) Blood pressure test Zebrafish preparation: Select adult zebrafish of similar age and weight, and randomly divide them into treatment group and control group to ensure that the basic conditions of each group are consistent.

[0050] Exposure treatment: The zebrafish in the treatment group were placed in water containing diglycerides, while the control group was placed in water under the same conditions but without the drug. The drug concentration was set according to the safe and effective range determined in preliminary experiments.

[0051] Blood pressure monitoring: Blood pressure of the two groups of zebrafish was measured and recorded using non-invasive methods at multiple time points before and after the start of the experiment (e.g., daily / every few days).

[0052] Behavioral and health status observation: Simultaneously observe and record the swimming speed, feeding behavior, and changes in body color of zebrafish to assess the effects of drugs on the overall physiological state of zebrafish.

[0053] Data collection and processing: All blood pressure measurement data were compiled, and statistical software was used to perform significance analysis of differences.

[0054] The experimental results, as shown in Table 3 above, confirm that the triple sustained-release system of pepper seed oil has a significant auxiliary effect in lowering blood pressure.

[0055] Experimental Conclusion: After a two-week observation period, the results showed that the average blood pressure of the zebrafish in the experimental group decreased significantly after treatment, and the difference was significant compared with the control group (P<0.01). This indicates that Zhongjiao Zhenxuan pepper kernel oil has an effective antihypertensive effect in the zebrafish model. Meanwhile, no obvious behavioral abnormalities or deterioration of health were observed in the zebrafish during the observation period, suggesting that Zhongjiao Zhenxuan pepper kernel oil has good safety and tolerability under the experimental conditions.

[0056] (2) Blood lipid test First, zebrafish were fed egg yolk powder (dissolved in the aquatic water) to establish a hyperlipidemia model. Then, the zebrafish were divided into three groups: a normal control group (not fed egg yolk powder), a model control group, and a group treated with Zhongjiao Zhenxuan pepper kernel oil. After a period of treatment with Zhongjiao Zhenxuan pepper kernel oil, fat-specific staining was performed on the zebrafish to observe changes in lipid accumulation. Simultaneously, a triglyceride kit was used to detect changes in triglyceride levels in the zebrafish.

[0057] The test results are as follows Figure 2 As shown in the zebrafish blood lipid phenotype diagram, the orange-red lipid droplets represent blood lipids. Combined with... Figure 3 As shown, the accumulation of blood lipids in the group taking triglycerides was significantly reduced compared to the model control group. Figure 3 The triglyceride content (mmol / gprot) of zebrafish was compared with that of the model control group. Specific data are shown in Table 4. The biochemical indicators show that the overall triglyceride content of zebrafish in the lipid-lowering agent group was significantly lower than that in the model control group. This demonstrates that the triple sustained-release system of pepper seed oil has a significant auxiliary lipid-lowering effect.

[0058] A comparative experiment involving 30 zebrafish in each group showed that the blood lipid accumulation and overall triglyceride levels in the group treated with Zhongjiao Zhenxuan pepper kernel oil were significantly reduced compared to the model control group. This experiment confirms that Zhongjiao Zhenxuan pepper kernel oil has a significant lipid-lowering effect.

[0059] (3) Blood glucose test A study on the hypoglycemic effect of Zhongjiao Zhenxuan pepper kernel oil was conducted using 6-month-old male zebrafish, divided into four groups: a control group (breeding water), a model group (1% glucose), a low-dose group (1% glucose + 66 mg / L Zhongjiao Zhenxuan pepper kernel oil), a high-dose group (1% glucose + 200 mg / L Zhongjiao Zhenxuan pepper kernel oil), and a positive control group (1% glucose + 5 mg metformin), with 10 zebrafish in each group. The zebrafish were cultured in 1% glucose (the control group used breeding water) for 7 days, with the water changed daily. From day 8, the treatment groups received Zhongjiao Zhenxuan pepper kernel oil extract (final concentrations of 66 and 200 mg / L, respectively) in their breeding water, while the positive control group received 5 mg metformin. This was continued for 7 consecutive days, with the water changed daily. After 7 days, remove the fish, wash away any residual glucose with clean water, place it in ice water (0℃), and remove it after it has stiffened. Dry the surface of the fish, cut off the tail 3-4 mm forward from the tail fin, and use the test strip to absorb blood from the tail. Use a blood glucose meter to measure the blood glucose level.

[0060] Experimental Results (Hypoglycemic Effect of Zhongjiao Selected Pepper Seed Oil): After 7 days of continuous culture in a 1% glucose solution, the blood glucose level of the model group zebrafish was 4.80 mmol / L, approximately 1.7 times that of the control group (P<0.05), indicating a successful zebrafish hyperglycemia model. The blood glucose level of the positive control group zebrafish was 3.13 mmol / L, a decrease of 34.8% compared to the model group (P<0.05), but still higher than the normal blood glucose level of 2.82 mmol / L in the control group. The blood glucose levels of the low-dose group and the high-dose group were 3.81 mmol / L and 3.02 mmol / L, respectively, representing decreases of 20.6% and 37.1% compared to the model group. The blood glucose level of the high-dose group was lower than that of the positive control group. (See Table 5 and...) Figure 4 As shown in the figure. This experiment successfully established a zebrafish hyperglycemia model and verified the hypoglycemic effect of Zhongjiao Zhenxuan pepper kernel oil. It confirms that the triple-release system pepper kernel oil has a significant auxiliary hypoglycemic effect.

[0061] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. The same or similar parts between the various embodiments can be referred to each other.

[0062] Although preferred embodiments of the present application have been described, those skilled in the art, upon learning the basic inventive concept, can make other changes and modifications to these embodiments. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments as well as all changes and modifications falling within the scope of the embodiments of the present application.

[0063] Finally, it should be noted that in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or terminal device that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or terminal device. Without further limitations, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or terminal device that includes said element.

[0064] The above provides a detailed description of the triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol, as well as its preparation method. Specific examples have been used to illustrate the principles and implementation methods of this application. The descriptions of the above embodiments are only for the purpose of helping to understand the method and core ideas of this application. At the same time, for those skilled in the art, there will be changes in the specific implementation methods and application scope based on the ideas of this application. Therefore, the content of this specification should not be construed as a limitation of this application.

Claims

1. A triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol (the "three highs") using pepper seed oil, characterized in that... The triple sustained-release system comprises, from the inside out: A pH-responsive gastric protective layer, through a sodium alginate-chitosan complex, responds to gastric acid-triggered expansion to form a gel barrier; Enzyme-responsive enteric coating, β-cyclodextrin-trypsin substrate peptide conjugate, responds to intestinal trypsin to specifically cleave peptide chains, releasing 30% of the active substance; A time-release colon-targeting layer, with acrylic resin Eudragit FS30D encapsulating nanoemulsion, continuously releases 65% of the active substances from the colon over 0–24 hours; The active substances include 20-30% α-linolenic acid, 6-8% nervonic acid, 2-3% hydroxy-α-sanshool from pepper seed oil, and the balance being fatty acids. Through synergistic regulation of the AMPK / mTOR pathway, they achieve the function of assisting in lowering blood pressure, blood sugar, and cholesterol.

2. The triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol as described in claim 1, characterized in that, The pH-responsive gastric protective layer, through a sodium alginate-chitosan complex, responds to gastric acid-triggered expansion to form a gel barrier, comprising: A pH-responsive gastric protective layer, consisting of a sodium alginate-chitosan complex with a mass ratio of 2:1 to 3:1, expands in response to gastric acid to form a gel barrier, with an α-linolenic acid sustained-release rate ≤5% / h.

3. The triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol as described in claim 1, characterized in that: The fatty acids include linoleic acid, oleic acid, and saturated fatty acids.

4. The triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol as described in claim 1, characterized in that: The preparation steps of the triple sustained-release system for assisting in lowering blood pressure, blood sugar, and cholesterol include: Nanoemulsion preparation steps: Nervonic acid nanoemulsion is prepared by high-pressure homogenization; Slow-release coating process: Using a fluidized bed bottom spraying process, under an environment with an inlet air temperature of 35-45℃, an atomization pressure of 0.7-1.0 bar, and a feed rate of 3-5 mL / min, the coating is performed using the acrylic resin Eudragit FS30D, resulting in a coating weight gain of 20%-25%. Product molding steps: Soft capsules are filled using a mass ratio of gelatin:glycerin:water of 5-4:3-2:3-2, with a disintegration time of 30-40 minutes.

5. The triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol as described in claim 4, characterized in that, The preparation of nervonic acid nanoemulsion by high-pressure homogenization includes: Nervonic acid nanoemulsions were prepared by high-pressure homogenization under an environment of 100MPa to 120MPa for 1 to 3 cycles. The particle size of the nervonic acid nanoemulsions was 80±5nm, the loading rate was ≥90 to 95%, and the zeta potential was -30mV to -35mV.

6. The triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol as described in claim 4, characterized in that, It also includes the preparation of active substance powders, specifically including: Mix 30g of pepper seed oil with an α-linolenic acid content of 20% with 5g of nervonic acid concentrate; Add 0.5% tocopherol and 0.3% rosmarinic acid compound antioxidant to the mixture; Nanoparticles were prepared using a W / O / W double emulsion method, and the active substance powder was obtained by spray drying.

7. The triple-release system for assisting in lowering blood pressure, blood sugar, and cholesterol as described in claim 4, characterized in that, The sustained-release coating step involves using a fluidized bed bottom spraying process, with an inlet air temperature of 35–45°C, an atomization pressure of 0.7–1.0 bar, and a feed rate of 3–5 mL / min, to coat the material using Eudragit FS30D acrylic resin. The coating weight gain is 20%–25%, and includes: Under conditions of bed temperature of 38℃, atomization pressure of 1.0 bar, and feed rate of 3 mL / min, Eudragit FS30D was sequentially coated to achieve a weight gain of 15%, β-cyclodextrin-peptide complex to achieve a weight gain of 7%, and sodium alginate layer to achieve a weight gain of 3%, with encapsulation efficiency ≥90%~95% and burst release rate <3%~5%.

8. A method for preparing pepper seed oil using a triple-release system to assist in lowering blood pressure, blood sugar, and cholesterol, characterized in that... Includes the following steps: (1) Preparation of active substance powder: 30g of pepper kernel oil with 20% α-linolenic acid content was mixed with 5g of nervonic acid concentrate, and 0.5% tocopherol and 0.3% rosmarinic acid composite antioxidant were added. Nanoparticles were prepared by W / O / W double emulsion method and spray dried to obtain active substance powder; (2) Nanoemulsion preparation: The active substance powder from step (1) is homogenized under high pressure and circulated for 1 to 3 cycles at 100 MPa to 120 MPa to prepare nervonic acid nanoemulsion with a particle size of 80 ± 5 nm. (3) Slow-release coating: Using fluidized bed bottom spraying process, under the conditions of inlet air temperature of 35-45℃, atomization pressure of 0.7-1.0 bar, and feed rate of 3-5 mL / min, acrylic resin Eudragit FS30D is used for coating, and the coating weight gain is 20%-25%; (4) Product molding: Soft capsules are filled using a mass ratio of gelatin: glycerin: water of 5-4: 3-2: 3-2, with a disintegration time of 30-40 minutes.

9. The preparation method according to claim 8, characterized in that, The specific conditions for the sustained-release layer coating are as follows: bed temperature 38℃, atomization pressure 1.0 bar, feed rate 3 mL / min, and sequential coating with acrylic resin Eudragit FS30D to achieve a weight gain of 15%, β-cyclodextrin-peptide complex to achieve a weight gain of 7%, and sodium alginate layer to achieve a weight gain of 3%.

10. An application of pepper seed oil based on the triple sustained-release system for assisting in lowering blood pressure, blood sugar, and cholesterol as described in any one of claims 1-7, characterized in that, The triple-release system provides sustained release in stages within the human digestive tract, specifically including: In the stomach: the release time was 2 hours, the release medium was 0.1N HCl, and the release rate was 8.2%. Small intestine stage: release time was 4 hours, release medium was pH 6.8 phosphate buffer, and release rate was 37.5%. Colonic phase: The release time was 18 hours, the release medium was pH 7.4 phosphate buffer and β-glucosidase, and the release rate was 54.3%.

Images