Method for entrapment of silymarin and escin composition by low-energy method and application of silymarin and escin composition in anti-allergic and anti-inflammatory products

Abstract

The invention discloses a method for entrapping a silymarin and escin composition by a low-energy method and application of the silymarin and escin composition in anti-allergic and anti-inflammatory products. The method comprises the following steps: mixing silymarin, escin and a solvent, heating and stirring until the silymarin, escin and the solvent are dissolved, adding an emulsifier, slowly dropwise adding grease, finally adding the balance of water, and cooling. The invention provides a method for preparing a stable and transparent nano-emulsion by comprehensively utilizing a phase composition method and a phase transformation method without the aid of strong shearing effects such as high-pressure homogenization, ultrasound or microjet and the like, and provides a method for preparing the stable and transparent nano-emulsion by applying a pharmaceutical preparation to wounds of patients and facilitating efficient and mild relieving of inflammation and discomfort.

Description

Technical Field

[0001] The invention belongs to the technical field of preparation of anti-allergic and anti-inflammatory products, and particularly relates to a method for encapsulating a silymarin and aescin composition by a low-energy method and application thereof in anti-allergic and anti-inflammatory products. Background Art

[0002] Silymarin (C 25 H 22 O 10 ) is a flavonoid lignin compound extracted from the fruit of the Asteraceae plant Silybum marianum. It is insoluble in water and oil and has a very low bioavailability. The molecule contains multiple functional groups such as phenolic hydroxyl groups. Studies have shown that silymarin is a safe and well-tolerated plant-derived drug with anti-inflammatory, antioxidant, anti-cancer and antiviral pharmacological activities. Aescin is a type of triterpenoid saponin. The basic structure is based on oleanane-type pentacyclic triterpenoids as the parent nucleus. It can significantly inhibit the exudation of inflammatory tissues, reduce its permeability, and has a significant effect on edema and damage of various tissues. However, it is also insoluble in water and oil and has a very low bioavailability.

[0003] How to improve the bioavailability of silymarin or aescin so as to effectively and gently relieve inflammation is a technical problem to be solved. Summary of the invention

[0004] The purpose of this section is to summarize some aspects of embodiments of the present invention and to briefly introduce some preferred embodiments.

[0005] As one aspect of the present invention, the present invention provides a method for encapsulating silymarin and aescin composition by a low-energy method, which comprises the following steps:

[0006] Step 1: Mix silymarin, aescin and solvent, heat and stir until dissolved;

[0007] Step 2: Add an emulsifier to the solution obtained in step 1 and stir evenly;

[0008] Step 3: Add oil dropwise to the solution obtained in step 2;

[0009] Step 4: adding water to the solution obtained in step 3, and cooling to room temperature to obtain an encapsulated composition emulsion;

[0010] Wherein, the oil includes one of lauroyl sarcosine isopropyl ester, hexyldecyl myristoyl methylamino propionate, and diisopropyl adipate.

[0011] As a preferred embodiment of the method for encapsulating silymarin and aescin composition by low-energy method of the present invention: in step 1, the solvent comprises one or more of dipropylene glycol, butylene glycol, hexylene glycol and 1,3-propylene glycol.

[0012] As a preferred embodiment of the method for encapsulating silymarin and aescin composition by low-energy method of the present invention: in step 1, the heating temperature is 78-80°C.

[0013] As a preferred embodiment of the method for low-energy encapsulation of silymarin and aescin composition described in the present invention: in step 2, the emulsifier includes one or more of Bacillus fermentation product, polyglycerol-10 myristate, polyglycerol-10 stearate, polyglycerol-6 caprylate, and sucrose laurate.

[0014] As a preferred embodiment of the method for encapsulating silymarin and aescin composition by low-energy method of the present invention: in step 2, the stirring is carried out at 73-76°C.

[0015] As a preferred embodiment of the method for encapsulating silymarin and aescin composition by low-energy method described in the present invention, the following ingredients are used: 6-8 parts of silymarin, 2-5 parts of aescin, 20-70 parts of solvent, 1-8 parts of emulsifier, 5-10 parts of oil, and the balance is water to 100 parts.

[0016] As a preferred embodiment of the method for encapsulating silymarin and aescin composition by low-energy method of the present invention: in step 2, the emulsifier is composed of Bacillus fermentation product and polyglycerol-6 caprylate.

[0017] As a preferred embodiment of the method for encapsulating silymarin and aescin composition by low-energy method of the present invention, the polyglycerol-6 caprylate is 1-3 parts and the Bacillus fermentation product is 0.5-5 parts.

[0018] Beneficial effects of the present invention: The present invention provides a low-energy dual-encapsulation method of silymarin and aescin and a mild wall material encapsulation into a nanoemulsion, which is used for applying a medical preparation to a patient's wound, and is conducive to efficiently and gently relieving inflammation and discomfort. The low-energy method of the present invention provides a method for preparing a stable and transparent nanoemulsion by comprehensively utilizing a phase composition method and a phase transition method without the aid of a strong shearing action such as high-pressure homogenization, ultrasound or microfluidization. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the technical solution of the embodiment of the present invention, the following briefly introduces the drawings required for describing the embodiment, wherein:

[0020] Figure 1 This is the particle size distribution diagram measured by Zeta potential and nanoparticle size analyzer.

[0021] Figure 2 It is the a* value of skin at different test times.

[0022] Figure 3 It is the blood flow density at different test time and depth.

[0023] Figure 4 This is a diagram of the encapsulation composition emulsion prepared in Example 1. DETAILED DESCRIPTION

[0024] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with specific embodiments.

[0025] The low-energy method encapsulation silymarin and aescin composition of the present invention comprises, by weight of raw materials, 3-8 parts of silymarin, 1-5 parts of aescin (CAS: 6805-41-O), 20-70 parts of dipropylene glycol, 1-3 parts of polyglycerol-6 caprylate, 0.5-5 parts of bacillus fermentation product, 5-10 parts of hexyldecyl myristoyl methylaminopropionate, and the balance of water.

[0026] The preparation method of the silymarin and aescin encapsulation composition is as follows:

[0027] Step 1: Mix silymarin, aescin and dipropylene glycol and heat to 78-80°C, and stir with a glass rod until dissolved;

[0028] Step 2: Add polyglycerol-6 caprylate, a fermentation product of Bacillus, to the dissolved solution in step 1, stir evenly with a glass rod and cool to 73-76°C;

[0029] Step 3: Slowly add hexyldecyl myristoyl methylamino propionate to step 2, stirring with a glass rod while adding;

[0030] Step 4: Quickly add the remaining water to step 3 and cool to room temperature.

[0031] Embodiment 1:

[0032] The encapsulation composition of the present invention is calculated by weight of raw materials: 8 parts of silymarin, 2 parts of aescin, 50 parts of dipropylene glycol, 2 parts of polyglycerol-6 caprylate (CAS No.: 51033-35-3), 2.5 parts of bacillus fermentation product, 5 parts of hexyldecyl myristoyl methylaminopropionate, and 100 parts of deionized water.

[0033] Step 1: Heat silymarin, aescin and dipropylene glycol to 80°C and stir until dissolved;

[0034] Step 2: Add polyglycerol-6 caprylate, Bacillus fermentation product (Beijing Yanwei Technology Co., Ltd., Evoglory TM Stir well and cool to 74℃;

[0035] Step 3: slowly drop hexyldecyl myristoyl methylamino propionate into the solution obtained in step 2, stirring rapidly while dropping, and the dropping speed is 0.02 g / s;

[0036] Step 4: Rapidly add the remaining water to step 3 and cool to room temperature to obtain an encapsulated composition emulsion.

[0037] Experimental results: The obtained packaging carrier is a transparent viscous liquid, and its particle size is measured by Zeta potential and nanoparticle size analyzer; its high and low temperature stability is measured by placing it in a high and low temperature cycle (-15℃~48℃) constant temperature box. Figure 1 The particle size distribution diagram measured by Zeta potential and nanoparticle size analyzer shows that the nanoemulsion has a relatively concentrated particle size distribution and good dispersion. It can be seen from the figure that the average particle size of the nanoemulsion is between 50-55nm.

[0038] Stability test results: No significant changes after one month of circulation.

[0039] Patch test: Patch test method: Select qualified patch test equipment, use the closed patch test method, place 0.020-0.025mL (0.020-0.025g) of the test substance in the patch tester, and apply it to the curved side of the subject's forearm with a low-allergenic tape. Remove the test substance after 24 hours, and observe the skin reaction 30 minutes after removing the test substance patch tester after the indentation disappears. If the result is negative, observe again 24h and 48h after the patch test. Record the reaction results according to the skin adverse reaction grading standard table (as shown in the following table).

[0040]

[0041] Experimental results:

[0042] The results of human skin patch test are shown in the following table. O of the 30 people had adverse skin reactions.

[0043] Table 1 Patch test results

[0044]

[0045] In vitro hyaluronidase inhibition assay:

[0046] Hyaluronidase is strongly correlated with inflammation and allergy, and is a participant in type I allergic reactions. Many anti-allergic drugs have a strong inhibitory effect on hyaluronidase activity. Drugs that affect the release of histamine from mast cells can regulate hyaluronidase activity. There is a good direct correlation between the inhibition of hyaluronidase activity and the inhibition of histamine release from mast cells. In this experiment, the encapsulated composition was diluted into a 50% (w / w) aqueous solution as a test sample, and four sample tubes A, B, C, and D were set up, including tube A (50U / mL hyaluronidase + test sample + 0.5mg / mL sodium hyaluronate (molecular weight 10W Da)), tube B (acetic acid buffer solution + test sample + acetate buffer solution), tube C (50U / mL hyaluronidase + deionized water + 0.5mg / mL sodium hyaluronate), and tube D (acetic acid buffer + deionized water + acetate buffer). After reacting in a 37°C water bath, acetylacetone solution was added under alkaline conditions and reacted in a boiling water bath for 15min, then ice bathed for 10min, and then returned to room temperature and p-dimethylaminobenzaldehyde solution was added for color development. After sufficient shaking, anhydrous ethanol was added to make the volume 8mL, mixed, and placed at room temperature for 30min. The absorbance was measured at 530nm, and the hyaluronidase inhibition rate was calculated according to the following formula. Deionized water was used instead of the test sample for the negative control.

[0047] Hyaluronidase inhibition rate % = {(CD)-(AB)} / (CD)×100%

[0048] The results of the significant difference P value analysis are shown in Table 2, which shows that the present carrier has a strong inhibitory effect on the activity of hyaluronidase.

[0049] Table 2 Hyaluronidase inhibition rate and difference analysis

[0050] Test items Negative control Test samples P-value Significance of difference Inhibition rate % 1.17±O.15 90.2±0.65 <0.001 Very significant

[0051] Human efficacy evaluation:

[0052] Measure the changes in skin redness a* value and blood flow velocity within a certain period of time after the skin is stimulated by capsaicin. After the subject's visit, wash the curved side of the left and right forearms and wipe them clean with a face towel; balance in a constant temperature and humidity laboratory for 30 minutes, enter the internal test room, use a marker to mark the test areas of each group, and each test area is at least 1 cm apart. Drinking water, walking, and touching the arm are prohibited during this period; use the SkinColorCatch probe to detect the a* value of each test area, measure 3 times and take the average value, which is recorded as the initial value; use a pipette to draw 30μL of capsaicin solution and place it in the spot tester, and apply it to each test area for 15min; remove the spot tester, balance for 5min, detect the a* value, and use VISIA to take skin photos at the same time, and use a skin structure imager to take subcutaneous blood flow density, which is recorded as after stimulation (Omin); use a pipette to draw 25μL of test sample, use a latex finger cot to evenly apply the sample to the test area, apply it evenly until absorbed, and start timing;

[0053] At 15min, 30min and 60min, the a* value was measured and VISIA photos were taken; at 30min and 60min, the subcutaneous blood flow density was photographed; and the statistical data were collected.

[0054] The a* value can reflect the change in redness of the skin surface after stimulation. Figure 2 It can be seen that compared with the initial value, the a* value of the sample increased significantly after capsaicin stimulation (Omin) (p<0.001), indicating that capsaicin can induce skin irritation. Compared with Omin, after the sample was tested with the encapsulated composition for 15min, 30min, and 60min, the a* values ​​of both groups showed a significant decrease.

[0055] The blood flow density of the sample group at 0min, 30min, and 60min after stimulation is as follows Figure 3 As shown ＊＊ , ＊＊＊ Distribution represents p<0.01, p<0.001). Compared with Omin (after stimulation), the blood flow density at subcutaneous 0.15, 0.20, 0.25, 0.30, 0.35, and 0.40 mm showed a significant decrease at 30min and 60min after the test sample was tested with the encapsulated composition.

[0056] This indicates that the silymarin and aescin composition encapsulated by the present invention has good anti-inflammatory, anti-allergic and soothing effects.

[0057] Comparative Example 1:

[0058] The raw material ratio is: by mass, 6 parts of silymarin, 2 parts of aescin, 50 parts of dipropylene glycol, 2.5 parts of Bacillus fermentation product, 5 parts of hexyldecyl myristoyl methylaminopropionate, and 100 parts of water. The preparation method is the same as that of Example 1 (except that polyglycerol-6 caprylate is not added).

[0059] Experimental results: a white emulsion was obtained, from which solids precipitated after standing, indicating that the encapsulation was not complete and the emulsification system was unstable.

[0060] Comparative Example 2:

[0061] The raw material ratio is: by mass, 6 parts of silymarin, 2 parts of aescin, 50 parts of dipropylene glycol, 2 parts of polyglycerol-6 caprylate, 5 parts of hexyldecyl myristoyl methylaminopropionate, and 100 parts of water. The preparation method is the same as that of Example 1 (only without adding the Bacillus fermentation product).

[0062] Experimental results: A translucent emulsion was obtained, and a small amount of solid precipitated after standing, indicating that the encapsulation was not complete and the emulsification system was unstable.

[0063] Comparative Example 3:

[0064] The raw material ratio is: by mass, 6 parts of silymarin, 2 parts of aescin, 50 parts of dipropylene glycol, 2 parts of polyglycerol-6 caprylate, 2.5 parts of Bacillus fermentation product, and 100 parts of water. The preparation method only omits step 3 of adding hexyldecyl myristoyl methylamino propionate dropwise, and the rest is the same as in Example 1.

[0065] Experimental results: Solids precipitated after adding water in the final step, indicating that the emulsified system was unstable.

[0066] Comparative Example 4:

[0067] The raw material ratios of the encapsulated composition are the same as those in Example 1, and the preparation process is:

[0068] Step 1: Heat silymarin, aescin and dipropylene glycol to 80°C and stir until dissolved;

[0069] Step 2: Add water, polyglycerol-6 caprylate and Bacillus fermentation product to the solution obtained in step 1, keep the temperature at 74°C and stir evenly;

[0070] Step 3: Slowly add hexyldecyl myristoyl methylamino propionate to the solution obtained in step 2, stirring rapidly at a rate of 0.02 g / s, and cool to room temperature after stirring evenly.

[0071] Experimental results: The obtained emulsion is white and opaque, indicating that the particle size is large. It breaks after standing, indicating that the emulsion system is unstable. Since water is added first, the system cannot form an intermediate phase oil gel, and the emulsifier has been dissolved in the water phase first, forming a large particle size emulsion, which eventually breaks and makes the emulsion unstable.

[0072] Comparative Example 5:

[0073] The raw material ratio is: by mass, 6 parts of silymarin, 2 parts of aescin, 50 parts of dipropylene glycol, 2 parts of polyglycerol-6 caprylate, 2.5 parts of Bacillus fermentation product, 5 parts of hydrogenated polyisobutylene, and 100 parts of water. The preparation method is the same as that of Example 1. Experimental results: A translucent emulsion is obtained, and a small amount of solid is precipitated after standing, indicating that the encapsulation is not complete and the emulsification system is unstable.

[0074] Comparative Example 6:

[0075] The raw material ratio is: by mass, 8 parts of silymarin, 2 parts of aescin, 50 parts of dipropylene glycol, polyglycerol-4 caprate, 2.5 parts of Bacillus fermentation product, 5 parts of hexyldecyl myristoyl methylaminopropionate, and 100 parts of water. The preparation method is the same as that of Example 1.

[0076] Experimental results: a white emulsion was obtained, from which solids precipitated after standing, indicating that the encapsulation was not complete and the emulsification system was unstable.

[0077] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A method for encapsulating a silymarin and aescin composition by a low-energy method, characterized in that: It consists of the following steps: Step 1: Mix silymarin, aescin and solvent, heat and stir until dissolved; Step 2: Add an emulsifier to the solution obtained in step 1 and stir evenly; Step 3: Add oil dropwise to the solution obtained in step 2; Step 4: adding water to the solution obtained in step 3, and cooling to room temperature to obtain an encapsulated composition emulsion; Wherein, the oil includes one of lauroyl sarcosine isopropyl ester, hexyldecyl myristoyl methylamino propionate, and diisopropyl adipate.

2. The method for encapsulating silymarin and aescin composition by low-energy method according to claim 1, characterized in that: In step 1, the solvent includes one or more of dipropylene glycol, butylene glycol, hexylene glycol, and 1,3-propylene glycol.

3. The method for encapsulating silymarin and aescin composition by low-energy method according to claim 1 or 2, characterized in that: In step 1, the heating temperature is 78-80°C.

4. The method for encapsulating silymarin and aescin composition by low-energy method according to claim 1 or 2, characterized in that: In step 2, the emulsifier includes one or more of Bacillus fermentation product, polyglycerol-10 myristate, polyglycerol-10 stearate, polyglycerol-6 caprylate, and sucrose laurate.

5. The method for encapsulating silymarin and aescin composition by low-energy method according to claim 1 or 2, characterized in that: In step 2, the stirring is carried out at 73-76°C.

6. The method for encapsulating silymarin and aescin composition by low-energy method according to claim 1 or 2, characterized in that: Calculated by weight, the composition includes 6 to 8 parts of milk thistle, 2 to 5 parts of aescin, 20 to 70 parts of solvent, 1 to 8 parts of emulsifier, 5 to 10 parts of oil, and the balance is water to 100 parts.

7. The method for encapsulating silymarin and aescin composition by low-energy method according to claim 6, characterized in that: In step 2, the emulsifier consists of bacillus fermentation product and polyglycerol-6 caprylate.

8. The method for encapsulating silymarin and aescin composition by low-energy method according to claim 7, characterized in that: The polyglycerol-6 caprylate is 1-3 parts and the bacillus fermentation product is 0.5-5 parts.

9. The encapsulated composition emulsion prepared by the method for encapsulating silymarin and aescin composition by a low-energy method according to claim 1.

10. Use of the encapsulated composition emulsion prepared by the method for encapsulating silymarin and aescin composition by a low-energy method according to claim 1 in anti-allergic and anti-inflammatory products.

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