Bisabolol nanoemulsion as well as preparation method and application thereof
By using specific oil phase and emulsifier in α-Red Mycohol to form nanoemulsion, the problem of insufficient bioavailability and biological activity is solved, significantly improving its anti-allergic and anti-inflammatory effects, and enhancing stability and sustained release characteristics.
Patent Information
- Application Number
- CN202411556129.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-04
- Publication Date
- 2024-12-27
AI Technical Summary
The lipophilicity and low water solubility of α-Redomycin limits its bioavailability and biological activity, resulting in poor efficacy in anti-allergic and anti-inflammatory.
A combination of specific types of oil phase and emulsifiers is used to form red mycoal nanoemulsions to improve its solubility and transdermal absorption, including caprylic caprylate triglyceride, triglyceride tri(ethylhexanoate) ester, and ethyl palmitate The combination of the oil phases such as hexyl ester and emulsifiers such as polyoxyethylene 40 hydrogenated castor oil and Tween-80 to form a nanoemulsion system, which significantly improves the bioavailability and biological activity of red mycohol.
It significantly improves the transdermal absorption rate and biological activity of erythrol, enhances its anti-allergic and anti-inflammatory effects, inhibits hyaluronidase, increases the content of IL-10, improves stability and sustained release characteristics .
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Figure CN119185202A_ABST
Abstract
Description
Technical Field
[0001] The present application relates to the field of biomedical technology, and in particular to a bisabolol nanoemulsion and a preparation method and application thereof. Background Art
[0002] The inflammatory response is one of the important mechanisms of the body's self-protection, which aims to remove damaged tissues or pathogens and promote wound healing. Inflammation can be divided into acute inflammation and chronic inflammation. The former is mainly caused by exogenous factors (microorganisms, viral infections, etc.), with rapid onset, often manifested as redness, swelling, pain and fever; the latter is mainly caused by endogenous factors (inflammatory mediators, etc.), with slow and continuous onset, which may eventually lead to tissue damage or fibrosis. These reaction signals are transmitted throughout the body through the blood and lymph, causing or aggravating the occurrence of related diseases. In the field of skin care, after the skin receives external stimuli, the skin will have a series of inflammatory reactions, such as sensitive skin redness, fever, red acne marks, etc.
[0003] α-Bisabolol is a monocyclic unsaturated sesquiterpene alcohol derived from plants such as chamomile, and is considered to be the main substance that makes chamomile work. The chemical formula of α-bisabolol is C15H260, and the structural formula is 2R)-6-methyl-2-[(1R)-4-methylcyclohex-3-en-1-yl]hept-5-en-2-ol. It is a colorless and transparent liquid, almost insoluble in water but slightly soluble in ethanol. Since α-bisabolol has multiple activities such as anti-inflammatory, analgesic, and blood circulation, it ranks second as an ingredient in sensitive skin care products.
[0004] Due to the lipophilicity and low water solubility of α-bisabolol, the bioavailability and biological activity of bisabolol are limited. Summary of the invention
[0005] The purpose of the present application is to overcome the deficiencies of the above-mentioned prior art and to provide a bisabolol nanoemulsion and a preparation method and application thereof.
[0006] To achieve the above purpose, the technical solution adopted by this application is:
[0007] The present application provides a bisabolol nanoemulsion, wherein the bisabolol nanoemulsion comprises an oil phase, an emulsifier, a co-emulsifier and bisabolol;
[0008] The oil phase comprises at least one of caprylic capric triglyceride, tri(ethylhexanoin)glyceride and ethylhexyl palmitate;
[0009] The emulsifier includes at least one of polyoxyethylene 40 hydrogenated castor oil, Tween-80, and castor oil polyoxyethylene (30) ether;
[0010] The mass ratio of the oil phase to the emulsifier is (1.5-5):(1-4.5).
[0011] Nanoemulsion is a thermodynamically stable system composed of an oil phase, an aqueous phase and a surfactant, and its average particle size is 1 to 100 nm. The present application adopts the above nanoemulsion system to solve the solubility problem of bisabolol and improve the bioavailability.
[0012] The present application adopts a specific type of oil phase and emulsifier, which can significantly improve the solubility of bisabolol, and has a sustained release and targeting effect, thereby improving the transdermal absorption and biological activity of bisabolol. The bisabolol nanoemulsion obtained in the present application is superior to bisabolol in terms of stability, transdermal absorption, anti-allergy, anti-inflammatory, etc.
[0013] As a preferred embodiment of the bisabolol nanoemulsion described in the present application, the oil phase is caprylic capric triglyceride, the emulsifier is Tween-80, and the mass ratio of caprylic capric triglyceride to Tween-80 is (1.5-2.5): (3.5-4.5).
[0014] As a preferred embodiment of the bisabolol nanoemulsion described in the present application, the oil phase is ethylhexyl palmitate, the emulsifier is polyoxyethylene 40 hydrogenated castor oil, and the mass ratio of ethylhexyl palmitate to polyoxyethylene 40 hydrogenated castor oil is (3-4): (1-2).
[0015] As a preferred embodiment of the bisabolol nanoemulsion described in the present application, the oil phase is caprylic capric triglyceride, the emulsifier is castor oil polyoxyethylene (30) ether, and the mass ratio of caprylic capric triglyceride to castor oil polyoxyethylene (30) ether is (3.5-5): (1-2.5).
[0016] As a preferred embodiment of the bisabolol nanoemulsion described in the present application, the oil phase is tri(ethylhexanoin)glyceryl, the emulsifier is polyoxyethylene 40 hydrogenated castor oil, and the mass ratio of tri(ethylhexanoin)glyceryl to polyoxyethylene 40 hydrogenated castor oil is (3.5-5):(1-2.5).
[0017] When the oil phase and the emulsifier are respectively matched with the above-mentioned types, a bisabolol nanoemulsion can be formed, which can effectively dissolve bisabolol, improve the transdermal absorption rate and biological activity of bisabolol, and thus enhance the anti-allergic and anti-inflammatory effects of bisabolol.
[0018] Specifically, the present application uses the above-mentioned types of oil phase and emulsifier respectively and combines them in a specific mass ratio, which can effectively inhibit hyaluronidase and increase the content of IL-10. Therefore, the bisabolol nanoemulsion prepared in the present application has good anti-allergic and anti-inflammatory effects.
[0019] When the oil phase and the emulsifier are combined in a mass ratio outside the above range, nanoemulsion cannot be formed, the dissolution of bisabolol cannot be promoted, and the transdermal absorption rate of bisabolol is low, resulting in poor anti-allergic and anti-inflammatory effects of bisabolol preparations.
[0020] As a preferred embodiment of the bisabolol nanoemulsion described in the present application, the co-emulsifier includes propylene glycol.
[0021] As a preferred embodiment of the bisabolol nanoemulsion described in the present application, the mass ratio of the bisabolol to the emulsifier is (0.5-1.5): (1-4.5).
[0022] When the mass ratio of bisabolol and the emulsifier is as mentioned above, a bisabolol nanoemulsion with better anti-allergic and anti-inflammatory effects can be obtained.
[0023] The present application also provides a method for preparing the above-mentioned bisabolol nanoemulsion, comprising the following steps:
[0024] Dissolve bisabolol in the oil phase, then add an emulsifier and an emulsifier assistant, mix and stir, slowly drop water into the oil phase, continue stirring at room temperature, and prepare a bisabolol nanoemulsion.
[0025] As a preferred embodiment of the method for preparing the bisabolol nanoemulsion described in the present application, the stirring speed is 2000-3000 r / min, and the stirring time is 30 min-60 min.
[0026] The present application also provides the use of the above-mentioned bisabolol nanoemulsion in the preparation of anti-allergic or anti-inflammatory products.
[0027] Compared with the prior art, the beneficial effects of this application are:
[0028] The present application provides a bisabolol nanoemulsion and a preparation method and application thereof. The present application adopts a specific type of oil phase and emulsifier, which can significantly improve the solubility of bisabolol, and has a sustained release and targeting effect, thereby improving the transdermal absorption and biological activity of bisabolol. The bisabolol nanoemulsion obtained in the present application not only has good stability and transdermal absorption rate, but also can better inhibit hyaluronidase and increase the content of IL-10, and is superior to bisabolol in anti-allergy, anti-inflammatory and other aspects. BRIEF DESCRIPTION OF THE DRAWINGS
[0029] Figure 1 This is the absorption result diagram of bisabolol nanoemulsion in the transdermal absorption test;
[0030] Figure 2 The transdermal absorption of bisabolol nanoemulsion and bisabolol solution in the transdermal absorption test;
[0031] Figure 3 This is the result diagram of hyaluronidase inhibition rate of bisabolol nanoemulsion and bisabolol solution;
[0032] Figure 4 This is a graph showing the effects of bisabolol nanoemulsion and bisabolol solution on IL-10. DETAILED DESCRIPTION
[0033] In order to better illustrate the purpose, technical solutions and advantages of the present application, the present application will be further described below in conjunction with the accompanying drawings and specific embodiments.
[0034] In the following examples and comparative examples, the experimental methods used are conventional methods unless otherwise specified, and the materials, reagents, etc. used are all commercially available unless otherwise specified, and the components and raw materials used in each parallel experiment are of the same type.
[0035] Example 1: Oil phase screening
[0036] In this example, the supersaturation method was used. Seven oil phases (10 g each) were placed in stoppered test tubes (three parallel copies), 2 g of bisabolol was added, and the mixture was placed in a 37°C constant temperature oscillator. After oscillation for 48 hours, the sample was centrifuged and the supernatant was diluted with methanol to an appropriate concentration (20 mg / g) as the test solution. The solubility of the drug in each excipient was determined by HPLC.
[0037] Solubility formula = 1-(M1-M2 / M1)*100%.
[0038] Note: M1 is the mass of supernatant, M2 is the mass of bisabolol.
[0039] The results are shown in Table 1.
[0040] Table 1
[0041]
[0042] From the conclusion of Table 1, it can be seen that isooctyl palmitate has the strongest ability to dissolve bisabolol, followed by caprylic capric triglyceride. The rest can effectively dissolve bisabolol.
[0043] Example 2: Emulsifier Screening
[0044] In this example, the appearance is used as an evaluation index to compare the compatibility between the oil phase (caprylic capric triglyceride, GTCC) and different emulsifiers and the appearance of the formed nanoemulsion, as shown in Table 2.
[0045] Table 2
[0046]
[0047] Conclusion: According to the compatibility, polyoxyethylene 40 hydrogenated castor oil, Tween-80 and castor oil polyoxyethylene (30) ether were subsequently studied.
[0048] Examples 3 to 14 and Comparative Examples 1 to 14, a bisabolol nanoemulsion and a preparation method thereof
[0049] The present embodiments 3 to 14 and comparative examples 1 to 14 provide a bisabolol nanoemulsion, and the specific formula is shown in Table 3-Table 4.
[0050] Dissolve bisabolol in the oil phase and mix well, then add emulsifier and co-emulsifier, stir at 2000r / min for half an hour, slowly drop water into the oil phase, continue stirring at room temperature for 1 hour, and prepare bisabolol nanoemulsion.
[0051] Table 3 (mass percentage)
[0052]
[0053]
[0054] Table 4
[0055]
[0056]
[0057] As can be seen from the above table, the formulations of Examples 3 to 14 of the present application can form nanoemulsions, while the formulations of Comparative Examples 1 to 14 cannot form nanoemulsions, because the present application needs to consider the compatibility of bisabolol and the oil phase, and the HLB value of the oil phase and the emulsifier must be balanced to form. When the oil phase and the emulsifier are respectively matched with the types of the present application, a bisabolol nanoemulsion can be formed, which can effectively dissolve bisabolol, improve the transdermal absorption rate and biological activity of bisabolol, and thus improve the anti-allergic and anti-inflammatory effects of bisabolol. When other types of oil phases and emulsifiers are used, and when they are matched with a mass ratio not within the above range, nanoemulsions cannot be formed, the dissolution of bisabolol cannot be promoted, and the transdermal absorption rate of bisabolol is low, resulting in poor anti-allergic and anti-inflammatory effects of bisabolol.
[0058] The following test examples use Example 1 as the test object to conduct stability, in vitro, and efficacy tests.
[0059] Test Example 1: Stability Evaluation
[0060] Zeta potential is defined as the electric potential of particles on the sliding plane in a liquid medium. Zeta potential indicates the surface charge and electrostatic repulsion or attraction between particles. Zeta potential affects the colloidal stability, aggregation behavior, flocculation rate, and interaction with biological membranes of lipid nanoparticles. In general, a higher Zeta potential (positive or negative) means higher electrostatic repulsion and better stability. PDI stands for Polydispersity Index, which is a parameter that describes the uniformity of particle size distribution. The smaller the PDI value, the more uniform the particle size distribution; conversely, the larger the PDI value, the more uneven the particle size distribution. Therefore, PDI is an important indicator to measure the uniformity of particle size distribution.
[0061] Method: Take 0.2 mL of the bisabolol nanoemulsion of Example 1, dilute it to 2 mL with deionized water, and use the Malvern nanoparticle size analyzer to measure the average particle size and dispersion index (PDI) of the nanoemulsion. The test conditions are as follows: the equilibrium time is set to 60 s, the scanning interval is 10 s, the temperature is set to 25 ° C, and each sample is analyzed in parallel 3 times. The indicators for evaluating the stability of the nanoemulsion according to the particle size, Zeta potential, and PDI are shown in Table 5.
[0062] Table 5
[0063]
[0064]
[0065] Test Example 2: Transdermal Absorption Test
[0066] method:
[0067] 1) Weigh bisabolol nanoemulsion, dissolve it in pure water, use pure water as blank, and scan at a wavelength of 185-400 nm.
[0068] 2) Using a modified Franz diffusion apparatus, pig skin was fixed between the drug administration pool and the receiving pool. The effective penetration area of the diffusion pool was 2.54 cm 2 , the stratum corneum faces the supply chamber, and the dermis faces the receiving chamber. Weigh 1 ml of sample (the bisabolol nanoemulsion or bisabolol solution of Example 1 (bisabolol concentration is 20 mg / g)), place it in the dosing pool, and apply it evenly on the ex vivo pig skin of the same size. Fix the upper and lower chambers, keep the temperature at (37±0.5)℃, set the diffusion cell speed to 200r / min, stir continuously, and take 2.0mL of receiving solution after 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12 hours, respectively, and replenish the same temperature and equal volume of receiving solution at the same time. Centrifuge the receiving solution, take the supernatant and filter it with a microporous filter membrane, measure the absorbance of the filtrate according to the given chromatographic conditions, and calculate the cumulative permeation per unit area Q.
[0069]
[0070] Where: Cn: drug mass concentration in the receiving solution at the nth sampling point;
[0071] V: volume of receiving pool;
[0072] S: effective diffusion area (2.54cm 2 ).
[0073] like Figure 1-Figure 2 As shown, in the transdermal absorption test, the transdermal absorption effect of the bisabolol nanoemulsion of the present application is better than that of the bisabolol solution.
[0074] Test Example 3: Hyaluronidase Inhibition Rate
[0075] Hyaluronic acid (HA) is a glycosaminoglycan present in the extracellular matrix, composed of repeating units of N-acetylglucosamine and glucuronic acid. When cancer, inflammation, and oxidative stress occur, the dynamic balance of the extracellular matrix is disrupted, and endogenous high molecular weight HA is rapidly degraded by hyaluronidase and reactive oxygen species, resulting in an increase in low molecular weight (≤250kDa) HA, stimulating the production of proinflammatory cytokines, and subsequently increasing the risk of disease. Therefore, the balance between the synthesis and degradation of HA is an important factor in maintaining health.
[0076] Hyaluronidase is a participant in type I allergic reactions. Hyaluronidase is strongly correlated with inflammation and allergy. Studies have reported that various drugs that release histamine from mast cells can regulate hyaluronidase activity. Some anti-allergic drugs have strong inhibitory effects on hyaluronidase activity. Therefore, inhibition of hyaluronidase activity is used as an indicator for studying anti-allergic effects.
[0077] method:
[0078] 1. Reagents and Instruments
[0079] 1. Hyaluronidase: concentration 500U / ml, freshly prepared and used, not overnight, use acetate buffer as solvent;
[0080] 2. Sodium hyaluronate: 0.5 mg / ml, prepared once, used multiple times, using acetate buffer as solvent;
[0081] 3. Buffer: Solution A (0.2 mol / L acetic acid 11.55 ml glacial acetic acid dissolved in 1 L distilled water) 4.8 ml solution B (0.2 mol / L sodium acetate 16.4 g anhydrous sodium acetate or 27.2 g sodium acetate trihydrate dissolved in 1 L distilled water) 45.2 ml, mix and dilute to 100 ml to prepare an acetate buffer solution with a pH of 5.6;
[0082] 4. Acetylacetone solution: Mix 50ml 1.0mol / L sodium carbonate solution and 3.5ml acetylacetone solution evenly (prepare immediately before use)
[0083] 5. P-DAB color developer: 0.8 g p-dimethylaminobenzaldehyde is dissolved in 15 ml concentrated hydrochloric acid and 15 ml anhydrous ethanol and mixed evenly.
[0084] 6. Calcium chloride solution CaCl2: 2.5 mol / L
[0085] 7. Sodium hydroxide solution NaOH: 5 mol / L
[0086] The reference concentration of the sample (the bisabolol nanoemulsion or bisabolol solution of Example 1) is 0.8 mg / ml.
[0087] 2. The test steps are shown in Table 6:
[0088] Table 6
[0089]
[0090]
[0091] Hyaluronidase inhibition rate (%) = [1-(AB) / (CD)] × 100%.
[0092] like Figure 3 As shown, the conclusion is: the hyaluronidase inhibition rate of bisabolol nanoemulsion is higher than that of bisabolol solution.
[0093] Test Example 4: Efficacy Test
[0094] Interleukin 10 (IL-10), also known as cytokine synthesis inhibitory factor (CSIF), is a pleiotropic cytokine that can exert immunosuppressive or immunostimulatory effects in various types of cells. In terms of inflammatory response, IL-10 can downregulate the expression of major histocompatibility antigen II on the surface of monocytes, reduce their antigen presentation function, downregulate T lymphocyte activity, and inhibit the activation, migration and adhesion of inflammatory cells; at the same time, IL-10 can also inhibit the synthesis and release of inflammatory factors.
[0095] experiment:
[0096] Cell culture: Mouse macrophage RAW264.7 cells were cultured in DMEM complete medium in a 5% CO2, 37°C cell culture incubator;
[0097] Cell grouping and treatment: RAW264.7 cells in the logarithmic growth phase were inoculated in a 6-well plate at a density of 5x10% cells / well. After 24 hours of attachment in a 37°C cell culture incubator, the original culture medium was discarded, washed twice with PBS, and stimulated with drugs for 24 hours respectively. The cells were collected for subsequent experiments. The groups were set as a blank control group (culture medium treatment), a DEX positive group (100ug / mL dex) and a bisabolol nanoemulsion drug treatment (0.5mg / ml, 0.65mg / ml, 0.8mg / ml, prepared with water).
[0098] like Figure 4 Compared with the bisabolol solution, the bisabolol nanoemulsion prepared in the present application has a greater effect on IL-10 and has a better effect of inhibiting inflammation.
[0099] Examples 2 to 9 were subjected to stability, in vitro, and efficacy tests, and the results were similar to those of Example 1.
[0100] The present application adopts a specific type of oil phase and emulsifier, which can significantly improve the solubility of bisabolol, and has a sustained release and targeting effect, thereby improving the transdermal absorption and biological activity of bisabolol. The bisabolol nanoemulsion obtained in the present application is superior to the simple bisabolol solution in terms of stability, transdermal absorption, anti-allergy, anti-inflammatory, etc.
[0101] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present application rather than to limit the scope of protection of the present application. Although the present application has been described in detail with reference to the preferred embodiments, ordinary technicians in this field should understand that the technical solution of the present application can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present application.
Claims
1. A bisabolol nanoemulsion, characterized in that: The bisabolol nanoemulsion comprises an oil phase, an emulsifier, a co-emulsifier and bisabolol; The oil phase comprises at least one of caprylic capric triglyceride, tri(ethylhexanoin)glyceride and ethylhexyl palmitate; The emulsifier includes at least one of polyoxyethylene 40 hydrogenated castor oil, Tween-80, and castor oil polyoxyethylene (30) ether; The mass ratio of the oil phase to the emulsifier is (1.5-5): (1-4.5).
2. The bisabolol nanoemulsion according to claim 1, characterized in that The oil phase is caprylic capric triglyceride, the emulsifier is Tween-80, and the mass ratio of caprylic capric triglyceride to Tween-80 is (1.5-2.5): (3.5-4.5).
3. The bisabolol nanoemulsion according to claim 1, characterized in that The oil phase is ethylhexyl palmitate, the emulsifier is polyoxyethylene 40 hydrogenated castor oil, and the mass ratio of ethylhexyl palmitate to polyoxyethylene 40 hydrogenated castor oil is (3-4): (1-2).
4. The bisabolol nanoemulsion according to claim 1, characterized in that The oil phase is caprylic acid capric acid triglyceride, the emulsifier is castor oil polyoxyethylene (30) ether, and the mass ratio of caprylic acid capric acid triglyceride to castor oil polyoxyethylene (30) ether is (3.5-5): (1-2.5).
5. The bisabolol nanoemulsion according to claim 1, characterized in that: The oil phase is tri(ethylhexanoic acid)glyceryl, the emulsifier is polyoxyethylene 40 hydrogenated castor oil, and the mass ratio of tri(ethylhexanoic acid)glyceryl to polyoxyethylene 40 hydrogenated castor oil is (3.5-5):(1-2.5).
6. The bisabolol nanoemulsion according to claim 1, characterized in that: The co-emulsifier includes propylene glycol.
7. The bisabolol nanoemulsion according to claim 1, characterized in that The mass ratio of the bisabolol to the emulsifier is (0.5-1.5): (1-4.5).
8. A method for preparing the bisabolol nanoemulsion according to any one of claims 1 to 7, characterized in that: The following steps are involved: Dissolve bisabolol in the oil phase, then add an emulsifier and an emulsifier assistant, mix and stir, slowly drop water into the oil phase, continue stirring at room temperature, and prepare a bisabolol nanoemulsion.
9. The method for preparing the bisabolol nanoemulsion according to claim 8, characterized in that: The stirring speed is 2000-3000 r / min, and the stirring time is 30-60 min.
10. Use of the bisabolol nanoemulsion according to any one of claims 1 to 7 in the preparation of anti-allergic or anti-inflammatory products.