Chrysanthemum-lactone-loaded camellia-seed oil nano-microemulsion as well as preparation method and application of parthenolide-loaded camellia-seed oil nano-microemulsion

Abstract

The invention belongs to the technical field of nano-carriers, and particularly relates to a parthenolide-loaded camellia-seed oil nano-microemulsion as well as a preparation method and application thereof. The nano microemulsion is prepared from the following raw materials in parts by weight: 3 to 8 parts of parthenolide, 17 to 23 parts of camellia-seed oil, 5 to 12 parts of caprylic / capric triglyceride, 2 to 8 parts of polyglycerol-10 myristate, 0.5 to 2 parts of tocopheryl acetate, 23 to 35 parts of glycerol, 0.3 to 0.8 part of lecithin, 5 to 15 parts of propylene glycol and 20 to 50 parts of water, and the mass ratio of the parthenolide to the camellia-seed oil is 1 to (3 to 6). The nano microemulsion prepared by the invention is high in encapsulation efficiency, stable in system and good in transdermal absorption amount, and the bioavailability of the parthenolide and the camellia-seed oil is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of nanocarriers, and particularly relates to a parthenolide-loaded camellia seed oil nano-microemulsion, a preparation method and an application thereof. Background Art

[0002] Sensitive skin is a highly reactive condition manifested under physiological or pathological conditions. It is often described as a syndrome in which the skin produces subjective symptoms such as stinging, burning, pain, itching, tightness, and tingling in response to routine stimuli. Although the skin may appear normal in appearance, it may sometimes be accompanied by objective signs such as erythema and scaling. The uncomfortable symptoms of sensitive skin result from the combined effects of neurovascular hyperreactivity caused by impaired barrier function and neurogenic inflammation. Currently, due to the influence of the environment and unfavorable living conditions, the number of patients with sensitive skin is increasing, and it has become a problem that urgently needs to be addressed.

[0003] Wild chamomile is a perennial herb known as the aspirin of the Middle Ages. It is commonly used to relieve fever, expel parasites, and reduce inflammation. The parthenolide it contains can reduce oxidative stress and cell damage, thereby helping to delay skin aging and promote skin cell regeneration. Parthenolide is a germarane-type sesquiterpene lactone extracted from wild chamomile. As a covalently reactive compound, it exhibits significant anti-inflammatory effects, as well as redox regulation and epigenetic activity. However, parthenolide is an oil-soluble ingredient and has poor stability at room temperature, which limits the application of such compounds in water-based sensitive skin care cosmetics.

[0004] The existing method to overcome the above-mentioned defects is to encapsulate the extracted parthenolide into nanocarriers to improve its solubility and stability while enhancing transdermal permeability. Nanoliposomes stand out among various nanocarriers due to their excellent biocompatibility and low toxicity, but parthenolide requires a large amount of oil to assist in dissolution, and a large amount of oil-soluble antioxidants must be added to the system to maintain the activity of the system. Nanoliposomes need to encapsulate hydrophobic active ingredients into a hydrophobic phospholipid bilayer to improve the solubility and bioavailability of the active ingredients. In this case, the effective encapsulation amount of parthenolide is low, and it cannot achieve the desired effect in a cost-effective manner.

[0005] For example, Chinese patent CN110292640A discloses the preparation and anti-tumor application of a PEGylated hydroxymethyl parthenolide nanoformulation. The nanoformulation comprises an amphiphilic conjugate formed by the reaction of hydrophobic hydroxymethyl parthenolide with a hydrophilic, carboxyl-terminated PEG. The molecular weight of the carboxyl-terminated PEG is between 500 and 50,000 g / mol. This amphiphilic conjugate can self-assemble in water to form spherical nanoparticles, whose particle size decreases with increasing PEG molecular weight. Compared to existing technologies, this invention achieves covalent conjugation of the hydrophobic anti-tumor drug hydroxymethyl parthenolide with a hydrophilic PEG, which can then self-assemble in water to form nanoparticles. This not only prolongs the drug's circulation time in the body but also allows it to enter and accumulate in tumor cells through the enhanced permeability and retention (EPR) effect, achieving a significant anti-tumor effect. However, the application of this patent on skin remains to be determined, and its irritation and safety profiles remain unknown.

[0006] For example, Chinese patent CN115590818A discloses a thermosensitive nanoliposome capable of achieving a stepped release of active ingredients and its application, comprising an anti-aging active ingredient, a liquid oil, a solid oil, a solvent, an emulsifier, and water. Based on the total weight percentage of the raw materials, the anti-aging active ingredient comprises 0.1-5%, the liquid oil content is 1-10%, the solid oil content is 1-5%, the solvent content is 2-40%, the emulsifier is 5-10%, and the balance is water to 100%, wherein the liquid oil is at least two of caprylic / capric triglyceride, hydrogenated polydecene, camellia seed oil, peony seed oil, sunflower seed oil, squalane, coconut oil, and jojoba oil. The nanoliposomes produced by this invention can achieve a stepped, slow release of active ingredients by temperature control between 20-50°C, and can be used in cosmetics of various dosage forms. However, this invention has complex ingredients and temperature control is difficult to achieve.

[0007] For example, Chinese patent CN 109498475 A discloses a microemulsion facial mask containing camellia seed oil and having repairing properties, and its preparation method. The mask comprises: 10-60% polyol; 0.5-10.0% lecithin; 0.05-5.0% camellia seed oil; 0.5-5.0% camellia and camellia leaf extract; 0.05-1.0% sterols; 0.005-0.5% ceramide compounds; 0.05-1.0% thickener; and 35-85% water. This invention uses lecithin as an emulsifier and a specific polyol as a solvent to formulate a microemulsion facial mask containing insoluble functional substances such as camellia seed oil, ceramides, and sterols. This allows these functional substances to quickly penetrate the skin when applied to the face using a facial mask cloth, repairing the skin barrier and providing excellent, long-lasting moisturizing and nourishing effects. The microemulsion mask has good stability and does not separate after centrifugation testing. However, the ingredients in this application are complex.

[0008] Therefore, there is an urgent need to develop a nano-microemulsion with simple ingredients that can improve the bioavailability of parthenolide and camellia seed oil. Summary of the Invention

[0009] To address the challenges of the existing technology, the present invention aims to provide a parthenolide-loaded camellia seed oil nano-microemulsion, as well as its preparation method and application. The parthenolide-loaded camellia seed oil nano-microemulsion prepared by the present invention effectively addresses the low encapsulation efficiency of parthenolide. It also addresses the poor water and fat solubility of parthenolide and camellia seed oil, the difficulty in using them in transparent systems, and the time-consuming and labor-intensive pretreatment process. This provides a theoretical basis for the application of parthenolide and camellia seed oil in soothing cosmetics.

[0010] To achieve the above object, the technical solution adopted by the present invention is:

[0011] In one aspect, the present invention provides a camellia seed oil nano-microemulsion loaded with parthenolide. The camellia seed oil nano-microemulsion loaded with parthenolide comprises the following raw materials, in parts by weight: 3-8 parts of parthenolide, 17-23 parts of camellia seed oil, 5-12 parts of caprylic / capric triglyceride, 2-8 parts of polyglycerol-10 myristate, 0.5-2 parts of tocopheryl acetate, 23-35 parts of glycerol, 0.3-0.8 parts of lecithin, 5-15 parts of propylene glycol, and 20-50 parts of water.

[0012] Preferably, the camellia seed oil nano-microemulsion comprises the following raw materials, by weight: 3-6 parts of parthenolide, 19-22 parts of camellia seed oil, 9-11 parts of caprylic / capric triglyceride, 3-5 parts of polyglycerol-10 myristate, 1-2 parts of tocopheryl acetate, 25-32 parts of glycerol, 0.4-0.6 parts of lecithin, 8-13 parts of propylene glycol and 20-30 parts of water.

[0013] As a preferred embodiment, the camellia seed oil nano-microemulsion loaded with parthenolide comprises the following raw materials in parts by weight: 5 parts of parthenolide, 20 parts of camellia seed oil, 10 parts of caprylic / capric triglyceride, 3 parts of polyglyceryl-10 myristate, 1 part of tocopheryl acetate, 30 parts of glycerol, 0.5 parts of lecithin, 10 parts of propylene glycol and 20.5 parts of purified water.

[0014] Preferably, the mass ratio of parthenolide to camellia seed oil is 1:3-6, preferably 1:4.

[0015] Furthermore, the present invention also provides a method for preparing camellia oil nano-microemulsion loaded with parthenolide, comprising the following steps:

[0016] (1) Mix parthenolide, camellia seed oil, caprylic / capric triglyceride, polyglyceryl-10 myristate, and tocopheryl acetate, heat in a water bath, and stir to dissolve uniformly to obtain a mixed solution 1;

[0017] (2) adding glycerol, lecithin, propylene glycol and water to the mixed solution 1 of step (1), heating in a water bath, stirring and dissolving uniformly, to obtain a mixed solution 2;

[0018] (3) using a homogenizer to perform shear homogenization on the mixed solution 2 obtained in step (2) to obtain a micron-sized crude emulsion;

[0019] (4) After returning to room temperature, the micron-sized coarse emulsion obtained in step (3) is subjected to micro-jet high-pressure homogenization treatment using a micro-jet homogenizer to obtain the camellia seed oil nano-microemulsion.

[0020] Preferably, the temperature of the constant temperature water bath in step (1) is 60-70°C.

[0021] Preferably, the stirring in step (1) is magnetic stirring, with a rotation speed of 130-150 r / min and a stirring time of 3-10 min.

[0022] Preferably, the purified water in step (2) is added to 100%.

[0023] Preferably, the temperature of the constant temperature water bath in step (2) is 60-70°C.

[0024] Preferably, the stirring in step (2) is carried out using a magnetic stirrer with a rotation speed of 130-150 r / min and a stirring time of 3-10 min.

[0025] Preferably, the shear homogenization treatment in step (3) is carried out for 10-20 min at a speed of 6000-8000 r / min.

[0026] Preferably, the homogenization pressure in the high-pressure homogenization treatment in step (4) is 25,000-30,000 psi, and the homogenization is performed for 3-5 cycles.

[0027] The present invention also provides an application of a parthenolide-loaded camellia seed oil nano-microemulsion in preparing cosmetics.

[0028] Preferably, the cosmetics include lotion, cream, essence water or facial cleanser.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] The invention firstly adopts a high shear homogenization method to mix and shear parthenolide with supercritical camellia seed oil to prepare colostrum, and then treats the microemulsion with a microjet homogenization method to prepare a parthenolide-loaded camellia seed oil nano-microemulsion. The invention not only solves the problem of low encapsulation efficiency of parthenolide by nanoliposomes, but also solves the problems of poor water solubility and fat solubility of parthenolide and camellia seed oil, difficulty in application in transparent systems, time-consuming and labor-intensive pretreatment, and poor stability, thereby providing a theoretical basis for the application of parthenolide and camellia seed oil in soothing cosmetics. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is a particle size diagram of the nano-microemulsion prepared in Example 1;

[0032] Figure 2 This is the potential diagram of the nano-microemulsion prepared in Example 1;

[0033] Figure 3 These are the cell scratch test images of the nano-microemulsions prepared in Example 1 and Comparative Example 4. DETAILED DESCRIPTION

[0034] The technical solution of the present invention is described clearly and completely below with specific embodiments. Obviously, the embodiments described are only some embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work are within the scope of protection of the present invention. The materials, reagents, etc. used, unless otherwise specified, are reagents and materials available from commercial sources.

[0035] Parthenolide was purchased from Wuhan Lanabai Pharmaceutical Chemical Co., Ltd.; camellia seed oil was purchased from Guangdong Yuewei Microbiology Technology Co., Ltd.; caprylic / capric triglyceride was purchased from Guangzhou Zhongran Biotechnology Co., Ltd.; polyglyceryl-10 myristate was purchased from INOLEX; tocopheryl acetate was purchased from BASF Group; lecithin was purchased from Guangzhou Keyun Biotechnology Co., Ltd.; and propylene glycol was purchased from Wanhua Chemical Group.

[0036] Example 1 A Camellia Seed Oil Nano-Microemulsion Loaded with Parthenolide

[0037] It is composed of the following raw materials:

[0038] 5 parts parthenolide, 20 parts camellia seed oil, 10 parts caprylic / capric triglyceride, 3 parts polyglyceryl-10 myristate, 1 part tocopheryl acetate, 30 parts glycerin, 0.5 parts lecithin, 10 parts propylene glycol and 20.5 parts water.

[0039] The preparation method is specifically as follows:

[0040] (1) 5 parts of parthenolide, 20 parts of camellia seed oil, 10 parts of caprylic / capric triglyceride, 3 parts of polyglycerol-10 myristate, and 1 part of tocopheryl acetate were mixed and stirred in a constant temperature water bath at 65°C and 150 r / min to obtain a mixed solution 1;

[0041] (2) adding 30 parts of glycerol, 0.5 parts of lecithin, 10 parts of propylene glycol and 20.5 parts of purified water to the mixed solution 1 of step (1) in a constant temperature water bath, stirring and dissolving uniformly to obtain a mixed solution 2;

[0042] (3) using a homogenizer to perform shear homogenization treatment on the mixed solution 2 obtained in step (2) at a speed of 7000 r / min for 15 min to obtain a micron-sized crude emulsion;

[0043] (4) After returning to room temperature, the micron-sized coarse emulsion obtained in step (3) was subjected to microfluidizer high-pressure homogenization treatment at a homogenization pressure of 25,000 psi for 4 cycles to obtain a camellia seed oil nano-microemulsion loaded with parthenolide.

[0044] Example 2 A camellia oil nano-microemulsion loaded with parthenolide and its preparation method

[0045] It is composed of the following raw materials:

[0046] 2 parts parthenolide, 17 parts camellia seed oil, 5 parts caprylic / capric triglyceride, 2 parts polyglyceryl-10 myristate, 0.5 parts tocopheryl acetate, 23 parts glycerin, 0.3 parts lecithin, 5 parts propylene glycol and 20 parts water.

[0047] The preparation method is specifically as follows:

[0048] (1) 3 parts of parthenolide, 17 parts of camellia seed oil, 5 parts of caprylic / capric triglyceride, 2 parts of polyglycerol-10 myristate, and 0.5 parts of tocopheryl acetate were mixed, heated in a water bath at 60° C., and stirred at a speed of 150 r / min to obtain a mixed solution 1;

[0049] (2) adding 23 parts of glycerol, 0.3 parts of lecithin, 5 parts of propylene glycol and 30 parts of water to the mixed solution 1 of step (1), heating in a water bath at 60° C., stirring and dissolving uniformly, to obtain a mixed solution 2;

[0050] (3) using a homogenizer to perform shear homogenization treatment on the mixed solution 2 obtained in step (2) at a speed of 6000 r / min for 10 min to obtain a micron-sized crude emulsion;

[0051] (4) After returning to room temperature, the micron-sized coarse emulsion obtained in step (3) was subjected to microfluidizer high-pressure homogenization treatment at a homogenization pressure of 30,000 psi for three homogenization cycles to obtain a camellia seed oil nano-microemulsion loaded with parthenolide.

[0052] Example 3 A camellia oil nano-microemulsion loaded with parthenolide and its preparation method

[0053] It is composed of the following raw materials:

[0054] 8 parts parthenolide, 23 parts camellia seed oil, 12 parts caprylic / capric triglyceride, 8 parts polyglyceryl-10 myristate, 2 parts tocopheryl acetate, 35 parts glycerin, 0.8 parts lecithin, 15 parts propylene glycol and 50 parts water.

[0055] The preparation method is specifically as follows:

[0056] (1) 8 parts of parthenolide, 23 parts of camellia seed oil, 12 parts of caprylic / capric triglyceride, 8 parts of polyglycerol-10 myristate, and 2 parts of tocopheryl acetate were mixed, heated in a water bath at 60° C., and stirred at a speed of 150 r / min to obtain a mixed solution 1;

[0057] (2) adding 35 parts of glycerol, 0.8 parts of lecithin, 15 parts of propylene glycol and 50 parts of water to the mixed solution 1 of step (1), heating in a water bath at 70° C., stirring and dissolving uniformly, to obtain a mixed solution 2;

[0058] (3) using a homogenizer to perform shear homogenization treatment on the mixed solution 2 obtained in step (2) at a speed of 8000 r / min for 20 min to obtain a micron-sized crude emulsion;

[0059] (4) After returning to room temperature, the micron-sized coarse emulsion obtained in step (3) was subjected to microfluidizer high-pressure homogenization treatment at a homogenization pressure of 25,000 psi for 5 cycles to obtain a camellia seed oil nano-microemulsion loaded with parthenolide.

[0060] Comparative Example 1 A camellia oil-loaded parthenolide nanoemulsion and its preparation method

[0061] The only difference from Example 1 is that 5 parts of parthenolide and 20 parts of camellia seed oil are replaced by 20 parts of parthenolide and 5 parts of camellia seed oil, that is, 20 parts of parthenolide, 5 parts of camellia seed oil, 10 parts of caprylic / capric triglyceride, 3 parts of polyglyceryl-10 myristate, 1 part of tocopheryl acetate, 30 parts of glycerin, 0.5 parts of lecithin, 10 parts of propylene glycol and 20.5 parts of water.

[0062] The preparation method is specifically as follows:

[0063] (1) 5 parts of parthenolide, 20 parts of camellia seed oil, 10 parts of caprylic / capric triglyceride, 3 parts of polyglycerol-10 myristate, and 1 part of tocopheryl acetate were mixed and stirred in a constant temperature water bath at 65°C and 150 r / min to obtain a mixed solution 1;

[0064] (2) adding 30 parts of glycerol, 0.5 parts of lecithin, 10 parts of propylene glycol and 20.5 parts of purified water to the mixed solution 1 of step (1) in a constant temperature water bath, stirring and dissolving uniformly to obtain a mixed solution 2;

[0065] (3) using a homogenizer to perform shear homogenization treatment on the mixed solution 2 obtained in step (2) at a speed of 7000 r / min for 15 min to obtain a micron-sized crude emulsion;

[0066] (4) After returning to room temperature, the micron-sized coarse emulsion obtained in step (3) was subjected to microfluidizer high-pressure homogenization treatment at a homogenization pressure of 25,000 psi for 4 cycles to obtain a camellia seed oil nano-microemulsion loaded with parthenolide.

[0067] Comparative Example 2: Camellia seed oil nano-microemulsion loaded with chondrolactone and its preparation method

[0068] The only difference from Example 1 is that 5 parts of parthenolide are replaced by 5 parts of chondrolactone, that is, 5 parts of chondrolactone, 20 parts of camellia seed oil, 10 parts of caprylic / capric triglyceride, 3 parts of polyglyceryl-10 myristate, 1 part of tocopheryl acetate, 30 parts of glycerol, 0.5 parts of lecithin, 10 parts of propylene glycol and 20.5 parts of water.

[0069] The preparation method is specifically as follows:

[0070] (1) 5 parts of chondrolactone, 20 parts of camellia seed oil, 10 parts of caprylic / capric triglyceride, 3 parts of polyglycerol-10 myristate, and 1 part of tocopheryl acetate were mixed and dissolved in a constant temperature water bath at 65°C and a speed of 150 r / min to obtain a mixed solution 1;

[0071] (2) adding 30 parts of glycerol, 0.5 parts of lecithin, 10 parts of propylene glycol and 20.5 parts of purified water to the mixed solution 1 of step (1) in a constant temperature water bath, stirring and dissolving uniformly to obtain a mixed solution 2;

[0072] (3) using a homogenizer to perform shear homogenization treatment on the mixed solution 2 obtained in step (2) at a speed of 7000 r / min for 15 min to obtain a micron-sized crude emulsion;

[0073] (4) After returning to room temperature, the micron-sized coarse emulsion obtained in step (3) was subjected to microfluidizer high-pressure homogenization treatment at a homogenization pressure of 25,000 psi for 4 cycles to obtain a camellia seed oil nano-microemulsion loaded with parthenolide.

[0074] Comparative Example 3 A camellia oil nano-microemulsion loaded with parthenolide and its preparation method

[0075] The only difference from Example 1 is that 5 parts of parthenolide and 20 parts of camellia seed oil are replaced by 12.5 parts of parthenolide and 12.5 parts of camellia seed oil, that is, 12.5 parts of parthenolide, 12.5 parts of camellia seed oil, 10 parts of caprylic / capric triglyceride, 3 parts of polyglyceryl-10 myristate, 1 part of tocopheryl acetate, 30 parts of glycerin, 0.5 part of lecithin, 10 parts of propylene glycol and 20.5 parts of water.

[0076] The preparation method is specifically as follows:

[0077] (1) 12.5 parts of parthenolide, 12.5 parts of camellia seed oil, 10 parts of caprylic / capric triglyceride, 3 parts of polyglycerol-10 myristate, and 1 part of tocopheryl acetate were mixed and dissolved in a constant temperature water bath at 65°C and a speed of 150 r / min to obtain a mixed solution 1;

[0078] (2) adding 30 parts of glycerol, 0.5 parts of lecithin, 10 parts of propylene glycol and 20.5 parts of purified water to the mixed solution 1 of step (1) in a constant temperature water bath, stirring and dissolving uniformly to obtain a mixed solution 2;

[0079] (3) using a homogenizer to perform shear homogenization treatment on the mixed solution 2 obtained in step (2) at a speed of 7000 r / min for 15 min to obtain a micron-sized crude emulsion;

[0080] (4) After returning to room temperature, the micron-sized coarse emulsion obtained in step (3) was subjected to microfluidizer high-pressure homogenization treatment at a homogenization pressure of 25,000 psi for 4 cycles to obtain a camellia seed oil nano-microemulsion loaded with parthenolide.

[0081] Comparative Example 4: A camellia oil nano-microemulsion loaded with parthenolide and its preparation method

[0082] The only difference from Example 1 is that all the parthenolide is replaced by camellia seed oil, that is, 25 parts of camellia seed oil, 10 parts of caprylic / capric triglyceride, 3 parts of polyglyceryl-10 myristate, 1 part of tocopheryl acetate, 30 parts of glycerin, 0.5 parts of lecithin, 10 parts of propylene glycol and 20.5 parts of water.

[0083] The preparation method is specifically as follows:

[0084] (1) 25 parts of camellia seed oil, 10 parts of caprylic / capric triglyceride, 3 parts of polyglycerol-10 myristate, and 1 part of tocopheryl acetate were mixed and dissolved uniformly in a constant temperature water bath at 65°C and a speed of 150 r / min to obtain a mixed solution 1;

[0085] (2) adding 30 parts of glycerol, 0.5 parts of lecithin, 10 parts of propylene glycol and 20.5 parts of purified water to the mixed solution 1 of step (1) in a constant temperature water bath, stirring and dissolving uniformly to obtain a mixed solution 2;

[0086] (3) using a homogenizer to perform shear homogenization treatment on the mixed solution 2 obtained in step (2) at a speed of 7000 r / min for 15 min to obtain a micron-sized crude emulsion;

[0087] (4) After returning to room temperature, the micron-sized coarse emulsion obtained in step (3) was subjected to microfluidizer high-pressure homogenization treatment at a homogenization pressure of 25,000 psi for 4 cycles to obtain a camellia seed oil nano-microemulsion loaded with parthenolide.

[0088] Comparative Example 5 Preparation method of camellia oil nano-microemulsion loaded with parthenolide

[0089] The only difference from Example 1 is that all the parthenolide is replaced by camellia seed oil, namely: 25 parts of parthenolide, 10 parts of caprylic / capric triglyceride, 3 parts of polyglyceryl-10 myristate, 1 part of tocopheryl acetate, 30 parts of glycerin, 0.5 parts of lecithin, 10 parts of propylene glycol and 20.5 parts of water.

[0090] The preparation method is specifically as follows:

[0091] (1) 25 parts of camellia seed oil, 10 parts of caprylic / capric triglyceride, 3 parts of polyglycerol-10 myristate, and 1 part of tocopheryl acetate were mixed and dissolved uniformly in a constant temperature water bath at 65°C and a speed of 150 r / min to obtain a mixed solution 1;

[0092] (2) adding 30 parts of glycerol, 0.5 parts of lecithin, 10 parts of propylene glycol and 20.5 parts of purified water to the mixed solution 1 of step (1) in a constant temperature water bath, stirring and dissolving uniformly to obtain a mixed solution 2;

[0093] (3) using a homogenizer to perform shear homogenization treatment on the mixed solution 2 obtained in step (2) at a speed of 7000 r / min for 15 min to obtain a micron-sized crude emulsion;

[0094] (4) After returning to room temperature, the micron-sized coarse emulsion obtained in step (3) was subjected to microfluidizer high-pressure homogenization treatment at a homogenization pressure of 25,000 psi for 4 cycles to obtain a camellia seed oil nano-microemulsion loaded with parthenolide.

[0095] Effect experiment

[0096] (1) In vitro skin retention performance test

[0097] Experimental methods:

[0098] The skin retention performance test of ex vivo pig skin was conducted with reference to GB / T 27818-2011 "In vitro test method for skin absorption of chemicals". The vertical Franz diffusion cell method was used. The dorsal and ventral skin of healthy Bama pigs aged one month (average thickness of about 750 μm) stored at -20°C was thawed naturally at room temperature. After being gently wiped clean with a phosphate buffer solution of pH 6.8-7.4, the initial transepidermal water loss value of the skin was measured using a transepidermal water loss meter. The initial transepidermal water loss value was <15 g / m 2 The skin stratum corneum was considered intact after 1 h. A vertical (Franz) diffusion cell was used as the diffusion device. The prepared ex vivo skin was fixed between the supply cell and the receiving cell, with the skin stratum corneum facing the supply cell and the dermis facing the receiving cell. 8 mL of phosphate buffer was added to the receiving cell to ensure close contact between the skin and the receiving solution without bubbles. The transdermal diffusion instrument was turned on, the water level in the tank was checked, the speed was set to 350 rpm, the temperature to 32°C, and preheated for 5 minutes. In each sample test group, a pressure of 0.2 g / cm 2 The nano-microemulsions prepared in Examples 1 to 3 and Comparative Examples 1 to 5 were added, followed by 100 mg of FITC. At 48 hours, the diffusion cell was removed, excess sample in the supply cell was aspirated and discarded, and the skin surface was cleaned with ultrapure water. Tissue blocks from the central, effectively permeated portion of the skin were removed and fixed in a tissue fixative and frozen. The tissue sections were then observed under a fluorescence microscope for the distribution of FITC fluorescence signals in the skin tissue. The samples were then transferred to a high-throughput tissue grinder and thoroughly ground into a skin homogenate. Extraction was performed with 1 mL of acetonitrile, followed by filtration and elution using a PursuitXRs C18 (500 g / 3 L) liquid chromatography column with a mobile phase consisting of a mixture of water and acetonitrile (40:60 by volume), a flow rate of 0.3 mL / min, an injection volume of 20 μL, a column temperature of 30°C, a chromatography time of 15 min, and a quantitative detection wavelength of 325 nm. The cumulative retention of parthenolide in the skin was calculated, as shown in Table 1.

[0099] Table 1 Cumulative retention of parthenolide in the skin

[0100] <![CDATA[Cumulative retention amount (μg·cm -1 )]]> Example 1 52.67 Example 2 49.56 Example 3 48.91 Comparative Example 1 25.87 Comparative Example 2 26.45 Comparative Example 3 32.43 Comparative Example 4 20.81 Comparative Example 5 18.52

[0101] As shown in Table 1, the cumulative retention of parthenolide in the skin of the nano-microemulsions prepared in Examples 1 to 3 was higher than that in Comparative Examples 1 to 5. Replacing parthenolide with chondrolactone, which has the same efficacy, also did not produce satisfactory results.

[0102] (2) Determination of particle size, PDI and Zeta potential

[0103] The average particle size, PDI and Zeta potential of the nano-microemulsions prepared in Examples 1 to 3 and Comparative Examples 1 to 5 were measured using a nanoparticle size analyzer. First, the nano-microemulsions prepared in Examples 1 to 3 and Comparative Examples 1 to 5 were diluted 100 times with ultrapure water. The test temperature was set to 25°C and the scattering angle was set to 90°. Each sample was scanned 3 times and the average value was used as the measured value. When measuring the Zeta potential, the sample was not diluted and was directly added to the Zeta potential cell for measurement. The results are shown in Table 2 and Table 3. Figure 1 、 Figure 2 shown.

[0104] Table 2 Particle size potential and dispersion index of each group

[0105] Particle size (nm) Potential (mV) PDI Example 1 40.33 -9.91 0.224 Example 2 48.21 -6.32 0.328 Example 3 45.88 -7.95 0.315 Comparative Example 1 99.54 -0.45 0.881 Comparative Example 2 95.03 -0.88 0.731 Comparative Example 3 82.17 -2.21 0.556 Comparative Example 4 170.25 0.74 1.073 Comparative Example 5 189.84 1.28 1.253

[0106] As shown in Table 2, the particle size, potential, and PDI of Examples 1 to 3 are much smaller than those of Comparative Examples 1 to 5. A high Zate potential (large absolute value) maintains a small particle size and low PDI through electrostatic repulsion, resulting in a narrow and uniform particle size distribution, which in turn achieves a stable system.

[0107] (3) Cell scratch test

[0108] HUVECs were plated at 1×10 5 Cells were seeded in 96-well plates, placed in a cell culture incubator at 37°C and 5% CO2 for 24 hours, and then mechanically scratched using a 96-well cell scratcher. A blank group (DMEM complete culture medium), a negative group (DMEM culture medium + HUVECs), a positive group (DMEM culture medium + HUVECs + growth factors), Example 1-Example 3 and Comparative Example 1-Comparative Example 5 groups were set up, the culture medium was removed, D-Hank's washed twice, and replaced with DMEM complete culture medium containing 5% fetal bovine serum according to the experimental grouping. The cells were placed in a cell culture incubator for continued culture, and photographed and recorded under an inverted microscope at 40 times, recorded as 0h; the cells were returned to the incubator for continued culture for 12h and 24h, and photographed and recorded again. The cell migration at different time points was observed and recorded under a phase contrast microscope, and the scratch healing rate was calculated using Image J software. The results are shown in Table 3 and Figure 3 shown.

[0109] Table 3 Scratch healing rate of each group

[0110] Healing rate (%) Blank group 0％ Negative group 38.3％±1.2* Positive group <![CDATA[84.1%±0.9 # *]]> Example 1 <![CDATA[87.2%±2.1 # *]]> Example 2 <![CDATA[84.4%±1.5 # *]]> Example 3 <![CDATA[83.1%±1.7 # *]]> Comparative Example 1 52.6％±1.8* Comparative Example 2 55.5％±2.2* Comparative Example 3 57.7％±1.3* Comparative Example 4 45.3％±1.1* Comparative Example 5 43.3％±1.4*

[0111] Note: Compared with the blank group*p<0.01; compared with the negative group # p<0.01.

[0112] As shown in Table 3, the healing rates of Examples 1 to 3 are significantly higher than those of Comparative Examples 1 to 5, which can effectively enhance the migration ability of skin cells over time and promote rapid skin repair.

[0113] (IV) Determination of encapsulation efficiency

[0114] The encapsulation efficiency was determined by petroleum ether extraction. Take 1 mL of sample solution, add 10 mL of petroleum ether, fully shake and stir at 30 ° C for 5 minutes (ultrasound or shake well), let it stand for 30 minutes, transfer the upper liquid to a rotary evaporation bottle, and add 10 mL of petroleum ether for extraction, and repeat twice. The upper liquid was vacuum evaporated at 50 ° C to remove petroleum ether and precipitate free parthenolide. Add 2 mL of chloroform to dissolve it again, and measure its absorbance at a wavelength of 205 nm. The content of free parthenolide was calculated according to the standard curve, as shown in Table 4. The encapsulation efficiency was calculated according to the following formula:

[0115]

[0116] Table 4 Encapsulation efficiency of each group

[0117] Encapsulation efficiency (%) Example 1 99.87 Example 2 97.41 Example 3 98.22 Comparative Example 1 79.88 Comparative Example 2 78.31 Comparative Example 3 82.15 Comparative Example 4 55.66 Comparative Example 5 45.66

[0118] The above results show that only the solution provided by the present invention can solve the problems of low encapsulation efficiency of parthenolide in nanoliposomes, poor water solubility and fat solubility of parthenolide and camellia seed oil, time-consuming and labor-intensive pretreatment, and poor stability.

[0119] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that although the technical solutions of the present invention may be modified or replaced by equivalents, this does not deviate from the essence and scope of the technical solutions of the present invention.

Claims

1. A camellia oil nano-microemulsion loaded with parthenolide, characterized in that: The invention comprises the following raw materials in parts by weight: 3-8 parts of parthenolide, 17-23 parts of camellia seed oil, 5-12 parts of caprylic / capric triglyceride, 2-8 parts of polyglycerol-10 myristate, 0.5-2 parts of tocopheryl acetate, 23-35 parts of glycerol, 0.3-0.8 parts of lecithin, 5-15 parts of propylene glycol and 20-50 parts of water, wherein the mass ratio of parthenolide to camellia seed oil is 1:3-6.

2. The camellia oil nano-microemulsion according to claim 1, characterized in that The invention comprises the following raw materials in parts by weight: 5 parts of parthenolide, 20 parts of camellia seed oil, 10 parts of caprylic / capric triglyceride, 3 parts of polyglycerol-10 myristate, 1 part of tocopheryl acetate, 30 parts of glycerol, 0.5 parts of lecithin, 10 parts of propylene glycol and 20.5 parts of water, wherein the mass ratio of parthenolide to camellia seed oil is 1:

4.

3. The method for preparing the camellia oil nano-microemulsion according to any one of claims 1 to 2, characterized in that: The steps include: (1) mixing parthenolide, camellia seed oil, caprylic / capric triglyceride, polyglyceryl-10 myristate, and tocopheryl acetate, heating, and stirring to obtain a mixed solution 1; (2) adding glycerol, lecithin, propylene glycol and water to the mixed solution 1 of step (1), heating and stirring to obtain a mixed solution 2; (3) homogenizing the mixed solution 2 obtained in step (2) to obtain a micron-sized crude emulsion; (4) After returning to room temperature, the micron-sized coarse emulsion obtained in step (3) is homogenized to obtain the camellia seed oil nano-microemulsion.

4. The preparation method according to claim 3, wherein: The heating in step (1) and step (2) is water bath heating, and the heating temperature is 60-70°C.

5. The preparation method according to claim 3, wherein: The stirring in step (1) is magnetic stirring, with a rotation speed of 130-150 r / min and a stirring time of 3-10 min.

6. The preparation method according to claim 3, wherein: The homogenization treatment in step (3) is a shear homogenization treatment, the shear homogenization treatment time is 10-20 minutes, and the rotation speed is 6000-8000r / min.

7. The preparation method according to claim 3, wherein: The homogenization treatment in step (4) is a jet high-pressure homogenization treatment, and the homogenization pressure is 25000-30000psi.

8. The preparation method according to claim 3, wherein: The homogenization treatment in step (4) requires 3-5 cycles of homogenization.

9. Use of the camellia seed oil nano-microemulsion according to any one of claims 1 to 2 or the camellia seed oil nano-microemulsion prepared by the preparation method according to any one of claims 3 to 8 in the preparation of cosmetics.

10. The use according to claim 9, characterized in that The cosmetics include lotion, cream, essence water or facial cleanser.

Images