Azelaic acid oiling agent as well as preparation method and application thereof

Abstract

The invention discloses an azelaic acid oiling agent as well as a preparation method and application thereof. The preparation method of the azelaic acid oiling agent comprises the following steps: S1, mixing oleic acid and triglyceride, introducing mixed gas of ozone and oxygen under a stirring condition, and reacting at 20-30 DEG C for 8-14 hours to obtain an intermediate product; and S2, introducing oxygen into the intermediate product at 70-95 DEG C, and reacting for 6-10 hours to obtain the azelaic acid oiling agent. According to the preparation technology disclosed by the invention, mutual dissolution of azelaic acid and oil is realized, and the prepared azelaic acid oil agent has the effects of relieving inflammation and protecting a pigment barrier, can be used for preventing and repairing acute and chronic light injury, especially plateau light injury, and can be used for relieving inflammation and color sedimentation and promoting fading of inflammation and color sedimentation. The traditional Chinese medicine composition can also be used for treating and preventing acute and chronic dermatitis and eczema skin diseases, and helping to repair and prevent a physical barrier and a pigment barrier of the skin; the product is an oil agent, has moistening and moisturizing effects, is convenient to use, has no obvious irritation and has light side effects.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to an azelaic acid oil preparation, its preparation method, and its application. Background Technology

[0002] Photodamage is a series of skin reactions and diseases caused by ultraviolet radiation and other forms of light exposure. It not only leads to skin aging and aesthetic problems but can also cause serious health issues, including acute photodamage (sunburn) and chronic photodamage (including photoaging, polymorphic light eruption (PLE), and chronic actinic dermatitis (CAD)). Sunburn, also known as solar dermatitis, solar edema, or solar erythema, is an acute phototoxic inflammatory reaction of the skin after ultraviolet radiation, manifesting as erythema, blisters, desquamation, and pigmentation, accompanied by pain and / or itching. Ultraviolet radiation is classified into long-wave ultraviolet (UVA), medium-wave ultraviolet (UVB), and short-wave ultraviolet (UVC) according to wavelength. UVA and UVB have a direct impact on human skin and can cause tanning. UVA causes photooxidation of melanin, while UVB irradiation can stimulate melanocytes in the skin. At the same dose, UVB causes much greater damage to the skin than UVA.

[0003] Plant oils possess various activities such as anti-inflammatory and antioxidant properties. In addition, their advantages of being green and environmentally friendly, safe to use, and easily absorbed by the skin make them ideal raw materials for further development and utilization as natural sunscreens.

[0004] Azelaic acid can be produced by Malassezia furfur and is found in whole grains, rye, and barley. It can also be formed through chemical synthesis and fermentation. Azelaic acid exhibits hypopigmentation properties because it acts as a competitive inhibitor of tyrosinase. It can inhibit abnormal melanocyte activity and reduce pigmentation. It can also inhibit the production of reactive oxygen species, thus exerting an anti-inflammatory effect. However, azelaic acid is insoluble in water and oil, has poor solubility, and a high melting point.

[0005] The prior art CN 109180462 A discloses a method for preparing nonanoic acid and azelaic acid from oleic acid. Oleic acid and glacial acetic acid are subjected to ozone oxidation and oxidative cracking reactions. The reactants are further extracted and purified to obtain high-purity nonanoic acid and azelaic acid, respectively; wherein the mass ratio of oleic acid to glacial acetic acid is 1:2-4.

[0006] The prior art CN 118084653 A relates to a high-purity azelaic acid suitable for pharmaceutical applications and a green synthesis method thereof. The method includes the following steps: mixing oleic acid and glacial acetic acid; introducing ozone into the reactants to induce a reaction, yielding oleic acid ozonide; introducing oxygen into the obtained oleic acid ozonide and heating to induce a reaction, yielding an oxidative cracking product; extracting the obtained oxidative cracking product with hot water, crystallizing, filtering, and drying to obtain a crude product; recrystallizing the crude product in a mixed solvent to obtain the final product, azelaic acid.

[0007] Both of the aforementioned existing technologies aim to obtain pure azelaic acid. The resulting azelaic acid is insoluble in water and oil, exhibiting poor solubility and a high melting point. Azelaic acid is widely used in the treatment of acne, melasma, rosacea, and hyperpigmentation, with an effective dose exceeding 10 wt%. Currently, clinically used azelaic acid formulations include creams, gels, and plasters. High concentrations of azelaic acid can cause a gritty sensation or easily lead to emulsion breakdown. Creams are commonly used in open-end (O / W) formulations, where azelaic acid is often added as a fine powder, which cannot achieve complete dissolution. In reported formulations, the incomplete dissolution of azelaic acid in the fine powder dispersed in the matrix reduces its bioavailability. In azelaic acid gels, to improve solubility, large amounts of oily components or solvents are typically added. However, excessive solvents and oily products can lead to poor skin permeability, poor usability, and significant side effects. Furthermore, existing formulations are too concentrated and highly irritating, making them unsuitable for use in acute or erosive skin diseases.

[0008] There is currently no azelaic acid oil formulation available. Developing suitable azelaic acid oil products and expanding the dosage form of this drug is of great significance. Summary of the Invention

[0009] The purpose of this invention is to provide an azelaic acid oil, its preparation method, and its application, in order to solve the solubility problem of azelaic acid and to develop the uses of the azelaic acid oil.

[0010] To achieve the above objectives, the present invention adopts the following technical solution: A method for preparing an azelaic acid oil includes the following steps: S1. Oleic acid and triglycerides are mixed and then a mixture of ozone and oxygen is introduced under stirring. The mixture is reacted at 20-30°C for 8-14 hours to obtain an intermediate product. S2. Oxygen is introduced into the intermediate product at 70℃-95℃ and the reaction is carried out for 6-10 hours to obtain azelaic acid oil.

[0011] In one preferred embodiment, the triglyceride is one or more of oleic acid triglyceride, dioleoyl palmitate triglyceride, dioleoyl stearate triglyceride, dioleoyl linoleate triglyceride, and linoleic acid triglyceride.

[0012] In one preferred embodiment, in step S1, the volume ratio of oleic acid to triglycerides is 5-20:80-95. Excessive oleic acid content results in poor product homogeneity and turbidity. Insufficient oleic acid content leads to a lower concentration of active ingredients and poorer therapeutic effect.

[0013] In one preferred embodiment, in step S1, the volume ratio of ozone to oxygen in the ozone-oxygen mixture is 3-5:95-97. Too low an ozone content will lead to reduced peroxidation efficiency; too high an ozone content will result in poor product uniformity and more byproducts.

[0014] In one preferred embodiment, in step S1, the rate at which the ozone and oxygen mixture is introduced is 0.5-1.5 L / min.

[0015] In one preferred embodiment, in step S1, the peroxide value of the intermediate product is 200-280 meq / kg.

[0016] In one preferred embodiment, in step S1, the ozone absorption capacity of the intermediate product is 80-144 mg / g.

[0017] In one preferred embodiment, the oxygen ventilation rate in step S2 is 0.2-0.7 L / kg.

[0018] In one preferred embodiment, in step S2, the azelaic acid content in the azelaic acid oil is 0.5-2.5 wt%.

[0019] In step S2, the reaction temperature is controlled between 70℃ and 95℃. If the temperature is too high, the process cost will be high and too many by-products will be generated. If the temperature is too low, the azelaic acid content will be too low and the effect will be poor.

[0020] Based on the same inventive concept, the present invention also claims protection for the azelaic acid oil prepared by the preparation method.

[0021] Based on the same inventive concept, the present invention also claims protection for the use of the azelaic acid oil in the preparation of medicaments for treating or preventing acute and chronic photodamage.

[0022] Based on the same inventive concept, this invention also claims protection for the use of the azelaic acid oil in the preparation of drugs for barrier repair, anti-inflammation, and pigmentation reduction.

[0023] This invention solves the solubility problem of azelaic acid in triglycerides by mixing and peroxidizing, followed by re-oxidation, to obtain a homogeneous azelaic acid oil, thus improving its bioavailability. Firstly, this invention eliminates the need for glacial acetic acid, as existing methods involve incomplete reactions with glacial acetic acid, which can cause skin irritation. Secondly, the azelaic acid oil prepared by this invention does not require additional purification steps; by controlling suitable reaction conditions, a product with good therapeutic effects can be obtained. This invention also produces triglycerides peroxide simultaneously with the synthesis of azelaic acid, combining the pigment-reducing effects of azelaic acid with the anti-inflammatory effects of triglycerides peroxide, achieving a synergistic effect greater than the sum of its parts.

[0024] Compared with the prior art, the beneficial effects of the present invention are as follows: 1. This invention provides an azelaic acid oil preparation that can be used to prevent and treat photodamage and has a barrier repair effect.

[0025] 2. The preparation technology of this invention achieves the miscibility of azelaic acid and oil. The prepared azelaic acid oil has the effect of relieving inflammation and can be used for the prevention and repair of photodamage.

[0026] 3. The triglycerides peroxide in the azelaic acid oil prepared by this invention have better breathability and usability than the general oily components added to the gel, and have virtually no side effects.

[0027] 4. The azelaic acid oil prepared in this invention can reduce pigmentation and promote pigment fading while having anti-inflammatory effects. It can also be used for barrier repair and prevention of pigmentation in other diseases such as atopic dermatitis and wounds. Attached Figure Description

[0028] Figure 1 The image shows the appearance of the azelaic acid oil obtained in Example 6.

[0029] Figure 2 The image shows the appearance of the azelaic acid oil obtained in Comparative Example 1.

[0030] Figure 3 The image shows the appearance of the azelaic acid oil obtained in Comparative Example 3.

[0031] Figure 4 Liquid chromatography-mass spectra for qualitative detection of azelaic acid content in Example 1.

[0032] Figure 5 High-performance liquid chromatography (HPLC) chromatogram for detecting the content of azelaic acid in Example 1.

[0033] Figure 6 A bar chart for high performance liquid chromatography (HPLC) detection of azelaic acid content in Examples 1, 2, 3, 4, 5, 6 and Comparative Examples 1 and 2.

[0034] Figure 7 The positive ion mode (M+ NH4) obtained by liquid chromatography-mass spectrometry analysis of the sample in Example 4 + Total ion chromatogram (TIC chromatogram).

[0035] Figure 8 To filter by m / z 1046 (C 57 H 104 O 15 [M+ NH4] + The extracted ion chromatogram (XIC chromatogram).

[0036] Figure 9 C 57 H 104 O 15 MS / MS spectrum of [M+ NH4]+ ions.

[0037] Figure 10 It is triglyceride peroxide (C 57 H 104 O 15 The molecular structural formula of ).

[0038] Figure 11 The image shows the appearance of an animal model of acute photodamage treated in Example 4. In the image, A represents the acute photodamage model group, B represents the triglyceride peroxide group, C represents the azelaic acid gel group, and D represents the azelaic acid oil group.

[0039] Figure 12 The images shown are confocal micrographs of the groups treated with acute photodamage in animal models in Example 4. In the images, A is the acute photodamage model group, B is the azelaic acid gel group, C is the triglyceride peroxide group, and D is the azelaic acid oil group.

[0040] Figure 13 This is a graph showing the clinical efficacy of Example 4 in treating acute photodamage. Detailed Implementation

[0041] This invention is not limited to the specific embodiments listed below. Those skilled in the art can implement this invention using various other specific embodiments based on the content disclosed herein. Any modifications or alterations made to the design structure and concept of this invention fall within the protection scope of this invention. It should be noted that, unless otherwise specified, the embodiments and features described in this invention can be combined with each other. Example 1

[0042] A method for preparing an azelaic acid oil includes the following steps: Step 1: Mix 5 parts oleic acid and 95 parts oleic acid triglycerides. Introduce an ozone / oxygen mixture into 350g of the oleic acid and oleic acid triglyceride mixture at a flow rate of 1L / min. The ozone concentration in the mixture is 3% (v / v). Stir at 400rpm and react at 20℃ for 8h to obtain an intermediate containing peroxidized oleic acid triglycerides and peroxidized oleic acid. The peroxide value of the intermediate at the reaction endpoint should be 200meq / kg, and the ozone absorption capacity should be 80mg / g.

[0043] Step 2: The reaction temperature is 70℃. Oxygen is introduced into the intermediate at a flow rate of 0.5 L / kg. The reaction is carried out for 8 hours to finally obtain azelaic acid oil. Example 2

[0044] A method for preparing an azelaic acid oil includes the following steps: Step 1: Mix 20 parts oleic acid and 80 parts dioleoyl palmitate triglyceride. Introduce an ozone / oxygen mixture at a flow rate of 1 L / min into 350 g of the oleic acid and dioleoyl palmitate triglyceride mixture. The ozone concentration in the mixture should be 3% (v / v). Stir at 400 rpm and react at 20°C for 8 h to obtain an intermediate containing dioleoyl palmitate triglyceride peroxide and oleic acid peroxide. The endpoint of the reaction is a peroxide value of 200 meq / kg. The ozone absorption capacity is 80 mg / g.

[0045] Step 2: The reaction temperature is 70℃. Oxygen is introduced into the intermediate at a flow rate of 0.5 L / kg. The reaction is carried out for 8 hours to finally obtain azelaic acid oil. Example 3

[0046] A method for preparing an azelaic acid oil includes the following steps: Step 1: Mix 5 parts oleic acid with 95 parts dioleoyl-stearic acid triglyceride. Introduce an ozone / oxygen mixture at a flow rate of 1 L / min into 350 g of the oleic acid and dioleoyl-stearic acid triglyceride mixture. The ozone concentration in the mixture should be 3% (v / v). Stir at 400 rpm and react at 20°C for 14 h to obtain an intermediate containing dioleoyl-stearic acid triglyceride peroxide and oleic acid peroxide. The peroxide value of the intermediate at the reaction endpoint should be 240 meq / kg. The ozone absorption capacity is 120 mg / g.

[0047] Step 2: The reaction temperature is 95℃. Oxygen is introduced into the intermediate at a flow rate of 0.5 L / kg. The reaction is carried out for 8 hours to finally obtain azelaic acid oil. Example 4

[0048] A method for preparing an azelaic acid oil includes the following steps: Step 1: Mix 20 parts oleic acid and 80 parts oleic triglyceride. Introduce an ozone / oxygen mixture at a flow rate of 1 L / min into 350 g of the oleic acid and oleic triglyceride mixture. The ozone gas concentration in the mixture should be 5% (v / v). Stir at 400 rpm and react at 20°C for 14 h to obtain an intermediate containing peroxidized oleic triglyceride and peroxidized oleic acid. The peroxide value of the intermediate at the reaction endpoint should be 280 meq / kg. The ozone absorption capacity is 144 mg / g.

[0049] Step 2: The reaction temperature is 95℃. Oxygen is introduced into the intermediate at a flow rate of 0.5 L / kg. The reaction is carried out for 8 hours to finally obtain azelaic acid oil.

[0050] Example 5 A method for preparing an azelaic acid oil includes the following steps: Step 1: Mix 5 parts oleic acid and 95 parts linoleic acid triglycerides. Introduce an ozone / oxygen mixture at a flow rate of 1 L / min into 350 g of the oleic acid and linoleic acid triglyceride mixture. The ozone gas concentration in the mixture should be 3% (v / v). Stir at 400 rpm and react at 20°C for 14 h to obtain an intermediate containing peroxide-treated linoleic acid triglycerides and peroxide-treated oleic acid. The peroxide value of the intermediate at the reaction endpoint should be 260 meq / kg. The ozone absorption capacity is 130 mg / g.

[0051] Step 2: The reaction temperature is 95℃. Oxygen is introduced into the intermediate at a flow rate of 0.4 L / kg. The reaction is carried out for 10 h to finally obtain azelaic acid oil. Example 6

[0052] A method for preparing an azelaic acid oil includes the following steps: Step 1: Mix 20 parts of oleic acid and 80 parts of dioleoyl-linoleic acid triglyceride. Introduce an ozone / oxygen mixture at a flow rate of 1 L / min into 350 g of the oleic acid and dioleoyl-linoleic acid triglyceride mixture. The ozone gas concentration in the mixture should be 4% (v / v). Stir at 400 rpm and react at 20°C for 14 h to obtain an intermediate containing dioleoyl-linoleic acid triglyceride peroxide and oleic acid peroxide. The peroxide value of the intermediate at the reaction endpoint should be 270 meq / kg. The ozone absorption capacity is 135 mg / g.

[0053] Step 2: The reaction temperature was 95℃. Oxygen was introduced into the intermediate at a flow rate of 0.6 L / kg, and the reaction was allowed to proceed for 8 hours to finally prepare azelaic acid oil. The appearance of the obtained azelaic acid oil is as follows. Figure 1 As shown. The azelaic acid oils obtained in Examples 1-5 also had a clear, transparent appearance without impurities, as shown. Figure 1 As shown.

[0054] Comparative Example 1 A method for preparing an azelaic acid oil includes the following steps: Step 1: Mix 40 parts oleic acid and 60 parts oleic triglyceride. Introduce an ozone / oxygen mixture at a flow rate of 1 L / min into 350 g of the oleic acid and oleic triglyceride mixture. The ozone gas concentration in the mixture is 3% (v / v). Stir at 400 rpm and react at 20°C for 8 h to obtain an intermediate containing peroxidized oleic triglyceride and peroxidized oleic acid. The peroxide value of the intermediate at the reaction endpoint is 200 meq / kg. The ozone absorption capacity is 80 mg / g.

[0055] Step 2: Increase the reaction temperature to 70℃, introduce oxygen into the intermediate at a rate of 0.5 L / kg, and react for 8 hours to finally obtain azelaic acid oil. The appearance of the obtained azelaic acid oil is as follows. Figure 2 As shown, it is turbid and has poor solubility.

[0056] Comparative Example 2 A method for preparing an azelaic acid oil includes the following steps: Step 1: Mix 2 parts oleic acid with 98 parts oleic triglyceride. Introduce an ozone / oxygen mixture at a flow rate of 1 L / min into 350 g of the oleic acid and oleic triglyceride mixture. The ozone concentration in the mixture is 3% (v / v). Stir at 400 rpm and react at 20°C for 8 h to obtain an intermediate containing peroxidized oleic triglyceride and peroxidized oleic acid. The peroxide value of the intermediate at the reaction endpoint is 200 meq / kg. The ozone absorption capacity is 80 mg / g.

[0057] Step 2: Increase the reaction temperature to 70℃, introduce oxygen into the intermediate at a rate of 0.5 L / kg, and react for 8 hours to finally obtain azelaic acid oil.

[0058] Comparative Example 3 A method for preparing an azelaic acid oil includes the following steps: Step 1: An ozone / oxygen mixture was introduced into 350g of triglycerides at a flow rate of 1L / min. The ozone concentration in the mixture was 3% (v / v). The mixture was stirred at 400rpm and reacted at 20℃ for 8h to prepare triglycerides peroxide with a peroxide value of 200 meq / kg. The ozone absorption capacity was 80mg / g.

[0059] Step 2: Increase the reaction temperature to 70℃, introduce oxygen into the intermediate at a rate of 0.5 L / kg, and react for 8 hours to finally obtain triglyceride peroxide.

[0060] Step 3: Mix 0.5 parts azelaic acid with 99.5 parts triglyceride peroxide at 300 rpm and a stirring temperature of 95°C to finally prepare the azelaic acid oil. The appearance of the obtained azelaic acid oil is as follows. Figure 3 As shown, it is turbid and has poor solubility.

[0061] Comparative Example 4 A method for preparing an azelaic acid oil includes the following steps: Step 1: Mix 5 parts oleic acid with 95 parts triglycerides. Introduce an ozone / oxygen mixture at a flow rate of 1 L / min into 350 g of the oleic acid and triglyceride mixture. The ozone gas concentration in the mixture should be 3% (v / v). Stir at 400 rpm and react at 20°C for 8 h to obtain an intermediate containing triglycerides peroxide and oleic acid peroxide. The peroxide value of the intermediate at the reaction endpoint should be 200 meq / kg. The ozone absorption capacity is 80 mg / g.

[0062] Step 2: The reaction temperature is 100℃. Oxygen is introduced into the intermediate at a flow rate of 0.5 L / kg. The reaction is carried out for 8 hours to finally obtain azelaic acid oil.

[0063] The prepared azelaic acid oil was subjected to qualitative and quantitative analysis of azelaic acid, qualitative analysis of the triglyceride peroxide component in the oil, and the performance of the azelaic acid oil was studied. The operating steps are as follows: 1. Qualitative detection of azelaic acid by liquid chromatography-mass spectrometry Qualitative analysis of the samples prepared in Example 1 was performed using a Shimadzu LCMS-8040 triple quadrupole liquid chromatography-mass spectrometry system. The chromatographic column used was a WomdaSil C18 (250 mm × 4.6 mm, 5 μm). The mobile phase A was 0.1% formic acid aqueous solution, and the mobile phase B was acetonitrile. The mobile phase flow rate was 1 mL / min, the injection volume was 10 µL, the column temperature was 40 °C, and the gradient elution program is shown in Table 1. The ion source was ESI, and the MRM parameters are shown in Table 2. The samples were extracted with boiling water, and the aqueous layer was diluted with 50% methanol-water for analysis. The detection results are as follows: Figure 4 As shown.

[0064] Table 1 Gradient elution time program

[0065] Table 2 MRM Parameters

[0066] Figure 4 The results show that azelaic acid is present in the azelaic acid oil prepared in Example 1. Similarly, azelaic acid is also present in the azelaic acid oil prepared in Examples 2-6.

[0067] 2. Quantitative liquid chromatography detection A Shimadzu LC-20A high-performance liquid chromatograph (SPD-16 UV-Vis detector, WondaSil C18-WR column, 5μm, 4.6×150mm) was used. The mobile phase flow rate was 1ml / min, and gradient elution was employed. The elution program is detailed in Table 3 below. The column temperature was 30℃. Azelaic acid showed a maximum absorption peak at a detection wavelength of 215nm. Mobile phase: Phase A 50mmol / L sodium dihydrogen phosphate, pH 3; Phase B acetonitrile. The sample was dissolved in isopropanol, and 20µL was injected. The azelaic acid content in Examples 1, 2, 3, 4, 5, and 6, and Comparative Examples 1 and 2 were quantitatively determined. The detection results of the azelaic acid oil in Example 1 are as follows: Figure 5 As shown.

[0068] Table 3 Gradient elution time program

[0069] The azelaic acid content in Examples 1, 2, 3, 4, 5, 6 and Comparative Examples 1 and 2 is as follows: Figure 6 As shown.

[0070] Comparative Example 4, due to excessively high temperature, produced a large amount of byproducts, resulting in a very low azelaic acid content of only 0.181%, far lower than that of Example 1. Comparative Examples 2 and 4, with their low azelaic acid content, showed inferior therapeutic effects compared to Example 1.

[0071] 3. Liquid chromatography-mass spectrometry (LC-MS) determination of triglycerides peroxide Triglycerides peroxide, glyceryl peroleate-palmitoyl peroxide, glyceryl peroleate-stearate, glyceryl peroleate-linoleic acid, and glyceryl peroleate-linoleic acid all belong to the category of triglycerides peroxide. This experiment, using Example 4 as a representative example, verifies through liquid chromatography-mass spectrometry (LC-MS) that triglycerides peroxide can be generated in the final product of the preparation process.

[0072] 100 μL of the sample prepared in Example 4 was dissolved in 300 μL of ice-cold methanol, vortexed for 30 s, and sonicated in an ice-water bath for 10 min. 1 mL of methyl tert-butyl ether (MTBE) was added, and the mixture was vortexed for 30 s. 300 μL of ultrapure water was added, and the mixture was vortexed. The sample was pre-cooled at 4 °C for 10 min, centrifuged at 14000 rpm at 4 °C for 10 min, and 400 μL of the supernatant was collected and evaporated to dryness under vacuum. 80 μL of dichloromethane-methanol reconstitution solution (1:1) was added to reconstitute the sample. 5 μL of the solution was analyzed using a Thermo QE HF-X mass spectrometer and a Thermo Vanquish chromatograph.

[0073] Testing conditions: Chromatographic conditions: A Thermo Accucore C30 (2.6 μm, 2.1 mm × 150 mm) column was used at a column temperature of 45 °C. Mobile phase A consisted of 40% water, 60% acetonitrile, 10 mM ammonium formate, and 0.1% formic acid. Mobile phase B consisted of 90% isopropanol, 10% acetonitrile, 10 mM ammonium formate, and 0.1% formic acid. The gradient elution table is shown below. Table 4 Gradient elution table

[0074] (2) Mass spectrometry conditions: ESI (Positive), ion source voltage 3500V, ion transmission tube temperature 350℃, evaporator temperature set to 350℃. Sheath gas flow rate, auxiliary gas flow rate and purge gas flow rate were set to 40 Arb, 10 Arb and 1 Arb respectively. S-lens RF intensity was set to 50% to optimize ion transmission efficiency.

[0075] Test results as follows Figure 7 , Figure 8 , Figure 9 As shown. Figure 10 It is triglyceride peroxide (C 57 H 104 O 15 The molecular structure of ) has a molecular weight of 1028. Figure 7 The sample was obtained in positive ion mode (M+ NH4) after liquid chromatography-mass analysis. + Total ion chromatogram (TIC chromatogram). Figure 8 The filter was based on m / z 1046 (C) 57 H 104 O 15 [M+ NH4] + Extraction ion chromatogram (XIC chromatogram). The figure shows triglycerides peroxide (C... 57 H 104 O 15 The peak appears around 22.54 min. Further analysis of C... 57 H 104 O 15 [M+NH4] + MS / MS analysis was performed, and the results are as follows: Figure 9 As shown in the figure, it can be seen that at m / z 383[(C 57 H 104 O 15 +NH4) -C 18 H 33 O5-C9H 18 O2-C9H 18 O2-NH3]、m / z 353[(C 57 H104 O 15 +NH4) -C 18 H 33 O5-C9H 18 O2-C 10 H 20 O3-NH3]、m / z 399[(C 57 H 104 O 15 +NH4)-C 18 H 33 O5-C9H 18 O-C9H 18 O2-NH3]、m / z 415[(C 57 H 104 O 15 +NH4)-C 18 H 33 O5-C9H 18 O-C9H 18 O-NH3] is C 57 H 104 O 15 [M+ NH4] + The presence of ion fragments, combined with the TIC spectrum and secondary mass spectrum, suggests that the sample contains triglycerides peroxide.

[0076] 4. Animal experiments with acute photodamage The prepared azelaic acid oil was subjected to acute photodamage animal experiments, as detailed below: 4.1 Experimental Objective To investigate the therapeutic effect of azelaic acid oil on acute photodamage, an ultraviolet-induced acute photodamage guinea pig model was established to explore its therapeutic effect on photodamage.

[0077] 4.2 Experimental Methods (1) Study subjects and grouping: 16 ordinary male guinea pigs, 6-8 weeks old (250-350g), were randomly divided into 4 groups: UVB model group, azelaic acid oil group, triglyceride peroxide group, and azelaic acid gel group, with 4 guinea pigs in each group. The feeding conditions such as light, temperature (24±1℃), and humidity were the same for all experimental guinea pigs in each group.

[0078] (2) Modeling method: The back of the guinea pig was shaved, with an area of ​​4cm×5cm. A broadband UVB phototherapy device (PL-S 9W / 12) was used to irradiate the shaved area on the back of each group of experimental guinea pigs, with a daily irradiation dose of 3.75J / cm. 2 Irradiation intensity 12mW / cm 2Irradiation time is 5 minutes, the distance between the light source and the shaved area skin is about 15cm, irradiate once a day, and observe with the naked eye for 2 consecutive days. When erythema and edema appear, it indicates that the modeling is successful.

[0079] (3) Intervention method: After successful modeling, the drug was applied to the back area exposed to light once a day for 5 consecutive days. The azelaic acid oil group was treated with the oil prepared in Example 4, the triglyceride peroxide group was treated with the oil prepared in Comparative Example 3, and the azelaic acid gel group was treated with Finacea azelaic acid gel (30g, azelaic acid content 15wt%). The UVB model group was not treated in any way.

[0080] 4.3 Experimental Results Experimental results are as follows Figure 11 As shown in the figure, A represents the acute photodamage model group (UVB model group), B represents the triglyceride peroxide group, C represents the azelaic acid gel group, and D represents the azelaic acid oil group. In the UVB model group, obvious erythema, crusting, and scaling were visible on the backs of the guinea pigs exposed to light. Compared to the model group, the azelaic acid oil group showed faster skin lesion recovery, with reduced crusting on the backs and significant reduction in inflammation. The azelaic acid gel group showed some relief compared to the model group, but the recovery was worse than that of the azelaic acid oil group.

[0081] like Figure 12 The results of confocal microscopy using reflection microscopy show that, in the image, A represents the acute photodamage model group (UVB model group), B represents the azelaic acid gel group, C represents the triglyceride peroxide group, and D represents the azelaic acid oil group. In the UVB model group, the epidermis was thickened, and numerous inflammatory cells were visible in the superficial dermis. Compared to the UVB model group, the triglyceride peroxide group and the azelaic acid oil group showed less papillary dermal edema and less inflammatory cell infiltration. However, the azelaic acid oil group showed significantly better results than the triglyceride peroxide group and the azelaic acid gel group.

[0082] 5. Clinical application observation experiment This experiment aims to observe whether azelaic acid oil and azelaic acid gel have protective and therapeutic effects on acute photodamage induced by simulated sunlight exposure.

[0083] Ten healthy female volunteers were recruited, and each volunteer signed an informed consent form. Inclusion criteria: no systemic diseases or skin conditions; no history of systemic or topical skin treatments; no history of photosensitivity. Before the experiment, the minimum erythema intensity (EI) was determined for each volunteer. The following procedures were performed on four sites on the volunteers' backs: Site 1 was treated with azelaic acid oil (Example 4); Site 2 was treated with azelaic acid gel (Finacea azelaic acid gel, 30g, 15% azelaic acid content, w / w); Site 3 was only irradiated using a sunlight simulator without the application of the test sample; and Site 4 was neither irradiated nor treated with the test sample. Each irradiated site was irradiated once daily for five consecutive days. The EI value was measured using a narrow-band reflectance spectrophotometer before the start of the experiment and after the last irradiation. Each site was tested three times, and the average value was taken.

[0084] EI values ​​reflect the degree of erythema on the skin. Before the study, EI values ​​fluctuated between 184.15 and 200. After the study, EI values ​​in sites 1-3 increased significantly, with sites 2 and 3 showing particularly significant increases. Sites 2 and 3 increased from 200 and 175.4 before treatment to 561.12 and 523.12, respectively. Site 1 showed a smaller increase, rising from 197.23 before treatment to 452.12. Furthermore, more significant pigmentation was observed in site 2 compared to site 1.

[0085] This demonstrates the protective effect of azelaic acid oil against acute photodamage induced by simulated sunlight exposure, while also reducing pigmentation.

[0086] 6. Clinical application and efficacy study of azelaic acid oil in the treatment of high-altitude photodamage. 6.1 Research Methods Study subjects: 25 patients diagnosed with acute photodamage in high-altitude environments.

[0087] Inclusion criteria for patients with acute photodamage: meeting the relevant diagnostic criteria of the "Guidelines for the Diagnosis and Treatment of Sunburn in Primary Care", the affected area is the exposed area, there is no cognitive impairment, good communication ability, high treatment compliance, and informed consent to treatment.

[0088] Exclusion criteria: Individuals who used or received related topical medications or treatments prior to this treatment, those exposed to known photosensitizers or photoallergens, or those with other diseases that cause photosensitivity symptoms (such as lupus erythematosus and rosacea).

[0089] Treatment: Routine skin cleansing and sun protection were performed, followed by topical application of azelaic acid oil (Example 4) twice daily. The patient's skin lesion recovery was assessed after two weeks of treatment.

[0090] Clinical efficacy grading: Cured means that the redness and swelling symptoms have completely subsided, the blisters have scabbed over and completely healed, and the pigmentation spots have faded or become difficult to observe; Significantly effective means that the redness, swelling, blisters and other symptoms have improved, and the pigmentation spots have decreased in size or color; Ineffective means that there is no significant change in the above indicators before and after medication.

[0091] 6.2 Research Results Of the 25 patients, 1 patient discontinued treatment midway, and 24 patients completed the treatment. After two weeks of treatment, the overall effective rate reached 91.66%, including 12 cases of complete recovery, 10 cases of significant improvement, and 2 cases of no effect. See details below. Figure 13 After two weeks of treatment, the scabs fell off, the skin ulcers healed, and there was no obvious pigmentation. Furthermore, the original pigmented lesions gradually returned to their normal skin color.

[0092] 6.3 Research Conclusions Azelaic acid oil is effective in treating acute photodamage, with an efficacy rate of up to 92%. The triglycerides peroxide in azelaic acid oil can effectively repair skin lesions and relieve inflammation. Simultaneously, azelaic acid can reduce pigment formation and repair the pigment barrier. As an oil, it has moisturizing and hydrating properties, preventing further deterioration of photodamage.

[0093] It should be noted that the above embodiments are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. Those skilled in the art can make other variations or modifications based on the above description. It is impossible to exhaustively list all possible implementations here. All obvious variations or modifications derived from the technical solutions of this invention are still within the scope of protection of this invention.

Claims

1. A method for preparing an azelaic acid oil, characterized in that, Includes the following steps: S1. Oleic acid and triglycerides are mixed and then a mixture of ozone and oxygen is introduced under stirring. The mixture is reacted at 20-30°C for 8-14 hours to obtain an intermediate product. S2. Oxygen is introduced into the intermediate product at 70℃-95℃ and the reaction is carried out for 6-10 hours to obtain azelaic acid oil.

2. The preparation method according to claim 1, characterized in that, The triglyceride is one or more of the following: oleic acid triglyceride, dioleoyl palmitate triglyceride, dioleoyl stearate triglyceride, dioleoyl linoleate triglyceride, and linoleic acid triglyceride.

3. The preparation method according to claim 1, characterized in that, In step S1, the volume ratio of oleic acid to triglycerides is 5-20:80-95.

4. The preparation method according to claim 1, characterized in that, In step S1, the volume ratio of ozone to oxygen in the ozone and oxygen mixture is 3-5:95-97.

5. The preparation method according to claim 1, characterized in that, In step S1, the rate at which the mixed gas of ozone and oxygen is introduced is 0.5-1.5 L / min.

6. The preparation method according to claim 1, characterized in that, In step S1, the peroxide value of the intermediate product is 200-280 meq / kg; the ozone absorption capacity of the intermediate product is 80-144 mg / g.

7. The preparation method according to claim 1, characterized in that, In step S2, the oxygen ventilation rate is 0.2-0.7 L / kg.

8. The preparation method according to any one of claims 1-7, characterized in that, The content of azelaic acid in azelaic acid oil is 0.5-2.5 wt%.

9. Azelaic acid oil prepared by the preparation method according to any one of claims 1-8.

10. The use of the azelaic acid oil preparation according to claim 9 in the preparation of medicaments for treating or preventing acute photodamage, chronic photodamage, barrier repair, anti-inflammation, or reducing pigmentation.

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