Peptide composition for treating allergic skin diseases and preparation method thereof

Abstract

The invention relates to the technical field of biological medicines, in particular to a peptide composition for treating allergic skin diseases, which comprises a lipid nanoparticle carrier, an active component, a stabilizer and deionized water, the lipid nanoparticle carrier is prepared from acetylated dioleoyl phosphatidyl glycerol, cholesterol and polyethylene glycol-100 stearate; the acetylated dioleoyl phosphatidyl glycerol is obtained by modifying dioleoyl phosphatidyl glycerol through an acetylation reagent; the active component is arginine / lysine polypeptide; the stabilizer is trehalose. According to the invention, acetylation-modified dioleoyl phosphatidyl glycerol is used for constructing a lipid nanoparticle carrier, and the lipid nanoparticle carrier is matched with an active component and a stabilizer to form the composition; the acetylated carrier can improve the stability and skin absorption efficiency of active ingredients and assist the active ingredients to play an anti-allergic role so as to treat allergic skin diseases, and meanwhile, the arginine / lysine polypeptide is cooperated with other components, so that the problem of facial contour aging caused by the skin diseases can be improved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine, in particular to a peptide composition for treating allergic dermatosis and a preparation method thereof. BACKGROUND

[0002] Allergic dermatosis is a common inflammatory skin disease in clinic, which covers atopic dermatitis, contact dermatitis, allergic eczema and other types. The core pathological feature of such diseases is the impairment of skin barrier function, which makes external allergens (such as dust mites, pollen, cosmetic ingredients) easily penetrate the epidermis to activate mast cells and eosinophils, release histamine, leukotrienes and other inflammatory mediators, and trigger acute symptoms such as erythema, pruritus and desquamation; and the long-term repeated inflammatory response further mediates the "inflammation-aging" cascade, becoming an important inducement of facial contour aging.

[0003] The facial contour aging caused by allergic dermatosis mainly includes: first, the sustained inflammation activates matrix metalloproteinases (MMP-1, MMP-3), accelerates the degradation of dermal layer type I and type III collagen, leads to the destruction of skin support structure, and causes contour relaxation problems such as apple muscle sagging and mandibular line blurring; second, inflammatory factors (such as TNF-α, IL-6) inhibit the activity of fibroblasts, reduce the synthesis of elastic fibers, and thus reduce the elasticity and tightness of the skin, further aggravating deep wrinkles such as command lines and puppet lines; third, after the impairment of skin barrier, the trans-epidermal water loss (TEWL) is significantly increased, and the water content in the dermal layer is reduced, leading to the dryness and lack of fullness of facial skin, further amplifying the visual effect of contour aging.

[0004] Current treatment methods for the above problems have certain limitations. On the one hand, traditional anti-allergic treatment mainly uses antihistamines and external glucocorticoids, which can short-term relieve inflammatory symptoms, but cannot repair the damaged skin barrier, nor reverse the collagen loss and elastic fiber damage that have already occurred, and thus cannot improve facial contour aging; and long-term use of glucocorticoids can easily cause skin atrophy and telangiectasia, and thus aggravate skin fragility and aging appearance. On the other hand, existing anti-aging skin care products mostly use retinol, vitamin C and hyaluronic acid as core ingredients, which can improve aging by promoting collagen production or supplementing water, but such ingredients are mostly irritating, which can easily cause secondary allergy to the damaged skin of patients with allergic dermatosis, and lack of targeting for "inflammation-mediated aging", thus cannot block the associated pathway of disease and aging from the source.

[0005] Peptide ingredients are gradually attracting attention in the field of sensitive skin anti-aging due to their advantages of low allergenicity, high biological activity and clear target. Previous studies have shown that cationic peptides such as arginine / lysine peptides can inhibit the release of inflammatory factors to play an anti-allergic effect, and at the same time, activate the TGF-β / Smad signaling pathway of fibroblasts to promote collagen synthesis. However, peptide molecules are easily degraded by skin proteases, and the transdermal absorption efficiency is insufficient, making it difficult to reach the dermis to play an anti-aging role. The existing technology often adds chemical penetration enhancers such as propylene glycol and azone to improve the transdermal depth of peptides to act on the dermis. For peptides that can stimulate collagen synthesis, the existing technology often adds chemical penetration enhancers such as propylene glycol and azone, but such penetration enhancers can damage the lipid barrier of the stratum corneum, leading to a decrease in the skin's water retention capacity, and long-term use can easily cause erythema and stinging in sensitive areas such as the mandibular margin and cheeks. Moreover, the penetration effect is unstable - the effective concentration of peptides actually reaching the dermis is insufficient, making it difficult to activate a sufficient number of fibroblasts to synthesize collagen, and unable to achieve long-term tightening of the contour.

[0006] Therefore, in view of the limitations of the above-mentioned prior art, it is of great significance to develop a peptide composition to fill the technical gap in the treatment of allergic skin diseases and the facial contour aging caused by them. SUMMARY

[0007] In view of the shortcomings of the prior art, the present application provides a peptide composition for treating allergic skin diseases and a preparation method thereof. The composition uses acetylated dioleoyl phosphatidylglycerol, cholesterol and polyethylene glycol-100 stearate to construct a lipid nanoparticle carrier, and combines arginine / lysine peptides as active ingredients and trehalose as a stabilizer. The acetylated carrier improves the transdermal efficiency and stability of the peptides. The peptides can inhibit the release of histamine and inflammatory reactions to treat allergic skin diseases, while activating fibroblasts to promote collagen synthesis, thereby improving the contour aging problems such as collagen loss and relaxation caused by repeated allergic skin diseases.

[0008] To achieve the above-mentioned purposes, the present application adopts the following technical solutions:

[0009] In a first aspect, the present application provides a peptide composition for treating allergic skin diseases, comprising a lipid nanoparticle carrier, active ingredients, a stabilizer and deionized water. The lipid nanoparticle carrier comprises acetylated dioleoyl phosphatidylglycerol, cholesterol and polyethylene glycol-100 stearate. The acetylated dioleoyl phosphatidylglycerol is obtained by modifying dioleoyl phosphatidylglycerol with an acetylation reagent. The active ingredient is arginine / lysine peptide. The stabilizer is trehalose.

[0010] In the present application, acetic anhydride acetylates the hydroxyl group on the phosphate group of the dioleoyl phosphatidylglycerol head, converting the hydrophilic hydroxyl group (-OH) into a more hydrophobic acetyl ester group (-OCOCH3). In an aqueous environment, the structure of the hydrophobic core is more compact through self-assembly and mutual aggregation, thus better coating the hydrophilic polypeptide. At the same time, the polyethylene glycol-100 stearate in the carrier reduces non-specific adsorption to skin cells through its electrically neutral and steric hindrance properties, thus protecting the carrier and promoting its transdermal delivery, sending the polypeptide to the inflammatory aggregation area in the dermis. Cholesterol fills the gaps between lipid molecules, enhancing the tightness and stability of the lipid membrane and preventing the leakage of the coated polypeptide.

[0011] In the inflammatory area microenvironment, the pH is acidic (pH ≈ 5.5-6.0), and the acidic conditions trigger the phosphate group (-O-PO3H 2- ) of the acetylated dioleoyl phosphatidylglycerol head to combine with H + , resulting in protonation (-O-PO3H - ) and a decrease in the negative charge of the phosphate group, thus destroying the structural stability of the lipid nanoparticle and triggering slight disassembly of the carrier, slowly releasing the coated polypeptide. At the same time, the amino group of the arginine / lysine polypeptide is protonated (-NH3 + ), and the positively charged polypeptide is more easily combined with the anion receptors on the surface of inflammatory cells (such as mast cells and macrophages), on the one hand inhibiting the release of inflammatory mediators such as histamine and TNF-α, and on the other hand penetrating the damaged stratum corneum to reach the dermis, activating the TGF-β / Smad pathway of fibroblasts, promoting the synthesis of type I and type III collagen, and improving facial contour relaxation and wrinkles.

[0012] Preferably, the mass ratio of the lipid nanoparticle carrier, active ingredient, stabilizer, and deionized water is (5-15):(0.1-2):(1-5):(80-90).

[0013] Preferably, in the lipid nanoparticle carrier, the mass ratio of acetylated dioleoyl phosphatidylglycerol, cholesterol, and polyethylene glycol-100 stearate is (3-5):(1-2):(0.5-1.5).

[0014] Preferably, the acetylation reagent is acetic anhydride, and the mass ratio of acetic anhydride to dioleoyl phosphatidylglycerol is 1:(5-7), and the acetylation degree of the modified acetylated dioleoyl phosphatidylglycerol is ≥85%.

[0015] Preferably, the trehalose is food grade, with a purity of ≥99.5% and a water content of ≤0.5%.

[0016] In a second aspect, the present application provides a method for treating allergic skin diseases using a peptide composition, comprising the following steps:

[0017] S1. Dioleoylphosphatidylglycerol is dissolved in an organic solvent, and an acetylation reagent is added under low temperature conditions and the mixture is stirred to react. After quenching, extraction, drying and rotary evaporation, acetylated dioleoylphosphatidylglycerol is obtained.

[0018] S2. Acetylated dioleoylphosphatidylglycerol, cholesterol, and polyethylene glycol-100 stearate are completely dissolved in ethanol at a certain mass ratio, and the mixture is stirred to obtain a first mixture; the first mixture is rapidly mixed and stirred with an acidic aqueous buffer, and then dialyzed and filtered to remove bacteria to obtain a lipid nanoparticle carrier.

[0019] S3. Arginine / lysine polypeptide is synthesized using a solid-phase synthesis method, wherein the solid-phase synthesis method includes, in sequence, deprotection, activation, cross-linking, cycling, elution, deprotection, purification and structural identification steps;

[0020] S4. The arginine / lysine polypeptide is used as the active ingredient and mixed with the lipid nanoparticle carrier, trehalose and deionized water in a predetermined ratio, and homogenized to obtain the peptide composition.

[0021] Preferably, in S1, the organic solvent is chloroform or dichloromethane; the mass ratio of dioleoylphosphatidylglycerol to the organic solvent is 1:(3.5-4.5); the low temperature is 2-5℃; the addition rate of the acetylation reagent is 0.8-1.2 mL / min; the stirring speed is 300-400 rpm and the time is 20-30 min.

[0022] Preferably, in S2, the acidic aqueous buffer solution includes any one of citrate-sodium citrate buffer, acetate-sodium acetate buffer, and sodium dihydrogen phosphate-disodium hydrogen phosphate buffer; the mass ratio of the first mixture to the acidic aqueous buffer solution is 1:(5-10); and the rapid mixing speed is 1000-1500 rpm.

[0023] Preferably, in step S4, the homogenization process uses a high-pressure homogenizer with a homogenization pressure of 30-60 MPa and a homogenization cycle of 3-5 times.

[0024] Compared with the prior art, the beneficial effects of this application are as follows:

[0025] This application provides a peptide composition for treating allergic skin diseases. In this application, acetic anhydride acetylates the hydroxyl group on the phosphate group of the dioleoylphosphatidylglycerol head, converting the hydrophilic hydroxyl group (-OH) into a more hydrophobic acetyl ester group (-OCOCH3). In an aqueous environment, this acetylation allows for self-assembly and aggregation, resulting in a more compact structure of the hydrophobic core and better encapsulation of the hydrophilic peptide. Simultaneously, polyethylene glycol-100 stearate in the carrier reduces non-specific adsorption to skin cells through its electroneutrality and steric hindrance properties, thereby protecting the carrier and promoting transdermal delivery of the peptide to the inflammatory accumulation area in the dermis. Cholesterol, by filling the gaps between lipid molecules, enhances the tightness and stability of the lipid membrane, preventing leakage of the encapsulated peptide.

[0026] The microenvironment in the inflammatory area is acidic (pH≈5.5~6.0). Acidic conditions trigger the formation of the phosphate group (-O-PO3) at the head of acetylated dioleoylphosphatidylglycerol. 2- It combines with H+ and undergoes protonation (-O-PO3H) - This process reduces the negative charge of the phosphate groups, disrupting the stability of the lipid nanoparticle structure and triggering a slight disassembly of the carrier, resulting in the slow release of the encapsulated peptide. Simultaneously, the amino groups of the arginine / lysine peptide are protonated (-NH3). + Positively charged peptides are more likely to bind to anion receptors on the surface of inflammatory cells (such as mast cells and macrophages). On the one hand, they inhibit the release of inflammatory mediators such as histamine and TNF-α, relieving allergy symptoms such as erythema and itching. On the other hand, peptides penetrate the damaged stratum corneum to reach the dermis, activate the TGF-β / Smad pathway of fibroblasts, promote the synthesis of type I and type III collagen, and improve facial contour laxity and wrinkles. Attached Figure Description

[0027] Figure 1 This is a schematic diagram of a method for treating allergic skin diseases using a peptide composition. Detailed Implementation

[0028] To make the technical problems, technical solutions, and beneficial effects to be solved by this application clearer, the application will be further described in detail below with reference to embodiments. However, this should not be construed as limiting the scope of this application to the following examples. All other embodiments obtained by those skilled in the art without creative effort without departing from the above-described methodological spirit of this application are within the scope of protection of this application.

[0029] In this application, the terminology used is for the purpose of describing particular embodiments only and is not intended to be limiting of this application.

[0030] The singular forms “for,” “or,” “a,” “any,” and “the” used in this application are intended to include the plural forms unless the context clearly indicates otherwise.

[0031] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0032] The following describes in detail, with reference to different embodiments, a method for preparing a peptide composition for treating allergic skin diseases according to this application.

[0033] Example 1

[0034] like Figure 1 As shown, this embodiment provides a method for preparing a peptide composition for treating allergic skin diseases, comprising the following steps:

[0035] S1. Dissolve dioleoylphosphatidylglycerol in chloroform at a mass ratio of 1:3.5 and stir until completely dissolved. Control the system temperature at 2°C, turn on the stirrer and adjust the speed to 300 rpm. Add acetic anhydride at a rate of 0.8 mL / min at a mass ratio of acetic anhydride to dioleoylphosphatidylglycerol of 1:4 and continue stirring for 30 min. After the reaction is completed, the mixture is quenched, extracted, dried and rotary evaporated in sequence to obtain acetylated dioleoylphosphatidylglycerol.

[0036] S2. Weigh acetylated dioleoylphosphatidylglycerol, cholesterol, and polyethylene glycol-100 stearate at a mass ratio of 3:1:0.5 and add them to anhydrous ethanol, wherein the mass ratio of acetylated dioleoylphosphatidylglycerol to anhydrous ethanol is 1:3; stir until all components are completely dissolved to obtain the first mixture. Using citrate-sodium citrate buffer, weigh the first mixture and citrate-sodium citrate buffer at a mass ratio of 1:5 and mix the first mixture with the buffer; then stir continuously at 1000 rpm for 15 min at 25°C, then transfer to a dialysis bag and dialyze with deionized water until no ethanol residue remains; after dialysis, filter sterilize using a microporous membrane to obtain the lipid nanoparticle carrier.

[0037] S3. Arginine / lysine polypeptides are synthesized using a solid-phase synthesis method, wherein the solid-phase synthesis method includes the following steps in sequence: deprotection, activation, cross-linking, cycling, elution, deprotection, purification, and structural identification.

[0038] S4. Weigh out the lipid nanoparticle carrier, arginine / lysine peptide, trehalose and deionized water in a mass ratio of 5:0.1:1:80 and mix them. Stir at room temperature until the trehalose is completely dissolved, then transfer to a high-pressure homogenizer and homogenize three times continuously at a homogenization pressure of 30 MPa to obtain a peptide composition.

[0039] Example 2

[0040] like Figure 1 As shown, this embodiment provides a method for preparing a peptide composition for treating allergic skin diseases, comprising the following steps:

[0041] S1. Dissolve dioleoylphosphatidylglycerol in chloroform at a mass ratio of 1:4 and stir until completely dissolved. Control the system temperature at 3°C, turn on the stirrer and adjust the speed to 350 rpm. Add acetic anhydride at a rate of 1.0 mL / min at a mass ratio of acetic anhydride to dioleoylphosphatidylglycerol of 1:6 and continue stirring for 25 min. After the reaction is completed, the mixture is quenched, extracted, dried and rotary evaporated in sequence to obtain acetylated dioleoylphosphatidylglycerol.

[0042] S2. Weigh acetylated dioleoylphosphatidylglycerol, cholesterol, and polyethylene glycol-100 stearate in a mass ratio of 4:1:1. Add anhydrous ethanol as a solvent, wherein the mass ratio of acetylated dioleoylphosphatidylglycerol to anhydrous ethanol is 1:4. Stir until all components are completely dissolved to obtain the first mixture. Using an acetate-sodium acetate buffer, weigh the first mixture and acidic aqueous buffer in a mass ratio of 1:8. Mix the first mixture and the buffer. Then, stir continuously at 1200 rpm for 20 minutes at 30°C. Transfer to a dialysis bag and dialyze with deionized water until no ethanol residue remains. After dialysis, filter sterilize using a microporous membrane to obtain the lipid nanoparticle carrier.

[0043] S3. Arginine / lysine polypeptides are synthesized using a solid-phase synthesis method, wherein the solid-phase synthesis method includes the following steps in sequence: deprotection, activation, cross-linking, cycling, elution, deprotection, purification, and structural identification.

[0044] S4. Weigh out the lipid nanoparticle carrier, arginine / lysine peptide, trehalose and deionized water in a mass ratio of 10:1:3:85 and mix them. Stir at room temperature until the trehalose is completely dissolved, then transfer to a high-pressure homogenizer and homogenize continuously for 4 times under a homogenization pressure of 40 MPa to obtain a peptide composition with stable structure and uniform dispersion.

[0045] Example 3

[0046] like Figure 1 As shown, this embodiment provides a method for treating allergic skin diseases using a peptide composition, comprising the following steps:

[0047] S1. Dissolve dioleoylphosphatidylglycerol in dichloromethane at a mass ratio of 1:4.5 and stir until completely dissolved. Control the system temperature at 5°C, turn on the stirrer and adjust the speed to 400 rpm. Add acetic anhydride at a rate of 1.2 mL / min at a mass ratio of acetic anhydride to dioleoylphosphatidylglycerol of 1:7 and continue stirring for 20 min. After the reaction is completed, the mixture is quenched, extracted, dried and rotary evaporated in sequence to obtain acetylated dioleoylphosphatidylglycerol.

[0048] S2. Weigh acetylated dioleoylphosphatidylglycerol, cholesterol, and polyethylene glycol-100 stearate at a mass ratio of 5:2:1. Add anhydrous ethanol as a solvent and stir until all components are completely dissolved to obtain the first mixture. Using sodium dihydrogen phosphate-disodium hydrogen phosphate buffer, weigh the first mixture and the sodium dihydrogen phosphate-disodium hydrogen phosphate buffer at a mass ratio of 1:10. Add the first mixture dropwise into the buffer. Then, stir continuously at 1500 rpm for 30 minutes at 35°C. Transfer the mixture to a dialysis bag and dialyze with deionized water until no ethanol residue remains. After dialysis, filter sterilize using a microporous membrane to obtain the lipid nanoparticle carrier.

[0049] S3. Arginine / lysine polypeptides are synthesized using a solid-phase synthesis method, wherein the solid-phase synthesis method includes the following steps in sequence: deprotection, activation, cross-linking, cycling, elution, deprotection, purification, and structural identification.

[0050] S4. Weigh out the lipid nanoparticle carrier, arginine / lysine peptide, trehalose and deionized water in a mass ratio of 15:2:5:90 and mix them. Stir at room temperature until the trehalose is completely dissolved, then transfer the mixture to a premix and send it to a high-pressure homogenizer. Homogenize continuously 5 times under a homogenization pressure of 60 MPa to obtain a peptide composition.

[0051] Comparative Example 1

[0052] A method for a peptide composition for treating allergic skin diseases, which differs from Example 2 in that soy lecithin is used instead of acetylated dioleoylphosphatidylglycerol.

[0053] Comparative Example 2

[0054] A method for treating allergic skin diseases using a peptide composition, which differs from Example 2 in that it uses unacetylated dioleoylphosphatidylglycerol.

[0055] Comparative Example 3

[0056] A method for treating allergic skin diseases using a peptide composition, which differs from Example 2 in that propionic anhydride is used instead of acetic anhydride.

[0057] Comparative Example 4

[0058] A method for a peptide composition for treating allergic skin diseases, which differs from Example 2 in that lipid nanoparticle carriers are not prepared.

[0059] Performance testing:

[0060] 1. Anti-allergy efficacy test: This test is divided into in vitro test and in vivo test. In vitro test uses rat peritoneal mast cells as a model, and uses ELISA to measure histamine release inhibition rate and flow cytometry to measure IL-4 / TNF-α secretion. In vivo test selects volunteers with allergic dermatitis, and after applying the samples of Examples 1-3 and Comparative Examples 1-4 in groups, the redness and swelling area is quantified.

[0061] 2. Transdermal efficiency test: Using a Franz diffusion cell combined with a simulated skin model (such as ex vivo pig skin), samples from Examples 1-3 and Comparative Examples 1-3 were applied to the surface of the model, and the content of arginine / lysine peptides in the receiving solution was detected after 24 hours. Transdermal rate = (total peptides in the receiving solution / total peptides in the applied sample) × 100% to evaluate the efficiency of active peptides in penetrating the stratum corneum of the skin and ensure that they can reach the skin target site to exert their efficacy.

[0062] 3. Stability test of active ingredients: The peptide compositions of Examples 1-3 and Comparative Examples 1-3 were placed at 45°C for accelerated storage for 3 months. The retention rate of arginine / lysine peptides was detected by high performance liquid chromatography (HPLC) to verify that the active ingredients of the compositions do not degrade or become ineffective during the shelf life and maintain stable efficacy.

[0063] 4. Skin Elasticity and Contour Firmness Test: Seventy volunteers aged 35-50 (half male, half female) were selected, all with sensitive skin and skin diseases leading to jawline laxity (elasticity value < 0.6). The volunteers were randomly divided into 6 groups of 10 each, corresponding to Examples 1, 2, 3, Comparative Examples 1, 2, and 3, respectively. Jawline skin elasticity was measured using a Cutometer® MPA580 before use, and after 7 and 14 days of use. The test compositions from the Example and Comparative Example groups were applied to the face (focusing on the jawline) after cleansing morning and evening, at a dosage of 0.5g per application. Other anti-aging products were discontinued during the treatment period. Note: The closer the elasticity value is to 1, the better the skin elasticity and the higher the firmness.

[0064] Table 1. Anti-allergy efficacy test data

[0065]

[0066] As shown in Table 1, the acetylation modification in the examples introduced negatively charged acetyl groups onto the phospholipid molecules. Mast cells and other immune cells have numerous positively charged receptors or membrane regions on their membrane surfaces. Through electrostatic adsorption, lipid nanoparticle carriers actively target and accumulate around effector cells (such as mast cells) in allergic reactions, achieving precise delivery. The nanoparticle structure can be more effectively taken up by cells through pathways such as endocytosis, thereby directly delivering high concentrations of active peptides into the cell, greatly improving the bioavailability and efficacy of the peptides.

[0067] Comparative Example 1: Soybean lecithin failed to form a stable, uniform nanoparticle structure, losing its active targeting capability and resulting in low drug delivery efficiency. Comparative Example 2: Unacetylated dioleoylphosphatidylglycerol, while capable of forming nanoparticles, lacked a targeting acetyl group, relying mainly on passive diffusion and failing to effectively accumulate around target cells. Comparative Example 3: Propionic anhydride modification introduced a propionyl group with greater steric hindrance, potentially altering the charge distribution, membrane fluidity, or binding mode of the phospholipid, resulting in slightly lower targeting efficiency and drug loading stability compared to the optimal acetic anhydride modification (Example 2). Comparative Example 4: Carrier-free "naked peptides" struggled to penetrate cell membranes and were easily degraded by extracellular enzymes, failing to effectively reach the target site, thus exhibiting the worst effect.

[0068] Table 2. Test data on transdermal efficiency and stability of active ingredients.

[0069]

[0070] As shown in Table 2, Examples 1-3, as the core solutions of this application, all achieved dermal peptide concentrations of 48.5-52.8 μg / cm² after 24 hours, with transdermal efficiencies of 19.4%-21.1%. After 3 months of accelerated treatment at 45°C, the peptide retention rate remained at 95.2%-96.8%. This is due to the acetylation modification giving dioleoylphosphatidylglycerol stronger membrane fusion activity and skin active transport recognition, combined with the structural stabilizing effect of cholesterol and the permeation-enhancing function of polyethylene glycol-100 stearate. This not only achieved efficient transdermal delivery of peptides but also ensured stable encapsulation of peptides by the carrier while maintaining the uniformity of system dispersion.

[0071] Compared to the examples, Comparative Example 1, which used soybean lecithin instead of acetylated dioleoylphosphatidylglycerol, had a 24-hour dermal peptide concentration of only 30 μg / cm³. 2The transdermal efficiency was as low as 8.9% because the structural compatibility between soybean lecithin and skin cell membranes is much lower than that of acetylated dioleoylphosphatidylglycerol, resulting in a lack of membrane fusion and active transport capabilities. This makes it impossible to efficiently deliver peptides and maintain the stability of the carrier system. Comparative Example 2 used unacetylated dioleoylphosphatidylglycerol, with a 24-hour dermal peptide concentration of 30.0 μg / cm² and a transdermal efficiency of 12.0%. The lack of acetylation modification led to a decrease in the membrane fusion activity of dioleoylphosphatidylglycerol, preventing it from rapidly penetrating the stratum corneum. Poor carrier dispersibility affects the enrichment of peptides in the dermis. In Comparative Example 3, propionic anhydride was used instead of acetic anhydride as the acetylation agent. Although the dermal peptide concentration of 32.5 μg / cm² and transdermal efficiency of 13.0% after 24 hours were close to those of Example 2, the peptide retention rate was only 90.5% after 3 months of accelerated storage at 45°C. The propionyl group introduced by propionic anhydride has a longer carbon chain than the acetyl group of acetic anhydride, which reduces the structural matching degree with human cell membrane phospholipid molecules. The carrier's protective ability for peptides is weakened, resulting in a large amount of peptide degradation during accelerated storage.

[0072] Table 3. Data from Skin Elasticity and Facial Firmness Tests

[0073]

[0074] As shown in Table 3, Examples 1-3 showed a significant increase in the elasticity value of the skin along the jawline, reaching 0.78-0.82 after 28 days, an increase of 0.24-0.26 compared to before use. Comparative Examples 1-3 showed only 0.65-0.70, an increase of 0.09-0.13. This difference is attributed to the high transdermal permeability (68%-70%) of the acetylated dioleoylphosphatidylglycerol carrier, which can precisely deliver the peptides to the dermis, activating fibroblasts to synthesize collagen and elastin; trehalose repairs the skin barrier and reduces collagen degradation. Example 2 had the highest elasticity value due to its optimal carrier ratio and homogenization parameters. Comparative Example 1 used soybean lecithin, resulting in poor carrier transdermal permeability (26%), making it difficult for the peptides to reach the target site; Comparative Example 2 did not use acetylated dioleoylphosphatidylglycerol, leading to insufficient carrier penetration; Comparative Example 3 suffered from carrier rupture due to propionic anhydride, causing peptide leakage. All these methods failed to effectively activate collagen synthesis, resulting in slow elasticity improvement.

[0075] In summary, the data in the table fully demonstrates that the peptide composition constructed by acetylated dioleoylphosphatidylglycerol in this application is irreplaceable in terms of peptide delivery efficiency, activity stability, and skin improvement effect. The comparative examples, due to the destruction of the core innovative features, cannot achieve the excellent performance of the examples.

[0076] The above results demonstrate and describe the basic principles and main features of this application, as well as its advantages.

[0077] Those skilled in the art should understand that this application is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of this application. Various changes and modifications can be made to this application without departing from the spirit and scope thereof, and all such changes and modifications fall within the scope of this application as claimed. The scope of protection of this application is defined by the equivalents of the appended claims.

Claims

1. A peptide composition for treating allergic skin diseases, characterized in that, include: The lipid nanoparticle carrier comprises an active ingredient, a stabilizer, and deionized water; the lipid nanoparticle carrier contains acetylated dioleoylphosphatidylglycerol, cholesterol, and polyethylene glycol-100 stearate; the acetylated dioleoylphosphatidylglycerol is obtained by modifying dioleoylphosphatidylglycerol with an acetylation reagent; the active ingredient is an arginine / lysine peptide; and the stabilizer is trehalose.

2. The peptide composition for treating allergic skin diseases according to claim 1, characterized in that, The mass ratio of the lipid nanoparticle carrier, active ingredient, stabilizer and deionized water is (5-15):(0.1-2):(1-5):(80-90).

3. The peptide composition for treating allergic skin diseases according to claim 1, characterized in that, In the lipid nanoparticle carrier, the mass ratio of acetylated dioleoylphosphatidylglycerol, cholesterol, and polyethylene glycol-100 stearate is (3-5):(1-2):(0.5-1.5).

4. The peptide composition for treating allergic skin diseases according to claim 1, characterized in that, The acetylation reagent is acetic anhydride, and the mass ratio of acetic anhydride to dioleoylphosphatidylglycerol is 1:(5-7).

5. A method for preparing a peptide composition for treating allergic skin diseases according to any one of claims 1-4, characterized in that, Includes the following steps: S1. Dioleoylphosphatidylglycerol is dissolved in an organic solvent, an acetylation reagent is added and the reaction is stirred. After quenching, extraction, drying and rotary evaporation, acetylated dioleoylphosphatidylglycerol is obtained. S2. Acetylated dioleoylphosphatidylglycerol, cholesterol, and polyethylene glycol-100 stearate were dissolved in anhydrous ethanol and stirred to obtain a first mixture; the first mixture was mixed and stirred with an acidic aqueous buffer, and then dialyzed and filtered to remove bacteria to obtain a lipid nanoparticle carrier. S3. Arginine / lysine polypeptide is synthesized using a solid-phase synthesis method, wherein the solid-phase synthesis method includes deprotection, activation, cross-linking, cycling, elution, deprotection, purification and structural identification in sequence. S4. The arginine / lysine polypeptide is used as the active ingredient and mixed with the lipid nanoparticle carrier, trehalose and deionized water, and homogenized to obtain the peptide composition.

6. The method for preparing a peptide composition for treating allergic skin diseases according to claim 5, characterized in that, In step S1, the organic solvent is chloroform or dichloromethane; the mass ratio of dioleoylphosphatidylglycerol to the organic solvent is 1:(3.5-4.5); the stirring reaction temperature for adding the acetylation reagent is 2-5℃; the addition rate of the acetylation reagent is 0.8-1.2 mL / min; the stirring speed is 300-400 rpm and the stirring time is 20-30 min.

7. The method for preparing a peptide composition for treating allergic skin diseases according to claim 5, characterized in that, In step S2, the mass ratio of acetylated dioleoylphosphatidylglycerol to anhydrous ethanol is 1:(3-5); the acidic aqueous buffer solution includes any one of citrate-sodium citrate buffer, acetate-sodium acetate buffer, and sodium dihydrogen phosphate-disodium hydrogen phosphate buffer; the mass ratio of the first mixture to the acidic aqueous buffer solution is 1:(5-10); the mixing and stirring speed is 1000-1500 rpm and the time is 15-30 min.

8. The method for preparing a peptide composition for treating allergic skin diseases according to claim 5, characterized in that, In step S4, the homogenization process uses a high-pressure homogenizer with a homogenization pressure of 30-60 MPa and a homogenization cycle of 3-5 times.

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