Polypeptide anti-inflammatory and antipruritic composition

Abstract

The invention discloses a polypeptide anti-inflammatory and antipruritic composition, and belongs to the technical field of anti-inflammatory and antipruritic. Comprising the following components in parts by weight: 3 to 15 parts of lysimachia christinae polypeptide composite extract, 5 to 15 parts of N-ethyl-L-menthyl formamide, 2 to 10 parts of copper tripeptide-1, 2 to 10 parts of bisabolol, 2 to 20 parts of laurocapram, 3 to 10 parts of tetrahydrodiferuloylmethane, 0.1 to 2 parts of sodium cocoyl methyl taurate, 1 to 5 parts of carboxymethyl cellulose sodium and 30 to 60 parts of propylene glycol. The composition prepared by the invention is free of hormones and antibiotics, has no obvious damage and stimulation effect on skin, is suitable for sensitive skin, reduces potential risks in a skin treatment process, has no biotoxicity, does not generate drug resistance, and has relatively high stability. According to the invention, solid-state fermentation is carried out on the lysimachia christinae hance through lactobacillus, so that the polypeptide composite extract with an anti-oxidation effect is obtained, and the polypeptide composite extract has the effects of preventing corrosion, inhibiting bacteria, resisting inflammation and relieving itching, has the characteristic of being easy to absorb as a small molecule peptide, is beneficial to improving the curative effect and reducing side effects, and can be used for cosmetics and medicines.

Description

Technical Field

[0001] The invention belongs to the technical field of anti-inflammatory and antipruritic, and in particular relates to a polypeptide anti-inflammatory and antipruritic composition. Background Art

[0002] Inflammation is the body's defense mechanism against external stimuli or damage, usually manifested as redness, swelling, heat, pain and other symptoms. It is the immune system's response to harmful stimuli such as tissue damage, infection or toxic chemicals, with the goal of eliminating irritants or pathogenic microorganisms and promoting tissue repair. However, long-term uncontrolled acute inflammation may progress to chronic inflammation, causing a variety of chronic diseases, and is often accompanied by adverse side effects.

[0003] Itch is a common skin symptom that can be caused by a variety of reasons, such as dry skin, medical diseases, neurological disorders, psychiatric conditions, irritation and allergic reactions. The itching sensation is unbearable and often leads to scratching, which can further damage the skin, creating a vicious cycle and increasing the risk of infection.

[0004] Peptide substances show potential in anti-inflammatory and antipruritic effects. They can participate in the regulation of the body's inflammatory response by regulating the secretion of cytokines and the synthesis of inflammatory mediators. For example, small peptides with a molecular weight of less than 1 kDa and an amino acid chain length between 2 and 20 residues usually have higher activity because they are more easily absorbed and penetrate into cells. However, there are some defects when peptides are used as drugs, such as stability problems, high costs, and possible bioavailability problems, which limit their widespread clinical application. Therefore, the development of new peptide anti-inflammatory and antipruritic compositions, aimed at improving their stability and bioavailability, reducing costs, while maintaining or enhancing their efficacy, is an important direction of current research. Summary of the invention

[0005] In view of the above situation, in order to overcome the defects of the prior art, the present invention provides a polypeptide anti-inflammatory and antipruritic composition, which has stable effects and can be used to prepare medicines and cosmetics.

[0006] In order to achieve the above object, the following technical scheme is adopted: The present invention provides a polypeptide anti-inflammatory and antipruritic composition, comprising the following components in parts by weight:

[0007] 3-15 parts of Lycopodiella ciliata polypeptide composite extract, 5-15 parts of N-ethyl-L-menthyl carboxamide, 2-10 parts of tripeptide-1 copper, 2-10 parts of bisabolol, 2-20 parts of laurocaprol, 3-10 parts of tetrahydrodiferuloylmethane, 0.1-2 parts of sodium cocoyl methyl taurate, 1-5 parts of sodium hydroxymethylcellulose and 30-60 parts of propylene glycol.

[0008] Preferably, the following components are included in parts by weight:

[0009] 8 parts of Lycopodiella ciliata polypeptide complex extract, 6 parts of N-ethyl-L-menthyl carboxamide, 6 parts of tripeptide-1 copper, 6 parts of bisabolol, 12 parts of laurocapram, 6 parts of tetrahydrodiferuloylmethane, 1.2 parts of sodium cocoyl methyl taurate, 2 parts of sodium hydroxymethylcellulose and 45 parts of propylene glycol.

[0010] Among them, the complex extract of Lycopodiella peptides has anti-inflammatory and skin health-promoting properties, which helps protect and repair the skin.

[0011] N-ethyl-L-menthyl carboxamide, which provides a long-lasting cooling sensation, helps provide a cooling sensation to the skin and helps relieve itching.

[0012] Tripeptide-1 Copper has the ability to boost collagen production and possesses antioxidant and anti-inflammatory properties to soothe irritated skin and reduce redness.

[0013] Bisabolol has anti-inflammatory, antibacterial, sedative and anti-sweat effects.

[0014] As a skin penetration enhancer, laurocapram can improve the skin absorption efficiency of other ingredients, thereby enhancing the therapeutic effect of the overall composition.

[0015] Tetrahydrodiferuloylmethane has antioxidant, anti-inflammatory and dispersant properties.

[0016] Sodium cocoyl methyl taurate, as an amino acid surfactant, has foaming and emulsifying effects and is mild in nature.

[0017] Sodium hydroxymethylcellulose has the functions of thickening, water retention, emulsification, etc., and helps the composition to have a gelling network structure.

[0018] Propylene glycol plays the role of moisturizing, lubricating the skin, and promoting drug penetration in the composition, and also has the functions of anti-oxidation and inhibiting bacterial growth.

[0019] Furthermore, the Lychnophora scabra polypeptide composite extract is prepared by the following steps:

[0020] (1) Wash the whole plant of Lychnophora japonica with distilled water, remove the leaves, dry it at 60°C to constant weight, then pound it into powder and pass it through a 60-mesh sieve;

[0021] (2) adding the sieved powder to a culture device, soaking it with ultrapure water, sterilizing it at 121° C. for 20 min, inoculating the Lactobacillus strain on the surface of the powder at an inoculum rate of 10%, and placing the culture device in an incubator at 37° C. for 48 h;

[0022] (3) The culture device after the culture is placed at 60° C. to dry for 48 hours, after adding anhydrous ethanol, extract at 37° C. for 1 hour, filter, pass the filtrate through a nanofiltration membrane, concentrate the retentate, and dry to obtain a complex extract of Lycopodiella ciliata polypeptides.

[0023] Furthermore, the ultrapure water in step (2) makes the moisture content of the powder be 50%-60%.

[0024] Furthermore, the mass of the anhydrous ethanol in step (3) is 5-20 times the mass of the feed liquid of the culture device.

[0025] Furthermore, the molecular weight cut-off of the nanofiltration membrane is 3-5 kDa.

[0026] Furthermore, the dosage form of the composition can be any one or a combination of solid, semi-solid or liquid for external use.

[0027] The present invention provides use of a polypeptide composition in preparing an anti-inflammatory and antipruritic composition. The composition can be used in preparing cosmetics.

[0028] The present invention provides an application of a polypeptide composition in preparing an anti-inflammatory and antipruritic composition: the composition can be used in preparing medicines.

[0029] The beneficial effects of the present invention are:

[0030] (1) The composition prepared by the present invention is hormone-free and antibiotic-free, has no obvious damage or irritation to the skin, is suitable for use on sensitive skin, reduces potential risks during skin treatment, has no biological toxicity, does not produce drug resistance, and has high stability;

[0031] (2) The present invention obtains a polypeptide composite extract with antioxidant effect by solid-state fermentation of Lychnophora by lactobacillus. Lychnophora contains flavonoid compounds such as orientin, isoorientin, vitexin, and isovitexin, as well as sesquiterpenes, triterpenes, lignans, and sterol compounds. The polypeptide composition obtained by fermentation of Lychnophora has the effects of antiseptic, antibacterial, anti-inflammatory and antipruritic, and as a small molecule peptide, it has the property of being easily absorbed. Therefore, the composition of the present invention has a high bioavailability, helps to improve the therapeutic effect and reduce side effects, and can be used in cosmetics and medicines. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The safety test results of a polypeptide anti-inflammatory and antipruritic composition of the present invention;

[0033] Figure 2 The anti-inflammatory effect test results of a polypeptide anti-inflammatory and antipruritic composition of the present invention;

[0034] Figure 3The results are the test results of the antipruritic effect of a polypeptide anti-inflammatory and antipruritic composition of the present invention.

[0035] The accompanying drawings are used to provide further understanding of the present invention and constitute a part of the specification. They are used to explain the present invention together with the embodiments of the present invention and do not constitute a limitation of the present invention. DETAILED DESCRIPTION

[0036] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0037] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein may be applied to the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only and are not intended to limit the content of this application.

[0038] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are purchased from commercial channels unless otherwise specified.

[0039] Example 1

[0040] A Lychnophora polypeptide compound extract

[0041] The Lychnophora scabra polypeptide composite extract is prepared by the following steps:

[0042] (1) Wash the whole plant of Lychnophora japonica with distilled water, remove the leaves, dry it at 60°C to constant weight, then pound it into powder and pass it through a 60-mesh sieve;

[0043] (2) adding the sieved powder to a culture device, soaking it with ultrapure water, sterilizing it at 121° C. for 20 min, inoculating the Lactobacillus strain on the surface of the powder at an inoculum rate of 10%, and placing the culture device in an incubator at 37° C. for 48 h;

[0044] (3) The culture device after the culture is placed at 60° C. to dry for 48 hours, after adding anhydrous ethanol, extract at 37° C. for 1 hour, filter, pass the filtrate through a nanofiltration membrane, concentrate the retentate, and dry to obtain a complex extract of Lycopodiella ciliata polypeptides.

[0045] The ultrapure water in step (2) makes the moisture content of the powder reach 50%.

[0046] The mass of anhydrous ethanol in step (3) is 5 times the mass of the feed liquid of the culture device.

[0047] The molecular weight cut-off of the nanofiltration membrane is 3-5 kDa.

[0048] Example 2

[0049] A Lychnophora polypeptide compound extract

[0050] The preparation process of the Lychnophora scabra polypeptide composite extract is the same as that in Example 1.

[0051] The ultrapure water in step (2) makes the moisture content of the powder reach 60%.

[0052] The mass of anhydrous ethanol in step (3) is 20 times the mass of the feed liquid of the culture device.

[0053] The molecular weight cut-off of the nanofiltration membrane is 3-5 kDa.

[0054] Example 3

[0055] A Lychnophora polypeptide compound extract

[0056] The preparation process of the Lychnophora scabra polypeptide composite extract is the same as that in Example 1.

[0057] The ultrapure water in step (2) makes the moisture content of the powder reach 55%.

[0058] The mass of anhydrous ethanol in step (3) is 12 times the mass of the feed liquid of the culture device.

[0059] The molecular weight cut-off of the nanofiltration membrane is 3-5 kDa.

[0060] Example 4

[0061] A polypeptide anti-inflammatory and antipruritic composition

[0062] The composition comprises the following components in parts by weight:

[0063] 3 parts of Lycopodiella Cerifera polypeptide complex extract, 5 parts of N-ethyl-L-menthyl carboxamide, 2 parts of tripeptide-1 copper, 2 parts of bisabolol, 2 parts of laurocaprol, 3 parts of tetrahydrodiferuloylmethane, 0.1 parts of sodium cocoyl methyl taurate, 1 part of sodium hydroxymethylcellulose, and 30 parts of propylene glycol.

[0064] Example 5

[0065] A polypeptide anti-inflammatory and antipruritic composition

[0066] The composition comprises the following components in parts by weight:

[0067] 15 parts of Lycopodiella ciliata polypeptide complex extract, 15 parts of N-ethyl-L-menthyl carboxamide, 10 parts of tripeptide-1 copper, 10 parts of bisabolol, 20 parts of laurocaprol, 10 parts of tetrahydrodiferuloylmethane, 2 parts of sodium cocoyl methyl taurate, 5 parts of sodium hydroxymethylcellulose and 60 parts of propylene glycol.

[0068] Example 6

[0069] A polypeptide anti-inflammatory and antipruritic composition

[0070] The composition comprises the following components in parts by weight:

[0071] 8 parts of Lycopodiella ciliata polypeptide complex extract, 6 parts of N-ethyl-L-menthyl carboxamide, 6 parts of tripeptide-1 copper, 6 parts of bisabolol, 12 parts of laurocapram, 6 parts of tetrahydrodiferuloylmethane, 1.2 parts of sodium cocoyl methyl taurate, 2 parts of sodium hydroxymethylcellulose and 45 parts of propylene glycol.

[0072] Experimental Example 1

[0073] Safety testing of the composition

[0074] The test method is as follows:

[0075] (1) Sample treatment: 100 μL of the composition sample obtained in Example 6 was pipetted, and 100 μL of DMSO was added to prepare a 50% (v / v) DMSO stock solution, which was then diluted with serum-free MEM to 0.1%, 0.3%, 1.0%, 3.0%, 10.0%, 30%, and 50%, for a total of 7 concentrations. Control group: serum-free MEM;

[0076] (2) Routine cell culture: The cell suspension was inoculated into a 96-well cell culture plate at a density of 0.8 to 1.0 × 10 4 / well, culture for 18-24h, discard the original culture medium in the well, add 100μL of test samples of different concentrations to each well, return to the incubator and incubate for 24±1h. Take out the culture plate, add 20μL MTT solution to each well, incubate in the incubator for 3-4h; remove the liquid in the well, add 100μL DMSO to each well, place it on an oscillator for 10-15min, and measure the absorbance at a wavelength of 570nm on an enzyme reader.

[0077] (3) Data analysis: The cell activity of the negative control group was taken as 100%, and the relative cell activity of each group was calculated. The results are shown in Figure 1 .

[0078] have Figure 1 It can be seen that the relative activity of the composition of the present invention at a concentration of 0.3-50% has no significant difference with that of the control group on the cells in each group, indicating that the composition of the present invention has no toxicity to cells when used at a concentration of less than 50%.

[0079] Experimental Example 2

[0080] Composition anti-inflammatory effect test

[0081] By using LPS-induced human keratinocytes as an in vitro inflammatory cell model, the inflammatory factors (IL-1α, IL-6, IL-8, PGE2) secreted in the cell supernatant were detected by ELISA kits to evaluate the anti-inflammatory efficacy of the test samples. (IL-1α is a ubiquitous and key proinflammatory cytokine, and is also a pyrogen component, which has the effect of causing heat and mediating inflammation; IL-6 is a binding inflammatory factor. When its concentration is high, it will cause damage to tissues and cause inflammatory reactions. Its secretion amount is one of the indicators to measure the severity of inflammation; IL-8 is a chemokine that can stimulate the chemotaxis of neutrophils, T lymphocytes and eosinophils, promote neutrophil degranulation, release elastase, and cause damage; PGE2 is a metabolite of arachidonic acid, which is mainly synthesized by cyclooxygenase. PGE2 has the function of dilating blood vessels, causing erythema, and can also induce clinical symptoms of inflammatory pain). The results are shown in Figure 2 .

[0082] Depend on Figure 2 It can be seen that the composition of the present invention has a significant inhibitory effect on IL-1α, IL-6, IL-8 and PGE2 when the concentration is 1.0-30.0%.

[0083] Experimental Example 3

[0084] Antipruritic effect test of the composition

[0085] (1) Establishment of itch model:

[0086] Experimental animal groups: 8 SPF-grade ICR male mice were weighed and randomly divided into a normal group (NOR) of 3 mice and a model group (AEW) of 5 mice.

[0087] Preparation of the skin of the modeling area: After anesthesia with 15% urethane intraperitoneally (0.1 ml / 20 g body weight), the hair of about 2 cm×3 cm area on the back of the animal's neck was shaved with an electric shaver and the animal was placed in a cage for overnight.

[0088] AEW treatment: Starting from the second day after shaving, mice in each group were treated accordingly: mice in the model group were dipped in a proper amount of AE solution with a cotton ball and applied to the skin of the depilated area for 15 seconds, and then a cotton ball of the same size dipped in distilled water was applied to the same area for 30 seconds. The above steps were performed twice a day, at 9 am and 5 pm, respectively, for 5 days. The mice in the normal control group were smeared with double distilled water, and the time frequency was the same as that of the model group.

[0089] Itch behavior detection: On the 6th day of the experiment, each mouse was placed individually in a transparent observation container of 20×10×10 cm. After 15 minutes of adaptation to the environment and elimination of the nervousness and anxiety caused by the unfamiliar environment, the behavior was observed again. The scratching behavior of the mice was recorded by video within 1 hour, and the number of scratches of each group of mice was counted in a blind manner. Each time the mouse raised its hind paw and then put it down, or raised it and then licked its hind paw with its mouth, it was counted as one scratch. Scratching of non-modeling parts or front paws was not counted.

[0090] (2) Verification of antipruritic effect:

[0091] The mice with successful modeling were divided into: SADBE group (deionized water), HAC group (hydrocortisone acetonide), and CH group (the composition obtained in Example 6), with 5-10 mice in each group.

[0092] Dosage method: Before each use, take out the Chinese medicine extract stored at 4°C in advance and rewarm it in a water bath (water bath temperature: 37°C, 3min).

[0093] HAC group: Topical hydrocortisone acetonide, 100 μL per mouse once a day. No need to wipe.

[0094] CH group: 200 μL each time, apply the solution evenly with disposable gloves, wipe off the solution with a sterile alcohol cotton ball dipped in warm water after 30 minutes, 3 times a day, 4 hours between each times. Continuous medication for 7 days.

[0095] SADBE group: Deionized water was applied topically, 200 μL each time. The deionized water was evenly applied with disposable gloves and wiped off with a sterile alcohol cotton ball dipped in warm water after 30 minutes. This was done 3 times a day, with an interval of 4 hours between each application.

[0096] (3) Mouse scratching behavior assessment: The scratching behavior of mice was counted every 1 hour. Figure 3 .

[0097] Depend on Figure 3 It can be seen that the composition of the present invention can significantly reduce the number of scratching of mice, and thus has an antipruritic effect.

[0098] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

[0099] The present invention and its implementation methods are described above, which is not restrictive. The drawings are only one of the implementation methods of the present invention, and the actual application is not limited thereto. In short, if ordinary technicians in the field are inspired by it and design methods and embodiments similar to the technical solution without creativity without departing from the purpose of the invention, they should all fall within the protection scope of the present invention.

Claims

1. A polypeptide anti-inflammatory and antipruritic composition, characterized in that: The composition comprises the following components in parts by weight: 3-15 parts of Lycopodiella ciliata polypeptide composite extract, 5-15 parts of N-ethyl-L-menthyl carboxamide, 2-10 parts of tripeptide-1 copper, 2-10 parts of bisabolol, 2-20 parts of laurocaprol, 3-10 parts of tetrahydrodiferuloylmethane, 0.1-2 parts of sodium cocoyl methyl taurate, 1-5 parts of sodium hydroxymethylcellulose and 30-60 parts of propylene glycol. The Lychnophora scabra polypeptide composite extract is prepared by the following steps: (1) Wash the whole plant of Lychnophora japonica with distilled water, remove the leaves, dry it at 60°C to constant weight, then pound it into powder and pass it through a 60-mesh sieve; (2) adding the sieved powder to a culture device, soaking it with ultrapure water, sterilizing it at 121° C. for 20 min, inoculating the Lactobacillus strain on the surface of the powder at an inoculum rate of 10%, and placing the culture device in an incubator at 37° C. for 48 h; (3) The culture device after the culture is placed at 60° C. to dry for 48 hours, after adding anhydrous ethanol, extract at 37° C. for 1 hour, filter, pass the filtrate through a nanofiltration membrane, concentrate the retentate, and dry to obtain a complex extract of Lycopodiella ciliata polypeptides.

2. The polypeptide anti-inflammatory and antipruritic composition according to claim 1, characterized in that: The composition comprises the following components in parts by weight: 8 parts of Lycopodiella ciliata polypeptide complex extract, 6 parts of N-ethyl-L-menthyl carboxamide, 6 parts of tripeptide-1 copper, 6 parts of bisabolol, 12 parts of laurocapram, 6 parts of tetrahydrodiferuloylmethane, 1.2 parts of sodium cocoyl methyl taurate, 2 parts of sodium hydroxymethylcellulose and 45 parts of propylene glycol.

3. The polypeptide anti-inflammatory and antipruritic composition according to claim 1, characterized in that: The ultrapure water in step (2) makes the moisture content of the powder be 50%-60%.

4. The polypeptide anti-inflammatory and antipruritic composition according to claim 1, characterized in that: The mass of the anhydrous ethanol in step (3) is 5-20 times the mass of the feed liquid of the culture device.

5. The polypeptide anti-inflammatory and antipruritic composition according to claim 1, characterized in that: The molecular weight cut-off of the nanofiltration membrane is 3-5 kDa.

6. The polypeptide anti-inflammatory and antipruritic composition according to claim 1, characterized in that: The dosage form of the composition can be any one or a combination of solid, semi-solid or liquid for external use.

7. The polypeptide anti-inflammatory and antipruritic composition according to claim 2, characterized in that: The dosage form of the composition can be any one or a combination of solid, semi-solid or liquid for external use.

8. The polypeptide anti-inflammatory and antipruritic composition according to any one of claims 1, 2, 6, and 7, characterized in that The composition is used in preparing cosmetics.

9. The polypeptide anti-inflammatory and antipruritic composition according to any one of claims 1, 2, 6, and 7, characterized in that The composition is used in preparing medicines.

Images