Composition of cyclic peptide and recombinant collagen as well as preparation method and application of composition

Abstract

The invention discloses a cyclic peptide composition, a preparation method thereof and application of the cyclic peptide composition in preparation of medical products and / or cosmetics. The cyclic peptide composition comprises a cyclic peptide and 21-type collagen, wherein the amino acid sequence of the cyclic peptide is as shown in SEQ ID NO. 1; the amino acid sequence of the type 21 collagen is as shown in SEQ ID NO. 2. The cyclopeptide composition disclosed by the invention is used for preparing medical products and / or cosmetics for resisting light aging and resisting aging, especially light aging, and the cyclopeptide and the type 21 collagen have a synergistic effect, so that an unexpected synergistic effect is achieved.

Description

Technical Field

[0001] The invention belongs to the field of bioengineering, and specifically relates to a cyclic peptide composition and a preparation method and application thereof. Background Art

[0002] Skin aging mainly includes natural aging and photoaging. Compared with objective natural aging, photoaging refers to skin aging caused by ultraviolet radiation. Relevant studies have shown that among the factors of skin aging, 80% of exogenous skin aging comes from sunlight exposure. When the skin is exposed to ultraviolet light for a long time, cells will produce excessive reactive oxygen species (ROS), which will cause a series of reactions to the skin; the most typical reaction is the inflammation caused by exposure to ultraviolet light, which will affect the synthesis of collagen and its precursors such as type I and type III procollagen, and destroy the formation of collagen while degrading collagen.

[0003] Collagen is a glycoprotein. It is divided into more than 28 types of collagen according to different tissue locations, physiological functions, and molecular structures. The common types are type I, type II, type III, type V, and type XI. The collagen in human skin is type I collagen and type III collagen. Type I collagen is mainly found in adult skin, tendons, and bone tissues, while type III collagen is mainly found in infant skin or vascular endothelium and intestines. Type I collagen and type III collagen are closely related to the skin injury repair process and repair quality. Many studies have shown that collagen is the most abundant protein in mammalian tissues, accounting for 30% of the total protein content of the body; it is also an important component of the extracellular matrix, maintaining the structure and function of tissues and organs such as skin, cartilage, tendons, ligaments, and internal organs.

[0004] Among various collagens, type 21 collagen is a non-fibrous collagen that is relatively rare in the adult human body, accounting for less than 1% of the total collagen. Current studies have confirmed that the type 21 collagen gene is involved in the formation of collagen in various tissues such as the heart and aorta, and is related to vascular remodeling and hypertension. However, compared with other more well-known types of collagen, the research on type 21 collagen is relatively small.

[0005] The inventor of the present application has applied for a cyclic peptide in advance, specifically a cyclic peptide disclosed in Chinese patent publication number CN116693612A, which can effectively prevent and treat atopic dermatitis, but no comprehensive research has been conducted on the other effects of the cyclic peptide. Therefore, it is of great significance to increase research and continue to develop a cyclic peptide composition that can both resist inflammation and anti-aging. Summary of the invention

[0006] In view of the defects of the prior art, the present invention provides a cyclic peptide composition and a preparation method thereof, and the use of the cyclic peptide composition in the preparation of medical products and / or cosmetics. The cyclic peptide composition provided by the present invention not only has anti-inflammatory effects, but also has anti-aging effects, and can effectively delay photoaging; the cyclic peptide is used in combination with type 21 collagen, and the obtained composition has significantly better anti-inflammatory and anti-aging effects, especially photoaging effects, than the cyclic peptide or type 21 collagen alone, and achieves unexpected synergistic effects.

[0007] To achieve the above object, the present invention provides a cyclic peptide composition, which comprises a cyclic peptide and type 21 collagen, wherein the amino acid sequence of the cyclic peptide is shown in SEQ ID NO.1; the amino acid sequence of type 21 collagen is shown in SEQ ID NO.2.

[0008] Furthermore, the concentration ratio of the cyclic peptide to type 21 collagen is 1:250 to 1:25.

[0009] Furthermore, the concentration of the cyclic peptide is 0.1 to 1 ppm.

[0010] Furthermore, the concentration of type 21 collagen is 25 to 100 ppm.

[0011] Furthermore, the concentration ratio of the cyclic peptide and type 21 collagen is 0.5ppm:50ppm.

[0012] Furthermore, the concentration ratio of the cyclic peptide and type 21 collagen is 0.75ppm:25ppm.

[0013] Furthermore, the cyclic peptide is formed by the two first and last cysteines forming a disulfide bond to form a ring.

[0014] Furthermore, the amino acid sequence of type 21 collagen comprises an amino acid sequence having at least 90% sequence identity with the amino acid sequence shown in SEQ ID No.1.

[0015] The present invention also provides a method for preparing the cyclic peptide composition, the preparation method comprising: mixing the cyclic peptide with type 21 collagen, wherein the concentration ratio of the cyclic peptide to the type 21 collagen is 1:250 to 1:25.

[0016] Furthermore, the method for producing type 21 collagen comprises the following steps:

[0017] (1) Construction of genetically engineered Pichia pastoris;

[0018] (2) Fermentation culture of genetically engineered Pichia pastoris;

[0019] (3) Induction and expression of recombinant human collagen;

[0020] (4) Purification of recombinant human collagen.

[0021] The present invention also provides an application of the cyclic peptide composition in the preparation of medical products and / or cosmetics.

[0022] Furthermore, the medical product and / or cosmetic is used for anti-inflammatory and / or anti-aging.

[0023] The cyclic peptide composition provided in the present application, its preparation method, and its application in the preparation of medical products and / or cosmetics have the following beneficial technical effects:

[0024] 1. The cyclic peptide composition of the present invention has a significant effect on anti-inflammatory and anti-aging. From the comparative experiment, it can be seen that the composition of the mixture of cyclic peptide and type 21 collagen is more excellent than the single use of cyclic peptide or type 21 collagen. The cyclic peptide and type 21 collagen have a synergistic effect and achieve an unexpected synergistic effect.

[0025] 2. The cyclic peptide in the present invention is a protein that plays a key role in inflammatory aging. It inhibits the activity of HMGB1 and reduces the inflammatory response, thereby counteracting the collagen degradation caused by photoaging.

[0026] 3. The type 21 collagen regeneration signal peptide in the present invention, as an external collagen signal peptide, can stimulate the regeneration of endogenous collagen in the skin and promote skin self-repair and regeneration.

[0027] 4. The cyclic peptide and type 21 collagen in the cyclic peptide composition of the present invention have different molecular mechanisms and signal pathways for anti-inflammatory and anti-aging effects, but both have anti-aging and anti-inflammatory effects. It is worth noting that the anti-aging and anti-inflammatory effects of 1+1>2 brought about by the superposition of these two components are unexpected in experimental screening.

[0028] The concept, specific structure and technical effects of the present invention will be further described below in conjunction with the accompanying drawings to fully understand the purpose, characteristics and effects of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 This is the experimental result of type I collagen content in this application.

[0030] Figure 2 It is the experimental result of IL1 beta content in this application. DETAILED DESCRIPTION

[0031] The following describes the embodiments of the present application through specific examples, and those skilled in the art can easily understand other advantages and effects of the present application from the contents disclosed in this specification. The present application can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present application. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0032] It should be noted that, despite the numerical ranges and parameter approximations shown in the broad scope of the present invention, the numerical values ​​shown in the specific embodiments are recorded as accurately as possible. However, any numerical value is bound to contain a certain error, which is caused by the standard deviation in their respective measurements. In addition, all ranges disclosed in the present invention should be understood to cover any and all sub-ranges contained therein; for example, the range of "1 to 10" recorded should be considered to include any and all sub-ranges between the minimum value 1 and the maximum value 10 (including the endpoints); that is, all sub-ranges starting with a minimum value of 1 or greater, such as 1 to 6.1, and sub-ranges ending with a maximum value of 10 or less, such as 5.5 to 10. In addition, any reference referred to as "incorporated herein" should be understood to be incorporated in its entirety. It is only for the convenience of narration, not to limit the scope of the application that can be implemented, and the change or adjustment of its relative relationship, without substantial change in the technical content, should also be regarded as the scope of the application that can be implemented.

[0033] The present invention relates to a cyclic peptide composition for use in medical products and / or cosmetics with anti-inflammatory and / or anti-aging effects, wherein the cyclic peptide composition comprises a cyclic peptide and type 21 collagen, the amino acid sequence of the cyclic peptide is shown in SEQ ID NO.1; the amino acid sequence of type 21 collagen is shown in SEQ ID NO.2.

[0034] In some embodiments, the cyclic peptide is formed by disulfide bonds between the first and last cysteines.

[0035] In other embodiments, modifications such as hydroxylation, carboxylation, carbonylation, methylation, acetylation, phosphorylation, esterification or glycosylation can be performed on the amino acid side chain groups, the amino terminus or the carboxyl terminus of the cyclic peptide to obtain derivatives of the cyclic peptide without affecting its biological activity.

[0036] The amino acid sequence of the cyclic peptide is CIPRFYLFYC.

[0037] The amino acid sequence of type 21 collagen is selected from SEQ ID NO.2: GKPGLQGPKGDPGLPGNPGYPGQPGQDGKPGYQGIAGTPGVPGSPGIQGARGLPGYKGEPGRDGDKGDRGLPGFPGLHGMPGSKGEMGAKGDKGSPGFYGKKGAKGEKGNAGFP GLPGPAGEPGRHGKDGLMGSPGFKGEAGSPGAPGQDGTRGEPGIPGFPGNRGLMGQKGEIGPPGQQGKKGAPGMPGLMGSNGSPGQPGTPGSKGSKGEPGIQGMPGASGLKGEPGATG.

[0038] In other embodiments, the amino acid sequence of type 21 collagen comprises an amino acid sequence that has at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100% or higher sequence identity with the amino acid sequence shown in SEQ ID No. 1 above.

[0039] In the present invention, the concentration of the cyclic peptide is 0.1-1 ppm, and the concentration of type 21 collagen is 25-100 ppm.

[0040] In some embodiments, the concentration of the cyclic peptide is: 0-10ppm, preferably 0.1ppm-1ppm, more preferably: 0.1ppm, 0.2ppm, 0.3ppm, 0.4ppm, 0.5ppm, 0.6ppm, 0.7ppm, 0.75ppm, 0.8ppm, 0.85ppm, 0.9ppm, 0.95ppm, 1ppm, most preferably 1ppm.

[0041] In some embodiments, the concentration of type 21 collagen is: in the range of 0 to 150 ppm, preferably 25 to 100 ppm; more preferably: 25 ppm, 30 ppm, 35 ppm, 40 ppm, 45 ppm, 50 ppm, 55 ppm, 60 ppm, 65 ppm, 70 ppm, most preferably: 100 pm.

[0042] The present invention relates to a protein encoding gene vector, a host cell of the protein encoding gene or the vector; and uses of recombinant human collagen, the vector, and the host cell in preparing cosmetics, health products or medicines.

[0043] In another aspect, the present invention relates to a method for producing the above-mentioned type 21 collagen, comprising the following steps:

[0044] (1) Construction of genetically engineered Pichia pastoris;

[0045] (2) Fermentation culture of genetically engineered Pichia pastoris;

[0046] (3) Induction and expression of recombinant human collagen;

[0047] (4) Purification of recombinant human collagen.

[0048] In some embodiments, the construction steps of the above-mentioned Pichia pastoris genetically engineered bacteria are:

[0049] (1) Activate yeast GS115 and streak culture on YPD plates;

[0050] (2) Selecting a single clone of Pichia pastoris GS115 activated on a YPD plate and culturing it in YPD liquid with constant temperature shaking;

[0051] (3) Pipette 100uL of bacterial solution and inoculate it into 100mL YPD liquid. Cultivate at 30℃ and 220rpm for 12-13h until OD 600 =1.3~1.5;

[0052] (4) Observe under a microscope for bacterial contamination, divide the bacterial cells into sterile 50-mL centrifuge tubes, and place on ice for 10 to 15 minutes;

[0053] (5) Centrifuge and discard the supernatant, and resuspend the cells in sterile water;

[0054] (6) Centrifuge and discard the supernatant, resuspend the cells with sorbitol, and then aliquot to obtain competent cells;

[0055] (7) The vector is transferred into competent cells by electroporation and then cultured on plates until clones are produced.

[0056] In some embodiments, the fermentation and culturing steps of the genetically engineered Pichia pastoris are as follows:

[0057] (1) Pick a single colony from the YPD plate and place it in a conical flask containing 50 mL of BMGY. Cultivate at 30°C and 220 rpm for 24 h. Measure the OD 600 The value confirms the end of cultivation;

[0058] (2) Centrifuge for 10 min, collect the cells, wash once with BMMY, centrifuge again, and dilute the cells to OD 600 =1.0.

[0059] In other embodiments, the induction and expression steps of the above recombinant human collagen are as follows:

[0060] (1) The diluted bacterial solution was transferred to a 500 mL conical flask and the yeast was induced to express protein at a temperature of 29°C and a rotation speed of 220 rpm; methanol was then added every 24 hours to make the final concentration of methanol in the culture medium 1%, and the induction culture was continued for 84 hours;

[0061] (2) Microscopic examination to see if there is any bacterial contamination and to measure the pH of the fermentation liquid, which must be less than 6;

[0062] (3) Transfer the fermentation broth to a 500 mL round-bottom centrifuge tube and centrifuge at 15,000 g at 4°C for 20 min to collect the supernatant;

[0063] (4) Take out 1 mL of supernatant and add corresponding loading buffer, mix well, incubate at 95°C for 10 min, and store at -20°C.

[0064] In other embodiments, the purification steps of the above recombinant human collagen are as follows:

[0065] (1) The obtained fermentation broth supernatant was transferred to a conical flask, NaCl was added to make the final concentration of NaCl 150 mM, and NaOH or HCl was used to adjust the pH of the fermentation broth supernatant to 8.0;

[0066] (2) Centrifuge at 15,000 g for 30 min to ensure that the supernatant is clear and transparent, and equilibrate the column with 10 nickel column bed volumes of binding buffer;

[0067] (3) Load the sample at a rate of 1 mL / min and collect the column liquid while loading the sample;

[0068] (4) Elute unbound proteins and impurities with 5 nickel column bed volumes of binding buffer;

[0069] (5) Elute the target protein with elution buffer (500 mM imidazole);

[0070] (6) Preparation of protein detection samples: Add the collected purified protein, supernatant before column and liquid after column into the corresponding loading buffer, mix well, incubate at 95°C for 10 min, and store at -20°C.

[0071] Example 1: Synthesis process of collagen

[0072] 1. Construction of genetically engineered Pichia pastoris

[0073] pPic9k was used as a vector to construct a recombinant plasmid, and the gene sequence of the type 21 collagen fragment GKPGLQGPKGDPGLPGNPGYPGQPGQDGKPGYQGIAGTPGVPGSPGIQGARGLPGYKGEPGRDGDKGDRGLPGFPGLHGMPGSKGEMGAKGDKGSPGFYGKKGAKGEKGNAGFPGLPGPAGEPGRHGKDGLMGSPGFKGEAGSPGAPGQDGTRGEPGIPGFPGNRGLMGQKGEIGPPGQQGKKGAPGMPGLMGSNGSPGQPGTPGSKGSKGEPGIQGMPGASGLKGEPGATG was inserted to obtain the type 21 collagen vector.

[0074] Activate yeast GS115 and streak culture on YPD plate, select single clone of Pichia pastoris GS115 activated on YPD plate and culture in YPD liquid, shake and culture at constant temperature; pipette 100uL bacterial solution and inoculate it in 100mL YPD liquid, culture at 30℃ and 220rpm for 12-13h, until OD 600 =1.3~1.5; observe under a microscope for bacterial contamination, divide the bacteria into sterile 50ml centrifuge tubes and place on ice for 10~15min.

[0075] After two centrifugations, discard the supernatant and resuspend the cells with sterile water; after two more centrifugations, discard the supernatant and resuspend the cells with sorbitol and then aliquot to obtain competent cells; transfer the vector into the competent cells by electroporation and then culture on YPD plates until clones are produced.

[0076] 2. Fermentation of genetically engineered Pichia pastoris

[0077] A single colony was picked from the YPD plate and placed in a conical flask containing 50 mL of BMGY. The culture was carried out at 30 °C and 220 rpm for 24 h. The OD 600 The culture was completed; centrifuged for 10 min, the cells were collected, washed once with BMMY, centrifuged again, and the cell concentration was diluted to OD 600 =1.0.

[0078] 3. Induction and expression of recombinant human collagen

[0079] Transfer the diluted bacterial solution to a 500mL conical flask, set the temperature to 29℃ and the rotation speed to 220rpm to induce yeast to express protein; then add methanol every 24h to make the final concentration of methanol in the culture medium to 1% and continue inducing culture for 84h; check under a microscope to see if there is any bacterial contamination, and measure the pH of the fermentation liquid, which must be less than 6; transfer the fermentation liquid to a 500mL round-bottom centrifuge tube, centrifuge at 15000g, 4℃, for 20min, and take the supernatant; take out 1mL of supernatant and add the corresponding Loading buffer, mix well, incubate at 95℃ for 10min, and store at -20℃.

[0080] 4. Purification of recombinant human collagen:

[0081] Transfer the obtained fermentation broth supernatant to a conical flask, add NaCl to make the final concentration of NaCl 150mM, adjust the pH of the fermentation broth supernatant to 8.0 with NaOH or HCl; centrifuge at 15000g for 30min to ensure that the supernatant is clear and transparent, balance the column with 10 nickel column bed volumes of binding buffer; load the sample at a speed of 1mL / min, and collect the column liquid while loading the sample. Elute the unbound protein and impurity protein with 5 nickel column bed volumes of binding buffer; elute the target protein with elution buffer (500mM imidazole).

[0082] Preparation of protein detection samples: Add the collected purified proteins, supernatant before column penetration, and liquid after column penetration into the corresponding loading buffer, mix well, incubate at 95℃ for 10min, and store at -20℃.

[0083] The above method was used to successfully prepare and purify type 21 recombinant humanized collagen, and the Pichia pastoris expression system was used to obtain a high-purity, high-activity target protein.

[0084] After sequencing confirmation, the amino acid sequence of type 21 collagen was obtained as shown in SEQ ID NO.2:

[0085] GKPGLQGPKGDPGLPGNPGYPGQPGQDGKPGYQGIAGTPGVPGSPGIQGARGLPGYKGEPGRDGDKGDRGLPGFPGLHGMPGSKGEMGAKGDKGSPGFYGKKGAKGEKGNAGFPGL PGPAGEPGRHGKDGLMGSPGFKGEAGSPGAPGQDGTRGEPGIPGFPGNRGLMGQKGEIGPPGQQGKKGAPGMPGLMGSNGSPGQPGTPGSKGSKGEPGIQGMPGASGLKGEPGATG

[0086] The sequence of the cyclic peptide (CIPRFYLFYC) can be obtained by chemical synthesis or synthetic biology fermentation followed by cyclization.

[0087] Example 2: Experiment on photoaging and anti-aging

[0088] 2.1 Experimental samples and groups

[0089] Group A: 100ppm recombinant type 21 collagen (also known as rh-col21);

[0090] Group B: 1ppm cyclic peptide (also known as cIY8);

[0091] Group C: 50ppm recombinant type 21 collagen (also known as rh-col21) + 0.5ppm cyclic peptide (also known as cIY8);

[0092] Group D: 25ppm recombinant type 21 collagen (also known as rh-col21) + 0.75ppm cyclic peptide (also known as cIY8);

[0093] The amino acid sequence of type 21 collagen is shown in SEQ ID NO.2, which is prepared by the above method. The amino acid sequence of the cyclic peptide is shown in SEQ ID NO.1.

[0094] Experimental groups:

[0095] Set up blank control group, negative control group, positive control group and sample group;

[0096] Set up more than 3 replicate wells for each group.

[0097] 2.2 Experimental procedures (1) Cell culture:

[0098] Dilute HFF-1 cells to 1 × 10 5 The inoculation density is pcs / mL;

[0099] Inoculate 1 mL / well of cell dilution into a 12-well plate;

[0100] Incubate at 37°C, 5% CO2, 95% RH for 24±2 hours.

[0101] Liquid preparation:

[0102] Sample: Dissolve the test sample in cell culture medium as required;

[0103] Positive control: thaw and dilute TGF-β1 stock solution to 0.1 μg / mL.

[0104] (2) UVA-induced modeling:

[0105] When the cell fusion degree reaches more than 80%, add the corresponding reagents to each group:

[0106] Blank control group: add 1 mL PBS;

[0107] Negative control group: add 1 mL PBS;

[0108] Positive control group: add 1 mL of PBS containing 100 ng / mL TGF-β1;

[0109] Sample group: Add samples containing corresponding concentrations.

[0110] That is: Group A: 100ppm recombinant type 21 collagen (also known as rh-col21);

[0111] Group B: 1ppm cyclic peptide (also known as cIY8);

[0112] Group C: 50ppm recombinant type 21 collagen (also known as rh-col21) + 0.5ppm cyclic peptide (also known as cIY8);

[0113] Group D: 25ppm recombinant type 21 collagen (also known as rh-col21) + 0.75ppm cyclic peptide (also known as cIY8);

[0114] Irradiate cells with UVA light (5 J / cm 2 ), only the blank control group was not induced by UVA.

[0115] The 12-well plate was placed in an incubator (37° C., 5% CO 2 , 95% RH) and cultured for 24±2 hours.

[0116] (3) Type I collagen detection:

[0117] Detect the standard by ELISA and draw a standard curve;

[0118] The type I collagen content in the supernatant of each group was detected;

[0119] The information of the ELISA kit used to detect the content of type I collagen is as follows:

[0120] Kit name: Human Collagen Type I (Col I) ELISA Kit; Manufacturer: CUSABIO.

[0121] Table 1: Experimental results of type I collagen content

[0122]

[0123] The experimental results are shown in Table 1 and Figure 1 As shown in the figure, the effects of Group C and Group D are significantly better than those of Group A and Group B. The synergy is calculated using the widely used Burgi formula:

[0124]

[0125] In the formula It refers to the effect of half dose of A and B taken together, while EA and EB are the effects of single dose of A and B respectively. The q calculated by the above formula is greater than 1, which proves that recombinant type 21 collagen and cyclic peptide have synergistic effects and achieve unexpected synergistic effects.

[0126] Example 3: Anti-inflammatory Experiment

[0127] (1) Cell culture:

[0128] Dilute RAW264.7 cells to 1.8 × 10 6 The inoculation density is pcs / mL;

[0129] Inoculate into 12-well plates at 1 mL / well;

[0130] Incubate at 37°C, 5% CO2, 95% RH for 24±2 hours.

[0131] Experimental groups:

[0132] Set up blank control group, negative control group, positive control group and sample group;

[0133] Set up more than 3 replicate wells for each group.

[0134] (2) Liquid preparation:

[0135] Sample: Dissolve the test sample in cell culture medium to prepare a concentration of 0.3%;

[0136] Positive control: Dexamethasone stock solution was thawed and diluted to 10 μg / mL.

[0137] (3) Sample addition:

[0138] When the cell fusion degree reaches more than 80%, add the corresponding reagents to each group:

[0139] Blank control group: added DMEM high-glucose medium;

[0140] Negative control group: DMEM high-glucose medium containing 500 ng / mL L lipopolysaccharide (LPS) was added;

[0141] Positive control group: DMEM high-glucose medium containing 10 μg / mL dexamethasone and 500 ng / mL LPS was added;

[0142] Sample group: DMEM high-glucose medium containing 500 ng / mL LPS and samples of corresponding concentrations was added;

[0143] The 12-well plate was returned to the incubator and cultured for 24 ± 2 hours.

[0144] (4) Content detection:

[0145] Detect the standard by ELISA and draw a standard curve;

[0146] The IL1 beta content in the supernatant of each group was detected.

[0147] (5) Data analysis:

[0148] The actual content of IL1 beta in each group was calculated based on the standard curve;

[0149] The data of each group were compared to evaluate the anti-inflammatory effect of the samples.

[0150] (6) Data Recording:

[0151] Data such as cell count, reagent addition amount, and ELISA test results were recorded, and the anti-inflammatory effect of the sample was evaluated based on the inhibition rate of IL1 beta.

[0152] The kit used to detect IL1 beta is as follows:

[0153] IL1 beta detection kit manufacturer: Lianke; Product name: Mouse IL1 beta ELISA Kit; used to quantitatively detect the content of IL1 beta protein from mice.

[0154] Table 2: IL1 beta content experimental results

[0155]

[0156] The experimental results show that (see Figure 2 Table 2): Compared with the negative control group, the IL1 beta content of the sample group was significantly reduced, and the sample had a significant anti-inflammatory effect. And based on the calculation of Burgi's formula method, the following conclusion can also be drawn: Type 21 collagen and cyclic peptide have a synergistic effect in reducing IL1 beta, achieving an unexpected synergistic effect.

[0157] The above description is only a preferred embodiment, which is only used as an example and does not limit the combination of features necessary to implement the present invention. The titles provided are not intended to limit the various embodiments of the present invention. Terms such as "comprising", "including" and "including" are not intended to be limiting. In addition, unless otherwise specified, plural forms are included when there is no numeral modification, and "or" and "or" mean "and / or". Unless otherwise defined herein, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art.

[0158] All publications and patent applications cited in this specification are incorporated herein by reference, as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference. In addition, any theory, mechanism, proof or discovery described herein is intended to further enhance the understanding of the application, and is not intended to limit the application to such theory, mechanism, proof or discovery in any way. Although the application has been shown and described in detail in the accompanying drawings and the preceding description, the application should be considered illustrative rather than restrictive.

Claims

1. A cyclic peptide composition, characterized in that The cyclic peptide composition comprises a cyclic peptide and type 21 collagen, wherein the amino acid sequence of the cyclic peptide is shown as SEQ ID NO.1; and the amino acid sequence of the type 21 collagen is shown as SEQ ID NO.

2.

2. The cyclic peptide composition according to claim 1, characterized in that The concentration ratio of the cyclic peptide to type 21 collagen is 1:250 to 1:

25.

3. The cyclic peptide composition according to claim 1, characterized in that The concentration of the cyclic peptide is 0.1-1 ppm.

4. The cyclic peptide composition according to claim 1, characterized in that The concentration of the type 21 collagen is 25 to 100 ppm.

5. The cyclic peptide composition according to claim 1, characterized in that The concentration ratio of the cyclic peptide and type 21 collagen is 0.5ppm:50ppm.

6. The cyclic peptide composition according to claim 1, characterized in that The concentration ratio of the cyclic peptide and type 21 collagen is 0.75ppm:25ppm.

7. The cyclic peptide composition according to claim 1, characterized in that The cyclic peptide is formed by two cysteines at the head and tail forming a disulfide bond to form a ring.

8. The cyclic peptide composition according to claim 1, characterized in that The amino acid sequence of type 21 collagen comprises an amino acid sequence having at least 90% sequence identity with the amino acid sequence shown in SEQ ID No.

1.

9. The method for preparing the cyclic peptide composition according to any one of claims 1 to 8, characterized in that: The preparation method comprises mixing the cyclic peptide with type 21 collagen, wherein the concentration ratio of the cyclic peptide to type 21 collagen is 1:250 to 1:

25.

10. Use of the cyclic peptide composition according to any one of claims 1 to 8 in the preparation of medical products and / or cosmetics.

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