Polypeptide for promoting skin repair and application thereof

A nucleic acid and selected technology, applied in skin repairing peptides and its application fields, can solve the problems of insufficient stability and dispersibility of cosmetics, achieve the effects of promoting collagen expression, anti-oxidation, good stability, and promoting skin repair Effect

Inactive Publication Date: 2021-11-26

四川丽妍工坊生物科技有限公司

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AI-Extracted Technical Summary

Problems solved by technology

[0009] These polypeptides all have the function of promoting skin repair, but they have many shortcomings in the application of pharmaceuticals and cosmetic products, such as the inability to apply to the...

Abstract

The invention discloses a polypeptide capable of promoting skin repair and application thereof, and relates to the technical field of biological medicine, in particular to a polypeptide capable of promoting skin repair, the proportion of hydrophobic amino acid in the polypeptide capable of promoting skin repair is 16-30%, the isoelectric point of the polypeptide is 0.55-1.00, the sequence length is 7-14aa, and the polypeptide capable of promoting skin repair has a skin repair function. By reasonably designing the proportion of hydrophobic amino acids, isoelectric points and appropriate peptide chain length, the polypeptide can maintain good stability in both acidic and alkaline solvents, has strong functions of promoting skin repair, promoting collagen expression, resisting oxidation and the like, and the polypeptide can be applied to preparation of cosmetics and medicines with the function of promoting skin repair.

Application Domain

Cosmetic preparationsPeptide/protein ingredients +4

Technology Topic

PeptideAmino acid +5

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Examples

Experimental program(3)
Comparison scheme(1)
Effect test(7)

Example Embodiment

[0048] The embodiment of the present invention also provides a preparation method of a polypeptide, which comprises: cultivating the host cell as described in the foregoing embodiments, or artificially synthesizing the polypeptide as described in any of the foregoing embodiments.

[0049] Conditions for culturing host cells can be obtained based on conventional techniques, as long as the host cells can express polypeptides, and will not be repeated here.

[0050] The embodiments of the present invention also provide the use of the polypeptide as described in any of the foregoing embodiments or the isolated nucleic acid as described in the foregoing embodiments in the preparation of a drug or cosmetic that promotes repair of damaged skin.

[0051] The embodiments of the present invention also provide the use of the polypeptide as described in any of the foregoing embodiments or the isolated nucleic acid as described in the foregoing embodiments in the preparation of drugs or cosmetics that promote skin collagen proliferation and/or anti-aging.

[0052] The embodiment of the present invention also provides a cosmetic, which includes: the polypeptide as described in any of the foregoing embodiments or the isolated nucleic acid as described in the foregoing embodiments.

[0053] In some embodiments, the cosmetic can also include its adjuvant, such as at least one of polyol, glycerin, animal oil, mineral oil and vegetable oil; the mineral oil is further preferably liquid vaseline or paraffin oil; the vegetable oil is further preferably crocodile Pear oil, soybean oil or macadamia nut oil; animal oil is further preferably lanolin.

[0054] In some embodiments, the adjuvant may also include substances that make the cosmetic shape, stabilize, or have color, fragrance, and antiseptic effects, such as surfactants, flavors or fragrances, pigments, pigments, and preservatives.

[0055] In some embodiments, the cosmetics may also include other commonly used cosmetic raw materials, such as at least one of other proteins, other polypeptides, plant extracts, cytokines and vitamins.

[0056] The embodiment of the present invention also provides a pharmaceutical composition, which comprises: the polypeptide as described in any of the foregoing embodiments or the isolated nucleic acid as described in the foregoing embodiments.

[0057] The drug also includes medically acceptable carriers such as lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl Cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylparaben, propylparaben, talc, magnesium stearate and mineral oil etc.

Example Embodiment

[0059] Example 1

[0060] The present embodiment provides skin repair promoting polypeptide RW3, which is a linear polypeptide, and the primary structure of the complete amino acid sequence is: Met-Pro-Phe-Arg-Met-Gly-Ile-Cys-Thr-Met-Asn (MPFRMGICTMN ).

[0061] The preparation method of the skin-promoting peptide RW3, the specific steps are as follows:

[0062] 1. According to the designed amino acid sequence: Met-Pro-Phe-Arg-Met-Gly-Ile-Cys-Thr-Met-Asn, the crude polypeptide was synthesized by solid phase synthesis;

[0063] 2. Desalting and purifying the crude polypeptide by HPLC reverse-phase column chromatography, and identifying its purity until the purity of the polypeptide is not less than 95%;

[0064] HPLC purification and identification method: Dissolve 0.1mg of the sample to be tested in 1mL ultrapure water containing 0.1% trifluoroacetic acid. If there are undissolved impurities, filter them with a 0.45μm filter membrane. Mobile phase A is 0.1% trifluoroacetic acid - water, the mobile phase B is 0.1% trifluoroacetic acid-acetonitrile, after the baseline is stable, start to load the sample, the sample volume is 50 μL; the chromatographic column is a silica gel alkyl bonded phase C18 column (4.6mm×300mm, colloidal particle size 5 μm , pore size is 100A), adopt binary mobile phase gradient elution system, carry out gradient elution, promptly within 30min, the content of mobile phase B in eluent increases linearly from 0%-80%, flow rate 1mL /min, detection wavelength 215nm, measured at 25°C.

[0065] 3. The molecular weight was determined to be 1300.6Da by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF);

[0066] Method: Dissolve the purified polypeptide in deionized water to make a solution of 1 μmol/mL, take 10 μL and an equal volume of saturated matrix solution (dissolve α-cyano-4-hydroxycinnamic acid in 0.1% trifluoroacetic acid 50% acetonitrile solution, to make a saturated solution, centrifuge, get the supernatant) and measure after mixing.

[0067] 4. The isoelectric point of the purified polypeptide was determined to be 8.25 by isoelectric focusing electrophoresis, and the amino acid sequence structure of the purified polypeptide was determined by an automatic amino acid sequencer, which was determined to be Met-Pro-Phe-Arg-Met-Gly-Ile- Cys-Thr-Met-Asn.

[0068] 5. The acid-base stability of the skin-repairing peptide RW3.

[0069] The stability of RW3 was investigated in the case of strong acid-base hydrolysis. Acid degradation conditions: 1N HCl/room temperature/30min; alkaline degradation conditions 1N NaOH/room temperature/30min. Under the above conditions, 1 mg/ml of RW3 was treated. Use the aforementioned HPLC identification method to identify the stability of RW3 in strong acid and strong alkali solutions.

[0070] experiment such as figure 1 , at room temperature, 1mg/ml RW3 was incubated in 1M HCl for 30 minutes, and the sample was diluted 10 times to determine the chromatographic behavior, compared with the control phase ( figure 1 In A) the specific chromatographic behavior has not changed, and no new impurity peaks ( figure 1 Medium B). RW3 was incubated in 1M NaOH for 30 minutes at room temperature, and the chromatographic behavior was measured. Sample dilution 10 times measures chromatographic behavior, compared with contrast, chromatographic behavior does not change, does not appear new impurity peak ( figure 1 Middle C). It shows that the designed skin repairing peptide RW3 has good stability in strong acid and strong alkali environment.

[0071] 6. The dispersibility of the skin repair promoting peptide RW3 in lipid substances.

[0072] Take the determination of the homogeneity and dispersion of polypeptides in lanolin in RW3 as an example. Take 1000g of lanolin and 1000mg of RW3 polypeptide, and mix them evenly in a SHW/R mobile high-shear emulsifier at room temperature at a stirring speed of 120 rpm for 30 minutes. After mixing, aliquot into 5ml tubes. The theoretical content of RW3 under this condition is 1mg/g. Take and pack into 20 tubes, accurately weigh an appropriate amount of 1g of the mixture (approximately equivalent to RW3 0.1mg 10ml) into a measuring bottle, add 20% ethanol solution to dissolve (ultrasound to dissolve if necessary) and dilute to the mark, accurately measure 2ml with Folin Phenol was used for protein quantification. Experiments show that the average content of RW3 in 20 samples is 0.95±0.08 mg/g, and the content range of each sample is within 90% of the theoretical content.

[0073] It shows that the designed skin repair peptide RW3 has good dispersibility in lipid environment.

[0074] The skin repair-promoting peptide RW3 can be used in the preparation of repair and regeneration drugs for damaged epidermis, as well as in the development of skin collagen proliferation, anti-aging and anti-oxidation skin care products.

Example Embodiment

[0075] Example 2-8

[0076] Examples 2-8 respectively provide a polypeptide and its preparation method. The preparation method is roughly the same as that of Example 1, the difference is that the sequence is different. The amino acid sequence of the polypeptide provided by Examples 2-8 is as follows:

[0077] RW0: AMFRMGICTMN (SEQ ID No. 1);

[0078] RW1: AMFRMGMCTMN (SEQ ID No. 2);

[0079] RW2: APFRMGICTMN (SEQ ID No. 3);

[0080] RW4: MPFRMGMCTMN (SEQ ID No. 5);

[0081] RW5: MPFRMGMCTMM (SEQ ID No. 6);

[0082] RW6: PFRMGMC™ (SEQ ID No. 7);

[0083] RW7: PFRMGMCTMN (SEQ ID No. 8).

PUM

Description & Claims & Application Information

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Classification and recommendation of technical efficacy words

good stability

Polypeptide for promoting skin repair and application thereof

AI-Extracted Technical Summary

Problems solved by technology

Abstract

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Examples

Example Embodiment

Example Embodiment

Example Embodiment

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Description & Claims & Application Information

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