PEPTIDE HAVING PROTECTIVE ACTIVITY AGAINST CELL DAMAGE CAUSED BY PARTICULATE MATTER, AND USES FOR SAME

Abstract

The present application relates to a peptide having protective activity against cell damage caused by particulate matter, and to uses for the peptide.

Claims

1. A peptide consisting of the amino acid sequence of SEQ ID NO: 1.

2. A composition for preventing cell damage caused by particulate matter, the composition comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

3. The composition of claim 2, wherein the cell damage is damage to skin cells, respiratory cells, or corneal cells.

4. The composition of claim 2, wherein the peptide inhibits the activation of the aryl hydrocarbon receptor (AHR).

5. The composition of claim 2, wherein the peptide inhibits the activity of cytochrome P450.

6. The composition of claim 2, wherein the particulate matter comprises polycyclic aromatic hydrocarbons (PAHs).

7. A cosmetic composition for preventing skin cell damage caused by particulate matter, the cosmetic composition comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

8. The cosmetic composition of claim 7, wherein the composition is for use of ameliorating skin aging, an increase in skin wrinkles, damage to skin elasticity, damage to skin color, pigment hyperplasia, melasma, freckles, dry skin, skin inflammation, contact dermatitis, atopic dermatitis, or psoriasis.

9. The cosmetic composition of claim 7, wherein the composition has at least one formulation selected from the group consisting of solution, suspension, emulsion, gel, lotion, essence, cream, powder, soap, shampoo, rinse, mask pack, surfactant-containing cleanser, cleansing foam, cleansing water, oil, liquid foundation, cream foundation, and spray.

10. A pharmaceutical composition for treating or preventing a disease caused by particulate matter, the pharmaceutical composition comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

11. The pharmaceutical composition of claim 10, wherein the disease caused by particulate matter is a disease selected from the group consisting of atopic dermatitis, contact dermatitis, seborrheic dermatitis, acne, xeroderma, psoriasis; allergic rhinitis, acute bronchitis, chronic bronchitis, emphysema, pulmonary function insufficiency, asthma, bronchiectasis, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, cystic fibrosis, bronchiolitis, acute upper respiratory infections, pneumonia, nasosinusitis, pharyngitis, tonsillitis, laryngitis; xerophthalmia, allergic conjunctivitis, glaucoma, cataracts, macular degeneration, retinal hemorrhage, retinal detachment, retinitis pigmentosa, senile macular degeneration, diabetic retinopathy; cancer, immunotoxicity, peripheral nervous system damage, central nervous system damage, endocrinopathy, reproductive organ disorders, developmental disorders of infants and toddlers, anemia, arrhythmia, heart attack, angina, and myocardial infarction.

12. The pharmaceutical composition of claim 10, wherein the peptide inhibits the activation of the aryl hydrocarbon receptor (AHR).

13. The pharmaceutical composition of claim 10, wherein the peptide inhibits the activity of cytochrome P450.

14. The pharmaceutical composition of claim 10, wherein the particulate matter comprises polycyclic aromatic hydrocarbons (PAHs).

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0062] FIG. 1 is a schematic view illustrating signaling mechanisms mediated by the aryl hydrocarbon receptor (AHR).

[0063] FIG. 2 shows a result of a Western blot experiment showing that an increase in the nuclear translocation of AHR induced by the treatment of particulate matter in human keratinocyte HaCaT is reduced in a concentration-dependent manner by the treatment of a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

[0064] FIG. 3 shows a result of a P450 activity measurement experiment showing that an increase in activity of cytochrome P450 induced by the treatment of particulate matter in human keratinocyte HaCaT is reduced in a concentration-dependent manner by the treatment of a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

[0065] FIG. 4 shows a result of a P450 activity measurement experiment showing that an increase in activity of cytochrome P450 induced by the treatment of particulate matter in human alveolar epithelial cell A549 is reduced in a concentration-dependent manner by the treatment of a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

MODE FOR CARRYING OUT THE INVENTION

[0066] Hereinafter, the present application will be described in detail with reference to examples. However, the following examples specifically illustrate the present application, and the contents of the present application are not limited by the following examples.

Examples

Preparation Example 1: Preparation of Peptide

[0067] Peptides having the amino acid sequence of SEQ ID NO: 1 shown in Table 1 below were synthesized using an automatic peptide synthesizer (Liberty, CEM Corporation, U.S.), and these synthesized peptides were purified by C18 reverse phase-high performance liquid chromatography (HPLC) (U-3000, Thermo fisher scientific, U.S.). Pursuit XRs C18 (250*4.65 mm 100 ?, Agilent, U.S.) was used as a column.

TABLE-US-00001 TABLE 1 SEQ ID NO Amino acid sequence of peptide 1 WYM

Experimental Example 1: Nuclear Translocation Test of AHR

[0068] Polyaromatic hydrocarbons (PAHs) present in particulate matter act as a ligand of AHR, thereby promoting nuclear translocation of AHR when the PAHs penetrate into cells and binds to AHR. In order to confirm the effect of the synthesized peptide in inhibiting cell damage caused by particulate matter, an experiment was performed for proving whether the peptide inhibits the nuclear translocation of AHR, which is usually observed at the time of ligand-AHR binding, after the cells were treated with particulate matter.

[0069] HaCaT cells, which are human keratinocytes, were seeded into a 6-well plate at a density of 5?10.sup.5 cells/well. After overnight incubation, the media were replaced with serum-free media. The keratinocytes, HaCaT were pretreated with the synthesized peptides for 1 hour at each concentration (20 ?M, 100 ?M, and 200 ?M). Then, the cells were treated with 50 ?g/cm.sup.2 of urban particulate matter (Sigma, NIST1648A) for 1 hour. After keratinocytes were collected, cell nucleus extracts were prepared, and western blotting was performed. Western blotting was performed using anti-AHR antibodies (Cell Signaling Technology) according to a protocol known in the art.

[0070] As a result of the experiment, as can be seen from the results of the western blotting shown in FIG. 2, it was confirmed that the nuclear translocation of AHR in the particulate matter-treated cells (NC) was rapidly increased compared to the control cells (Con) which were not treated with the particulate matter. In addition, it was confirmed that in the cells treated with the peptide of the amino acid sequence of SEQ ID NO: 1, the level of the nuclear translocation of AHR increased by the treatment of particulate matter was significantly reduced. In addition, the increase in the nuclear translocation of AHR by the treatment of particulate matter was reduced in a manner dependent on the concentration of the treated peptide when the cells were treated with the peptide.

Experimental Example 2: Analysis of Cytochrome P450 Activity

[0071] When polyaromatic hydrocarbons (PAHs) present in particulate matter penetrate into cells and bind to AHR, it promotes nuclear translocation of AHR. It is known that the AHR translocated into the nucleus forms a complex AHR/ARNT with an AHR nuclear translocator (ARNT), and this complex act as a transcription factor to promote the expression of cytochrome P450 (CYPs). In order to confirm the effect of the synthesized peptide in inhibiting cell damage caused by particulate matter, an experiment was performed for confirming whether the peptide inhibits the activity of cytochrome P450 after the cells were treated with particulate matter.

[0072] First, HaCaT cells, which are human keratinocytes, were seeded into a 96-well plate at a density of 1?10 4 cells/well. After overnight incubation, the media were replaced with serum-free media. The keratinocytes, HaCaT were pretreated with the synthesized peptides for 1 hour at each concentration (20 ?M, 100 ?M, and 200 ?M). Then, the cells were treated with 50 ?g/cm.sup.2 of urban particulate matter (Sigma, NIST1648A) for 24 hours. After the keratinocytes were collected, the activity of cytochrome P450 was measured using P450-Glo? Assay kit (Promega).

[0073] As a result of the experiment, as can be seen from the results of measuring the activity of cytochrome P450 shown in FIG. 3, it was confirmed that the activity of cytochrome P450 was rapidly increased in the particulate matter-treated cells (NC) compared to the control cells (Con) which were not treated with the particulate matter. In addition, it was confirmed that in the cells treated with the peptide of the amino acid sequence of SEQ ID NO: 1, the activity of the cytochrome P450 increased by the treatment of particulate matter was reduced. In addition, the increase in the activity of cytochrome P450 by the treatment of particulate matter was reduced in a concentration dependent manner of the treated peptide when the cells were treated with the peptide.

[0074] In addition, in order to confirm whether the peptide of the present invention can suppress respiratory damage caused by particulate matter, an experiment for measuring the activity of cytochrome P450 was also performed on A549 cells, which are human alveolar epithelial cells.

[0075] First, human alveolar epithelial cells, A549 cells were seeded into a 96-well plate at a density of 1.2?10 4 cells/well. After overnight incubation, the media were replaced with serum-free media. The alveolar epithelial cells, A549 cells were pretreated with the synthesized peptides for 1 hour at each concentration (20 ?M, 100 ?M, and 200 ?M). Then, the cells were treated with 50 ?g/cm.sup.2 of urban particulate matter (Sigma, NIST1648A) for 24 hours. After the treated cells were collected, the activity of cytochrome P450 was measured using P450-Glo? Assay kit (Promega).

[0076] As a result of the experiment, as can be seen from the results of measuring the activity of cytochrome P450 shown in FIG. 4, it was confirmed that the activity of cytochrome P450 was rapidly increased in the particulate matter-treated cells (NC) compared to the control cells (Con) which were not treated with particulate matter, and it was confirmed that the activity of cytochrome P450 increased by the treatment of particulate matter was reduced in the cells treated with the peptide of the amino acid sequence of SEQ ID NO: 1. In addition, the increase in the activity of cytochrome P450 by the treatment of particulate matter was reduced in a manner dependent on the concentration of the treated peptide when the peptide was treated.

[0077] Although the representative embodiments of the present application have been exemplarily described, the scope of the present application is not limited to the specific embodiments as described above, and a person skilled in the art can change the present application within the scope described in the claims of the present application.

[Sequence List Free Text]

[0078] SEQ ID NO: 1: Trp Tyr Met