Anti-Inflammatory Peptides and Composition Comprising the Same

a technology of anti-inflammatory peptides and compositions, applied in the direction of peptide sources, transferases, immunological disorders, etc., can solve the problems of increased capillary permeability and concentration, increased blood flow rate, and abnormal biological defense response excessively,

Inactive Publication Date: 2017-09-28
GEMVAX & KAEL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0035]In another embodiment of the composition, the above-mentioned composition is a cosmetic composition for improving or preventing skin inflammation.

Problems solved by technology

However, inflammatory diseases can result if an abnormal biological defense response occurs excessively.
The released histamine and kinin will result in anglectasis, increased capillary permeability and concentration of macrophages at the inflammation site, and it causes increased blood flow rate, edema, immunocyte and antibody migration, pain and heat generation.
These drugs do not fundamentally cure inflammation, and they have side effects such as hypersensitivity reaction, and deterioration of immune system,
However, problems have arisen in anti-inflammation substances that had been developed previously.
Diverse categories of anti-inflammatory drugs including Non-steroidal Anti-inflammatory Drugs (NSAIDs) and Steroidal Anti-inflammatory Drugs (SAIDs) have been developed; but not only do these drugs often bear side effects upon use, they also do not fundamentally cure the inflammation.
As one example, in acute or chronic inflammations such as chronic rheumatoid arthritis, not only do non-steroidal anti-inflammatory drugs suppress COX-2 enzyme activity, they are also known to suppress COX-1 activity, causing side effects such as gastrointestinal disorders.

Method used

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  • Anti-Inflammatory Peptides and Composition Comprising the Same
  • Anti-Inflammatory Peptides and Composition Comprising the Same
  • Anti-Inflammatory Peptides and Composition Comprising the Same

Examples

Experimental program
Comparison scheme
Effect test

example 1

Synthesis of PEP-1 and Measurement of Anti-Inflammatory Activities of PEP-1 (SEQ ID NO: 1)

Experiment 1. Synthesis of PEP-1 (SEQ ID NO: 1)

[0109]A peptide comprised of 16 amino acids with the chemical structure 1 as below having the sequence SEQ ID: 1 (PEP-1) derived from human telomerase was synthesized:

[0110]SEQ ID NO: 1 (PEP-1) was synthesized according to the existing method of solid phase peptide synthesis. In detail, the peptides were synthesized by coupling each amino acid from C-terminus through Fmoc solid phase peptide synthesis, SPPS, using ASP485 (Peptron, Inc., Daejeon ROK). Those peptides with their first amino acid at the C-terminus being attached to resin were used as follows:[0111]NH2-Lys(Boc)-2-chloro-Trityl Resin[0112]NH2-Ala-2-chloro-Trityl Resin[0113]NH2-Arg(Pbf)-2-chloro-Trityl Resin

[0114]All the amino add materials to synthesize the peptide were protected by Fmoc at the N-terminus, and the amino acid residues were protected by Trt, Boc, t-Bu (t-butylester), Pbf (...

experiment 2-1

sis

[0129]The level of nitric oxide (NO) was measured in Raw 264.7 cell (1×106 cell / ml) using Griess reagent system (Promega, USA). Culture medium of 50 μl was added to a 96-well plate and Griess reagent I (NED) solution and Griess reagent II (Sulfanilamide solution) are added in the same amount. After 10 min incubation of cells with the reagents, the optical density at 540 nm was measured within 30 min using a microplate reader (Molecular Devices, USA). The concentration of NO was calculated by using a standard curve (0˜100 μM) of sodium nitrite.

[0130]As shown in Table 3 below, stimulation of cells with LPS increased the expression of NO, but in co-treatment with LPS and Pep1, the expression level of NO mentioned above decreased. NO is produced during inflammation, and the result showing Pep1 reduced NO level to 65% of the control strongly support the anti-inflammatory effect of Pep1.

TABLE 3The measurement of anti-inflammatory effect of humantelomerase derived PEP 1NO ExpressionDecr...

experiment 2-2

tokine Inhibitory Effect

[0131]To investigate the effect of PEP1 on inhibiting pro-inflammatory cytokine production RAW 264.7 cell were pre-treated with PEP 1 at a concentration of 5 μg / mL challenged with LPS at a concentration of 1 μg / mL, and cells were further incubated for 24 hr. The supernatant samples containing cell culture medium was collected and analyzed for the cytokine levels using ELISA kits (eBloscience, San Diego).

[0132]96 wells plates were coated with 100 μl of capture antibodies (diluted in coating buffer to the concentration recommended by manufacturer's protocol) overnight at 4° C. Then, after washing the plates 5 times, 200 μL of assay diluents was added to each well and incubated for 1 hr at room temperature for blocking. After washing each well with wash buffer five times, cell culture sample or each cytokine standard protein sample was diluted and 100 μl of each added into each well. The plate containing samples were incubated overnight at 4° C.

[0133]Then, after...

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Abstract

The present invention relates to a peptide with anti-inflammatory activity, wherein the peptide comprises any one amino acid sequence of SEQ ID NO:1 to SEQ ID NO: 161, the peptide has above 80% homology of amino acid sequence with above-mentioned sequences, or the peptide is the fragment of the above-mentioned peptides. The present invention also relates to an inflammatory composition comprising the above mentioned peptides. According to the present invention, a peptide that has at least one amino acid sequence of SEQ ID NO:1 to SEQ ID NO: 161 has outstanding efficacy in both suppressing inflammation and in prophylactic means. Therefore, the composition comprising the peptides of this invention can be used as anti-inflammatory pharmaceutical compositions or as cosmetic compositions, in turn, treating and preventing a variety of different types of inflammatory diseases.

Description

FIELD OF THE INVENTION[0001]The present invention relates to anti-inflammatory peptides and compositions comprising the same.BACKGROUND OF THE INVENTION[0002]Inflammation is a type of biological defense as a means of protecting the body from damage of biological tissues that could be caused by external physical stimuli, chemical stimuli such as exposure to various aHergens, or invasion of microorganisms including bacteria, fungi and viruses.[0003]The Cyclooxygenase(COX) pathway or Lipoxygenase(LOX) Pathway can used for signaling inflammation, which produce prostaglandin, thromboxane, etc. Once the inflammatory signal is delivered, one of many changes that happen in the body is the expansion of the blood vessel for increased blood supply around the inflammation to concentrate blood cells such as neutrophils required for the inflammatory response. However, inflammatory diseases can result if an abnormal biological defense response occurs excessively. To prevent this, drugs that suppre...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N9/12A61K38/45A23L33/18
CPCC12Y207/07049A23L33/18A61K38/45C12N9/1276A23V2002/00A61K38/00A61P1/00A61P11/00A61P13/00A61P15/00A61P17/00A61P19/00A61P25/00A61P29/00A61P31/00A61P35/00A61P37/00A61P43/00A61P9/00Y02A50/30C07K14/435C07K14/47
Inventor KIMKIM, KYUNG HEELEE, KYU-YONGKOH, SEONG-HOKIM, BUM JOONPARK, HYUN-HEEHUH, SUNG JINLEE, WOO JINJIANG, HWANHA, JUNG SOON
Owner GEMVAX & KAEL
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