Novel Peptides
A pharmacology and composition technology, applied in the field of novel peptides or their derivatives or their salts, can solve the problems of unknown combination of angiotensin-converting enzyme fibroblasts and collagen adhesion promoting activities, etc., to restore tension and elasticity , Prevent the reduction of bonding ability and enhance the effect of bonding
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Embodiment 1
[0087] Embodiment 1: the preparation of peptide
[0088] 1-1. DLDVF (Asp-Leu-Asp-Val-Phe) peptide:
[0089] (1) Synthesis of peptides:
[0090] The title peptide was synthesized by a solid-phase synthesis method using the Fmoc method using an automatic peptide synthesizer (manufactured by Shimadzu Corporation: PSSM8). The specific procedure is as follows: First, after the C-terminal of Fmoc-Phe(1-Trt)-OH is bound to the resin for solid-phase synthesis, the protective group (Fmoc) is removed by piperidine treatment, and then the resin is neutralized. · After washing, introduce Fmoc-Val-OH into the N-terminus of Phe. Next, the protective group (Fmoc) was removed by piperidine treatment, and after neutralizing and washing the resin again, Fmoc-Asp(OtBu)-OH was introduced into the N-terminal of Val. Next, the protective group (Fmoc) was removed by piperidine treatment, and after neutralizing and washing the resin again, Fmoc-Leu-OH was introduced into the N-terminal of Asp. N...
Embodiment 2
[0103] Example 2: Determination of angiotensin-converting enzyme inhibitory activity
[0104] The angiotensin-converting enzyme inhibitory activity was measured using the various peptides prepared in Example 1 above.
[0105] Specifically, test solutions were prepared so that the concentration of each peptide was 1 mg / mL, and 1-fold, 10-fold, 100-fold, and 1000-fold dilutions were used to measure angiotensin-converting enzyme inhibitory activity. The angiotensin-converting enzyme inhibitory activity was evaluated by using the ACE Kit-WST kit (manufactured by Dojin Chemical Research Institute) to detect 3-hydroxybutyryl-Gly-Gly-Gly (3HB-GGG ), and measure the absorbance at 450 nm with a microplate reader. The ACE Kit-WST kit was used according to the attached protocol. The results are shown below.
[0106] [Table 1]
[0107] peptide
[0108] DLDV
[0109] As shown in the above results, it was revealed that any of the three test peptides exhibited high an...
Embodiment 3
[0110] Example 3: Evaluation of Fibroblast Adhesion to Collagen Coatings
[0111] The effect of promoting adhesion of fibroblasts to the collagen coating was evaluated using various peptides prepared in Example 1 above.
[0112] Specifically, first, 2 × 10 4 Fibroblasts (クラボウ: KF-4009) from human normal skin were inoculated at a concentration of cells / well. Next, 200 μl of Dulbecco’s modified Eagle’s medium (D-MEM; FCS10%·AB1%) diluted so that the concentration of each peptide was 5 μg / ml was added to each well. % carbon dioxide and 95% air environment for 20 hours of cultivation. On the other hand, cells to which 200 μl of D-MEM medium to which no peptide was added were prepared as a control. After 4 hours from the start of the culture, the supernatant of each well was washed with PBS, and the remaining amount of collagen coating adhered to and not washed was measured in each well using the Cell Counting Kit-8 kit (manufactured by Dojin Chemical Research Institute of Co., ...
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