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Polypeptide analogue with hair growth effect as well as preparation method and application

A technology for hair growth and peptides, applied in the field of peptides, can solve problems such as unsatisfactory effects, achieve the effects of shortening the synthesis cycle, inhibiting downstream signals, and facilitating automation

Pending Publication Date: 2020-12-22
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hair growth products currently on the market are unsatisfactory

Method used

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  • Polypeptide analogue with hair growth effect as well as preparation method and application
  • Polypeptide analogue with hair growth effect as well as preparation method and application
  • Polypeptide analogue with hair growth effect as well as preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Val-Gly-Ser-Tyr-Leu-Glu-Ser-Asp-Leu-NH 2 (SEQ.ID NO:2) microwave-facilitated solid-phase synthesis

[0041] (1) Swelling of the resin

[0042] Weigh 50 mg of Fmoc-Rink amide-MBHA Resin (Sigma Company) (substitution amount: 0.4 mmol / g), swell with 7 mL of DCM for 30 min, remove DCM by suction filtration, then swell with 10 mL of NMP for 30 min, and finally rinse with NMP and 7 mL of DCM respectively.

[0043] (2) Microwave promotes removal of Fmoc protecting group

[0044] Put the swollen resin into the reactor, add 7mL of 25% piperidine / NMP (V / V) solution containing 0.1M HOBT, react in the microwave reactor for 1min, the microwave power is 15W, and the reaction temperature is controlled at 50°C Within the time period, use an air compressor to compress the air to cool, and filter the solution after the reaction; add 7 mL of 25% piperidine / NMP (V / V) solution containing 0.1M HOBT and react in a microwave reactor for another 4 minutes, and the microwave power is 25W , the ...

Embodiment 2

[0056] Binding experiment of FGF5 and FGF5 inhibitor polypeptide (FGF-OPI)

[0057] In order to determine whether the FGF5 inhibitor polypeptide has an activating effect on FGFR, we analyzed the affinity of the FGF5 inhibitor polypeptide and FGFR1c. We used BIAcore T200 system (GE Healthcare, NJ, USA) to carry out surface ion resonance SPR experiments, the buffer system is HBS-EP buffer (10mM HEPES-NaOH, pH 7.4, 150mMNaCl, 3mM EDTA and 0.005% [v / v ] polysorbate 20), the temperature is 25°C.

[0058] Since the D2-D3 region of FGFR1c is sufficient to exert ligand binding ability, we immobilized FGFR1c (D2-D3 region) on the CM5 chip through amino coupling reaction to obtain the FGFR1c chip. Solvent effects were subtracted from the control channel as a control. FGF5 inhibitor polypeptides (25uM, 50uM, 100uM, 200uM, 400uM) were flowed through the chip one by one (including the experimental channel and the control channel). The chip was regenerated by flowing regeneration solutio...

Embodiment 3

[0061] Evaluation of the blocking effect of FGF5 inhibitor (FGF-OPI) on downstream signaling pathways

[0062] In order to study the inhibitory effect of FGF5 inhibitors on FGF5, the epidermal cell Hcat was used to stimulate the relevant downstream signaling molecular mechanisms, such as figure 2 As shown, the results of Western blot showed that FGF5 can significantly enhance the expression of p-FRS2, p-Erk2 and p-AKT downstream, and after adding inhibitor polypeptides, the corresponding phosphorylation indicators of these signaling pathways were weakened in a dose-dependent manner. reduce. It shows that the FGF5 inhibitor polypeptide can effectively inhibit the activity of FGF5.

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PUM

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Abstract

The invention discloses a polypeptide with a hair growth effect. The structure of the polypeptide has the following form: Val-Xaa1-Xaa2-Tyr-Leu-Glu-Xaa3-Asp-Leu, the polypeptide is prepared by microwave promoted Fmoc / tBu orthogonal protection solid-phase synthesis, the polypeptide synthesis efficiency is greatly improved, and the synthesis period is shortened. The fibroblast growth factor 5 (FGF 5) inhibitor polypeptide analogue prepared by the microwave-promoted solid-phase synthesis technology is high in yield, short in synthesis period, easy in crude product purification, low in productioncost and easy in industrial automatic production. The prepared FGF 5 inhibitor polypeptide is suitable for being used as an active ingredient for treating alopecia and promoting hair growth.

Description

technical field [0001] The invention relates to a polypeptide, in particular to a polypeptide analogue with hair growth function, its preparation method and application. Background technique [0002] With the increase of life pressure, endocrine disorders, mental trauma and other reasons cause more and more hair loss people. The hair growth products currently on the market are all unsatisfactory. There is a broad market demand for hair growth agents that generally exert effects on hair thinning and hair loss. [0003] Hair growth has a certain periodicity, hair does not grow continuously at a certain speed, but repeats the growth period and rest period at the same time. Studies have found that fibroblast growth factor 5 (FGF-5) also plays a biologically active role in the hair cycle. FGF-5 has the function of inhibiting the growth of hair, making the hair cycle enter the catagen phase from the growth phase, resulting in hair loss. Studies have found that FGF-5S, a short-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06A61K38/08A61K8/64A61Q7/00A61Q5/00A61P17/14
CPCC07K7/06A61K8/64A61Q7/00A61Q5/00A61P17/14A61K38/00
Inventor 范兰兰王蓓蓓杨选鑫胡振宇金杉杉阮成鑫
Owner WENZHOU MEDICAL UNIV
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