Preparation and Application of Myristoyl Pentapeptide-17

General description

Myristoyl Pentapeptide-17, also known as eyelash peptide, is a kind of pentapeptide-17 that can effectively promote the growth of eyelashes. It has significant effects in activating the growth of keratin gene, enhancing the absorption of nutrients by hair follicles, and enhancing the length, thickness and quantity of eyelashes. It fundamentally solves the problem of eyelashes shortage and scarcity. This peptide is currently a relatively safe and effective eyelash promoting ingredient, which can be widely used in mascara, eyelash care lotion, shampoo and hair care products and other hair enhancing products.

Fig. 1 The structure of Myristoyl Pentapeptide-17.

Preperation

(1) The first protective amino acid is connected to the resin;

a1. In the polypeptide reactor with sand core, the resin after swelling and activation is added successively, then the resin is drained, washed, and drained under reduced pressure; The protection mode of solid phase peptide synthesis is much simpler than that of liquid phase peptide synthesis. The protective amino acids are groups used for α-amino group protection in solid phase peptide synthesis, which are usually Boc(tert-butyloxy), Moz(p-methoxy benzyl oxygen) and Fmoc(9-fluorenylmethoxy carbonyl).

b1. Add the deprotective agent, deprotective group reaction, after complete deprotection, filter, wash the resin, and test; It is a conventional technique in this field to add a deprotective agent for the deprotection reaction. Under mild conditions, it is usually possible to add piperidine for the deprotection reaction. A 20% solution of piperidine-DMF was added to the peptide resin at 28 °C. Add 20% piperidine-DMF solution, reaction 30 min, two times;

c1. Add the first protective amino acid, shrinkage agent and organic solvent and mix them evenly. Place the polypeptide reactor on a shaker for condensation reaction or inject nitrogen or inert gas into the reaction system to volatilize the volatile solvent, and keep the reaction system temperature at 25℃; Finally, add the help shrinkage mixture to start the condensation reaction of protective amino acid and resin, reaction temperature 22-28℃, and detection; After the condensation reaction is complete, the precipitate is filtered and washed.

(2) Linking the second protective amino acid

a2. After washing in step (1), add organic solvent and piperidine to the sand core reaction vessel containing resin for deprotective agent reaction; After removing the protection completely, the resin was filtered, washed and tested.

b2. add the second protective amino acid, shrinkage agent, organic solvent and mix evenly, put the polypeptide reactor in the shaker for condensation reaction or into the reaction system nitrogen or inert gas to volatilize the volatile solvent, and maintain the reaction system temperature at 15℃; Finally, adding the help shrinkage agent to start the condensation reaction of protective amino acid and resin, the reaction temperature is 25℃, and detection; After the condensation reaction is complete, the resin is filtered and washed.

(3) Lys-Leu-Ala-Lys(Boc)-Lys(Boc)-Rink amide resin was synthesized

The products obtained after washing in step (2) were deprotected, washed and tested in the same way as in step (a1). In accordance with step (b1), amino acids Fmoc-Ala-OH, Fmoc-Leu-OH and FmocLys-OH were successively connected in the same way. Fmoc-Lys-Leu-Ala-Lys(Boc)-Lys(Boc)-Rink amide resin was obtained, and the resin was deprotected, washed and tested in the same way as in step (a1). Lys-Leu-Ala-Lys(Boc)-Lys (Boc)-Rink amide resin was obtained.

(4) Myristoyl - Lys (Boc) - Leu - Ala - Lys (Boc) - Lys (Boc) - Rink amide resin

The products obtained after washing, add myristicyl chloride, shrinkage agent, organic solvent and mix evenly, put the polypeptide reactor in the shaker for condensation reaction or into the reaction system to make the volatile solvent volatile, and maintain the reaction system temperature at 25℃; Finally, adding the help shrinkage agent to start the condensation reaction of protective amino acid and resin, the reaction temperature is 28℃, and detection; After the condensation reaction is complete, the resin is filtered, washed and drained under pressure.

(5) Cleavage of peptide chains

In the drained synthetic peptide resin to add cutting solution, in the fume hood at room temperature stirring cracking, the use of sand core funnel filter to remove the resin, filtrate collection, filtrate by ether extraction placed in the dry bottle dry, until it becomes a white powder, that is, polypeptide crude.

(6) Detection and purification of crude peptides

The synthetic peptides were detected by reverse high performance liquid chromatography (HPLC), and the purity of the peptides in the synthetic peptides was analyzed. HPLC detection method: the peptide sample was placed in the EP tube, TFA and water were added, and the peptide sample was dissolved in the ultrasonic vibration machine. The C18 column was used for detection, the column temperature was 45℃, and the HPLC analysis was performed. Phase B: acetonitrile, 0.1% TFA, elution gradient: solubility of phase B from 5% to 95%, elution rate was 1ml/min, elution time was 35min, detection wavelength was 215 nm [1].

Application

Promote hair growth

Hair not only has aesthetic effect, but also has biological function and physiological function, which will affect people's psychology and social interaction. With the rapid development of social economy and the acceleration of people's pace of life, the phenomenon of hair loss caused by various reasons is increasing day by day. Alopecia and alopecia have become common clinical skin diseases at home and abroad. According to the health index issued by the relevant departments of the state, the incidence of alopecia in the Chinese adult population is as high as 40%, and the age of onset tends to be younger. Studies have shown that people with hair loss experience both sleep quality and mood problems. There is a growing demand for hair care products. Hair care products have become the fastest growing consumer category in recent years. The development of safe and effective topical products for alopecia has long been a research hotspot. Biopeptides developed in recent years, such as Biotinoyl Tripeptide?1 and Myristoyl Pentapeptide?17, also promote scalp microcirculation, Inhibition of 5α-reductase activity and significant activation of keratin [2].

References

[1] Li H, Zhou Z, Xu C, et al. New myristoyl pentapeptide-17 useful in the field of food, medicine and cosmetics for promoting hair growth, preferably the growth of eyelashes[P]. Faming Zhuanli Shengqing, CN110204596, 2019.

[2] Liu M, Yang A. Composition useful in anti-hair loss hair-growing product e.g. shampoo, hair conditioner, hair care essential oil for preventing human hair loss comprises biotin tripeptide-1, myristoyl pentapeptide-17, glossy privet fruit extract[P]. Faming Zhuanli Shengqing, CN112870333, 2021.

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