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WO2019039630A1 - Peptide bioactif dérivé de follistatine et son utilisation - Google Patents

Peptide bioactif dérivé de follistatine et son utilisation Download PDF

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Publication number
WO2019039630A1
WO2019039630A1 PCT/KR2017/009296 KR2017009296W WO2019039630A1 WO 2019039630 A1 WO2019039630 A1 WO 2019039630A1 KR 2017009296 W KR2017009296 W KR 2017009296W WO 2019039630 A1 WO2019039630 A1 WO 2019039630A1
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WIPO (PCT)
Prior art keywords
peptide
hair
growth
skin
composition
Prior art date
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Ceased
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PCT/KR2017/009296
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English (en)
Korean (ko)
Inventor
안인숙
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Genecellpharm Co Ltd
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Genecellpharm Co Ltd
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Priority to CN201780086789.9A priority Critical patent/CN110312731B/zh
Publication of WO2019039630A1 publication Critical patent/WO2019039630A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/04—Linear peptides containing only normal peptide links
    • C07K7/06—Linear peptides containing only normal peptide links having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0625—Epidermal cells, skin cells; Cells of the oral mucosa
    • C12N5/0627—Hair cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/998—Proteins not provided for elsewhere

Definitions

  • the present invention relates to novel uses of bioactive peptides derived from growth factors.
  • the animals have a molting period in which the hairs are totally replaced by the seasons, but in humans, some of the whole hairs are constantly replaced every day, so that a similar number is always maintained.
  • the hair cycle can be divided into three periods of growth period, regenerative period and pausing period.
  • anagen is a period when active division of cells occurs in the dermal papilla and new hair growth is accelerated. During this period, hair grows .
  • the period of hair appearance is approximately 3 to 5 years for males and 4 to 6 years for females, and about 80 to 85% of hair as a whole is the growth period.
  • the catagen is about 3 to 4 weeks when cell division is gradually stopped.
  • the taloge is a period when the hair is separated from the capillaries by the atrophy of the papillary hair and is simply put on the scalp. It is about 3 months. The hair in the pauses is easily separated from the fur by physical stimulation.
  • hair loss refers to the shortening of hair growth rate in the growing period and the increase of hair in the retrograde period or rest period in such a cycle, resulting in abnormal abundance of hair.
  • Regular hair growth requires hair regeneration and stem cell activation, which is known to be caused by the production and activation of Wnt / ⁇ -catenin.
  • the DKK-1 mentioned above is known to be an antagonist of Wnt / ⁇ -catenin, and the above signal system is very important for controlling hair growth.
  • TGF- ⁇ transforming growth factor- ⁇
  • TGF- ⁇ transforming growth factor- ⁇
  • polystatin It has been known through amino acid sequence analysis of polystatin that it consists of four domains: an N-terminal domain, a pol statin domain I, a pol statin domain II, and a pol statin domain III (Shimasaki, Proc. Natl. 1988, 85; 12, p4218-4222).
  • the double pol statin domain is known to be involved in the binding of actin.
  • HDP human dermal papilla
  • various growth factors that regulate hair growth and regression are produced and regulated from the dermal papilla cells of the hair follicles.
  • actin is also expressed in dermal papilla cells, inducing the formation of hair follicles and activating the hair growth cycle (Sung, Experimental dermatology. 2017, 26, 2, p108-115).
  • actin inhibits the entry of hair into the systolic phase, and the proteins expressed by actinin-pol statin control play an important role in the growth and differentiation of hair, and new cells can be produced.
  • actibin regulates epidermal proliferation and cleavage, and plays an important signaling role in the regulation of epidermal homeostasis.
  • the herbal remedy for the treatment or prevention of alopecia that has been developed so far includes female hormone-based preparations such as minoxidil, finasteride, tricosaccharide, etc. for promoting blood circulation, enhancing hair follicle function, scalp moisturizing and male hormone suppression, (trichosaccharide), etc., but there are many limitations in use due to adverse effects such as sexual dysfunction, possibility of birth of deformed baby, and general hair growth.
  • female hormone-based preparations such as minoxidil, finasteride, tricosaccharide, etc. for promoting blood circulation, enhancing hair follicle function, scalp moisturizing and male hormone suppression, (trichosaccharide), etc.
  • the present inventors newly identified that a specific site of polystin acts as an actin binding motif, and the peptide containing the motif competitively binds at binding sites such as actin and inhibits the binding of natural polystin and actin Thereby promoting the growth of hair.
  • the present inventors also intend to provide the possibility of utilization of the peptide for treating alopecia as well as various diseases associated with the signal system.
  • the present invention has been made keeping in mind the above problems occurring in the prior art, and it is an object of the present invention to provide a physiologically active peptide having excellent biostability and excellent hair growth promoting effect and skin condition improving effect.
  • a physiologically active peptide consisting of at least one amino acid sequence selected from the group consisting of SEQ ID NOS: 1-7.
  • the peptide is capable of promoting the growth of fibroblasts, keratinocytes, or hair follicle cells.
  • the peptide may activate growth factors of dermal papilla cells.
  • R 1 is a peptide consisting of one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 1 to 3
  • R 2 is a peptide consisting of the amino acid sequence of SEQ ID NO: 4.
  • A may be a compound represented by the following formula (2).
  • n is an integer of 1? N? 18.
  • A may be 6-aminohexanoic acid.
  • the physiologically active peptide construct may facilitate the growth of fibroblasts, keratinocytes, or hair follicle cells.
  • the physiologically active peptide construct can activate growth factors of dermal papilla cells.
  • composition for promoting hair loss prevention and hair growth comprising the peptide or the peptide structure as an active ingredient.
  • composition comprising the peptide or the peptide structure as an active ingredient for improving wrinkles, improving skin elasticity, improving skin moisturization, or regenerating skin.
  • a cosmetic composition comprising the peptide or the peptide structure as an active ingredient.
  • an external preparation for skin comprising the peptide or the peptide structure as an active ingredient.
  • a pharmaceutical composition for preventing or treating hair loss comprising the peptide or the peptide structure as an active ingredient.
  • an excellent skin penetration rate can be realized by using a small-sized peptide, thereby improving the hair and skin condition improving effect.
  • the peptides can be used for various purposes such as medicines, quasi-drugs, health functional foods, and cosmetics.
  • Figure 1 shows the results of HPLC analysis of peptides according to one embodiment of the present invention.
  • 2A is a graph showing the effect of promoting cell growth in keratinocytes treated with a peptide according to an embodiment of the present invention.
  • FIG. 2B is a graph showing a cell growth promoting effect in a fibroblast treated with a peptide according to an embodiment of the present invention.
  • FIG. 2C is a graph showing a cell growth promoting effect in a hair follicle cell treated with a peptide according to an embodiment of the present invention.
  • FIG. 3A is a microscopic examination of the effect of promoting cell growth in a hair follicular cell treated with a peptide according to an embodiment of the present invention through a three-dimensional solid cell culture method.
  • FIG. 3B shows the sizes of hair follicle cell spheroids in 3-dimensional solid cells treated with peptides according to an embodiment of the present invention.
  • Figure 4 shows the increase of SMAD2 phosphorylation according to the peptide treatment according to an embodiment of the present invention.
  • FIG. 5 is a graph showing an increase in signal transduction for activating hair follicles according to the peptide treatment according to an embodiment of the present invention.
  • FIG. 6 is a graph showing an effect of promoting self-assembly of a papilla according to an embodiment of the present invention and an increase in the expression of a hair growth factor through immunostaining.
  • FIG. 7 shows hair growth observed at a site of a mouse treated with a peptide according to an embodiment of the present invention.
  • the terminology used herein is intended to encompass all commonly used generic terms that may be considered while considering the functionality of the present invention, but this may vary depending upon the intent or circumstance of the skilled artisan, the emergence of new technology, and the like. Also, in certain cases, there may be a term selected arbitrarily by the applicant, in which case the meaning thereof will be described in detail in the description of the corresponding invention. Therefore, the term used in the present invention should be defined based on the meaning of the term, not on the name of a simple term, but on the entire contents of the present invention.
  • One aspect of the present invention provides a physiologically active peptide comprising at least one amino acid sequence selected from the group consisting of SEQ ID NOS: 1-7.
  • the present inventors have identified the in vivo physiological activities of the peptides, and in particular, they have clarified the skin condition improving effect and the hair growth promoting effect of the peptides.
  • physiologically active peptide is a concept that collectively refers to a peptide having a predetermined physiological action in vivo, and it is a concept that regulates genetic expression and physiological function, It can play a role of correcting it.
  • the peptide may be composed of the amino acid sequence of SEQ ID NOS: 1 to 7, or may include the amino acid sequence as a part thereof.
  • the peptide refers to a polymer composed of one or more amino acids linked by amide bonds (or peptide bonds).
  • the peptide may refer to peptides or fragments thereof that exhibit wrinkle-improving and skin elasticity-improving effects.
  • the general rules for naming the peptides may be based on three letter or single letter amino acid codes, unless specifically indicated otherwise.
  • the center of the amino acid structure is represented by a three-letter code (e.g., Ala, Lys), and a D-stereotype (e.g., D-Ala, D-Lys) It is possible to assume an L-steric form.
  • the amino acid residue constituting the peptide may be a natural or non-natural amino acid residue.
  • the peptide randomly partially synthesizes various sites of the pol statin protein to first search for a binding site for the receptor protein, and then optimizes the amino acid sequence of the predicted site. Screening for the peptide having the highest activity among the candidate peptides It was completed.
  • the peptide comprises an actin binding motif of native polystin and can promote the activity of actin by binding to actin and interfering with its binding to native pol statin.
  • the peptide has fibroblast, keratinocyte, or hair follicle cell growth promoting ability, and can transmit an actin signal even in the presence of native polystin.
  • the peptides can increase the expression of Wnt5, FGF7, Versican, Lef1, the major genes of hair follicles, and increase the size of the dermal papilla cells through three dimensional solid cell culture.
  • the Wnt ligand inhibits hair loss by activating the beta-catenin signaling mechanism in cells.
  • the Wnt binds to the follicular receptors of hair follicle cells to activate dermal papilla cells and inhibit hair follicle cell destruction materials including TGF-b, DKK-1 and BMP, thereby inhibiting hair loss. That is, the peptide significantly increases the amount of Wnt produced and promotes the growth of hair follicle cells, thereby achieving excellent hair loss prevention and hair growth promoting effect.
  • the peptide competes with the natural polystin protein to inhibit the function of the natural pol statin and is superior in stability and skin permeability as compared to natural pol statin protein.
  • the peptide promotes the growth of fibroblasts or keratinocytes to impart elasticity to the skin, thereby suppressing the formation of wrinkles. Furthermore, the peptide may promote hair follicle growth and induce hair growth.
  • the keratinocytes and fibroblasts in the skin may be damaged and degenerated due to various causes such as mental stress, external harmful factors, and endogenous aging.
  • the damage of keratinocytes and fibroblasts due to internal and external stress may lower the density of skin cells and interfere with the synthesis of collagen, which can deepen the formation of skin wrinkles.
  • the collagen is a major component of the connective tissue and is a component mainly distributed in the bones and skin. It is a light protein that maintains the structure and shape of the skin and provides firm strength and elasticity. When the skin is aged, exposed to ultraviolet rays, heat, etc., collagen synthesis ability decreases and collagen amount can be significantly reduced. When the amount of the collagen decreases, the shape of the muscle can not be maintained, so that a skin wrinkle can be generated.
  • the peptides originate from the body of polystatin and are high in skin permeability due to their small molecular weight and size, and are excellent in promoting skin growth.
  • the peptide promotes the growth of fibroblast and keratinocyte and promotes collagen synthesis, it can grow the stratum corneum, epithelial layer and dermal layer of the skin and improve wrinkles, skin elasticity, skin aging, skin moisturization, .
  • the peptide can be produced biologically by extracting a protein in vivo and treating it with proteolytic enzymes to lower molecular weight or by using a gene recombination and protein expression system or by a chemical synthesis method using a peptide synthesizer or the like .
  • the peptides can be prepared according to chemical synthesis methods known in the art, particularly solid-phase synthesis techniques (Merrifield, J. Amer. Chem. 1963, Soc. 85: 2149- 54; Stewart, et al., Solid Phase Peptide Synthesis, 2nd ed., Pierce Chem. 1984, Co .: Rockford, 111).
  • a gene encoding a fusion partner and a fusion protein composed of the peptide may be prepared through gene manipulation, and the host microorganism may be transformed with the gene and expressed in the form of a fusion protein in the host microorganism.
  • the fusion protein can be cleaved and separated using proteolytic enzymes or compounds, and the peptide can be obtained.
  • a DNA sequence encoding an amino acid residue that can be cleaved by a proteolytic enzyme, such as Factor Xa or an enterokinase, a compound such as CNBr or hydroxylamine can be inserted between the fusion partner and the gene of the peptide .
  • the N or C-terminal of the peptide may be bonded with an acetyl group, a fluorenylmethoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group or polyethylene glycol (PEG).
  • the stability of the peptide can be remarkably improved by the modification.
  • the stability is a concept including not only in vivo stability but also storage stability.
  • the protecting group can protect the peptide of the present invention from attack of a protein cleaving enzyme in vivo.
  • the peptide may comprise a functional equivalent of a peptide comprising the amino acid sequence.
  • Said "functional equivalent” means that at least 40%, preferably at least 80%, more preferably at least 90%, more preferably at least 95% sequence identity with said amino acid sequence as a result of addition, substitution or deletion of amino acids Means a protein having homology and having a physiological activity substantially equivalent to the above amino acid sequence.
  • the "substanti homogenous physiological activity" means a skin condition improving activity such as anti-inflammatory activity due to the structural and functional homology of the peptide.
  • the sequence homology is determined by comparing the two optimally arranged sequences with the comparison region, and some nucleic acid sequences in the comparison region may be added or deleted.
  • R 1 is a peptide consisting of one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 1 to 3
  • R 2 is a peptide consisting of the amino acid sequence of SEQ ID NO: 4.
  • the peptide construct comprises a peptide consisting of the amino acid sequence of one of SEQ ID NOS: 1 to 3, and a peptide consisting of the amino acid sequence of SEQ ID NO: 4, wherein the peptide can be linked by a linker.
  • the linker may be a flexible linker, specifically a peptide linker or a non-peptide linker.
  • non-peptide linker means a biocompatible linker in which two or more repeating units are bonded, and the repeating units may be connected to each other through any covalent bond other than a peptide bond.
  • the non-peptide linker may be selected from the group consisting of polyethylene glycol (PEG) homopolymer, polypropylene glycol homopolymer, ethylene glycol-propylene glycol copolymer, polyoxyethylated polyol, polyvinyl alcohol, polysaccharide, dextran, polyvinyl ethyl Ether, biodegradable polymer, lipid polymer, chitin, hyaluronic acid, or a combination thereof, but is not limited thereto.
  • PEG polyethylene glycol
  • the A may be a compound represented by the following formula (2), preferably 6-aminohexanoic acid.
  • n is an integer of 1? N? 18.
  • the 6-aminohexanoic acid may be represented by the following general formula (3), and may further serve as a linker for the peptide and further increase the physiological activity of the peptide.
  • composition for promoting hair loss prevention and hair growth comprising the peptide or the peptide structure as an active ingredient.
  • &quot hair loss " refers to a state in which hair is normally broken at a site where hair should be present, and generally means that the hair of the scalp is missing. Unlike colorless and thick skin, the Virgin Mary can cause cosmetic problems when it is missing.
  • the hair loss can be classified into two types, that is, a case where the hair is clinically formed and a case where the hair is not formed.
  • the hair loss (scarring) in which a scar is formed can not be regenerated due to the destruction of the hair follicle, Scarring) maintains the hair follicles so that the hair can be regenerated after the symptoms disappear.
  • the hair loss may include alopecia areata, alopecia totalis, alopecia universalis, androgenic alopecia, telogen effluvium, anagen effluvium, or chemotherapy- But is not limited to, chemotherapy-induced alopecia.
  • the peptide can promote the growth of hair follicle cells, hair growth can be fundamentally promoted and hair growing can be effectively protected.
  • &quot preventing hair loss " or " promoting hair growth " can be used in a similar sense, which may have the same meaning as another term used in the art, hair loss improvement, wool promotion and hair growth promoting.
  • composition comprising the peptide or the peptide structure as an active ingredient for improving wrinkles, improving skin elasticity, improving skin moisturization, or regenerating skin.
  • the " wrinkles” refers to the scarring caused by degeneration of the skin, and may be caused by a cause of genes, a reduction of collagen and elastin present in the skin dermis, and an external environment.
  • the " wrinkle improvement” refers to a phenomenon that inhibits or inhibits the generation of wrinkles on the skin, or alleviates the already generated wrinkles.
  • the " skin elasticity” means elasticity realized by elastic fibers composed of elastin existing in the dermal layer. Since the elastic fibers have a low elastic modulus like rubber, they are easily deformed by external force, It can be easily restored to its original state.
  • the elastic fibers are embedded in microfibrils in an amorphous matrix called elastin and the elastin is a protein composed of a unique amino acid found only in elastic fibers such as desmosine and isodesmosine derived from lysine .
  • the desmosine and isodesmosin form cross-links within the long peptide chain, and the structure can confer rubber-like properties to elastin.
  • the " improvement of skin elasticity” means that elastic fibers composed of elastin are present together with collagen, and elastic elasticity is maintained or improved in a state where elastin and collagen are sufficiently present.
  • a cosmetic composition or a composition for external application for skin comprising the peptide or the peptide structure as an active ingredient.
  • the external preparation for skin or the cosmetic composition may be formulated by a conventional method.
  • Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA can refer to, and in the formulation of the cosmetic composition, International cosmetic ingredient dictionary, 6th ed., The cosmetic, Toiletry and Fragrance Association, Inc., Washington, 1995, which is incorporated herein by reference.
  • the external preparation for skin or the composition for cosmetics may contain a conventional acceptable carrier such as oil, water, a surfactant, a moisturizer, a lower alcohol, a thickener, a chelating agent, a pigment, a preservative and a flavoring agent. But is not limited thereto.
  • the composition may further include a base component that can be compounded with general external scalp and hair treatment agents.
  • the composition may contain a solubilizer, a surfactant, a moisturizer, a thickener, a pH adjuster, an antiseptic, an antioxidant, , A bactericide, an anti-inflammatory agent, an antimicrobial agent, a solvent, a coloring agent, or a flavoring agent.
  • the composition may be formulated as a composition for treating hair, such as an emulsion, a cream, a paste, a gel, a lotion, a pack, a lotion, a powder, a spray or a soap, , A hair mousse, a hair gel, a hair soap, a hair shampoo, a hair rinse, a hair pack, and a hair treatment, and may be specifically formulated into a liquid hair shampoo.
  • a composition for treating hair such as an emulsion, a cream, a paste, a gel, a lotion, a pack, a lotion, a powder, a spray or a soap, , A hair mousse, a hair gel, a hair soap, a hair shampoo, a hair rinse, a hair pack, and a hair treatment, and may be specifically formulated into a liquid hair shampoo.
  • components other than the above-mentioned essential ingredients can be appropriately selected and blended by those skilled in the art depending on the use or purpose of the other external preparation.
  • composition of the composition is a paste, cream or gel, an animal fiber, a plant fiber, a wax, a paraffin, a starch, a tracer, a cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, Can be used.
  • lactose When the formulation of the composition is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as the carrier component.
  • lactose talc
  • silica aluminum hydroxide
  • calcium silicate or polyamide powder may be used as the carrier component.
  • chlorofluorohydrocarbons propane, And may further include propellants such as dimethyl ether.
  • a solvent When the formulation of the composition is a solution or an emulsion, a solvent, a solvent or an emulsifier may be used as the carrier component.
  • the solvent include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan can be used.
  • the carrier component may be a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, and the like.
  • a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, and the like.
  • auxiliary agent and the mixing ratio thereof can be appropriately selected so as not to affect the desirable properties of the product according to the present invention.
  • a pharmaceutical composition for preventing or treating hair loss comprising the peptide or the peptide structure as an active ingredient.
  • the " prevention " means any action that reduces the frequency or extent of the pathological event. Prevention can be complete or partial. In this case, hair loss symptoms in the subject may be reduced as compared with the case where the composition is not used.
  • &quot means any clinical intervention to alter the natural course of a subject or cell to be treated, and may be performed during or during the course of a clinical pathology.
  • the desired therapeutic effect may be to prevent the occurrence or recurrence of the disease, to alleviate the symptoms, to reduce all direct or indirect pathological consequences of the disease, to prevent metastasis, to reduce the rate of disease progression, Relieving or temporarily alleviating the condition, or improving the prognosis.
  • compositions can be administered in the form of oral delivery, parenteral delivery.
  • the composition may be administered systemically or topically, and the administration may include oral administration and parenteral administration.
  • the composition may be formulated with a suitable amount of a pharmaceutically acceptable vehicle or carrier to provide a suitable dosage form.
  • the composition may further comprise a carrier, an excipient and a diluent used in the preparation of the pharmaceutical composition.
  • the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, But are not limited to, cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.
  • composition may be formulated in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or the like, external preparation, suppository and sterilized injection solution.
  • the solid preparation for oral administration may be a tablet, a pill, a powder, a granule, a capsule, etc.
  • the solid preparation may be prepared by adding to the compound and its fractions at least one or more excipients such as starch, calcium carbonate, sucrose, lactose , Or gelatin.
  • excipients such as starch, calcium carbonate, sucrose, lactose , Or gelatin.
  • lubricants such as magnesium stearate and talc may be used.
  • the liquid preparation for oral administration may be a suspension, a solution, an emulsion, a syrup, etc.
  • various excipients such as wetting agents, sweeteners, fragrances and preservatives may be used.
  • the preparation for parenteral administration may be a sterilized aqueous solution, a non-aqueous solvent, a suspension, an emulsion, a lyophilized preparation, or a suppository.
  • the non-aqueous solutions and suspensions may include injectable esters such as propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and ethyl oleate.
  • the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, and glycerogelatin.
  • the pharmaceutical composition may be administered to a subject in a pharmaceutically effective amount.
  • the " pharmaceutically effective amount " means an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to the medical treatment, and the effective dosage level will depend on the type of disease, severity, activity of the drug, Sensitivity, time of administration, route of administration and rate of excretion, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts.
  • the pharmaceutical composition may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiply. It is desirable to administer an amount that allows for all of the above factors to be maximally effected in a minimal amount without side effects, which can be readily determined by one skilled in the art.
  • N-Diisopropyl carbodiimde (DIC, 48 ⁇ l) as a carboxyl group activator and N-hydrobenzotriazole (HOBt, 40 mg) as an additive additive were added to 1.5 mL of anhydrous DMF organic solvent, ) was added and mixed and activated for about 20 minutes.
  • the activated solution and the resin were reacted for 2 hours with stirring, and then the reaction solution was filtered.
  • the resin was washed several times with DMF and methanol, and the kaiser test was performed.
  • the above-mentioned activated solution was made and participated. If the result of the test is negative, it is mixed with 3 mL of piperidine solution dissolved at a concentration of 20 wt% in DMF solvent and reacted for about 15 minutes to remove the Fmoc group at the N-terminus. Then, the amino acid of the following sequence is activated by the same method, ≪ / RTI >
  • the N-terminal Fmoc group was removed using 20 wt% piperidine / DMF.
  • the resin was washed several times with DMF and methanol and then dried.
  • the peptide bound to the resin was reacted with cleavage solution (trifluoroacetic acid (80%), triisopropylsilane (5%), thioanisole (5%), 1,2-ethanedithiol (5% )
  • cleavage solution trifluoroacetic acid (80%), triisopropylsilane (5%), thioanisole (5%), 1,2-ethanedithiol (5% )
  • the side chain protecting group of the amino acid was removed and the peptide was isolated from the resin.
  • Trifluoroacetic acid of the peptide solution was removed by passing through nitrogen gas, diethyl ether cooled to -20 ° C was added, and the precipitated peptide was centrifuged (3,000 rpm, 20 min ).
  • the obtained peptide was purified using reverse phase HPLC (C-18 column C18-120A, 50 mm * 250 mm) (Eluent: water / acetonitrile in 0.1% TFA (gradient): Flow Rate 50 mL / min). A substance of 1308 molecular weight was identified using an ESI-Mass mass spectrometer.
  • Example 1 Gly-Asn-Cys-Trp-Leu SEQ ID NO: 1
  • Example 2 Gly-Asn-Met-Trp-Leu SEQ ID NO: 2
  • Example 3 Gly-Asn-Ser-Trp-Leu SEQ ID NO: 3
  • Example 4 Asn-Thr-Leu-Phe-Lys SEQ ID NO: 4
  • Example 5 Gly-Asn-Cys-Trp-Leu-Asn-Thr-Leu-Phe-Lys SEQ ID NO: 5
  • Example 6 Gly-Asn-Met-Trp-Leu-Asn-Thr-Leu-Phe-Lys SEQ ID NO: 6
  • Example 7 Gly-Asn-Ser-Trp-Leu-Asn-Thr-Leu-Phe-Lys SEQ ID NO: 7
  • Example 8 Gly-Asn-Cys-Trp-Leu-Linker-Asn
  • Human epidermal keratinocytes HNK
  • human dermal fibroblasts nHDF
  • human dermal attenuate a human dermal papillae
  • human dermal papillae a human epidermal keratinocytes synthesized in the synthetic examples.
  • Cell growth potential of human dermal papilla was measured by MTT staining.
  • Human epidermal keratinocytes (Lonza, Switzerland) and human dermal fibroblasts, and human dermal papilla cells with DMEM (Dulbecco's modified Eagle's medium , gibco, USA) , each containing 10% FBS to the medium, 37C, 5% CO 2 conditions In an incubator.
  • DMEM Dulbecco's modified Eagle's medium , gibco, USA
  • the cultured cell lines were separated from the bottom of the culture dish with 1% trypsin solution and centrifuged to obtain cell precipitate only.
  • the cells were resuspended in DMEM containing no FBS and cultured in a 96-well tissue culture plate at 3 x 10 3 cells / well for 24 hours at 37 ° C and 5% CO 2 .
  • the medium was exchanged with the same culture medium except for the serum completely. Then, a blank sample and a synthetic peptide for standardization were dissolved in 10% DMSO in a sterilized state. Then, 1 ng / mL, 10 ng / mL, mL, 10 ⁇ g / mL and 100 ⁇ g / mL for 48 hours under the same conditions as above. Viability was measured using MTT after the culture was completed.
  • Example 1 116 155 215
  • Example 2 117 153 221
  • Example 3 118 160 218
  • Example 4 117 156 212
  • Example 5 120 154 224
  • Example 6 117 156 213
  • Example 7 118 155 216
  • Example 8 119 158 223 Control group 100 100 100
  • the peptide of Example 8 promoted the growth of keratinocytes, fibroblasts, and hair follicle cells in a concentration-dependent manner.
  • the parental spermoid was formed through three-dimensional stereoculture of dermal papilla cells (Sung, J. Invest. Dermatol. 2012, 132; 237) Thereby effectively promoting spironoid autogenesis.
  • the peptide not only has excellent stability to the skin or human body, but also promotes the growth of fibroblasts that synthesize collagen effectively, thus making it possible to utilize the peptide as a cosmetic, especially for improving skin wrinkles.
  • the cultured human dermal papilla cells were treated with the peptides of the example, and after 48 hours, the amount of phosphorylation of SMAD2, which is a signal of activin regulated by polystatin, and the expression level of hair growth factor were measured.
  • Example 1 division VCAN mRNA FGF7 mRNA WNT5a mRNA LEF1 mRNA
  • Example 2 2.0 2.5 1.4 2.7
  • Example 2 2.1 2.4 1.5 2.6
  • Example 3 2.0 2.5 1.4 2.8
  • Example 4 1.9 2.6 1.5 2.7
  • Example 5 2.1 2.7 1.3 3.0
  • Example 6 1.9 2.6 1.4 2.7
  • Example 7 2.2 2.5 1.5 2.9
  • Example 8 2.3 3.0 1.6 3.3
  • Example 8 when the peptide construct of Example 8 was treated with the expression of intracellular native polystin, the phosphorylation of SMAD2 was increased through activation of actibin. The expression of ⁇ -catenin, a major factor, was increased.
  • the peptides of the Examples were evaluated to increase the expression of versican, a papillary cell spe- land self-assembly activity factor.
  • the dermal papilla cells were plated in 96-well low-adherent tissue culture plates at 1 ⁇ 10 4 cells per well and cultured at 37 ° C. and 5% CO 2 for 48 hours.
  • a blank for capturing the standard and the peptide construct of Example 8 were added to the medium in sterilized condition in 10% DMSO.
  • dermal papilloma cell sphere was immobilized in 2% formaldehyde, PBS, and immunofluorescence staining was performed.
  • Example 8 The results showed that the self-assembly activity factor activated by the peptide structure of Example 8 promoted the self-assembly of the papillary spheredoids, and the peptides of the Examples increased the hair growth promoting activity of the papillary cells, May be implemented.
  • the hair growth promoting effect of the Examples and Comparative Examples was evaluated after removing the hair of the mouse (C57BL / 6) whose hair was in the resting period and about 50 days old.
  • mice with the characteristics of body weight, size, etc. were divided into 9 groups of 10 rats and individually cultured to measure the degree of hair growth.
  • Example 1 194.1 ⁇ 10.1
  • Example 2 195.7 ⁇ 11.2
  • Example 3 193.2 ⁇ 11.5
  • Example 4 188.3 ⁇ 10.6
  • Example 5 193.4 ⁇ 11.4
  • Example 6 196.5 ⁇ 12.1
  • Example 7 194.9 ⁇ 11.4
  • Example 8 212.1 ⁇ 12.2 Control group 69.5 ⁇ 4.5
  • the hair growth promoting effect by the peptides of the Examples is very excellent, so that hair health can be improved and hair loss can be suppressed.
  • the medium was replaced with DMEM (Dulbecco's Modified Eagle's Medium, Sigma), and the peptide was treated at a concentration of 50 mg / L and then cultured.
  • the control group was prepared by adding purified water without treating the above peptides.
  • the amount of collagen in each test sample culture fluid was calculated based on the amount of type I collagen in the control group (100%). The measurement results are shown in Table 5 below.
  • HaCaT cells cultured for 48 hours were treated with the peptides of Examples 1 and 2, and after 72 hours, the concentrations of laminin and hyaluronic acid, which are markers of skin wrinkle improvement, were measured. At this time, the concentration was measured using a Laminin ELISA kit and a Hyaluronic acid ELISA kit.
  • Example 8 153 Control group 100 Hyaluronic acid Example 1 145 Example 2 143 Example 3 147 Example 4 145 Example 5 148 Example 6 150 Example 7 151 Example 8 153 Control group 100
  • SEQ ID NO: 1 Physiologically active peptide 1 derived from polystatin
  • SEQ ID NO: 2 Physiologically active peptide 2 derived from polystatin
  • SEQ ID NO: 3 Physiologically active peptide 3 derived from polystatin
  • SEQ ID NO: 4 Physiologically active peptide derived from polystatin 4
  • SEQ ID NO: 5 Polystatin-derived physiologically active peptide 5
  • SEQ ID NO: 6 Polystatin-derived physiologically active peptide 6
  • SEQ ID NO: 7 Polystatin-derived physiologically active peptide 7

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Abstract

La présente invention porte sur un peptide bioactif dérivé du facteur de croissance et concerne un peptide bioactif composé d'au moins une séquence d'acides aminés choisie dans le groupe constitué par SEQ ID NO : 1 à 7.
PCT/KR2017/009296 2017-08-22 2017-08-25 Peptide bioactif dérivé de follistatine et son utilisation Ceased WO2019039630A1 (fr)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20040096592A (ko) * 2002-02-21 2004-11-16 와이어쓰 폴리스타틴 도메인을 포함하는 단백질
KR20090032148A (ko) * 2001-04-24 2009-03-31 더 존스 홉킨스 유니버시티 근육량을 증가시키기 위한 폴리스타틴의 용도
KR20150111347A (ko) * 2013-01-25 2015-10-05 샤이어 휴먼 지네틱 테라피즈 인크. 듀켄씨 근이영양증의 치료에서의 폴리스타틴
KR20170005891A (ko) * 2014-06-04 2017-01-16 악셀레론 파마 인코포레이티드 폴리스타틴 폴리펩티드를 이용한 장애의 치료방법 및 치료를 위한 조성물
KR20170085386A (ko) * 2016-01-14 2017-07-24 고려대학교 산학협력단 폴리스타틴에 특이적으로 결합하는 핵산 앱타머 및 그 용도

Family Cites Families (7)

* Cited by examiner, † Cited by third party
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US7507787B2 (en) * 2001-12-03 2009-03-24 The University Of British Columbia Effectors of innate immunity
EP2392659B1 (fr) * 2002-06-17 2015-01-14 Thrasos Innovation, Inc. Composés associés au TDF de domaine unique et analogues associés
EP2319526A1 (fr) * 2004-06-17 2011-05-11 Thrasos Therapeutics, Inc. Composés apparentés au TDF et leur analogues
CN101724632B (zh) * 2009-07-15 2012-11-07 吉林大学 牛蛙皮肤活性肽及其基因和在制药中的应用
WO2014200910A2 (fr) * 2013-06-10 2014-12-18 Iogenetics, Llc Procédés bio-informatiques de détermination de liaisons peptidiques
KR101632948B1 (ko) * 2014-05-13 2016-06-27 (주)케어젠 항염증, 골 형성 및 발모 촉진 활성을 갖는 펩타이드 및 이의 용도
US9597274B2 (en) * 2015-06-17 2017-03-21 Avon Products, Inc. Peptides and their use in the treatment of skin

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20090032148A (ko) * 2001-04-24 2009-03-31 더 존스 홉킨스 유니버시티 근육량을 증가시키기 위한 폴리스타틴의 용도
KR20040096592A (ko) * 2002-02-21 2004-11-16 와이어쓰 폴리스타틴 도메인을 포함하는 단백질
KR20150111347A (ko) * 2013-01-25 2015-10-05 샤이어 휴먼 지네틱 테라피즈 인크. 듀켄씨 근이영양증의 치료에서의 폴리스타틴
KR20170005891A (ko) * 2014-06-04 2017-01-16 악셀레론 파마 인코포레이티드 폴리스타틴 폴리펩티드를 이용한 장애의 치료방법 및 치료를 위한 조성물
KR20170085386A (ko) * 2016-01-14 2017-07-24 고려대학교 산학협력단 폴리스타틴에 특이적으로 결합하는 핵산 앱타머 및 그 용도

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