JP2018108974A - Hair restorer - Google Patents

Hair restorer Download PDF

Info

Publication number
JP2018108974A
JP2018108974A JP2017000752A JP2017000752A JP2018108974A JP 2018108974 A JP2018108974 A JP 2018108974A JP 2017000752 A JP2017000752 A JP 2017000752A JP 2017000752 A JP2017000752 A JP 2017000752A JP 2018108974 A JP2018108974 A JP 2018108974A
Authority
JP
Japan
Prior art keywords
hair
fulvic acid
acid
humus
humus extract
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
JP2017000752A
Other languages
Japanese (ja)
Other versions
JP6233999B1 (en
Inventor
金川シーラ
Sheila KANAGAWA
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Style & Value Japan Inc
Original Assignee
Style & Value Japan Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Style & Value Japan Inc filed Critical Style & Value Japan Inc
Priority to JP2017000752A priority Critical patent/JP6233999B1/en
Application granted granted Critical
Publication of JP6233999B1 publication Critical patent/JP6233999B1/en
Priority to PCT/JP2017/046295 priority patent/WO2018128104A1/en
Publication of JP2018108974A publication Critical patent/JP2018108974A/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/02Medicinal preparations containing materials or reaction products thereof with undetermined constitution from inanimate materials
    • A61K35/10Peat; Amber; Turf; Humus
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/06Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/08Solutions
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/10Dispersions; Emulsions
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/10Dispersions; Emulsions
    • A61K9/107Emulsions ; Emulsion preconcentrates; Micelles
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/14Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/14Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/20Pills, tablets, discs, rods
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/48Preparations in capsules, e.g. of gelatin, of chocolate
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/70Web, sheet or filament bases ; Films; Fibres of the matrix type containing drug
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00Drugs for dermatological disorders
    • A61P17/14Drugs for dermatological disorders for baldness or alopecia
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q7/00Preparations for affecting hair growth

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Dispersion Chemistry (AREA)
  • Dermatology (AREA)
  • Mycology (AREA)
  • Botany (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Medical Informatics (AREA)
  • Microbiology (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Birds (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Nutrition Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Cosmetics (AREA)
  • Medicinal Preparation (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

PROBLEM TO BE SOLVED: To provide hair restorers containing humus extract and having excellent hair restoring effect.SOLUTION: The hair restorer of the present invention comprises an effective amount of humus extract containing fulvic acid having an infrared absorption peak at wavenumbers 3362, 2875, 1675, 1559, 1360, 1200, 1047 and 835 cmin the infrared absorption spectrum in the FT-IR method, and a cosmetically, dermatology and/or pharmaceutically acceptable additive or food additive.SELECTED DRAWING: Figure 5

Description

本発明は、育毛剤に関し、より詳細には腐植抽出物を含有する育毛剤に関する。   The present invention relates to a hair restorer, and more particularly to a hair restorer containing a humus extract.

毛髪の成長は、毛母細胞の増殖と分化によって起こり、そして毛乳頭細胞によって調節されていることが知られている(非特許文献1)。毛乳頭細胞の集合体である毛乳頭は、毛髪の成長期に大きく成長することから、毛乳頭細胞の増殖が、太くてしっかりした毛髪を形成する重要な役割を果たしていると考えられる。   It is known that hair growth occurs by proliferation and differentiation of hair matrix cells and is regulated by hair papilla cells (Non-patent Document 1). The dermal papilla, which is an aggregate of dermal papilla cells, grows greatly during the growth phase of the hair. Therefore, it is considered that the proliferation of dermal papilla cells plays an important role in forming thick and firm hair.

毛乳頭細胞の増加は、EGFやFGFといった細胞増殖因子によって促進される、男性ホルモンの一種であり、動物試験等で細胞増殖抑制物質として知られているジヒドロテストステロン(DHT)の産生抑制等が考えられる。したがって、育毛促進方法として、従来、EGF、FGF等の細胞増殖因子を活性化させる方法、DHT等の細胞増殖抑制物質を抑制させる方法等が検討されている。後述の実施例で陽性対照として使用した、抗アンドロゲン薬として知られるフィナステリド(Finasteride)は、EGFやFGFの産生促進というよりも、5αリダクターゼ阻害剤として作用が強いため、阻害作用の結果として、毛乳頭細胞に含まれるアンドロゲンレセプターへ関与し、細胞増殖を示したと考えられている。ミノキシジルもまた、フィナステリドと同様の作用機序と思われる。   The increase in dermal papilla cells is a type of male hormone that is promoted by cell growth factors such as EGF and FGF, and is considered to suppress the production of dihydrotestosterone (DHT), which is known as a cell growth inhibitor in animal studies. It is done. Therefore, conventionally, methods for activating cell growth factors such as EGF and FGF, methods for suppressing cell growth inhibitors such as DHT, and the like have been studied as methods for promoting hair growth. Finasteride, which is known as an antiandrogen used as a positive control in Examples described later, is more effective as a 5α-reductase inhibitor than EGF or FGF production promotion. It is thought to be involved in the androgen receptor contained in the papillary cells and showed cell proliferation. Minoxidil also appears to have a similar mechanism of action as finasteride.

特許文献1には、腐植土抽出物を含むことを特徴とする育毛剤が開示されている。特許文献1でによれば、人体に優しく、かつ、使用感、保湿効果に優れた育毛剤を提供可能であるとされる。特許文献2には、腐植土抽出物質含有水性液を用いて増毛剤・育毛剤を製造することが開示されている。特許文献1及び特許文献2の実施例で使用された腐植土抽出液は、長崎県北高来郡森山町唐比西名で採取された腐植土の抽出物である。この腐植土抽出物が、後述の比較例2で使用した市販の育毛剤に含まれている。   Patent Document 1 discloses a hair growth agent characterized by containing a humus soil extract. According to Patent Document 1, it is said that it is possible to provide a hair restorer that is gentle to the human body and that has an excellent feeling of use and a moisturizing effect. Patent Document 2 discloses that a hair thickener and a hair restorer are produced using an aqueous liquid containing a humus extract material. The humus soil extract used in the examples of Patent Document 1 and Patent Document 2 is an extract of humus soil collected from Kara Nishina, Moriyama-machi, Kitakaku-gun, Nagasaki Prefecture. This humus soil extract is contained in the commercially available hair restorer used in Comparative Example 2 described later.

勝岡憲生、「毛成長誘導の主役は毛乳頭細胞?毛包幹細胞?」、第74回日本皮膚科学会東京支部学術大会(1)会長講演、(http://medical.radionikkei.jp/maruho_hifuka_pdf/maruho_hifuka-110825.pdf)Norio Katsuoka, “Hair papillary cell or hair follicle stem cell is the leading role in hair growth induction?”, The 74th Annual Meeting of the Japanese Dermatological Association Tokyo Branch (1) President's Lecture, (http://medical.radionikkei.jp/maruho_hifuka_pdf/ maruho_hifuka-110825.pdf)

特開2006−327978(育毛剤)JP 2006-327978 (hair restorer) 特許第3507347(腐植土抽出物質含有水性液)Patent No. 3507347 (humus soil extractable aqueous solution)

腐植は、一般に、約1億年前の多くの栄養素が含まれた肥沃な土壌が石油や石炭に変性することなく、有機物として残存したものである。より詳細には、前記時代に生息した植物が、氷河期に入ると地下に埋没し、バクテリア等の微生物によってピートモスに変化し、その上に新しい有機物が積み重なって、再度、バクテリア等で分解され、時間の経過とともに堆積層を形成したものである。腐植は、土壌中で植物残渣や微生物遺体の中の炭水化物及びタンパク質が微生物によって分解され、さらにその分解産物が縮合してできたフルボ酸やフミン酸といった無定形有機高分子重合体や、多種類のミネラルを含有する。フミン酸は、アルカリに可溶であって酸に不溶な赤褐色ないし黒褐色無定形高分子有機酸であり、フルボ酸は、アルカリ及び酸に可溶な無定形高分子有機酸である。   In general, humus is a fertilized soil containing many nutrients approximately 100 million years old that remains as organic matter without being transformed into oil or coal. More specifically, when the plant that inhabited the period entered the ice age, it was buried underground, changed to peat moss by microorganisms such as bacteria, and new organic matter accumulated on top of it, and again decomposed by bacteria etc. A deposited layer is formed as the process proceeds. Humus is an amorphous organic polymer such as fulvic acid or humic acid, which is formed by the decomposition of carbohydrates and proteins in plant residues and microbial remains in the soil by microorganisms, and the decomposition products are condensed. Contains minerals. Humic acid is a red-brown or black-brown amorphous high-molecular organic acid that is soluble in alkali and insoluble in acid, and fulvic acid is an amorphous high-molecular organic acid that is soluble in alkali and acid.

フルボ酸やフミン酸のような無定形重合高分子有機酸は、腐植の産地の土壌やその生成年代期等の要因によって変化する。腐植の効能もまた、無定形重合高分子有機酸の構造や物性の相違によって変化すると考えられる。   Amorphous polymeric organic acids such as fulvic acid and humic acid vary depending on factors such as the soil of the humus production area and the age of its generation. The effect of humus is also considered to change due to the difference in the structure and physical properties of the amorphous polymer organic acid.

そこで、本発明の課題は、腐植抽出物質を含有し、育毛作用に優れた育毛剤を提供することである。   Then, the subject of this invention is providing the hair restoring agent which contains humus extract substance and was excellent in the hair growth effect | action.

本発明者は、上記課題を鋭意検討した結果、多種多様な分子構造と置換基を有するフルボ酸の中でも特定の赤外線吸収に基づく構造を有するフルボ酸には、従来、フィナステリドのように育毛作用を発揮し、さらに、従来公知の腐植抽出物含有育毛剤と対比しても優れた育毛作用に発揮することを発見し、本発明を完成させた。すなわち、本発明は、FT−IR法における赤外線吸収スペクトルにおいて波数3362,2875,1675,1559,1360,1200,1047及び835cm−1に赤外線吸収ピークを有するフルボ酸を含有する腐植抽出物を有効量含み、そして化粧品学的、皮膚病学的及び/又は薬学的に許容される添加剤あるいは食品添加物を含んでなる育毛剤を提供する。本明細書において、「育毛剤」は、育毛効果のほかに発毛効果を含む意味で使用される。 As a result of intensive studies on the above problems, the inventor of the present invention has hitherto been known to have a hair growth action like finasteride on a fulvic acid having a structure based on specific infrared absorption among fulvic acids having various molecular structures and substituents. Furthermore, the present invention was completed by discovering that it exhibited an excellent hair growth action even when compared with a conventionally known humus extract-containing hair restorer. That is, the present invention provides an effective amount of a humus extract containing fulvic acid having infrared absorption peaks at wave numbers 3362, 2875, 1675, 1559, 1360, 1200, 1047 and 835 cm −1 in the infrared absorption spectrum in the FT-IR method. And a hair restorer comprising cosmetic, dermatological and / or pharmaceutically acceptable additives or food additives. In the present specification, the “hair growth agent” is used in the meaning including a hair growth effect in addition to the hair growth effect.

前記腐植抽出物は、より特定的には、図1に示す赤外線吸収スペクトルを示すフルボ酸を含有する。   More specifically, the humus extract contains fulvic acid having an infrared absorption spectrum shown in FIG.

前記腐植抽出物は、ナフタレン環構造を有さないことが好ましい。   It is preferable that the said humus extract does not have a naphthalene ring structure.

前記腐植抽出物中のフルボ酸の平均分子量は、4,000よりも小さいことが好ましい。   The average molecular weight of the fulvic acid in the humus extract is preferably less than 4,000.

本発明の育毛剤は、例えば経皮投与用である。   The hair restorer of the present invention is for example for transdermal administration.

前記経皮投与用の育毛剤の形態は、例えば水剤、液剤、乳剤、乳液、エマルション、クリーム、粉体、又はクレイパックである。   The form of the hair growth agent for transdermal administration is, for example, a liquid agent, a liquid agent, an emulsion, an emulsion, an emulsion, a cream, a powder, or a clay pack.

本発明の育毛剤は、例えば経口投与用である。   The hair restorer of the present invention is for oral administration, for example.

前記経口投与用の育毛剤の形態は、例えば散剤、顆粒剤、カプセル剤、丸剤、錠剤、チュアブル錠、ドロップ、水剤、シロップ、又はドリンク剤である。   Examples of the hair-restoring agent for oral administration are powders, granules, capsules, pills, tablets, chewable tablets, drops, liquids, syrups, or drinks.

特定の赤外線吸収波長によって規定されるフルボ酸を含有する腐植抽出物を含有することを特徴とする本発明の育毛剤によれば、フィナステリドのように育毛作用が発揮され、しかも、その効能は腐植抽出物を含有する公知の腐植物質含有育毛剤よりも顕著に高い。   According to the hair restorer of the present invention, which contains a humus extract containing a fulvic acid defined by a specific infrared absorption wavelength, the hair restoring action is exerted like finasteride, and its effect is humus It is significantly higher than known humic substance-containing hair restorers containing extracts.

実施例1で使用した腐植抽出物(製品名:ヒューミクル(登録商標))のフルボ酸分画のATR法に基づくFT−IR吸収スペクトルの図である。It is a figure of the FT-IR absorption spectrum based on the ATR method of the fulvic acid fraction of the humus extract (product name: Humicle (trademark)) used in Example 1. FIG. 比較例3で使用した段戸産の標準フルボ酸及び標準フミン酸のATR法に基づくFT−IR吸収スペクトル図である。It is a FT-IR absorption spectrum figure based on the ATR method of the standard fulvic acid and the standard humic acid of Danto used in Comparative Example 3. 実施例1で使用した腐植抽出物の4%水溶液の3次元蛍光スペクトル図である。2 is a three-dimensional fluorescence spectrum diagram of a 4% aqueous solution of humus extract used in Example 1. FIG. 比較例3で使用した段戸産フルボ酸の水溶液(濃度10μg/mL)及び段戸産フミン酸の水溶液(濃度10μg/mL)の3次元蛍光スペクトル図である。It is a three-dimensional fluorescence spectrum figure of the aqueous solution (concentration 10 microgram / mL) of the terraced fulvic acid used in the comparative example 3, and the aqueous solution (concentration 10 microgram / mL) of the terraced humic acid. 実施例1、並びに比較例1〜4において、X軸に腐植抽出物の濃度(対数目盛)を、そしてY軸にヒト毛乳頭細胞の増殖率をプロットした図である。増殖率100%は、コントロール(試験品添加無し)を意味する。図5から、本発明の育毛剤(実施例1)は、人工的に合成されたフルボ酸製品(比較例1)、腐植抽出物を含む既存の育毛剤製品(比較例2)や腐植抽出物標準品(比較例3及び4)よりも高い毛乳頭細胞増殖率を示すことがわかる。In Example 1 and Comparative Examples 1-4, it is the figure which plotted the density | concentration (logarithmic scale) of the humus extract to the X-axis, and the proliferation rate of the human hair papilla cell to the Y-axis. A growth rate of 100% means a control (no test article added). From FIG. 5, the hair restorer of the present invention (Example 1) is an artificially synthesized fulvic acid product (Comparative Example 1), an existing hair restorer product (Comparative Example 2) or a humus extract containing a humus extract. It can be seen that the hair papilla cell proliferation rate is higher than that of the standard product (Comparative Examples 3 and 4).

以下、本発明の実施の形態を詳細に説明する。本発明の育毛剤は、FT−IR法における赤外線吸収スペクトルにおいて波数3362,2875,1675,1559,1360,1200,1047及び835cm−1に赤外線吸収ピークを有するフルボ酸を含有する腐植抽出物を有効量含む。 Hereinafter, embodiments of the present invention will be described in detail. The hair restorer of the present invention effectively uses a humus extract containing fulvic acid having infrared absorption peaks at wave numbers 3362, 2875, 1675, 1559, 1360, 1200, 1047 and 835 cm −1 in the infrared absorption spectrum in the FT-IR method. Including quantity.

本発明の育毛剤には、腐植抽出物の中でも、上記した特定の赤外線吸収ピークを有する腐植抽出物を使用することを必須とする。腐植抽出物は、上記の赤外線吸収特性を有するものであれば、その由来を問わない。   In the hair restorer of this invention, it is essential to use the humus extract which has the above-mentioned specific infrared absorption peak among humus extracts. A humus extract will not ask | require the origin, if it has said infrared absorption characteristic.

上記赤外線吸収ピークを有する腐植抽出物を含んだ腐植を水(例えば蒸留水、イオン交換水、逆浸透圧水等)と混合することにより、腐植抽出物が水内に抽出される。その際、腐植を、水100重量部に対して通常、5〜35重量部、好ましくは15〜25重量部、特に好ましくは、17〜18重量部の割合で混合する。上記混合物を、室温で、通常、24〜96時間、好ましくは48〜72時間日間放置する。上記放置中、適宜、混合物を撹拌してもよい。次いで、上記混合物の上澄液又は濾液を腐植抽出物とする。液状の腐植抽出物のpHは、通常、3.0である。この液状の腐植抽出物を乾固して、粉末としてもよい。   By mixing the humus containing the humus extract having the infrared absorption peak with water (for example, distilled water, ion exchange water, reverse osmotic pressure water, etc.), the humus extract is extracted into the water. In that case, humus is normally mixed in 5-35 weight part with respect to 100 weight part of water, Preferably it is 15-25 weight part, Most preferably, it is a ratio of 17-18 weight part. The mixture is left at room temperature, usually for 24 to 96 hours, preferably 48 to 72 hours. During the standing, the mixture may be appropriately stirred. Next, the supernatant or filtrate of the above mixture is used as a humus extract. The pH of the liquid humus extract is usually 3.0. This liquid humus extract may be dried to a powder.

上記腐植抽出物中のフルボ酸の平均分子量は、通常、4,000よりも小さく、好ましくは3,500以下であり、特に好ましくは3,000以下であり、さらに好ましくは3,000〜1,000である。   The average molecular weight of the fulvic acid in the humus extract is usually less than 4,000, preferably 3,500 or less, particularly preferably 3,000 or less, more preferably 3,000 to 1, 000.

前記腐植抽出物は、市販のものでもよく、日本では例えば株式会社スタイルアンドバリュージャパンより製品名「ヒューミクル(登録商標)HCフルボ酸パウダー」として入手可能である。ヒューミクルの成分分析結果(試料湿重量中の含有量)を、表1に示す。   The humus extract may be commercially available, and is available in Japan under the product name “Humicle (registered trademark) HC fulvic acid powder” from, for example, Style and Value Japan. Table 1 shows the component analysis results (content in the sample wet weight).

ヒューミクルの4%水溶液の全有機炭素(TOC)の測定、及びフルボ酸分画の全有機炭素(TOC)測定の結果から、ヒューミクル中の炭素のほぼ全量が有機炭素で占められ、そして有機炭素のほぼ全量がフルボ酸を構成していることが確認された。そして、ヒューミクル中のフルボ酸含有量は、約3%と推定された。   From the results of the total organic carbon (TOC) measurement of the 4% aqueous solution of Humicle and the total organic carbon (TOC) measurement of the fulvic acid fraction, almost all of the carbon in the Humicle is occupied by organic carbon, and the organic carbon It was confirmed that almost the entire amount constituted fulvic acid. The fulvic acid content in the fumicle was estimated to be about 3%.

ヒューミクルのアルカリ(0.1N水酸化ナトリウム)及び酸(塩酸、pH2)可溶成分を抽出することにより、フルボ酸を分画した。ヒューミクル又はフルボ酸分画の物性試験方法とその分析結果を以下に示す。   Fulvic acid was fractionated by extracting the fumaric alkali (0.1N sodium hydroxide) and acid (hydrochloric acid, pH 2) soluble components. The physical property test method and analysis results of the fumicle or fulvic acid fraction are shown below.

(1)3次元分光蛍光光度法
ヒューミクルの4%水溶液の3次元励起−蛍光スペクトル(測定装置JASCO FP−8200)を図3に示す。比較のため、愛知県段戸森林土壌から抽出精製されたフルボ酸及びフミン酸(いずれも、日本腐植物質学会から「日本腐植物質学会 標準資料」として頒布されている、以下、それぞれ、段戸産フルボ酸及び段戸産フミン酸という)の3次元蛍光スペクトルを図4に示す。ヒューミクル及び段戸産フルボ酸の3次元励起−蛍光スペクトルには、フルボ酸由来の蛍光の特徴である、励起250nm付近−蛍光430nm付近のピーク及び、励起320nm付近−蛍光430nm付近のピークが出現している。
(1) Three-dimensional spectrofluorimetry The three-dimensional excitation-fluorescence spectrum (measuring device JASCO FP-8200) of a 4% aqueous solution of Humicle is shown in FIG. For comparison, fulvic acid and humic acid extracted and refined from Danto forest soil in Aichi Prefecture (both are distributed as “Standard Material of the Japanese Humic Society” by the Japanese Humic Society, respectively) FIG. 4 shows three-dimensional fluorescence spectra of fulvic acid and corrugated humic acid. In the three-dimensional excitation-fluorescence spectrum of Humicle and Danto's fulvic acid, there are peaks of excitation around 250 nm-fluorescence near 430 nm and excitation around 320 nm-fluorescence near 430 nm, which are characteristic of fluorescence derived from fulvic acid. ing.

(2)GPC−UV/TC分析
GPC−UV/TC分析法は、水中溶存有機物を分子の大きさや排除の程度によって高速に分離し、分離した溶存分子をUV検出器及びTC検出器(Total Carbon Analyzer)でモニタリングする手法である。以下GPC−TC装置:
GPC:Shodex GPC SYSTEM 21
分離カラム TOSO HW−50S
溶離液 30mM NaHPO
UV吸収(220nm)検出器:GL7450(GL サイエンス製)
TC検出器:LCT−100(東レエンジニアリング製)
を用いて、ヒューミクル及び段戸産フルボ酸の分子量を測定した。
(2) GPC-UV / TC analysis The GPC-UV / TC analysis method separates dissolved organic substances in water at high speed according to the size of molecules and the degree of exclusion, and separates the separated dissolved molecules into a UV detector and a TC detector (Total Carbon). (Analyzer) is a monitoring method. The following GPC-TC device:
GPC: Shodex GPC SYSTEM 21
Separation column TOSO HW-50S
Eluent 30 mM Na 2 HPO 4
UV absorption (220 nm) detector: GL7450 (manufactured by GL Sciences)
TC detector: LCT-100 (manufactured by Toray Engineering)
Was used to measure the molecular weights of the fumicle and the fulvic acid from Dando.

両者のTCクロマトグラムでは、段戸産フルボ酸のピーク位置が51分であるのに対して、ヒューミクル4%溶解液では54分であった。ピーク位置の保持時間を、使用した分離カラムの多糖類やPEG(ポリエチレングリコール)の排除特性で換算すると、段戸フルボ酸の分子量は、約4,000であり、ヒューミクル溶解液中の有機物の分子量は、約1,000〜3,000と推定された。   In both TC chromatograms, the peak position of Danfu fulvic acid was 51 minutes, whereas it was 54 minutes for the 4% humicule solution. When the retention time at the peak position is converted into the exclusion characteristics of the polysaccharide and PEG (polyethylene glycol) of the separation column used, the molecular weight of the terraced fulvic acid is about 4,000, and the molecular weight of the organic matter in the Humicle solution Was estimated to be about 1,000 to 3,000.

ピークトップとなった保持時間でのTC出力値(mV)とUV(220nm)吸収での出力値(mV)、及びUV出力値/TC出力値(単位有機物量(炭素量)当たりUV吸収の強さ)を表2に示す。   TC output value (mV) at the retention time at the peak top, output value (mV) at UV (220 nm) absorption, and UV output value / TC output value (intensity of UV absorption per unit organic substance amount (carbon amount)) The results are shown in Table 2.

表2から、ヒューミクル溶解液のUV/TC値は、段戸産フルボ酸溶解液のものにほぼ近い。有機物の分子構造に由来する紫外吸光(220nm)に類似性があることは、両者の分子構造の骨格や官能基の種類に類似性があることを示している。   From Table 2, the UV / TC value of the Humicle solution is nearly similar to that of the Danto fulvic acid solution. The similarity in ultraviolet absorption (220 nm) derived from the molecular structure of organic matter indicates that there is similarity in the skeleton and functional group types of both molecular structures.

(3)フーリエ変換赤外分光光度計法(FT−IR)
前記フルボ酸分画のATR法(Attenuated Total Reflection、全反射吸収法)にて赤外線吸収スペクトルを取得した。具体的には、前記フルボ酸分画をDax−8樹脂(Spelco社製)に施用し、そこに吸着した有機物をアルカリ性溶液で脱着した。脱着した有機物を、さらにイオン交換樹脂に通して不要な塩類を取り除いた。得られた試料をATR法に基づくFT−IR分光分析(PerkinElmer Spectrum 65)に供した。
(3) Fourier transform infrared spectrophotometer method (FT-IR)
An infrared absorption spectrum was obtained by the ATR method (Attenuated Total Reflection, total reflection absorption method) of the fulvic acid fraction. Specifically, the fulvic acid fraction was applied to Dax-8 resin (manufactured by Spelco), and the organic matter adsorbed thereon was desorbed with an alkaline solution. The desorbed organic substance was further passed through an ion exchange resin to remove unnecessary salts. The obtained sample was subjected to FT-IR spectroscopic analysis (PerkinElmer Spectrum 65) based on the ATR method.

腐植抽出物(ヒューミクル)に含有されるフルボ酸の赤外線吸収スペクトルを図1に示す。比較のため、段戸産のフルボ酸及びフミン酸の赤外線吸収スペクトルを図2に示す。図1(ヒューミクル)及び図2(段戸産フルボ酸)から主要な赤外線吸収ピークをそれぞれ抽出し、その結果を波数で表3に示す。   FIG. 1 shows an infrared absorption spectrum of fulvic acid contained in the humus extract (humicicle). For comparison, FIG. 2 shows infrared absorption spectra of fulvic acid and humic acid produced in Danto. Major infrared absorption peaks were extracted from FIG. 1 (Humicle) and FIG. 2 (Stepbo fulvic acid), and the results are shown in Table 3 in terms of wave number.

ヒューミクルに含有されるフルボ酸及び段戸産フルボ酸は、共通して1200cm−1にカルボキシル基(C−O)吸収ピークを有するが、そのレベルは、ヒューミクルの方が段戸産フルボ酸よりも小さい。ヒューミクル中のフルボ酸が段戸産フルボ酸のスペクトルと明確に相違する点は、1559、1047及び835cm−1にトリ置換ベンゼンタイプのシグナルが観測され、そして3362及び1360cm−1にフェノールあるいはアルコールタイプのシグナルが観測されることである。 The fulvic acid contained in the fumicle and the fulvic acid produced from the terraced house have a carboxyl group (CO) absorption peak at 1200 cm −1 in common, but the level of the humicule is higher than that of the fulvic acid produced from the terraced house. small. That fulvic acid in Hyumikuru is clearly different spectrum of Danto producing fulvic acids are 1559,1047 and tri-substituted benzene type signals were observed at 835 cm -1, and phenol or an alcohol type 3362 and 1360 cm -1 Is observed.

赤外分光分析の結果は、ヒューミクル中のフルボ酸が段戸産フルボ酸よりもカルボキシル基が少なく、トリ置換ベンゼンタイプのフェノール化合物の構造を多いことを示唆する。発明を限定する意図ではないが、この分子構造の違いが、後述の実施例と比較例に示すように、毛乳頭細胞の増殖率の相違に反映したと考えられる。すなわち、本発明で特定する特定の赤外線吸収スペクトル特性を有する中のフルボ酸を含有する腐植抽出物が有効量添加された育毛剤は、腐植物質を含む従来の育毛剤よも優れた効能を有する。   The results of infrared spectroscopic analysis suggest that the fulvic acid in the humicle has fewer carboxyl groups than the dansted fulvic acid, and has more trisubstituted benzene type phenolic compounds. Although not intended to limit the invention, it is considered that this difference in molecular structure is reflected in a difference in the proliferation rate of hair papilla cells as shown in Examples and Comparative Examples described later. That is, the hair restorer to which an effective amount of a humus extract containing fulvic acid having the specific infrared absorption spectrum characteristic specified in the present invention is added has an effect superior to that of a conventional hair restorer containing humic substances. .

本発明の経皮用育毛剤には、上記の腐植抽出物以外に、化粧品学的、皮膚病学的及び/又は薬学的に許容される添加剤として汎用されるものが添加される。例えば、水;エチレングリコール、ポリエチレングリコール、プロピレングリコール、1,3−ブチレングリコール、1,4−ブチレングリコール、ジプロピレングリコール、グリセリン、ジグリセリン、ポリグリセリン、ペンチレングリコール、イソプレングリコール、グルコース、マルトース、フルクトース、キシリトール、ソルビトール、マルトトリオース、エリスリトール等の多価アルコール;メタノール、エタノール、プロピルアルコール、イソプロピルアルコール、ブチルアルコール、イソブチルアルコール等の低級アルコール;オレイン酸、イソステアリン酸、ラウリン酸、ミリスチン酸、パルミチン酸、ステアリン酸、ベヘン酸、ウンデシレン酸等の高級脂肪酸類;オリーブ油、トウモロコシ油、ツバキ油、マカデミアナッツ油、アボカド油、ナタネ油、ゴマ油、ヒマシ油、サフラワー油、綿実油、ホホバ油、ヤシ油、パーム油等の油脂;カルナバロウ、キャンデリラロウ、ミツロウ、ラノリン等のロウ類;ソルビトール、マンニトール、グルコース、ショ糖、ラクトース、トレハロース等の糖類;カラギーナン、キサンタンガム、ゼラチン、ペクチン、アガロース、アルギン酸塩、デキストリン、メチルセルロース、エチルセルロース、ヒドロキシプロピルセルロース、ヒドロキシエチルセルロース、カルボキシメチルセルロース、カルボキシビニルポリマー、ポリビニルアルコール、ポリビニルピロリドン、アラビアガム、カラヤガム、トラガントガム、タマリンドガム等の増粘剤;フェノキシエタノール、メチルパラベン、エチルパラベン、プロピルパラベン、ブチルパラベン、パラオキシ安息香酸エステル、安息香酸、サリチル酸とその塩類、ソルビン酸とその塩類、デヒドロ酢酸とその塩類、クロルクレゾール、ヘキサクロロフェン等の防腐剤;ラウロイル硫酸ナトリウム、モノオレイン酸ポリオキシエチレンソルビタン等の非イオン界面活性剤、アルキルサルフェート塩、ノルマルドデシルベンゼンスルホン酸ナトリウム等の陰イオン界面活性剤、ポリオキシエチレンドデシルモノメチルアンモニウムクロライド等の陽イオン界面活性剤;ステロイド系及び非ステロイド系抗炎症剤;ビタミンA、ビタミンD、ビタミンE、ビタミンF、ビタミンK等のビタミン類やジカプリル酸ピリドキシン、ジパルミチン酸ピリドキシン、ジパルミチン酸アスコルビル、モノパルミチン酸アスコルビル、モノステアリン酸アスコルビル等のビタミン誘導体;フラボノイド、カロテノイド等の抗酸化剤;スクワラン、スクワレン、流動パラフィン等の高級脂肪族炭化水素類;セラミド、セレブロシド、スフィンゴミエリン等のスフィンゴ脂質;コレステロール、フィトステロール等のステロール類;メチルポリシロキサン、メチルフェニルポリシロキサン、メチルシクロポリシロキサン、オクタメチルシクロテトラシロキサン、オクタメチルシクロペンタシロキサン、デカメチルシクロペンタシロキサン、メチルハイドロジェンポリシロキサン等のシリコーン類;パラアミノ安息香酸、パラアミノ安息香酸モノグリセリンエステル、アントラニル酸メチル、ホモメンチル−N−アセチルアントラニレート、パラメトキシケイ皮酸オクチル、エチル−4−イソプロピルシンナメート等の紫外線吸収剤;ベントナイト、スメクタイト、バイデライト、ノントロナイト、サポナイト、ヘクトライト、ソーコナイト、スチーブンサイト等の鉱物;ベンガラ、黄酸化鉄、黒酸化鉄、酸化コバルト、群青、紺青、酸化チタン、酸化亜鉛等の無機顔料;赤色202号、黄色4号、青色404号等の着色料;香料;香油等が挙げられる。   In addition to the above-mentioned humus extract, those commonly used as cosmetically, dermatologically and / or pharmaceutically acceptable additives are added to the hair growth agent for transdermal use of the present invention. For example, water; ethylene glycol, polyethylene glycol, propylene glycol, 1,3-butylene glycol, 1,4-butylene glycol, dipropylene glycol, glycerin, diglycerin, polyglycerin, pentylene glycol, isoprene glycol, glucose, maltose, Polyhydric alcohols such as fructose, xylitol, sorbitol, maltotriose, erythritol; lower alcohols such as methanol, ethanol, propyl alcohol, isopropyl alcohol, butyl alcohol, isobutyl alcohol; oleic acid, isostearic acid, lauric acid, myristic acid, palmitic acid Higher fatty acids such as acid, stearic acid, behenic acid, undecylenic acid; olive oil, corn oil, camellia oil, macadamia Oil, avocado oil, rapeseed oil, sesame oil, castor oil, safflower oil, cottonseed oil, jojoba oil, coconut oil, palm oil, etc .; waxes such as carnauba wax, candelilla wax, beeswax, lanolin; sorbitol, mannitol, glucose Sugars such as sucrose, lactose, trehalose; carrageenan, xanthan gum, gelatin, pectin, agarose, alginate, dextrin, methylcellulose, ethylcellulose, hydroxypropylcellulose, hydroxyethylcellulose, carboxymethylcellulose, carboxyvinyl polymer, polyvinyl alcohol, polyvinylpyrrolidone, Thickeners such as gum arabic, karaya gum, tragacanth gum, tamarind gum; phenoxyethanol, methyl paraben, ethyl paraben, propyl parabe , Butylparaben, paraoxybenzoic acid ester, benzoic acid, salicylic acid and its salts, sorbic acid and its salts, dehydroacetic acid and its salts, chlorcresol, hexachlorophene, etc .; preservatives such as sodium lauroyl sulfate, polyoxyethylene sorbitan monooleate Nonionic surfactants such as alkyl sulfate salts, anionic surfactants such as sodium normal dodecylbenzenesulfonate, and cationic surfactants such as polyoxyethylene dodecyl monomethylammonium chloride; steroidal and nonsteroidal anti-inflammatory agents Vitamins such as vitamin A, vitamin D, vitamin E, vitamin F, vitamin K, pyridoxine dicaprylate, pyridoxine dipalmitate, ascorbyl dipalmitate, ascorbyl monopalmitate, Vitamin derivatives such as ascorbyl nostearate; antioxidants such as flavonoids and carotenoids; higher aliphatic hydrocarbons such as squalane, squalene and liquid paraffin; sphingolipids such as ceramide, cerebroside and sphingomyelin; sterols such as cholesterol and phytosterols Silicones such as methylpolysiloxane, methylphenylpolysiloxane, methylcyclopolysiloxane, octamethylcyclotetrasiloxane, octamethylcyclopentasiloxane, decamethylcyclopentasiloxane, methylhydrogenpolysiloxane; paraaminobenzoic acid, paraaminobenzoic acid Acid monoglycerol ester, methyl anthranilate, homomenthyl-N-acetylanthranilate, octyl paramethoxycinnamate, ethyl -UV absorbers such as isopropyl cinnamate; minerals such as bentonite, smectite, beidellite, nontronite, saponite, hectorite, soconite, stevensite; bengara, yellow iron oxide, black iron oxide, cobalt oxide, ultramarine, bitumen, Inorganic pigments such as titanium oxide and zinc oxide; coloring agents such as red 202, yellow 4 and blue 404; fragrances; fragrance oils and the like.

本発明の経皮用育毛剤の形態の例は、水剤、液剤、乳剤、乳液、エマルション、クリーム、粉体、及びクレイパックである。皮膚への直接の効果を引き出すために、経皮用育毛剤は、水剤、液剤、乳剤、乳液、エマルション、又はクリームの形態が好ましい。   Examples of the form of the hair growth agent for transdermal use of the present invention are a liquid agent, a liquid agent, an emulsion, an emulsion, an emulsion, a cream, a powder, and a clay pack. In order to bring out a direct effect on the skin, the hair growth agent for transdermal use is preferably in the form of a solution, solution, emulsion, emulsion, emulsion or cream.

本発明の経皮用育毛剤は、シャンプー、リンス、リンスインシャンプー等の頭髪用入浴剤;ヘアミスト、ヘアスプレー、ヘアフォーム、ヘアリキッド等の頭髪用化粧品に含有させ、これらの入浴剤や化粧品にさらに育毛作用を付加してもよい。   The transdermal hair restorer of the present invention is contained in hair cosmetics such as shampoos, rinses and rinse-in shampoos; hair cosmetics such as hair mists, hair sprays, hair foams, hair liquids, and the like. A hair restoring action may be added.

本発明の経皮用育毛剤中の腐植抽出物(フルボ酸を約3%含有)の含有量は、有効量であればよく、通常、0.001〜20重量%でよく、好ましくは0.01〜20重量%であり、さらに好ましくは0.05〜15重量%であり、特に好ましくは0.1〜10重量%である。   The content of the humus extract (containing about 3% fulvic acid) in the hair growth agent for transdermal use of the present invention may be an effective amount, usually 0.001 to 20% by weight, preferably 0. It is 01-20 weight%, More preferably, it is 0.05-15 weight%, Especially preferably, it is 0.1-10 weight%.

本発明の経皮用育毛剤の1回当たりの適用量もまた、年齢、発毛状況、個人差等に応じて有効量であればよい。   The applied amount of the percutaneous hair restorer of the present invention may be an effective amount depending on age, hair growth status, individual differences and the like.

本発明の経口用育毛剤は、汎用の食品添加物を添加してもよい。例えば、経口投与の形態に応じて、汎用の賦形剤、崩壊剤、結合剤、滑沢剤、ビタミン、キサンチン誘導体、アミノ酸、pH調整剤、清涼化剤、懸濁化剤、粘稠剤、溶解補助剤、抗酸化剤、コーティング剤、可塑剤、界面活性剤、水、アルコール類、水溶性高分子、甘味料、矯味剤、酸味料、香料、着色剤等を発明の効果を損なわない質的及び量的範囲で添加することが可能である。   A general food additive may be added to the oral hair restorer of the present invention. For example, depending on the form of oral administration, general-purpose excipients, disintegrants, binders, lubricants, vitamins, xanthine derivatives, amino acids, pH adjusters, cooling agents, suspending agents, thickeners, Solubilizing agents, antioxidants, coating agents, plasticizers, surfactants, water, alcohols, water-soluble polymers, sweeteners, corrigents, acidulants, fragrances, coloring agents, etc. It is possible to add in the target and quantitative range.

本発明の経口用育毛剤の形態は、例えば、散剤、顆粒剤、カプセル剤、丸剤、錠剤、チュアブル錠、ドロップ、水剤、シロップ、及びドリンク剤のような通常の食品とは異なる経口投与剤に加工される。   The form of the oral hair restorer of the present invention is different from ordinary foods such as powders, granules, capsules, pills, tablets, chewable tablets, drops, liquids, syrups and drinks. It is processed into an agent.

本発明の経口用育毛剤中の腐植抽出物(フルボ酸を約3%含有)の含有量は、有効量であればよく、通常、0.1〜70重量%でよく、好ましくは0.1〜30重量%であり、より好ましくは0.1〜15重量%である。   The content of the humus extract (containing about 3% fulvic acid) in the oral hair restorer of the present invention may be an effective amount, usually 0.1 to 70% by weight, preferably 0.1. -30% by weight, more preferably 0.1-15% by weight.

本発明の経口用育毛剤の1日当たりの摂取量もまた、年齢、発毛状況、個人差等に応じて有効量であればよい。   The daily intake of the oral hair restorer of the present invention may also be an effective amount depending on age, hair growth status, individual differences and the like.

以下に、本発明の実施例及び比較例を示すことにより、本発明をより詳細に説明する。しかし、本発明は、以下の実施例に限定されるものではない。   Hereinafter, the present invention will be described in more detail by showing Examples and Comparative Examples of the present invention. However, the present invention is not limited to the following examples.

〔実施例1、比較例1〜4〕
本発明の育毛剤が育毛作用を有するか否かを、毛乳頭細胞の実験モデル系である毛乳頭細胞増殖効果試験(in vitro)で調べた。具体的には、毛乳頭細胞へ本発明の育毛剤を添加し、MTTアッセイによって毛乳頭細胞の細胞増殖促進率を測定し、さらに毛乳頭細胞増殖率を算出した。
[Example 1, Comparative Examples 1 to 4]
Whether or not the hair-restoring agent of the present invention has a hair-growing action was examined by a hair papilla cell proliferation effect test (in vitro), which is an experimental model system for hair papilla cells. Specifically, the hair growth agent of the present invention was added to the hair papilla cells, the cell growth promotion rate of the hair papilla cells was measured by MTT assay, and the hair papilla cell growth rate was calculated.

1.育毛剤試験品の準備
実施例1として、米国ユタ産の腐植の水性抽出物を乾固した腐植抽出物製品「ヒューミクル」(株式会社スタイルアンドバリュージャパン製)を用意した。ヒューミクルに含まれるフルボ酸分画の物性は、上記したとおりである。
1. Preparation of Hair Growth Agent Test Product As Example 1, a humus extract product “Humicle” (manufactured by Style and Value Japan Co., Ltd.) obtained by drying an aqueous extract of humus produced in Utah, USA was prepared. The physical properties of the fulvic acid fraction contained in the vehicle are as described above.

また、本発明の育毛剤の効果を評価するための陽性対照として、従来、育毛作用の知られているフィナステリドを用意した。また、コントロール(陰性対照)として、培地(試験品添加無し)を採用した。さらに、本発明を満たさない腐植物質を含む育毛剤と本発明の育毛剤とを対比するために、以下に示す腐植抽出物を含む製品、及び標準フルボ酸を用意した。
比較例1:果物や植物の発酵により得られたフルボ酸100%のM社製健康飲料
比較例2:長崎県北高来郡森山町唐比西産の腐植抽出物(フルボ酸)を含んだN社製清涼飲料水
比較例3:段戸産フルボ酸(腐植酸標準物質、日本腐植物質学会製)
比較例4:琵琶湖産フルボ酸(腐植酸標準物質、日本腐植物質学会製)
As a positive control for evaluating the effect of the hair restorer of the present invention, finasteride, which has been conventionally known for its hair restoring action, was prepared. As a control (negative control), a medium (no test product added) was employed. Furthermore, in order to compare the hair-restoring agent containing the humic substance which does not satisfy | fill this invention, and the hair-restoring agent of this invention, the product containing the humus extract shown below and the standard fulvic acid were prepared.
Comparative Example 1: 100% fulvic acid obtained by fermentation of fruits and plants, health drink manufactured by M Company Comparative Example 2: Company N containing humus extract (fulvic acid) from Karabi, Moriyama-cho, Kitakagi-gun, Nagasaki Prefecture Soft drink comparative example 3: Danto fulvic acid (humic acid reference material, manufactured by Japanese Society of Humic Substances)
Comparative example 4: Lake Biwa fulvic acid (humic acid reference material, manufactured by Japanese Humic Society)

2.ヒト頭髪毛乳頭細胞の基礎培養
試験用細胞には、ヒト頭髪毛乳頭細胞(タカラバイオ社製)を用いた。培養フラスコ(培養面積75cm)にヒト頭髪毛乳頭細胞培地及びヒト頭髪毛乳頭細胞懸濁液を培養液中の細胞の最終濃度が2.5×10cells/mL、全量10mLとなるように添加し、COインキュベーター(37℃、CO 5%)で、7日培養して必要細胞数を確保した。
2. Basic culture of human hair papilla cells Human hair papilla cells (manufactured by Takara Bio Inc.) were used as test cells. In a culture flask (culture area 75 cm 2 ), the human hair dermal papilla cell culture medium and the human hair dermal papilla cell suspension are adjusted to a final concentration of 2.5 × 10 4 cells / mL in the culture solution and a total volume of 10 mL. The cells were added and cultured in a CO 2 incubator (37 ° C., CO 2 5%) for 7 days to ensure the necessary number of cells.

3.試験品溶液の調製
実施例1及び比較例1〜4の育毛剤試験品を、それぞれ、試験品濃度が125、31.25、7.81、1.95、0.48、又は0.12μg/mlとなるように、順次、毛乳頭細胞培地で希釈した。陽性対照のFinasterideは、3ng/mLとなるように希釈した。
3. Preparation of test article solution The hair growth agent test articles of Example 1 and Comparative Examples 1 to 4, respectively, had a test article concentration of 125, 31.25, 7.81, 1.95, 0.48, or 0.12 μg / Diluted sequentially with hair papilla cell culture medium to a volume of ml. The positive control Finasteride was diluted to 3 ng / mL.

4.細胞増殖能評価試験
4.1 ヒト頭髪毛乳頭細胞の培養
培養したヒト頭髪毛乳頭細胞をTrypsin処理により培養フラスコから回収し、細胞数をカウントした。細胞を毛乳頭細胞培地で懸濁し、細胞濃度が1×10cells/mLの細胞懸濁液を調製した。調製した細胞懸濁液を試験プレート(96well micro plate)に0.2mLずつ、各wellに添加し、室温で20分静置した。COインキュベーター(37℃、CO 5%)にて18時間の前培養した。前培養後、試験プレート各ウェルの培地と調製した試験品溶液を置き換え、さらにCOインキュベーター(37℃、CO 5%)にて72時間培養した。72時間後、MTTアッセイによる生存率を確認した。※独立3回にて試験結果を算出した。
4). 4. Cell Proliferation Evaluation Test 4.1 Culture of human hair dermal papilla cells Cultured human hair dermal papilla cells were collected from the culture flask by trypsin treatment, and the number of cells was counted. Cells were suspended in a hair papilla cell medium to prepare a cell suspension having a cell concentration of 1 × 10 4 cells / mL. 0.2 mL of the prepared cell suspension was added to each well on a test plate (96 well micro plate), and allowed to stand at room temperature for 20 minutes. Pre-cultured for 18 hours in a CO 2 incubator (37 ° C., CO 2 5%). After the preculture, the medium in each well of the test plate was replaced with the prepared test solution, and further cultured in a CO 2 incubator (37 ° C., CO 2 5%) for 72 hours. After 72 hours, viability was confirmed by MTT assay. * Test results were calculated three times independently.

4.2 MTTアッセイ
培養終了後のプレートの培地を捨て、新たにMTT試薬含有培地(1mgMTT/mL培地)を0.1mLずつ各Wellに添加した。COインキュベーター(37℃、CO 5%)にて3時間培養し、呈色反応を行った。MTT可溶化溶液(イソプロパノール)を0.1mLずつ各wellに添加し、生細胞に取り込まれたMTTを抽出した。マイクロプレートリーダーを使用してMTT抽出液の吸光度を570nmで測定した。下記式に示すように、コントロール(試験品未処理細胞)の吸光度を100とした場合の百分率で細胞増殖率を算出した。
4.2 MTT assay The medium of the plate after completion of the culture was discarded, and 0.1 mL of MTT reagent-containing medium (1 mg MTT / mL medium) was newly added to each well. The color reaction was performed by culturing in a CO 2 incubator (37 ° C., CO 2 5%) for 3 hours. MTT solubilized solution (isopropanol) was added to each well in an amount of 0.1 mL to extract MTT taken up by living cells. The absorbance of the MTT extract was measured at 570 nm using a microplate reader. As shown in the following formula, the cell growth rate was calculated as a percentage when the absorbance of the control (untreated cells of the test product) was 100.

4.3 試験データ及び統計解析
得られたデータについて、分散分析を行い、等分散が確認された場合には統計解析として対応のないt検定を実施し、対照区との比較を実施した。試験結果を表4及び図5に示す。
4.3 Test data and statistical analysis An analysis of variance was performed on the obtained data. When equal variance was confirmed, an uncorresponding t-test was performed as a statistical analysis, and a comparison with a control group was performed. The test results are shown in Table 4 and FIG.

陽性対照である濃度(3ng/ml)のフィナステリドは、コントロール(細胞増殖率100%)に対して、106.8±0.6%の毛乳頭細胞増殖率を示し、毛乳頭細胞増加活性が確認された。   Finasteride at a concentration (3 ng / ml), which is a positive control, showed a hair papillary cell proliferation rate of 106.8 ± 0.6% relative to the control (cell growth rate 100%), confirming hair papillary cell increasing activity. It was done.

それに対して、本発明の育毛剤(実施例1)は、濃度0.48〜125μg/mlにおいて、107.4〜116.2%の細胞増殖率を示した。特に、実施例1の濃度1.95μg/mlでの毛乳頭細胞増殖率116.2%は、陽性対照に対して1.09倍(=116.2/106.8)上回る増殖活性を示した。   On the other hand, the hair restorer of the present invention (Example 1) showed a cell growth rate of 107.4 to 116.2% at a concentration of 0.48 to 125 μg / ml. In particular, the dermal papilla cell proliferation rate of 116.2% at a concentration of 1.95 μg / ml in Example 1 showed 1.09 times (= 116.2 / 106.8) greater proliferation activity than the positive control. .

次に、比較例3(段戸酸フルボ酸)及び比較例4(琵琶湖酸)は、日本腐植学会標準品のフルボ酸である。比較例3及び4を実施例1と対比すると、比較3及び4は、フルボ酸濃度0.48〜125μg/mlにおいて実施例1と似た傾向を示したが、その細胞増殖率は、99.8〜106.7%(比較例3)及び96.4〜102.7%(比較例4)と、実施例1よりも劣った。   Next, Comparative Example 3 (stepped acid fulvic acid) and Comparative Example 4 (Lake Lake acid) are fulvic acids that are standard products of the Japanese Humic Society. When Comparative Examples 3 and 4 were compared with Example 1, Comparatives 3 and 4 showed a tendency similar to Example 1 at a fulvic acid concentration of 0.48 to 125 μg / ml, but the cell growth rate was 99.99. 8-106.7% (Comparative Example 3) and 96.4-102.7% (Comparative Example 4), inferior to Example 1.

比較例1は、本発明に使用する腐植抽出物と違って、果物や植物を出発材料として人工的に合成したフルボ酸である。比較例2は、特許文献1及び2の実施例で使用された長崎県北高来郡森山町唐比西産の腐植抽出物と推定される。これらの比較例1及び2を、実施例1と対比すると、比較例1及び2の細胞増殖率は、それぞれ、86.1〜104.4%、及び94.2〜101.2%と実施例1よりも明らかに劣った。   Comparative Example 1 is a fulvic acid artificially synthesized using fruits and plants as starting materials, unlike the humus extract used in the present invention. Comparative Example 2 is presumed to be a humus extract produced in Karabi, Moriyama-cho, Moritaka-machi, Kitakagi-gun, Nagasaki Prefecture, which was used in Examples of Patent Documents 1 and 2. When these Comparative Examples 1 and 2 are compared with Example 1, the cell growth rates of Comparative Examples 1 and 2 are 86.1 to 104.4% and 94.2 to 101.2%, respectively. Obviously inferior to 1.

以上の結果から、特定の腐植抽出物を含む本発明の育毛剤は、毛乳頭細胞増殖活性が知られているフィナステリド、腐植抽出物標準品、及び腐植抽出物を含む既存の育毛剤製品よりも高い毛乳頭細胞増殖活性があることが判明した。したがって、本発明の育毛剤は、優れた育毛作用を有する育毛剤としての産業上の利用性が期待される。   From the above results, the hair restorer of the present invention containing a specific humus extract is more effective than finasteride, a humus extract standard product, and an existing hair restorer product containing a humus extract, which are known to have hair papilla cell proliferation activity. It was found to have high hair papilla cell proliferation activity. Accordingly, the hair growth agent of the present invention is expected to be industrially useful as a hair growth agent having an excellent hair growth action.

Claims (8)

FT−IR法における赤外線吸収スペクトルにおいて波数3362,2875,1675,1559,1360,1200,1047及び835cm−1に赤外線吸収ピークを有するフルボ酸を含有する腐植抽出物を有効量含み、そして化粧品学的、皮膚病学的及び/又は薬学的に許容される添加剤あるいは食品添加物を含んでなる育毛剤。 An effective amount of a humus extract containing fulvic acid having infrared absorption peaks at wave numbers 3362, 2875, 1675, 1559, 1360, 1200, 1047 and 835 cm −1 in the infrared absorption spectrum in the FT-IR method, and cosmetics A hair restorer comprising a dermatologically and / or pharmaceutically acceptable additive or food additive. 前記腐植抽出物は、図1に示す赤外線吸収スペクトルを示すフルボ酸を含有する、請求項1に記載の育毛剤。   The hair restoration agent according to claim 1, wherein the humus extract contains fulvic acid having an infrared absorption spectrum shown in FIG. 1. 前記腐植抽出物は、ナフタレン環構造を有さないことを特徴とする、請求項1又は2に記載の育毛剤。   The hair restoring agent according to claim 1 or 2, wherein the humus extract does not have a naphthalene ring structure. 前記腐植抽出物中のフルボ酸の平均分子量は、4,000よりも小さい、請求項1〜3のいずれか一項に記載の育毛剤。   The hair restoring agent according to any one of claims 1 to 3, wherein an average molecular weight of fulvic acid in the humus extract is less than 4,000. 経皮投与用である、請求項1〜4のいずれか一項に記載の育毛剤。   The hair restorer according to any one of claims 1 to 4, which is for transdermal administration. 水剤、液剤、乳剤、乳液、エマルション、クリーム、粉体、又はクレイパックの形態である、請求項5に記載の育毛剤。   The hair-restoring agent according to claim 5, which is in the form of a liquid preparation, liquid preparation, emulsion, emulsion, emulsion, cream, powder, or clay pack. 経口投与用である、請求項1〜4のいずれか一項に記載の育毛剤。   The hair restorer according to any one of claims 1 to 4, which is for oral administration. 散剤、顆粒剤、カプセル剤、丸剤、錠剤、チュアブル錠、ドロップ、水剤、シロップ、又はドリンク剤の形態である、請求項7に記載の育毛剤。   The hair restorer according to claim 7, which is in the form of a powder, granule, capsule, pill, tablet, chewable tablet, drop, solution, syrup, or drink.
JP2017000752A 2017-01-05 2017-01-05 Hair restorer Active JP6233999B1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP2017000752A JP6233999B1 (en) 2017-01-05 2017-01-05 Hair restorer
PCT/JP2017/046295 WO2018128104A1 (en) 2017-01-05 2017-12-25 Hair growth promoter

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP2017000752A JP6233999B1 (en) 2017-01-05 2017-01-05 Hair restorer

Publications (2)

Publication Number Publication Date
JP6233999B1 JP6233999B1 (en) 2017-11-22
JP2018108974A true JP2018108974A (en) 2018-07-12

Family

ID=60417525

Family Applications (1)

Application Number Title Priority Date Filing Date
JP2017000752A Active JP6233999B1 (en) 2017-01-05 2017-01-05 Hair restorer

Country Status (2)

Country Link
JP (1) JP6233999B1 (en)
WO (1) WO2018128104A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2019019108A (en) * 2017-07-20 2019-02-07 学校法人 中村産業学園 5α-REDUCTASE INHIBITOR, AND COMPOSITION FOR PREVENTIVE TREATMENT OF ALOPECIA OR HAIR GROWTH PROMOTION, AND METHOD FOR INHIBITING 5α-REDUCTASE IN SCALP AND HAIR GROWTH PROMOTION METHOD

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN112972313A (en) * 2021-03-08 2021-06-18 德利宝(广州)香料有限公司 Preparation method of modified humus extract and shampoo composition containing modified humus extract

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006327978A (en) * 2005-05-25 2006-12-07 Myubio Co Ltd Hair grower
WO2007102813A1 (en) * 2006-03-07 2007-09-13 Advantage Marketing, Inc. Compositions and methods for human use containing fulvic acid
JP2013107857A (en) * 2011-11-22 2013-06-06 Funao Kurosawa Skin external preparation

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006327978A (en) * 2005-05-25 2006-12-07 Myubio Co Ltd Hair grower
WO2007102813A1 (en) * 2006-03-07 2007-09-13 Advantage Marketing, Inc. Compositions and methods for human use containing fulvic acid
JP2013107857A (en) * 2011-11-22 2013-06-06 Funao Kurosawa Skin external preparation

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2019019108A (en) * 2017-07-20 2019-02-07 学校法人 中村産業学園 5α-REDUCTASE INHIBITOR, AND COMPOSITION FOR PREVENTIVE TREATMENT OF ALOPECIA OR HAIR GROWTH PROMOTION, AND METHOD FOR INHIBITING 5α-REDUCTASE IN SCALP AND HAIR GROWTH PROMOTION METHOD
JP2022008886A (en) * 2017-07-20 2022-01-14 学校法人 中村産業学園 5α-REDUCTASE INHIBITOR, AND COMPOSITION FOR PREVENTIVE TREATMENT OF ALOPECIA OR HAIR GROWTH PROMOTION, AND METHOD FOR INHIBITING 5α-REDUCTASE IN SCALP AND HAIR GROWTH PROMOTION METHOD
JP7046304B2 (en) 2017-07-20 2022-04-04 学校法人 中村産業学園 A method for producing a 5α-reductase inhibitor, and a method for producing a composition for preventing and treating alopecia or promoting hair growth.

Also Published As

Publication number Publication date
JP6233999B1 (en) 2017-11-22
WO2018128104A1 (en) 2018-07-12

Similar Documents

Publication Publication Date Title
TW201438756A (en) External dermal composition for anti-ageing and method for producing the same
JP4831849B2 (en) Active oxygen scavenger and its use
JP6233999B1 (en) Hair restorer
JP2008063266A (en) Anti-aging agent, bleaching agent, anti-oxidizing agent and anti-inflammatory agent
JP6120342B1 (en) Beauty composition
WO2023094517A1 (en) A topical composition
JP2009209063A (en) Skin preparation for external use, or skin cosmetic
JP4155430B2 (en) Skin aging inhibitor and use thereof
JP2010018545A (en) Active oxygen scavenger, and skin care preparation for external use, composition for oral cavity and food
JP5465037B2 (en) Anti-aging agent, antioxidant, whitening agent, immunostimulant, external preparation for skin and functional oral composition
JP2008303180A (en) Skin-lightening agent, skincare preparation for external use, and functional oral composition
JP2008247841A (en) Skin-lightening agent and melanin generation inhibitor
JP5025201B2 (en) Moisturizer, anti-aging agent, whitening agent, anti-inflammatory agent, and antioxidant
JP6739944B2 (en) External preparation for skin or cosmetics
JP2013224318A (en) Active oxygen scavenger, skin care preparation, composition for oral cavity and food product
JP2003113066A (en) Cosmetic
KR102572519B1 (en) A cosmetic composition for skin whitening containing loess extract
KR101028701B1 (en) Cosmetic compostions comprising Camellia japonica stamen with pollen and its extract for improving skin
JP7454203B2 (en) Skin external preparations or internal preparations
JPH08333270A (en) Cell activator
JP5610846B2 (en) Anti-aging agent, antioxidant, anti-inflammatory agent, whitening agent, moisturizer, external preparation for skin and functional oral composition
JP2006347934A (en) Glutathione production accelerator
EP2571483B1 (en) A personal care composition
KR20220088317A (en) Anti-aging composition comprising fermented propolis
WO2013149791A1 (en) A personal care composition

Legal Events

Date Code Title Description
A521 Request for written amendment filed

Free format text: JAPANESE INTERMEDIATE CODE: A523

Effective date: 20170825

TRDD Decision of grant or rejection written
A01 Written decision to grant a patent or to grant a registration (utility model)

Free format text: JAPANESE INTERMEDIATE CODE: A01

Effective date: 20171002

A61 First payment of annual fees (during grant procedure)

Free format text: JAPANESE INTERMEDIATE CODE: A61

Effective date: 20171023

R150 Certificate of patent or registration of utility model

Ref document number: 6233999

Country of ref document: JP

Free format text: JAPANESE INTERMEDIATE CODE: R150

R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250

R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250

R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250

R250 Receipt of annual fees

Free format text: JAPANESE INTERMEDIATE CODE: R250