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US20170216385A1 - Longevity gene expression enhancer - Google Patents

Longevity gene expression enhancer Download PDF

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Publication number
US20170216385A1
US20170216385A1 US15/500,687 US201515500687A US2017216385A1 US 20170216385 A1 US20170216385 A1 US 20170216385A1 US 201515500687 A US201515500687 A US 201515500687A US 2017216385 A1 US2017216385 A1 US 2017216385A1
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Prior art keywords
sirtuin
extract
japonicum thunb
activating agent
peucedanum japonicum
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US15/500,687
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Inventor
Kaedeko FUKADA
Tatsuya Hasegawa
Hiroshi Mori
Yoshiharu Suzuki
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Shiseido Co Ltd
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Shiseido Co Ltd
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Assigned to SHISEIDO COMPANY, LTD. reassignment SHISEIDO COMPANY, LTD. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: FUKADA, Kaedeko, HASEGAWA, TATSUYA, MORI, HIROSHI, SUZUKI, YOSHIHARU
Publication of US20170216385A1 publication Critical patent/US20170216385A1/en
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/17—Preparation or pretreatment of starting material involving drying, e.g. sun-drying or wilting
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration

Definitions

  • the present invention provides a sirtuin-1 activating agent, and a formulation for suppressing cell senescence caused by oxidative stress, that contains the agent.
  • sirtuins genes for NAD + -dependent deacetylases known as sirtuins, as longevity genes.
  • the Sir2 gene was first identified in yeast, and in experimental systems using lower animals such as yeast and nematodes, it has been reported that lifespan is shortened by deletion of Sir2, while lifespan is lengthened when it is overexpressed (NPLs 1 and 2).
  • the Sir2 gene is conserved in mammals, with sirtuin-1 to sirtuin-7 having been identified. Much attention among these has been focused on sirtuin-1, as it is the one with a structure and function most resembling yeast Sir2.
  • sirtuin-1 The protein target of sirtuin-1 has been identified, and research suggests that it is involved in intracellular metabolism, energy consumption, and inflammation and stress response pathways.
  • activation of the sirtuin-1 gene is thought to have effects for arteriosclerosis, diabetes, heart disease, cancer, diabetes complications, neuropathic pain, microvascular dysfunction, life extension, mitochondrial disease, mitochondrial myopathy, neurodegenerative diseases (such as Alzheimer's disease, amyotrophic lateral sclerosis and Parkinson's disease), chronic obstructive pulmonary disease (COPD) and psoriasis (NPLs 3 and 4).
  • sirtuin activating substances there are known polyphenol compounds, which include resveratrol found in grape peels and the like or fisetin found in strawberries and apples, and non-polyphenol compounds such as SRT1720.
  • polyphenol compounds which include resveratrol found in grape peels and the like or fisetin found in strawberries and apples
  • non-polyphenol compounds such as SRT1720.
  • the major pharmaceutical company GlaxoSmithKline plc of England is vigorously conducting development of sirtuin-1 activating substances, and for example, SRT2104 which has been reported to be effective for prevention of type 2 diabetes, osteoporosis, sarcopenia and skeletal muscle atrophy (Reference 5) is currently in the clinical trial stage.
  • Peucedanum japonicum Thunb. plant body and its extract has action of inhibiting disaccharidase and efficacy for prevention and treatment of diabetes as well as countering obesity (PTL 1), has a cell-activating effect, antioxidant effect and melanin production inhibiting effect (PTL 2), exhibits inhibitory action against heparanase activity and is effective for preventing and improving wrinkles (PTL 3), and promotes ceramide synthesis (PTL 4).
  • Peucedanum japonicum Thunb. also known as “longevity grass”, is a plant of which it has been traditionally said in Okinawa Prefecture, that “eating one serving extends the life by a day”.
  • Sirtuin genes have already been known as longevity genes for over 15 years. However, absolutely no previous reports exist regarding whether the plant body or extract of Peucedanum japonicum Thunb. has a sirtuin-1 activating effect, and finding the connection between these has not been an easy task. Since traditional accounts regarding Peucedanum japonicum Thunb. and longevity do not constitute scientific evidence, it has not been easy to establish a connection between them.
  • a sirtuin-1 activating agent consisting of a plant body or solvent extract of Peucedanum japonicum Thunb.
  • sirtuin-1 activating agent according to (1) or (2), wherein the plant body of Peucedanum japonicum Thunb. is the leaves, stem or entire plant.
  • a formulation for suppressing cell senescence due to oxidative stress comprising a sirtuin-1 activating agent consisting of a plant body or solvent extract of Peucedanum japonicum Thunb.
  • sirtuin-1 activating agent for production of a formulation for suppressing cell senescence due to oxidative stress, wherein the sirtuin-1 activating agent comprises a plant body or solvent extract of Peucedanum japonicum Thunb.
  • a method for suppressing cell senescence due to oxidative stress wherein a sirtuin-1 activating agent consisting of a plant body or solvent extract of Peucedanum japonicum Thunb. is administered to a subject in need of suppression of cell senescence due to oxidative stress.
  • a method for prevention or treatment of a disease selected from among anti-arteriosclerosis, anti-diabetes, heart disease, anti-cancer, diabetes complications, neuropathic pain, microvascular dysfunction, life extension, mitochondrial disease, mitochondrial myopathy, neurodegenerative disease (such as Alzheimer's disease, amyotrophic lateral sclerosis or Parkinson's disease), chronic obstructive pulmonary disease, (COPD) and psoriasis, wherein a sirtuin-1 activating agent consisting of a plant body or solvent extract of Peucedanum japonicum Thunb. is administered to a subject in need of treatment for such a disease.
  • a sirtuin-1 activating agent consisting of a plant body or solvent extract of Peucedanum japonicum Thunb. is administered to a subject in need of treatment for such a disease.
  • a sirtuin-1 activating agent of the invention is able to activate sirtuin-1. According to the invention it is possible to provide an ingestion formulation containing a sirtuin-1 activating agent.
  • FIG. 1 shows evaluation of Western blotting of sirtuin-1 expression.
  • FIG. 2 shows the cell senescence-inhibiting effect of Peucedanum japonicum Thunb. against oxidative stress.
  • Administration of a sirtuin-1 activating agent of the invention is able to activate sirtuin-1.
  • Cell senescence due to oxidative stress can be suppressed by activation of the sirtuin-1 gene.
  • Cell senescence due to oxidative stress means damage to intracellular proteins, lipids, DNA and the like caused by peroxides and free radicals produced by oxidative stress including exposure to chemical agents produced by smoking, excessive eating or drinking or active oxygen generation, overfatigue, or exposure to ultraviolet rays or radiation, whereby an irreversible condition results in which cell replication is halted or proliferation becomes impossible, and it is distinguished from the phenomenon of senescence due to reduced cell proliferation activity as occurs with aging.
  • sirtuin-1 gene is known to be associated with cell senescence due to oxidative stress, and for example, it has been reported that resveratrol and statins can activate the sirtuin-1 gene and suppress cell senescence due to oxidative stress (NPLs 8 and 9). Activation of the sirtuin-1 gene is therefore useful for suppression of cell senescence due to oxidative stress.
  • Activation of the sirtuin-1 gene is thought to be useful for prevention or treatment of anti-arteriosclerosis, anti-diabetes, heart disease, anti-cancer, diabetes complications, neuropathic pain, microvascular dysfunction, life extension, mitochondrial disease, mitochondrial myopathy, neurodegenerative diseases (such as Alzheimer's disease, amyotrophic lateral sclerosis and Parkinson's disease), chronic obstructive pulmonary disease (COPD) and psoriasis.
  • anti-arteriosclerosis anti-diabetes
  • heart disease anti-cancer
  • diabetes complications neuropathic pain, microvascular dysfunction, life extension
  • mitochondrial disease mitochondrial myopathy
  • neurodegenerative diseases such as Alzheimer's disease, amyotrophic lateral sclerosis and Parkinson's disease
  • COPD chronic obstructive pulmonary disease
  • psoriasis psoriasis.
  • Sirtuin-1 gene activation means promoted expression of the sirtuin-1 gene, and for example, it means promoted sirtuin-1 gene expression when a sirtuin-1 activating agent has been administered, compared to a non-administered state (control), with a statistically significant difference where the significance level is 5% (by Student's t test, for example).
  • activation of the sirtuin-1 gene may mean that when a sirtuin-1 activating agent has been administered, the sirtuin-1 gene expression is promoted by, for example, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 100% or more, 200% or more, 300% or more, 400% or more or 500% or more, compared to a non-administered state (control).
  • the sirtuin-1 activating agent of the invention contains an orally ingestible extract of Peucedanum japonicum Thunb. as an active ingredient.
  • the Peucedanum japonicum Thunb. (alternate name: “longevity grass”) to be used for the invention is a plant of Umbelliferae Peucedanum L. which is distributed throughout Ishikawa Prefecture, west of the Kanto region (Honshu island), Shikoku, Kyushu, Okinawa and the Korean peninsula, China, Taiwan and the Philippines, and is a perennial herb that grows in well sunlit seashore areas (Newly Revised Makino's Color Illustrated Compendium of Oriental Drugs, p. 368 (2002)).
  • Peucedanum japonicum Thunb. may be used either in raw or dried form, but it is preferably used as a dry powder or extract from the viewpoint of ease of use and formulation.
  • the method for obtaining dry powder may be shredding and pulverizing of the entire plant or parts thereof (leaves, flowers, roots, etc.), followed by drying, or shredding or pulverizing of the dried plant to obtain a dry powder.
  • Another method that may be employed is shredding and pulverization of the plant, followed by fermentation or enzyme treatment, and then drying and pulverizing to the desired particle size, as necessary.
  • the extraction method for the extract to be used for the invention may be solvent extraction.
  • solvent extraction the entire plant or a portion thereof (leaves, flowers, roots, etc.) are dried, if necessary, further shredded and crushed if necessary, and then extracted using an aqueous extraction agent such as cold water, warm water or hot water at the boiling point or lower or a water-containing organic solvent, or an organic solvent such as methanol, ethanol, 1,3-butanediol or ether, at ordinary temperature or with heating.
  • an aqueous extraction agent such as cold water, warm water or hot water at the boiling point or lower or a water-containing organic solvent, or an organic solvent such as methanol, ethanol, 1,3-butanediol or ether, at ordinary temperature or with heating.
  • the extraction process is not limited to solvent extraction and may be carried out by any ordinary method known in the field.
  • the form of the extract does not have to be the extract itself, as it may be in a form obtained by appropriate dilution or concentration by an
  • the sirtuin-1 activating agent of the invention may be administered by ingestion or topical application, but it is preferably ingested.
  • the content of the Peucedanum japonicum Thunb. plant body or its solvent extract may be appropriately determined depending on the type of plant, the purpose, the form and the method of use. It is preferably prepared so that the amount of plant body or solvent extract consumed per day per adult is about 0.16 g to about 16 g (as dry mass). It is more preferably prepared to about 0.8 to about 8 g (as dry mass).
  • the active ingredient of the invention is preferably added in an amount allowing the sirtuin-1 activating effect to be satisfactorily exhibited.
  • sirtuin-1 activating agent of the invention When the sirtuin-1 activating agent of the invention is to be added to an ingestion formulation, it may be used in combination with desired additives selected as necessary. Excipients or the like may be added as additives.
  • Excipients may be any that are commonly used when preparing desired dosage forms, and examples include starches such as wheat starch, rice starch, corn starch, potato starch, dextrin, cyclodextrin and the like, crystalline celluloses, saccharides such as lactose, glucose, sugar, reduced maltose, rice jelly, fructooligosaccharides or emulsified oligosaccharides, and sugar alcohols such as sorbitol, erythritol, xylitol, lactitol and mannitol. Any of these excipients may be used alone or in combinations of two or more.
  • the ingestion form may be selected as an appropriate form, such as liquid, solid, granular, particulate, paste or gel form.
  • Peucedanum japonicum Thunb. dry powder was immersed in 70% ethanol and stirred overnight at room temperature. The filtrate was concentrated under reduced pressure and collected, and for the experiment it was dissolved in DMSO and evaluated.
  • Human umbilical vein endothelial cells (HUVEC) (LONZA, Md.) were seeded in a 6-well Collagen-Coated Microplate and cultured to subconfluence. They were then incubated for 48 hours in culture medium (EBM2) containing Peucedanum japonicum Thunb. extract (final concentration: 10 ⁇ g/mL) and resveratrol (5 ⁇ M), the protein was collected with Phospho Safe Extraction Reagent (EMD chemicals, CA), and the protein concentration was measured using a BCA kit (Pierce Biotechnology, Rockford, Ill.).
  • EBM2 culture medium
  • EMD chemicals Phospho Safe Extraction Reagent
  • the sample was prepared to an equivalent protein concentration, and evaluation was conducted by Western blotting using anti-sirtuin-1 (Santa Cruz Biotechnology, CA) and anti- ⁇ -actin (Sigma Aldrich, St. Louis, Mo.) as primary antibodies and anti-rabbit (GE Healthcare, Buckinghamshire, UK) and anti-mouse (GE Healthcare, Buckinghamshire and UK) as the respective secondary antibodies. Student's t-test was used as the statistical significance test.
  • Peucedanum japonicum Thunb. is known to contain chlorogenic acid, rutin and hesperidin, but it has been reported that chlorogenic acid has no sirtuin-1 activating effect (PLoS One, 2014, 9(2), e89166), while in the case of rutin, quercetin that is produced by conversion from rutin in the body has been reported to have a sirtuin-1 expression inhibiting effect (J. Pharmacol. Sci., 2008, 108, 364-71). On the other hand, hesperidin which is found in trace amounts in Peucedanum japonicum Thunb. plant body has been reported to have an effect of activating sirtuin-1 at 5 ⁇ M (Biosci. Biotechnol.
  • the cell senescence-inhibiting effect of Peucedanum japonicum Thunb. against oxidative stress induced by addition of H 2 O 2 was evaluated in terms of increased activity of the senescence marker SA ⁇ -Gal (Senescence-associated (3-galactosidase). Cells that have undergone cell senescence have increased SA ⁇ -Gal activity and are stained blue upon addition of substrate.
  • SA ⁇ -Gal Senescence-associated (3-galactosidase
  • HUVEC were cultured to subconfluence in culture medium (EBM2) (Lonza, Basal, Switzerland) in a 100 mm/Collagen-Coated Dish.
  • EBM2 culture medium
  • H 2 O 2 final concentrations: 0 and 100 ⁇ M
  • EBM2 culture medium
  • Peucedanum japonicum Thunb. extract final concentration: 10 ⁇ g/mL
  • Culturing was then carried out for 5 days, and a Cellular Senescence Assay Kit (Millipore, Mass.) was used for evaluation. Student's t-test was used as the statistical significance test.

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JP2014158163A JP5666053B1 (ja) 2014-08-01 2014-08-01 長寿遺伝子発現増強剤
JP2014-158163 2014-08-01
PCT/JP2015/071679 WO2016017767A1 (fr) 2014-08-01 2015-07-30 Activateur de l'expression d'un gène lié à la longévité

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JP6631877B2 (ja) * 2015-03-31 2020-01-15 株式会社東洋新薬 抗老化剤
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CN112755027A (zh) * 2021-01-26 2021-05-07 合肥工业大学 次乌头碱及其结构类似物作为Sirt3抑制剂和诱导体内外氧化应激的应用
JP7727312B2 (ja) * 2021-07-12 2025-08-21 丸善製薬株式会社 ＳＡ－β－ｇａｌ発現抑制剤及びオートファジー誘導剤

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KR20220113554A (ko) 2022-08-12
EP3175860A4 (fr) 2018-01-24
US20180344791A1 (en) 2018-12-06

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