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WO2009035169A1 - Peptide having anti-hypertensive activity - Google Patents

Peptide having anti-hypertensive activity Download PDF

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Publication number
WO2009035169A1
WO2009035169A1 PCT/JP2008/067052 JP2008067052W WO2009035169A1 WO 2009035169 A1 WO2009035169 A1 WO 2009035169A1 JP 2008067052 W JP2008067052 W JP 2008067052W WO 2009035169 A1 WO2009035169 A1 WO 2009035169A1
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WO
WIPO (PCT)
Prior art keywords
peptide
hyp
gly
seq
sequence number
Prior art date
Application number
PCT/JP2008/067052
Other languages
French (fr)
Japanese (ja)
Inventor
Toru Hayakawa
Ai Egusa
Tomomi Kouguchi
Koji Iwai
Yoshihisa Takahata
Takashi Ohmori
Original Assignee
Nippon Meat Packers, Inc.
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Application filed by Nippon Meat Packers, Inc. filed Critical Nippon Meat Packers, Inc.
Publication of WO2009035169A1 publication Critical patent/WO2009035169A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0821—Tripeptides with the first amino acid being heterocyclic, e.g. His, Pro, Trp
    • C07K5/0823—Tripeptides with the first amino acid being heterocyclic, e.g. His, Pro, Trp and Pro-amino acid; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17—Amino acids, peptides or proteins
    • A23L33/175—Amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/12—Antihypertensives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/14—Vasoprotectives; Antihaemorrhoidals; Drugs for varicose therapy; Capillary stabilisers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06026—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atom, i.e. Gly or Ala
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/06034—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms
    • C07K5/06043—Leu-amino acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06017—Dipeptides with the first amino acid being neutral and aliphatic
    • C07K5/0606—Dipeptides with the first amino acid being neutral and aliphatic the side chain containing heteroatoms not provided for by C07K5/06086 - C07K5/06139, e.g. Ser, Met, Cys, Thr
    • C07K5/06069—Ser-amino acid
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06008—Dipeptides with the first amino acid being neutral
    • C07K5/06078—Dipeptides with the first amino acid being neutral and aromatic or cycloaliphatic
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06104—Dipeptides with the first amino acid being acidic
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/06—Dipeptides
    • C07K5/06139—Dipeptides with the first amino acid being heterocyclic
    • C07K5/06165—Dipeptides with the first amino acid being heterocyclic and Pro-amino acid; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
    • C07K5/0806—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing 0 or 1 carbon atoms, i.e. Gly, Ala
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
    • C07K5/0808—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
    • C07K5/081—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing O or S as heteroatoms, e.g. Cys, Ser
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0819—Tripeptides with the first amino acid being acidic
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C07K5/1002—Tetrapeptides with the first amino acid being neutral
    • C07K5/1005—Tetrapeptides with the first amino acid being neutral and aliphatic
    • C07K5/101—Tetrapeptides with the first amino acid being neutral and aliphatic the side chain containing 2 to 4 carbon atoms, e.g. Val, Ile, Leu
    • 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
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to a peptide having an antihypertensive effect. More specifically, the present invention relates to a peptide comprising a specific amino acid sequence, having an antihypertensive action, a blood vessel preservation action and the like, and a functional food or drug containing the peptide.
  • Angiotensin Converting Enzyme also referred to herein as “ACE” converts angiotensin I (inactive form) to angiotensin II (active form), which has a strong vasoconstrictor action. It breaks down bradykinin, which has vasodilator action, into three inactive peptides.
  • ACE inhibitors substances that inhibit the enzyme activity of angiotensin converting enzyme
  • angiotensin converting enzyme such as captopril and enalapril that inhibit ACE activity (the enzyme activity of angiotensin converting enzyme) have been conventionally used as antihypertensive drugs. .
  • ACE inhibitors obtained from foods or food ingredients can be expected as low-toxic and highly safe antihypertensives and health-oriented foods, and can be ingested in daily eating habits. So far, ACE inhibitors have been reported among many natural products and enzymatic degradation products such as food. In recent years, food-derived ingredients such as sea bream, bonito, and seaweed have been found to have an effect of suppressing an increase in blood pressure, and some are commercially available as foods suitable for people with high blood pressure (for example, 2-3 1 1 4 9 4 gazette, JP 2 0 0 0-4 7 9 9 gazette). There are very few ACE inhibitors derived from meat and meat products.
  • ACE inhibitors derived from meat and meat products are not well known, and even after degradation in the digestive tract, substances (peptides) that have a sufficiently high blood pressure rise inhibitory effect (peptides) It is hardly found. Because of these problems, the present inventors have studied ACE inhibitory substances derived from meat and meat products. As a result, protease degradation products of chicken and pork collagen and peptides contained in the degradation products are found. It was also found that it has an excellent antihypertensive effect even in vivo.
  • the low molecular fraction of the decomposed product has strong ACE inhibitory activity.
  • SHR spontaneously hypertensive rats
  • blood pressure decreased after 2 hours, and the increase in blood pressure was significantly suppressed even in a 4-week long-term administration study. It became clear.
  • the fraction was further fractionated by HPLC to obtain a peptide having a high ACE inhibitory activity, and its amino acid sequence was determined with a protein sequencer. As a result, a peptide having a specific amino acid sequence was obtained.
  • the peptide has a strong ACE inhibitory activity and a blood vessel preservation action. All of the peptides described in the present application are peptides that are not degraded by, for example, the digestive enzyme trypsin chymotrypsin, etc., and are not degraded by digestive enzymes in the digestion solution when ingested orally and retain their activity. It is a peptide.
  • the present invention is based on such findings, and provides a peptide having an antihypertensive action and / or a blood vessel preservation action, and a functional food or drug containing the peptide. Disclosure of the invention
  • Such a peptide has a blood pressure increase inhibitory action and / or a blood vessel preservation action.
  • the present invention is a functional food or drug having an antihypertensive action and / or a blood vessel preservation action, comprising at least one of the peptides of the present invention.
  • SEQ ID NO: 2, 3, 7 to 11, 1, 14, 16 and 18 to 20, more preferably containing at least one peptide consisting of the amino acid sequence represented by SEQ ID NOs: 2, 14, 16, and 19 Is preferred.
  • the present invention comprises the above-described configuration, and the peptide of the present invention comprises a peptide comprising the amino acid sequence represented by SEQ ID NOs: 1 to 23 shown below and an amino acid sequence comprising Hyp-Gly, Ala-Hyp or Pro-Hyp. It is a 2 to 6-mer peptide.
  • Sequence number 22 Ser-Hyp SEQ ID NO: 23: Ile-Hyp
  • Hyp 4-hydroxyl-L-proline residue.
  • the peptide of the present invention can be chemically synthesized according to a conventional peptide synthesis method.
  • collagen (or gelatin) derived from chicken or pig is subjected to enzymatic degradation using a protease according to a conventional method to obtain a collagen degradation peptide, and further subjected to enzymatic degradation as necessary.
  • Purify and isolate the resulting collagen-degrading peptide by conventional purification means such as ultrafiltration, reverse osmosis filtration, gel filtration, ion exchange ram chromatography, reversed-phase high-speed liquid chromatography, etc. Can be obtained.
  • the type of collagen and the collection site are not particularly limited, and various types of collagen can be used.
  • type I collagen derived from chicken and pig legs, skin, bones, tendons, intestines, etc. is used because of its abundant raw materials.
  • the collagen can be prepared according to a conventional method.
  • Gelatin may be used in place of collagen.
  • the protease is not particularly limited as long as it is a protease capable of enzymatically degrading collagen (gelatin), and any of acidic protease, neutral protease, and alkaline protease can be used.
  • an animal-derived protease For example, trypsin, chymotrypsin, pepsin, etc.
  • plant-derived proteases for example, papain, promeline, huisin, etc.
  • microorganism-derived protease, etc. trypsin is preferably used from the viewpoint of enzyme treatment efficiency.
  • the dipeptide in the peptides represented by SEQ ID NOs: 1 to 23 is a peptide consisting of the amino acid sequence represented by SEQ ID NOs: 4 to 7, 9 to 10, or 16 to 18, according to a conventional method, It can also be obtained by enzymatic degradation using carboxypeptidase or aminopeptidase under the optimal conditions.
  • carboxypeptidase examples include carboxypeptidase derived from a kidney or a microorganism, and examples thereof include carboxypeptidase A, B, P, and Y.
  • aminopeptidase examples include a kidney, a kidney derived from a microorganism, an aminopeptidase derived from a microorganism, and the like, and examples thereof include aminopeptidase.
  • the enzyme is deactivated by means such as heating, and then the conventional purification means ⁇ "For example, ultrafiltration, reverse osmosis filtration, gel filtration, ionic exchange ram chromatography, reverse phase high performance liquid chromatography, etc.
  • the target peptide can be obtained by purifying and isolating it.
  • the functional food of the present invention is a functional food having an antihypertensive action and / or a blood vessel preservation action, comprising at least one of the peptides of the present invention as an active ingredient.
  • functional food is a concept that includes ordinary food, beverages, confectionery, feed, and the like.
  • These functional foods are useful for the treatment and prevention of active ingredients as they are, added with various nutrients, or contained in foods and drinks for the purpose of suppressing blood pressure rise and / or maintaining blood vessels. It is eaten as (or food material).
  • an appropriate auxiliary agent for example, glucose, lactose, sucrose, starch, mannitol, dextrin, polyethylene glycol, hydroxyxetyl starch, ethylene glycol, amino acid, etc.
  • It may be formed into a form suitable for edible use, such as granules, granules, tablets, capsules, pastes, etc., and used for food.
  • Seafood processed foods such as chikuwa, confectionery such as snacks, dairy products such as breads, butter and powdered milk, soy products such as tofu and fried milk, etc.), water, fruit juice, milk, refreshing It may be used by adding to beverages such as beverages and dessert foods such as pudding and jelly. Further, it may be in the form of animal feed (including pet food).
  • the intake of the peptide of the present invention in the form of such functional food is appropriately selected and determined according to age, body weight, symptom, degree of disease, form of food, etc.For example, as the amount of peptide per day, 1. 0 mg to 10. O mg, preferably 2.0 mg to 8.0 mg, but since it has the advantage of not adversely affecting the body even if consumed in large amounts, it is more than that. You can take the amount of.
  • the peptide of the present invention is used as a drug having an antihypertensive action and / or a blood vessel preservation action, and in a method for treating hypertension comprising administering an effective amount of the peptide.
  • the peptide is used as an active ingredient, and if necessary, it is mixed with necessary ingredients in the preparation such as appropriate physiologically acceptable additives (for example, carriers, excipients, diluents, etc.) It can be obtained by preparing in an appropriate dosage form, and examples of the form include tablets, powders, granules, capsules and the like.
  • the dose can be appropriately determined according to the patient's symptoms, age, weight, etc., referring to the above intake.
  • the peptide of the present invention has an antihypertensive action and / or a blood vessel preservation action, and the effect is the method described in the following examples and the already known vascular endothelial function evaluation system.
  • the ACE inhibitory activity of the peptide consisting of the amino acid sequence represented by SEQ ID NOs: 1 to 23 was measured by the following method.
  • the peptides consisting of the amino acid sequences shown in SEQ ID NOs: 1 to 23 were prepared by an automatic peptide synthesizer (PSSM8 Shimadzu) by the Fmoc solid phase method. After deprotection, confirm the peptide structure using reverse-phase HPLC (PEGASIL-300, 20 X 250 mm; Senshu, Tokyo, Japan) and mass spectrometer ESI mass spectrometer LC-Q (Thermo Finnigan, San jose, CA). I went. Peptide synthesis reagents were purchased from Shimadzu (Kyoto, Japan).
  • the inhibitory activity against ACE was measured according to the method of Cheung et al. (Cheung et al. J. Biol. Chem. 1980, 255, 401-407).
  • lOOraM folic acid solution pH 8.3
  • 5mM Hip-HL 500mM NaCl
  • 20mU Usagi Lung ACE 20mU Usagi Lung ACE
  • test sample was added at 37 ° C for 30 minutes. Inn I did a cuvate.
  • the enzyme reaction was stopped by adding 1NHC1.
  • -Hip-HL was measured for the amount of hippuric acid released at an absorbance of 228 nm.
  • the inhibitor concentration at which the inhibition of ACE activity 50% was defined as IC 50.
  • Table 1 shows the results of the ACE inhibitory activity (IC 5 ) of the peptides consisting of the amino acid sequences represented by SEQ ID NOs: 1 to 23. As shown in Table 1, all the peptides consisting of the amino acid sequences represented by SEQ ID NOs: 1 to 23 had ACE inhibitory activity. In particular, peptides consisting of the amino acid sequences represented by SEQ ID NOs: 2, 3, 7 to: 11, 14, and 18 to 20 had extremely strong ACE inhibitory activity.
  • a 2 to 6-mer preferably 2 to 5-mer peptide having an amino acid sequence consisting of Hyp-Gly, Ala-Hyp or PnrHyp is effective as an ACE inhibitor. It is done.
  • Sequence number 18 Ser-Hyp-Gly 0. 305
  • L-NAME N-nitro-L-arginine methyl ester
  • a NO synthase inhibitor N-nitro-L-arginine methyl ester
  • drinking water lmg / ml in water, 8 weeks old
  • a peptide 5 g / kg / day
  • water containing NAME was used for drinking water throughout the breeding period.
  • the thoracic aorta was collected, immediately Krebs-Henseleit solution (118. 4mM NaCl, 4. 7 mM KCl, 2. 5mM CaCl 2, 1. 2mM KH 2 P0 4, 1. 2mM MgS0 4 - 7H 2 0, 25 mM NaHC0 3 , 11.6 mM C 6 H 12 0 6 .PH 7.4, 4 ° C), cut into a ring shape of about 3-4 mm, and installed in an organ yanbar 'microbus .
  • Krebs-Henseleit solution 118. 4mM NaCl, 4. 7 mM KCl, 2. 5mM CaCl 2, 1. 2mM KH 2 P0 4, 1. 2mM MgS0 4 - 7H 2 0, 25 mM NaHC0 3 , 11.6 mM C 6 H 12 0 6 .PH 7.4, 4 ° C
  • the organ chamber's microbus was filled with Kerbs-Henseleit solution (same as above, 37 ° C) and bubbled with a mixture of 95% 0 2 and 5% C0 2 so as not to impair the function of the blood vessel specimen. .
  • the blood vessel sample was set to be loaded with lg and left for about 60 minutes to stabilize. Thereafter, phenylene Refurin the After (final concentration 1 X 10- 7 M) was added to cause the blood vessel specimens are contraction, measuring the extension reaction upon addition of acetylcholine (final concentration 1 X 10- 6 M) Total
  • the degree of dilation was evaluated as a blood vessel preservation action, and the effect was shown in four stages (1: No effect 2: Slightly effective 3: Effective) 4: Significant effect).
  • Natural fruit juice (concentrated juice reduction) is mixed with 10 mg of the peptide of the present invention (peptide consisting of the amino acid sequence represented by SEQ ID NO: 2) per 20 O ml of the juice, and then sterilized according to a conventional method. Asse boutique packaging to obtain a juice product.
  • the sausage casing After mixing 10 mg of the peptide of the present invention (peptide consisting of the amino acid sequence shown in SEQ ID NO: 14) per 15 g of the paste, the sausage casing is filled according to a conventional method. Smoked, sterilized, packaged after cooling to obtain wiener sausage.
  • Peptide of the present invention (peptide consisting of the amino acid sequence represented by SEQ ID NO: 19) 6 According to a conventional method, mg was filled into a hard capsule to produce a capsule. Industrial applicability
  • the peptide of the present invention has strong ACE inhibitory activity, vascular endothelial damage recovery, function maintenance, and the like, and has a function of suppressing blood pressure elevation and / or blood vessel preservation. It can be used as a sex food or medicine. Since it is a low molecular weight compound, it has excellent absorbability and can maintain its effect even after being absorbed by the living body.
  • the functional food or drug of the present invention contains a peptide having the above-described action, and it is important to prevent and improve high blood pressure, which is a lifestyle-related disease, through daily life.
  • high blood pressure which is a lifestyle-related disease
  • the functional food or drug of the present invention contains a peptide having the above-described action, and it is important to prevent and improve high blood pressure, which is a lifestyle-related disease, through daily life.

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Abstract

Disclosed is a peptide having an inhibitory activity on an ACE (an angiotensin-converting enzyme) (i.e., an anti-hypertensive activity)/a blood vessel-protecting activity. Also disclosed is a functional food or a medicinal agent comprising the peptide. The peptide comprises an amino acid sequence depicted in any one of SEQ ID NOs: 1 to 23. The peptide has an anti-hypertensive activity and/or a blood vessel-protecting activity. The functional food or the medicinal agent comprises the peptide as an active ingredient, and can be ingested on a daily basis to meliorate hypertension or exhibit its blood vessel-protecting effect including the recovery of a disorder in an endothelium and the maintenance of the functions of an endothelium.

Description

明細書 血圧上昇抑制作用を有するぺプチド 技術分野  Peptide having blood pressure elevation inhibiting effect Technical Field
本発明は血圧上昇抑制作用を有するペプチドに関する。 より詳細には、 特定のァ ミノ酸配列からなり、 血圧上昇抑制作用、 血管保全作用などを有するペプチド及び 当該ペプチドを含有する機能性食品又は薬剤に関する。 背景技術  The present invention relates to a peptide having an antihypertensive effect. More specifically, the present invention relates to a peptide comprising a specific amino acid sequence, having an antihypertensive action, a blood vessel preservation action and the like, and a functional food or drug containing the peptide. Background art
高血圧患者は、 アメリカ · ョ一口ッパ · 日本で約 2億人いると言われ、 危険域の ヒ トを含めると 10億人に達すると推測されている。 高血圧発症のメカニズムは種々 存在するが、 レニン ·アンジォテンシン系と力リクレイン · キニン系が大きく関係 するといわれている。アンジォテンシン変換酵素（Angiotensin Converting Enzyme、 本明細書では 「ACE」 ともいう） は、 アンジォテンシン I (inactive form)を強い血 管収縮作用をもつアンジォテンシン II (active form)に変換すると共に、血管拡張作 用のあるブラジキニンを不活性な 3種のペプチドに分解する。 そこで、 ACE活性 （ァ ンジォテンシン変換酵素の酵素活性） を阻害するカプトプリルゃェナラプリルなど の ACE阻害物質 （アンジォテンシン変換酵素の酵素活性を阻害する物質） が高血圧 治療薬として、 従来から利用されている。  There are approximately 200 million people with hypertension in the United States, Japan, and Japan, and it is estimated that the number of hypertensive patients will reach 1 billion including those in danger zones. There are various mechanisms for the development of hypertension, but it is said that the renin-angiotensin system and the force lyrein / kinin system are closely related. Angiotensin Converting Enzyme (also referred to herein as “ACE”) converts angiotensin I (inactive form) to angiotensin II (active form), which has a strong vasoconstrictor action. It breaks down bradykinin, which has vasodilator action, into three inactive peptides. Therefore, ACE inhibitors (substances that inhibit the enzyme activity of angiotensin converting enzyme) such as captopril and enalapril that inhibit ACE activity (the enzyme activity of angiotensin converting enzyme) have been conventionally used as antihypertensive drugs. .
食品又は食品原料から得られる ACE阻害物質は低毒性で安全性の高い降圧剤、 健 康志向食品として期待でき、 日常の食生活において摂取可能となる。 これまで多く の天然物や食品等の酵素分解物の中から ACE阻害物質が報告されている。 近年、 ィ ヮシ、 カツォ、 ワカメなど食品由来成分に血圧の上昇を抑制する作用が見出されて おり、 一部は血圧が高めの人に適した食品として市販されている （例えば、 特開平 2 - 3 1 1 4 9 4公報、 特開 2 0 0 0— 4 7 9 9公報）。 し力、し、 食肉 ·食肉製品に 由来する ACE阻害物質は非常に少ない。  ACE inhibitors obtained from foods or food ingredients can be expected as low-toxic and highly safe antihypertensives and health-oriented foods, and can be ingested in daily eating habits. So far, ACE inhibitors have been reported among many natural products and enzymatic degradation products such as food. In recent years, food-derived ingredients such as sea bream, bonito, and seaweed have been found to have an effect of suppressing an increase in blood pressure, and some are commercially available as foods suitable for people with high blood pressure (for example, 2-3 1 1 4 9 4 gazette, JP 2 0 0 0-4 7 9 9 gazette). There are very few ACE inhibitors derived from meat and meat products.
また、 食品由来の ACE阻害物質の中には、 消化器官で分解されたりして活性が低 下するものもあり、 その有効性が疑問視されるものもある。 そのため、 通常の食事 における摂取、 つまり消化器官で分解などがされた後でも、 十分に高い血圧上昇抑 制作用を有する物質 （ペプチド） が望まれるが、 係る物質はほとんど見出されてい なかった。 In addition, some food-derived ACE inhibitors are degraded in the digestive tract and their activity decreases, and some of them are questionable for their effectiveness. For this reason, even after ingestion in a normal diet, that is, after decomposition in the digestive tract, a sufficiently high suppression of blood pressure increase is achieved. Substances (peptides) for production purposes are desired, but few such substances have been found.
上述のように、食肉 ·食肉製品に由来する ACE阻害物質はあまり知られておらず、 更に消化器官で分解などがされた後でも、 十分に高い血圧上昇抑制作用を有する物 質 （ペプチド） はほとんど見出されていない。 このような問題点から、 本願発明者 らは、 食肉 ·食肉製品に由来する ACE阻害物質を研究したところ、 鶏及び豚由来コ ラーゲンのプロテアーゼ分解物及びこの分解物に含有されているぺプチドが、 in vivoにおいても優れた血圧上昇抑制作用を有することを見出した。  As mentioned above, ACE inhibitors derived from meat and meat products are not well known, and even after degradation in the digestive tract, substances (peptides) that have a sufficiently high blood pressure rise inhibitory effect (peptides) It is hardly found. Because of these problems, the present inventors have studied ACE inhibitory substances derived from meat and meat products. As a result, protease degradation products of chicken and pork collagen and peptides contained in the degradation products are found. It was also found that it has an excellent antihypertensive effect even in vivo.
より具体的には、 鶏足又は豚皮等から抽出したコラーゲンを各種プロテアーゼで 分解し、 当該分解物の ACE阻害活性を測定したところ、 当該分解物の低分子画分は 強い ACE阻害活' 14を有し、 更に高血圧自然発症ラット（Spontaneously Hypertensive Rat, SHR)に投与したところ、 2時間後から血圧の低下が認められ、 4週間の長期投 与試験においても、 有意に血圧の上昇を抑制することが明らかとなった。 次いで、 当該画分をさらに HPLCで分画し、 高い ACE阻害活性を有する画分のぺプチドを得、 それについてプロテインシークェンサ一でアミノ酸配列を決定したところ、 特定の ァミノ酸配列を有するぺプチドが強い ACE阻害活性及び血管保全作用を有すること を見出した。 なお、 本願記載のペプチドは、 いずれも、 例えば、 消化酵素であるト リプシンゃキモトリプシン等では分解されないぺプチドであり、 経口摂取時に、 消 化液中の消化酵素では分解されず、 その活性を保持するぺプチドである。  More specifically, when collagen extracted from chicken legs or pig skin is decomposed with various proteases and the ACE inhibitory activity of the decomposed product is measured, the low molecular fraction of the decomposed product has strong ACE inhibitory activity. In addition, when administered to spontaneously hypertensive rats (SHR), blood pressure decreased after 2 hours, and the increase in blood pressure was significantly suppressed even in a 4-week long-term administration study. It became clear. Subsequently, the fraction was further fractionated by HPLC to obtain a peptide having a high ACE inhibitory activity, and its amino acid sequence was determined with a protein sequencer. As a result, a peptide having a specific amino acid sequence was obtained. It was found that the peptide has a strong ACE inhibitory activity and a blood vessel preservation action. All of the peptides described in the present application are peptides that are not degraded by, for example, the digestive enzyme trypsin chymotrypsin, etc., and are not degraded by digestive enzymes in the digestion solution when ingested orally and retain their activity. It is a peptide.
本発明は係る知見に基づくもので、血圧上昇抑制作用及び/又は血管保全作用を有 するぺプチド並びにそれを含有する機能性食品又は薬剤を提供するものである。 発明の開示  The present invention is based on such findings, and provides a peptide having an antihypertensive action and / or a blood vessel preservation action, and a functional food or drug containing the peptide. Disclosure of the invention
上記の課題を解決するためになされた本発明は、 （1)配列番号 1〜2 3で示される ァミノ酸配列からなるぺプチド； （2) Hyp-Gly, Ala-Hyp又は Pro-Hypからなるァ ミノ酸配列を有する 2〜6量体のペプチドである （以下、 上記 (1)及び (2)のペプチド を合わせて、 本発明のペプチドという）。 係るペプチドは血圧上昇抑制作用及び/又 は血管保全作用を有する。  The present invention has been made to solve the above problems. (1) A peptide comprising an amino acid sequence represented by SEQ ID NOs: 1 to 23; (2) consisting of Hyp-Gly, Ala-Hyp or Pro-Hyp It is a 2- to 6-mer peptide having an amino acid sequence (hereinafter, the peptides (1) and (2) above are collectively referred to as the peptide of the present invention). Such a peptide has a blood pressure increase inhibitory action and / or a blood vessel preservation action.
更に、 本発明は、 本発明のペプチドの少なくとも一種を含有する、 血圧上昇抑制 作用及び/又は血管保全作用を有する機能性食品又は薬剤である。特に、配列番号 2、 3、 7〜 1 1、 1 4、 1 6及び 1 8〜2 0、 より好ましくは配列番号 2、 1 4、 1 6及び 1 9で示されるアミノ酸配列からなるぺプチドの少なくとも一種を含有する ことが好ましい。 発明を実施するための最良の形態 Furthermore, the present invention is a functional food or drug having an antihypertensive action and / or a blood vessel preservation action, comprising at least one of the peptides of the present invention. In particular, SEQ ID NO: 2, 3, 7 to 11, 1, 14, 16 and 18 to 20, more preferably containing at least one peptide consisting of the amino acid sequence represented by SEQ ID NOs: 2, 14, 16, and 19 Is preferred. BEST MODE FOR CARRYING OUT THE INVENTION
本発明は上記の構成からなり、 本発明のペプチドは、 下記に示される配列番号 1 〜 2 3で示されるアミノ酸配列からなるペプチド及び Hyp-Gly、 Ala-Hyp 又は Pro-Hypからなるアミノ酸配列を有する 2〜 6量体のぺプチドである。  The present invention comprises the above-described configuration, and the peptide of the present invention comprises a peptide comprising the amino acid sequence represented by SEQ ID NOs: 1 to 23 shown below and an amino acid sequence comprising Hyp-Gly, Ala-Hyp or Pro-Hyp. It is a 2 to 6-mer peptide.
配列番号 1 ： Gly-Ala-Hyp-Gly-Leu-Hyp Sequence number 1: Gly-Ala-Hyp-Gly-Leu-Hyp
配列番号 2 ： Gly-Leu-Hyp-Gly-Pro Sequence number 2: Gly-Leu-Hyp-Gly-Pro
配列番号 3 ： Leu-Hy -GlyPro Sequence number 3: Leu-Hy-GlyPro
配列番号 4 ： Gly-Ala-Hyp Sequence number 4: Gly-Ala-Hyp
配列番号 5 ： Ala-Hyp-Gly Sequence number 5: Ala-Hyp-Gly
配列番号 6 ： Gly-LeirHyp Sequence number 6: Gly-LeirHyp
配列番号 7 ： Hyp-Gly-Pro Sequence number 7: Hyp-Gly-Pro
配列番号 8 ： Gly-Pro Sequence number 8: Gly-Pro
配列番号 9 ： Leu-Hyp-Gly Sequence number 9: Leu-Hyp-Gly
配列番号 10 ： Hyp-Gly-Leu Sequence number 10: Hyp-Gly-Leu
配列番号 11 ： Hyp-Gly Sequence number 11: Hyp-Gly
配列番号 12 ： Gly-Leu Sequence number 12: Gly-Leu
配列番号 13 ： GlyAla Sequence number 13: GlyAla
配列番号 14 ： Ala-Hyp Sequence number 14: Ala-Hyp
配列番号 15 ： Leu-Hyp Sequence number 15: Leu-Hyp
配列番号 16 ： Pro-Hyp-Gly Sequence number 16: Pro-Hyp-Gly
配列番号 17 ： Glu-Hyp-Gly Sequence number 17: Glu-Hyp-Gly
配列番号 18 ： Ser Hyp-Gly Sequence number 18: Ser Hyp-Gly
配列番号 19 ： Pro -Hyp Sequence number 19: Pro-Hyp
配列番号 20 ： Phe-Hyp Sequence number 20: Phe-Hyp
配列番号 21 ： Glu-Hyp Sequence number 21: Glu-Hyp
配列番号 22 ： Ser-Hyp 配列番号 23： Ile-Hyp Sequence number 22: Ser-Hyp SEQ ID NO: 23: Ile-Hyp
なお、 式中、 Hypは 4—ヒ ドロキシ一 L一プロリン残基を意味する。  In the formula, Hyp means 4-hydroxyl-L-proline residue.
本発明のぺプチドは、 慣用のぺプチド合成法に準じて化学的に合成することがで きる。 また、 鶏又は豚由来コラーゲン （又はゼラチン） を、 常法に準じてプロテア ーゼを用いた酵素分解に付してコラーゲン分解べプチドを得、 更に必要に応じて酵 素分解に付した後、 得られたコラーゲン分解ペプチドを慣用の精製手段、 例えば、 限外濾過、 逆浸透濾過、 ゲル濾過、 ィォン交換力ラムクロマトグラフィー、 逆相高 速液体クロマトグラフィーなどに付して精製 ·単離することにより得ることができ る。  The peptide of the present invention can be chemically synthesized according to a conventional peptide synthesis method. In addition, collagen (or gelatin) derived from chicken or pig is subjected to enzymatic degradation using a protease according to a conventional method to obtain a collagen degradation peptide, and further subjected to enzymatic degradation as necessary. Purify and isolate the resulting collagen-degrading peptide by conventional purification means such as ultrafiltration, reverse osmosis filtration, gel filtration, ion exchange ram chromatography, reversed-phase high-speed liquid chromatography, etc. Can be obtained.
上記で使用されるコラーゲンに関し、 コラーゲンのタイプ及び採取部位などは特 に限定されず、 種々のコラーゲンを使用することができる。 好適には、 原料が豊富 であることから、 鶏及び豚の足、 皮、 骨、 腱、 腸などに由来する I型コラーゲンが 使用される。 当該コラーゲンは常法に準じて調製することができる。 コラーゲンに 代えてゼラチンを使用してもよい。  Regarding the collagen used above, the type of collagen and the collection site are not particularly limited, and various types of collagen can be used. Preferably, type I collagen derived from chicken and pig legs, skin, bones, tendons, intestines, etc. is used because of its abundant raw materials. The collagen can be prepared according to a conventional method. Gelatin may be used in place of collagen.
また、 上記のプロテアーゼとしては、 コラーゲン （ゼラチン） を酵素分解できる プロテアーゼであれば特に限定はされず、 酸性プロテアーゼ、 中性プロテアーゼ、 アルカリ性プロテアーゼのいずれも使用することができ、 例えば、 動物由来プロテ ァーゼ （例えば、 トリプシン、 キモトリブシン、 ペプシン等）、 植物由来プロテア一 ゼ （例えば、 パパイン、 プロメリン、 フイシン等）、 微生物由来のプロテアーゼなど が挙げられ、 酵素処理効率の点からトリプシンが好適に使用される。  The protease is not particularly limited as long as it is a protease capable of enzymatically degrading collagen (gelatin), and any of acidic protease, neutral protease, and alkaline protease can be used. For example, an animal-derived protease (For example, trypsin, chymotrypsin, pepsin, etc.), plant-derived proteases (for example, papain, promeline, huisin, etc.), microorganism-derived protease, etc., trypsin is preferably used from the viewpoint of enzyme treatment efficiency.
また、 配列番号 1〜2 3で示されるペプチドの中のジペプチドは、 配列番号 4〜 7、 9〜1 0又は 1 6〜1 8で示されるアミノ酸配列からなるペプチドを、 常法に 準じ、 酵素の至適条件下で、 カルボキシぺプチダーゼ又はアミノぺプチダーゼを作 用させて酵素分解することにより得ることもできる。  In addition, the dipeptide in the peptides represented by SEQ ID NOs: 1 to 23 is a peptide consisting of the amino acid sequence represented by SEQ ID NOs: 4 to 7, 9 to 10, or 16 to 18, according to a conventional method, It can also be obtained by enzymatic degradation using carboxypeptidase or aminopeptidase under the optimal conditions.
カルボキシぺプチダーゼとしては、 睥臓、 微生物などに由来するカルボキシぺプ チダーゼが挙げられ、 例えば、 カルボキシぺプチダーゼ A、 B、 P、 Yなどが例示 できる。  Examples of the carboxypeptidase include carboxypeptidase derived from a kidney or a microorganism, and examples thereof include carboxypeptidase A, B, P, and Y.
また、 アミノぺプチダーゼとしては、 鸱臓、 微生物などに由来する陴臓、 微生物 などに由来するアミノぺプチダーゼが挙げられ、 例えば、 アミノぺプチダーゼ Μな どが例示できる。 酵素反応終了後、 加熱などの手段で酵素を失活させ、 次いで慣用 精製手段 Γ"例 えば、 限外濾過、 逆浸透濾過、 ゲル濾過、 ィオン交換力ラムクロマトグラフィー、 逆相高速液体クロマトグラフィーなどに付して精製 ·単離することにより目的ぺプ チドを得ることができる。 Examples of the aminopeptidase include a kidney, a kidney derived from a microorganism, an aminopeptidase derived from a microorganism, and the like, and examples thereof include aminopeptidase. After completion of the enzyme reaction, the enzyme is deactivated by means such as heating, and then the conventional purification means Γ "For example, ultrafiltration, reverse osmosis filtration, gel filtration, ionic exchange ram chromatography, reverse phase high performance liquid chromatography, etc. The target peptide can be obtained by purifying and isolating it.
後記実施例に示されるように、 Hyp-Gly、 Ala-Hyp又は Pro-Hypからなるァミノ 酸配列を有する 2〜 6量体、 好ましくは 2〜 5量体のぺプチドは ACE阻害剤として 有効であることが明らかとなり、 係るぺプチドも本発明のぺプチドに包含される。 本発明の機能性食品は、 本発明のぺプチドの少なくとも一種を有効成分として含 有することからなる、血圧上昇抑制作用及び/又は血管保全作用を有する機能性食品 である。 なお、 本明細書において、 機能性食品とは、 通常の食品、 飲料、 菓子類、 飼料などを含む概念である。  As shown in the examples below, 2- to 6-mer, preferably 2- to 5-mer peptides having an amino acid sequence consisting of Hyp-Gly, Ala-Hyp or Pro-Hyp are effective as ACE inhibitors. It becomes clear that such peptides are also included in the peptides of the present invention. The functional food of the present invention is a functional food having an antihypertensive action and / or a blood vessel preservation action, comprising at least one of the peptides of the present invention as an active ingredient. In this specification, functional food is a concept that includes ordinary food, beverages, confectionery, feed, and the like.
当該機能性食品は、 有効成分そのまま、 又は種々の栄養分を加えて、 若しくは飲 食品中に含有せしめて、 血圧上昇抑制及び/又は血管保全を目的として、 その治療及 び予防に有用な機能性食品 （又は食品素材） として食される。 例えば、 適当な助剤 (例えば、 グルコース、 乳糖、 ショ糖、 澱粉、 マンニトール、 デキス トリン、 ポリ エチレングリコール、 ヒ ドロキシェチルデンプン、 エチレングリコール、 アミノ酸 等） を添加した後、 慣用の手段を用いて、 食用に適した形態、 例えば、 顆粒状、 粒 状、 錠剤、 カプセル、 ペース ト等に成形して食用に供してもよく、 また種々の食品 (例えば、 ハム、 ソーセージ等の食肉加工食品、 かまぼこ、 ちくわ等の水産加工食 品、 スナック菓子等の菓子類、 パン類、 バター、 粉乳等の乳製品、 豆腐、 油揚げ等 の大豆製品など） に添加して使用されたり、 水、 果汁、 牛乳、 清涼飲料等の飲物、 プリン、 ゼリーなどのデザート食品に添加して使用してもよい。 また、 動物用の飼 料 （ペットフードなどを含む） の形態であってもよい。  These functional foods are useful for the treatment and prevention of active ingredients as they are, added with various nutrients, or contained in foods and drinks for the purpose of suppressing blood pressure rise and / or maintaining blood vessels. It is eaten as (or food material). For example, after adding an appropriate auxiliary agent (for example, glucose, lactose, sucrose, starch, mannitol, dextrin, polyethylene glycol, hydroxyxetyl starch, ethylene glycol, amino acid, etc.), using conventional means It may be formed into a form suitable for edible use, such as granules, granules, tablets, capsules, pastes, etc., and used for food. Seafood processed foods such as chikuwa, confectionery such as snacks, dairy products such as breads, butter and powdered milk, soy products such as tofu and fried milk, etc.), water, fruit juice, milk, refreshing It may be used by adding to beverages such as beverages and dessert foods such as pudding and jelly. Further, it may be in the form of animal feed (including pet food).
係る機能性食品の形態における、 本発明のペプチドの摂取量は、 年齢、 体重、 症 状、 疾患の程度、 食品の形態等により、 適宜選択 ·決定され、 例えば、 1 日当りぺ プチドの量として、 1. 0 mg〜10. O mg程度、 好ましくは 2. 0 mg〜8. 0 mgとされるが、 多量に摂取しても生体に悪影響を与えない利点を有 1"ることから、 それ以上の量を 摂取してもよレ、。  The intake of the peptide of the present invention in the form of such functional food is appropriately selected and determined according to age, body weight, symptom, degree of disease, form of food, etc.For example, as the amount of peptide per day, 1. 0 mg to 10. O mg, preferably 2.0 mg to 8.0 mg, but since it has the advantage of not adversely affecting the body even if consumed in large amounts, it is more than that. You can take the amount of.
また、 本発明のペプチドは、 血圧上昇抑制作用及び/又は血管保全作用を有する薬 剤として及び当該べプチドの有効量を投与することからなる高血圧症の治療法にお いても利用することができる。 係る製剤は、 当該ペプチドを有効成分とし、 必要に 応じて、 適宜の生理的に許容される添加剤 （例えば、 担体、 賦形剤、 希釈剤等） な どの製剤上必要な成分と混合し、 適宜な剤形の形態に調製することにより得られ、 係る形態としては錠剤状、 粉末状、 顆粒状、 カプセル剤状などが例示できる。 投与 量は、 患者の症状、 年齢、 体重などに応じて、 上記の摂取量を参考にして適宜決定 することができる。 In addition, the peptide of the present invention is used as a drug having an antihypertensive action and / or a blood vessel preservation action, and in a method for treating hypertension comprising administering an effective amount of the peptide. You can use it. In such preparations, the peptide is used as an active ingredient, and if necessary, it is mixed with necessary ingredients in the preparation such as appropriate physiologically acceptable additives (for example, carriers, excipients, diluents, etc.) It can be obtained by preparing in an appropriate dosage form, and examples of the form include tablets, powders, granules, capsules and the like. The dose can be appropriately determined according to the patient's symptoms, age, weight, etc., referring to the above intake.
本発明のペプチドは、 血圧上昇抑制作用及び/又は血管保全作用を有しており、 そ の効果は下記実施例に記載の方法及び既に知られてレ、る血管内皮機能評価システム The peptide of the present invention has an antihypertensive action and / or a blood vessel preservation action, and the effect is the method described in the following examples and the already known vascular endothelial function evaluation system.
(例えば、 特開 2 0 0 7— 2 0 9 4 9 2号公報など） を適用することにより確認す ることができる。 実施例 This can be confirmed by applying (for example, Japanese Patent Laid-Open No. 2 0 07-2 0 9 4 92 2). Example
以下、 実施例に基づいて本発明をより詳細に説明するが、 本発明はこれらの例に 限定されるものではない。  EXAMPLES Hereinafter, although this invention is demonstrated in detail based on an Example, this invention is not limited to these examples.
実施例 1 (ACE阻害活性試験） Example 1 (ACE inhibitory activity test)
配列番号 1〜 2 3で示されるアミノ酸配列からなるぺプチドの ACE阻害活性の測 定を以下の方法で行った。  The ACE inhibitory activity of the peptide consisting of the amino acid sequence represented by SEQ ID NOs: 1 to 23 was measured by the following method.
(1)材料  (1) Material
配列番号 1〜2 3で示されるアミノ酸配列からなるペプチドは、 Fmoc固相法によ り自動ペプチド合成機 （PSSM8 島津） を用いて作製した。 脱保護の後、 逆相 HPLC (PEGASIL-300, 20 X 250 mm; Senshu, Tokyo, Japan) 及び質量分析計 ESI mass spectrometer LC-Q (Thermo Finnigan, San jose, CA) でぺプチドの構造確認を行 つた。 ペプチド合成試薬は島津 （京都、 日本） より購入した。  The peptides consisting of the amino acid sequences shown in SEQ ID NOs: 1 to 23 were prepared by an automatic peptide synthesizer (PSSM8 Shimadzu) by the Fmoc solid phase method. After deprotection, confirm the peptide structure using reverse-phase HPLC (PEGASIL-300, 20 X 250 mm; Senshu, Tokyo, Japan) and mass spectrometer ESI mass spectrometer LC-Q (Thermo Finnigan, San jose, CA). I went. Peptide synthesis reagents were purchased from Shimadzu (Kyoto, Japan).
ゥサギ肺 ACE及び Hip- HL (Bz-Gly- His-Leu)はシグマ （St. Louis, MO, USA) から 購入した。 その他の試薬は和光純薬 （大阪、 日本） より購入した。  Usagi lung ACE and Hip-HL (Bz-Gly-His-Leu) were purchased from Sigma (St. Louis, MO, USA). Other reagents were purchased from Wako Pure Chemical (Osaka, Japan).
(2) ACE阻害活性測定  (2) ACE inhibitory activity measurement
ACEに対する阻害活性の測定は Cheungらの方法 (Cheung et al. J. Biol. Chem. 1980， 255, 401-407) に従って行った。  The inhibitory activity against ACE was measured according to the method of Cheung et al. (Cheung et al. J. Biol. Chem. 1980, 255, 401-407).
より具体的には、最終容量が 0. 25mlとなるように lOOraMホゥ酸溶液（pH8. 3)、 5mM Hip- HL、 500mM NaCl、 20mU ゥサギ肺 ACE及び試験試料を加え、 37°Cで 30分間イン キュベートをおこなった。 酵素反応は 1NHC1の添加で停止させた。 - Hip-HLの加_水分 解の程度は放出された馬尿酸 （Hippuric acid) 量を吸光値 228nmで測定した。 ACE 活性を 50%阻害するときの阻害剤濃度を IC50と定義した。 More specifically, lOOraM folic acid solution (pH 8.3), 5mM Hip-HL, 500mM NaCl, 20mU Usagi Lung ACE and test sample were added to a final volume of 0.25ml, and the test sample was added at 37 ° C for 30 minutes. Inn I did a cuvate. The enzyme reaction was stopped by adding 1NHC1. -Hip-HL was measured for the amount of hippuric acid released at an absorbance of 228 nm. The inhibitor concentration at which the inhibition of ACE activity 50% was defined as IC 50.
(3)結果 (3) Results
配列番号 1〜2 3で示されるアミノ酸配列からなるペプチドの ACE 阻害活性 (IC5。）の結果を表 1に示した。 表 1に示されるように、 配列番号 1〜23で示される アミノ酸配列からなるペプチドはいずれも ACE阻害活性を有していた。 特に配列番 号 2、 3、 7〜： 1 1、 14及び 18〜20で示されるアミノ酸配列からなるぺプチ ドが極めて強い ACE阻害活性を有していた。 Table 1 shows the results of the ACE inhibitory activity (IC 5 ) of the peptides consisting of the amino acid sequences represented by SEQ ID NOs: 1 to 23. As shown in Table 1, all the peptides consisting of the amino acid sequences represented by SEQ ID NOs: 1 to 23 had ACE inhibitory activity. In particular, peptides consisting of the amino acid sequences represented by SEQ ID NOs: 2, 3, 7 to: 11, 14, and 18 to 20 had extremely strong ACE inhibitory activity.
係るペプチドの構造から考察すると、 Hyp-Gly、 Ala-Hyp又は PnrHypからなる ァミノ酸配列を有する 2〜 6量体、 好ましくは 2〜 5量体のぺプチドは ACE阻害剤 として有効であると考えられる。  Considering from the structure of such a peptide, it is considered that a 2 to 6-mer, preferably 2 to 5-mer peptide having an amino acid sequence consisting of Hyp-Gly, Ala-Hyp or PnrHyp is effective as an ACE inhibitor. It is done.
表 1  table 1
配列番号及び配列 I C50 ( m g Zm 1 ) SEQ ID NO: and sequence IC 50 (mg Zm 1)
配列番号 1 Gly-Al a-Hyp-Gly-Leu-Hyp 1 3 1. 336 SEQ ID NO: 1 Gly-Al a-Hyp-Gly-Leu-Hyp 1 3 1. 336
配列番号 2 Gl y -し eu_Hyp - Gl y Pro 0. 079 SEQ ID NO: 2 Gl y-then eu_Hyp-Gl y Pro 0. 079
配列番号 3 し eu— Hy — ly - Pro 0. 582 SEQ ID NO: 3 and eu— Hy — ly-Pro 0. 582
配列番号 4 Gly-A la-Hyp 4. 51 7 SEQ ID NO: 4 Gly-A la-Hyp 4.5 1 7
配列番号 5 Ala-Hyp-Gly 0. 930 SEQ ID NO: 5 Ala-Hyp-Gly 0. 930
配列番号 6 Gly-Leu-Hyp 1. 122 SEQ ID NO: 6 Gly-Leu-Hyp 1. 122
配列番号 7 Hyp-Gly-Pro 0. 290 SEQ ID NO: 7 Hyp-Gly-Pro 0. 290
配列番号 8 Gly-Pro 0. 386 SEQ ID NO: 8 Gly-Pro 0. 386
配列番号 9 し eu— Hyp_Gly 0. 332 SEQ ID NO: 9 and eu—Hyp_Gly 0. 332
配列番号 10 ： Hyp - Gly_し eu 0, 299 SEQ ID NO: 10: Hyp-Gly_ eu 0, 299
配列番号 11 ： Hyp-Gly 0. 434 Sequence number 11: Hyp-Gly 0. 434
配列番号 12 ： Gly-Leu 0. 778 Sequence number 12: Gly-Leu 0. 778
配列番号 13 ： Gly-Ala 5. 1 22 Sequence number 13: Gly-Ala 5.12.2
配列番号 14 ： Ala-Hyp 0. 088 Sequence number 14: Ala-Hyp 0. 088
配列番号 15 ：し eu - Hyp 2. 182 SEQ ID NO: 15 then eu-Hyp 2. 182
配列番号 16 ： Pro - Hyp - Gly 1. 024 Sequence number 16: Pro-Hyp-Gly 1.024
配列番号 17 ： Glu - Hyp - Gly 1. 216 Sequence number 17: Glu-Hyp-Gly 1.216
配列番号 18 ： Ser-Hyp-Gly 0. 305 Sequence number 18: Ser-Hyp-Gly 0. 305
配列番号 19 ： Pro-Hyp 0. 063 Sequence number 19: Pro-Hyp 0.063
配列番号 20 ： Phe-Hyp 0. 375 Sequence number 20: Phe-Hyp 0. 375
配列番号 21 ： ulu-Hyp 2. 383 Sequence number 21: ulu-Hyp 2.383
配列番号 22 ： Ser-Hyp 1. 443 Sequence number 22: Ser-Hyp 1.443
配列番号 23 ： Ile-Hyp 3. 227 実施例 2 (血管保全作用試験） Sequence number 23: Ile-Hyp 3.227 Example 2 (Vessel Conservation Action Test)
馴化後の Wistar 系ラッ ト（雄、 8週齢）に NO 合成酵素阻害剤である L- NAME (N-nitro-L-arginine methyl ester)を飲用水に混合し投与（lmg/ml in water, 1週 間）し、 血管機能損傷モデルとした。 その後、 配列番号 1〜2 3で示されるアミノ酸 配列からなるペプチド （5 g/kg/day) を 8週間経口投与した。 また、 飼育期間全体 を通して、 飲用水にはい NAME (lmg/ml) を含有する水を使用した。  L-NAME (N-nitro-L-arginine methyl ester), a NO synthase inhibitor, is mixed with drinking water after administration (lmg / ml in water, 8 weeks old) 1 week) and used as a vascular function injury model. Thereafter, a peptide (5 g / kg / day) consisting of the amino acid sequence represented by SEQ ID NOs: 1 to 23 was orally administered for 8 weeks. In addition, water containing NAME (lmg / ml) was used for drinking water throughout the breeding period.
飼育期間の終了後、 胸部大動脈を採取し、 直ちに Krebs-Henseleit溶液 （118. 4mM NaCl, 4. 7 mM KCl, 2. 5mM CaCl2, 1. 2mM KH2P04, 1. 2mM MgS04 - 7H20, 25mM NaHC03, 11. 6mM C6H1206. PH 7. 4, 4°C)に浸漬しておき、 約 3-4瞧のリング状に切り出し、 オーガンチ ヤンバー 'マイクロバスに設置した。 この際、 オーガンチャンバ一'マイクロバス 内は Kerbs- Henseleit溶液 （同上、 37°C) で満たし、 血管標本の機能を損なわない よう 95% 02、 5% C02の混合気体でバブリングしておいた。 After the end of the feeding period, the thoracic aorta was collected, immediately Krebs-Henseleit solution (118. 4mM NaCl, 4. 7 mM KCl, 2. 5mM CaCl 2, 1. 2mM KH 2 P0 4, 1. 2mM MgS0 4 - 7H 2 0, 25 mM NaHC0 3 , 11.6 mM C 6 H 12 0 6 .PH 7.4, 4 ° C), cut into a ring shape of about 3-4 mm, and installed in an organ yanbar 'microbus . At this time, the organ chamber's microbus was filled with Kerbs-Henseleit solution (same as above, 37 ° C) and bubbled with a mixture of 95% 0 2 and 5% C0 2 so as not to impair the function of the blood vessel specimen. .
設置した血管標本に lgの負荷がかかるようセッティングし、 60分程度放置し、安 定化させた。 その後、 フエ二レフリン （終濃度 1 X 10—7M) を添加して血管標本を収 縮させたのちに、 アセチルコリン （終濃度 1 X 10— 6M) を添加した際の拡張反応を計 測し、 拡張の度合いを血管保全作用の評価とし、 その効果を 4段階 （1 ：効果無し 2 ：やや効果有り 3 ：効果有り 4 ：著しい効果有り） で示した。 The blood vessel sample was set to be loaded with lg and left for about 60 minutes to stabilize. Thereafter, phenylene Refurin the After (final concentration 1 X 10- 7 M) was added to cause the blood vessel specimens are contraction, measuring the extension reaction upon addition of acetylcholine (final concentration 1 X 10- 6 M) Total The degree of dilation was evaluated as a blood vessel preservation action, and the effect was shown in four stages (1: No effect 2: Slightly effective 3: Effective) 4: Significant effect).
その結果を表 2に示した。 表 2に示されるように、 配列番号 1〜2 3で示される ァミノ酸配列からなるぺプチドはいずれも血管保全作用を有しており、 特に配列番 号 2、 1 4、 1 6及び 1 9で示されるアミノ酸配列からなるペプチドは強い活性を 有していた。 The results are shown in Table 2. As shown in Table 2, all of the peptides consisting of the amino acid sequences represented by SEQ ID NOs: 1 to 23 have a vascular conservation action, and in particular, SEQ ID NOs: 2, 14, 16, and 19 The peptide consisting of the amino acid sequence shown by has a strong activity.
表 2 Table 2
配列番号及び配列 血管保全作用評価  Sequence number and sequence Evaluation of vascular conservation effect
配列番号 1 Gly-Ala-Hyp-Gly-Leu-Hyp 3 SEQ ID NO: 1 Gly-Ala-Hyp-Gly-Leu-Hyp 3
配列番号 2 Gly-Leu-Hyp-Gly-Pro 4 SEQ ID NO: 2 Gly-Leu-Hyp-Gly-Pro 4
配列番号 3 し eu- Hyp - Gly- Pro 3 SEQ ID NO: 3 and eu- Hyp-Gly- Pro 3
配列番号 4 Gly-Ala-Hyp 3 SEQ ID NO: 4 Gly-Ala-Hyp 3
配列番号 5 Ala- Hyp - Gly 3 SEQ ID NO: 5 Ala- Hyp-Gly 3
配列番号 6 Gly-Leu-Hyp 3 SEQ ID NO: 6 Gly-Leu-Hyp 3
配列番号 7 Hyp - Gly - Pro 3 SEQ ID NO: 7 Hyp-Gly-Pro 3
配列番号 8 Gly- Pro 3 SEQ ID NO: 8 Gly- Pro 3
配列番号 9 し eu - Hyp_Gly 3 SEQ ID NO: 9 and eu-Hyp_Gly 3
配列番号 10 ： Hyp- Gly - Leu 3 Sequence number 10: Hyp-Gly-Leu 3
配列番号 11 ： Hyp- Gly 3 Sequence number 11: Hyp-Gly 3
配列番号 12 ： Gly- Leu 3 Sequence number 12: Gly- Leu 3
配列番号 13 ： Gly-Ala 3 Sequence number 13: Gly-Ala 3
配列番号 14 ： Ala-Hyp 4 Sequence number 14: Ala-Hyp 4
配列番号 15 ： Leu-Hyp 3 Sequence number 15: Leu-Hyp 3
配列番号 16 ： Pro - Hyp - Gly 4 Sequence number 16: Pro-Hyp-Gly4
配列番号 17 ： Glu- Hyp - Gly 3 Sequence number 17: Glu- Hyp-Gly 3
配列番号 18 ： Ser - Hyp - Gly 3 Sequence number 18: Ser-Hyp-Gly3
配列番号 19 ： Pro-Hyp 4 Sequence number 19: Pro-Hyp 4
配列番号 20 ： Phe-Hyp 3 Sequence number 20: Phe-Hyp 3
配列番号 21 ： Glu - Hyp 2 Sequence number 21: Glu-Hyp 2
配列番号 22 ： Ser - Hyp 2 Sequence number 22: Ser-Hyp2
配列番号 23 ： l ie— Hyp 2 Sequence number 23: l ie— Hyp 2
コントロール （ペプチド投与せず） 1 製造例 1 Control (no peptide administration) 1 Production Example 1
天然果汁 （濃縮果汁還元） に、 当該果汁 2 0 O m l当り本発明のぺプチド （配列 番号 2で示されるアミノ酸配列からなるペプチド） 1 0 m gを混合した後、 常法に 準じて殺菌し、 ァセブティック包装して、 果汁製品を得た。  Natural fruit juice (concentrated juice reduction) is mixed with 10 mg of the peptide of the present invention (peptide consisting of the amino acid sequence represented by SEQ ID NO: 2) per 20 O ml of the juice, and then sterilized according to a conventional method. Asse boutique packaging to obtain a juice product.
製造例 2 Production example 2
ウインナソーセージ用練り肉に、 当該練り肉 1 5 g当り本発明のペプチド （配列 番号 1 4で示されるアミノ酸配列からなるペプチド） 1 0 m gを混合した後、 常法 に準じてソーセージケーシングに充填し、 燻煙し、 殺菌し、 冷却後に包装し、 ウイ ンナソーセージを得た。  After mixing 10 mg of the peptide of the present invention (peptide consisting of the amino acid sequence shown in SEQ ID NO: 14) per 15 g of the paste, the sausage casing is filled according to a conventional method. Smoked, sterilized, packaged after cooling to obtain wiener sausage.
製造例 3 Production Example 3
本発明のペプチド （配列番号 1 9で示されるアミノ酸配列からなるペプチド） 6 m gを常法に準じてハードカプセルに充填し、 カプセル剤を製造した。 産業上の利用可能性 Peptide of the present invention (peptide consisting of the amino acid sequence represented by SEQ ID NO: 19) 6 According to a conventional method, mg was filled into a hard capsule to produce a capsule. Industrial applicability
実施例に示されるよう、 本発明のぺプチドは強い ACE阻害活性及び血管内皮の障 害回復、 機能保持などの作用を有しており、 血圧上昇抑制作用及び/又は血管保全作 用を有する機能性食品又は薬剤として利用することができる。 特に低分子であるか ら吸収性に優れ、 生体に吸収された後においても効果を持続することができるとい う特長を有している。  As shown in the Examples, the peptide of the present invention has strong ACE inhibitory activity, vascular endothelial damage recovery, function maintenance, and the like, and has a function of suppressing blood pressure elevation and / or blood vessel preservation. It can be used as a sex food or medicine. Since it is a low molecular weight compound, it has excellent absorbability and can maintain its effect even after being absorbed by the living body.
また、 本発明の機能性食品又は薬剤は、 上記の作用を有するペプチドを含有して おり、 生活習慣病である高血圧などは、 日常の生活を通して、 予防 ·改善すること が重要であり、 本発明のペプチドを食事などにより うまく摂取することで、 対象者 の QOL (Quality of life) を損なうことなく、 血圧の正常化、 血管保全などを達成 できるという特長を有する。  In addition, the functional food or drug of the present invention contains a peptide having the above-described action, and it is important to prevent and improve high blood pressure, which is a lifestyle-related disease, through daily life. By taking these peptides well with meals, etc., it is possible to achieve normalization of blood pressure and blood vessel preservation without impairing the QOL (Quality of life) of the subject.

Claims

請求の範囲 The scope of the claims
1. 配列番号 1〜 23で示されるァミノ酸配列からなるぺプチド。 1. A peptide comprising an amino acid sequence represented by SEQ ID NOs: 1 to 23.
2. Hyp-Gly, Ala-Hyp又は Pro-Hy からなるァミノ酸配列を有する 2〜 6量体 のぺプチド。  2. A 2 to 6-mer peptide having an amino acid sequence consisting of Hyp-Gly, Ala-Hyp or Pro-Hy.
3. 請求項 1又は 2記載のぺプチドの少なくとも一種を含有する、 血圧上昇抑制作 用及び/又は血管保全作用を有する機能性食品又は薬剤。  3. A functional food or drug having an antihypertensive action and / or a blood vessel preservation action, comprising at least one peptide according to claim 1 or 2.
4. ペプチドが、 配列番号 2、 3、 7〜： L l、 14、 16及び 18〜 20で示され るアミノ酸配列からなるぺプチドである請求項 3記載の血圧上昇抑制作用及び/又 は血管保全作用を有する機能性食品又は薬剤。  4. The antihypertensive action and / or blood vessel according to claim 3, wherein the peptide is a peptide consisting of the amino acid sequence represented by SEQ ID NOs: 2, 3, 7 to: L1, 14, 16, and 18 to 20. A functional food or drug having a protective action.
5. 請求項 1又は 2記載のぺプチドの少なくとも一種の有効量を投与することか らなる高血圧症の治療方法。  5. A method for treating hypertension, comprising administering an effective amount of at least one of the peptides according to claim 1 or 2.
6. ペプチドが、 配列番号 2、 3、 7-1 1, 14、 16及び 18〜20で示され るアミノ酸配列からなるぺプチドである請求項 5記載の高血圧症の治療方法。  6. The method for treating hypertension according to claim 5, wherein the peptide is a peptide consisting of an amino acid sequence represented by SEQ ID NOs: 2, 3, 7-1 1, 14, 16, and 18 to 20.
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Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2012100612A (en) * 2010-11-12 2012-05-31 Nippon Meat Packers Inc Cataract-preventing food
WO2012102308A1 (en) * 2011-01-27 2012-08-02 新田ゼラチン株式会社 Therapeutic agent and preventive agent for diabetes
US20130123167A1 (en) * 2009-03-11 2013-05-16 Kanazawa Medical University Drug inhibiting the progression of atherosclerosis, preventive drug, blood cholesterol-lowering drug, functional food, and specific health food
JP2014001182A (en) * 2012-06-20 2014-01-09 Nitta Gelatin Inc Therapy or preventive of disease associated with bone, cartilage or skin
JP2014141450A (en) * 2012-12-26 2014-08-07 Nitta Gelatin Inc Elastin production promoter
CN104271144A (en) * 2012-12-13 2015-01-07 新田明胶株式会社 Myoblast differentiation accelerator
JP2015059087A (en) * 2013-09-17 2015-03-30 新田ゼラチン株式会社 Therapeutic or preventive agent for osteoarthritis or osteoporosis
CN107586318A (en) * 2017-05-25 2018-01-16 青岛大学 A kind of blood pressure lowering peptide and preparation method thereof
JP2019094271A (en) * 2017-11-20 2019-06-20 株式会社ニッピ Angiotensin-converting enzyme inhibitor, food, beverage, and supplement

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08231588A (en) * 1995-03-01 1996-09-10 Nissin Food Prod Co Ltd Angiotensinase inhibiting peptide and its production
JP2001106699A (en) * 1999-10-05 2001-04-17 Suetsuna Yoko New hexapeptide and angiotensin-converting enzyme inhibitor
JP2003137807A (en) * 2001-11-01 2003-05-14 Miyagi Kagaku Kogyo Kk Collagen-producing promoter, cosmetic, food and pharmaceutical containing the same and external preparation for preventing or improving dermatosis
JP2005206469A (en) * 2004-01-20 2005-08-04 National Fisheries Univ Novel hexapeptide and angiotensin converting enzyme inhibitors
JP2006160625A (en) * 2004-12-03 2006-06-22 Yoshihiro Futamura Water-soluble extract derived from bee larva having protease inhibitory action, method for producing the same, and product containing the same
JP2007222117A (en) * 2006-02-27 2007-09-06 Kao Corp Packaged liquid seasoning

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08231588A (en) * 1995-03-01 1996-09-10 Nissin Food Prod Co Ltd Angiotensinase inhibiting peptide and its production
JP2001106699A (en) * 1999-10-05 2001-04-17 Suetsuna Yoko New hexapeptide and angiotensin-converting enzyme inhibitor
JP2003137807A (en) * 2001-11-01 2003-05-14 Miyagi Kagaku Kogyo Kk Collagen-producing promoter, cosmetic, food and pharmaceutical containing the same and external preparation for preventing or improving dermatosis
JP2005206469A (en) * 2004-01-20 2005-08-04 National Fisheries Univ Novel hexapeptide and angiotensin converting enzyme inhibitors
JP2006160625A (en) * 2004-12-03 2006-06-22 Yoshihiro Futamura Water-soluble extract derived from bee larva having protease inhibitory action, method for producing the same, and product containing the same
JP2007222117A (en) * 2006-02-27 2007-09-06 Kao Corp Packaged liquid seasoning

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
KOUGUCHI T ET AL: "Hypotensive effects and safety of intake of lactic acid beverage containing chicken collagen hydrolysate in subjects with mild hypertension od high-normal blood pressure", JPN. PHARMACOL. THR., vol. 36, no. 6, 20 June 2008 (2008-06-20), pages 561 - 575 *
OSHIMA G: "Peptide inhibitors of angiotensin I-converting enzyme in digests of gelatin bu bacterial collagenase", BIOCHIM. BIOPHYS. ACTA, vol. 566, 1979, pages 128 - 137 *
SAIGA A ET AL: "Angiotensin I-converting enzyme-inhibitory peptides obtained from chicken collagen hydrolysate", J. AGR. FOOD CHEM., vol. 56, no. 20, 23 September 2008 (2008-09-23), pages 9586 - 9591 *
SAIGA A: "Action mechanism of an angiotensin I-converting enzyme inhibitory peptide derived from chicken breast extract", J. AGR. FOOD CHEM., vol. 54, no. 3, 2006, pages 942 - 945 *
SAIGA A: "Angiotensin I-converting enzyme inhibitory peptides in a hydrolyzed chicken breast muscle extract", J. AGRIC. FOOD CHEM., vol. 51, 2003, pages 1741 - 1745 *

Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130123167A1 (en) * 2009-03-11 2013-05-16 Kanazawa Medical University Drug inhibiting the progression of atherosclerosis, preventive drug, blood cholesterol-lowering drug, functional food, and specific health food
US9339524B2 (en) * 2009-03-11 2016-05-17 Jellice Co., Ltd. Drug inhibiting the progression of atherosclerosis, preventive drug, blood cholesterol-lowering drug, functional food, and specific health food
JP2012100612A (en) * 2010-11-12 2012-05-31 Nippon Meat Packers Inc Cataract-preventing food
CN103347530B (en) * 2011-01-27 2015-04-01 新田明胶株式会社 Therapeutic agent and preventive agent for diabetes
WO2012102308A1 (en) * 2011-01-27 2012-08-02 新田ゼラチン株式会社 Therapeutic agent and preventive agent for diabetes
CN103347530A (en) * 2011-01-27 2013-10-09 新田明胶株式会社 Therapeutic agent and preventive agent for diabetes
JPWO2012102308A1 (en) * 2011-01-27 2014-06-30 新田ゼラチン株式会社 Diabetes treatment or prevention
JP5612131B2 (en) * 2011-01-27 2014-10-22 新田ゼラチン株式会社 Diabetes treatment or prevention
TWI507203B (en) * 2011-01-27 2015-11-11 Nitta Gelatin Kk The use of a collagen peptide mixture for the manufacture of a therapeutic or prophylactic agent for diabetes mellitus
US9061003B2 (en) 2011-01-27 2015-06-23 Nitta Gelatin Inc. Therapeutic or preventive agent for diabetes
JP2014001182A (en) * 2012-06-20 2014-01-09 Nitta Gelatin Inc Therapy or preventive of disease associated with bone, cartilage or skin
CN104271144A (en) * 2012-12-13 2015-01-07 新田明胶株式会社 Myoblast differentiation accelerator
US10632177B2 (en) 2012-12-13 2020-04-28 Nitta Gelatin Inc. Myoblast differentiation promoter
JP2014141450A (en) * 2012-12-26 2014-08-07 Nitta Gelatin Inc Elastin production promoter
JP2015059087A (en) * 2013-09-17 2015-03-30 新田ゼラチン株式会社 Therapeutic or preventive agent for osteoarthritis or osteoporosis
CN107586318A (en) * 2017-05-25 2018-01-16 青岛大学 A kind of blood pressure lowering peptide and preparation method thereof
CN107586318B (en) * 2017-05-25 2021-01-05 青岛大学 Antihypertensive peptide and preparation method thereof
JP2019094271A (en) * 2017-11-20 2019-06-20 株式会社ニッピ Angiotensin-converting enzyme inhibitor, food, beverage, and supplement
JP7048270B2 (en) 2017-11-20 2022-04-05 株式会社ニッピ Angiotensin-converting enzyme inhibitors, foods, beverages, and supplements

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