JPWO2009057287A1 - 破骨細胞形成抑制用食品素材 - Google Patents
破骨細胞形成抑制用食品素材 Download PDFInfo
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- JPWO2009057287A1 JPWO2009057287A1 JP2009538926A JP2009538926A JPWO2009057287A1 JP WO2009057287 A1 JPWO2009057287 A1 JP WO2009057287A1 JP 2009538926 A JP2009538926 A JP 2009538926A JP 2009538926 A JP2009538926 A JP 2009538926A JP WO2009057287 A1 JPWO2009057287 A1 JP WO2009057287A1
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- Prior art keywords
- protein fraction
- milk protein
- osteoclast formation
- milk
- amino acid
- Prior art date
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Classifications
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- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4702—Regulators; Modulating activity
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- A—HUMAN NECESSITIES
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Abstract
Description
本発明の乳タンパク質画分又は乳タンパク質画分分解物は、破骨細胞形成抑制作用を有するので、骨疾患の予防又は治療、骨強化等を目的とした破骨細胞形成抑制剤として、また、骨疾患の予防又は治療用もしくは破骨細胞形成抑制用医薬品、飲食品及び飼料の有効成分として有用である。
(1) 乳由来であること。
(2) ソディウムドデシルサルフェート−ポリアクリルアミドゲル(SDS-PAGE)電気泳動で,分子量80,000〜85,000ダルトンの範囲のタンパク質を含有する画分であること。
(3) 構成アミノ酸組成中に塩基性アミノ酸を16〜18重量%含有し、かつ、塩基性アミノ酸/酸性アミノ酸比が0.65〜0.85の範囲であること。
(4) 破骨細胞形成抑制作用を有すること。
(B)前記乳タンパク質画分をタンパク質分解酵素で分解して得られる乳タンパク質画分分解物。
(C) (A)又は(B)に記載の乳タンパク質画分又は乳タンパク質画分分解物を含有する破骨細胞形成抑制剤。
(D) (A)又は(B)に記載の乳タンパク質画分又は乳タンパク質画分分解物を配合した破骨細胞形成抑制用医薬品。
(E) (A)又は(B)に記載の乳タンパク質画分又は乳タンパク質画分分解物を配合した破骨細胞形成抑制用飲食品。
(F) (A)又は(B)に記載の乳タンパク質画分又は乳タンパク質画分分解物を配合した破骨細胞形成抑制用飼料。
下記の方法(特許第3112637号参照)により、骨強化が認められる市販品の乳タンパク質画分を調製した。
陽イオン交換樹脂のスルホン化キトパール(富士紡績製) 0.5リットルを充填した直径10cmのカラムを脱イオン水で充分洗浄した。このカラムに未殺菌脱脂乳 50リットルを流速100 ml /minで通液した後、このカラムを脱イオン水で充分洗浄し、0.95M塩化ナトリウムを含む 0.05Mリン酸緩衝液(pH 7.0) 2.5リットルを通液して樹脂に吸着したタンパク質を溶出した。そして、この溶出液を逆浸透(RO)膜処理により、脱塩し、濃縮した後、凍結乾燥して粉末状の乳タンパク質画分を得た。この操作を2回繰り返して104gのタンパク質画分を得た。このタンパク質画分の等電点は 7.0〜8.5 であって、このタンパク質画分に含まれる塩基性アミノ酸は17.8%であった。
骨芽細胞が産生する破骨細胞形成抑制因子(OPG)は、生理的な破骨細胞の分化抑制因子として知られている。そこで、参考例1及び実施例1で得られた各乳タンパク質画分について、破骨細胞形成抑制作用として、骨芽細胞におけるOPGの産生促進作用を調べた。ヒト骨芽細胞MG63を2×104 cells/mlになるようにMEM‐E培地〔GIBCO社製、ウシ胎児血清10%含有、非必須アミノ酸溶液(GIBCO社製)1%含有、ペニシリン‐ストレプトマイシン溶液(GIBCO社製)1%含有〕に懸濁した後、200μl/wellずつ96wellプレートに播種した。これを5%炭酸ガスインキュベーター中で4日間培養し、培地を吸引除去後、上記MEM‐E培地を160μl/well、MEM‐E培地で1.0mg/ml又は0.1mg/mlの濃度に溶解した参考例1及び実施例1で得られたタンパク質画分を40μl/well添加して、さらに4日間培養した。この培養上清を回収し、培養上清中のOPG濃度をヒトOPG‐ELISA測定キット(BIOMEDICA社製)を用いて測定した。また、試料溶液の代わりにMEM‐E培地のみで培養したものを対照とした。
表1に結果を示す。
参考例1及び実施例2で得られた各タンパク質画分について、動物実験により骨強化作用を調べた。動物実験には4週齢のWistar系雌ラットを用いた。1週間の予備飼育後、卵巣摘出手術を施し、その後、カルシウム欠乏食で5週間飼育して動物実験に供した。なお、卵巣を摘出し、カルシウム欠乏食で5週間飼育したラットは、明らかに骨粗鬆症状態にあった。この骨粗鬆症状態を惹起したラットを1群7匹ずつ、乳タンパク質画分無添加の対照群(A群)、参考例1で得られた乳タンパク質画分0.1重量%投与群(B群)、実施例2で得られた乳タンパク質画分0.1重量%投与群(C群)の3試験群に分け、それぞれ表1に示す試験飼料で4ヶ月飼育した。なお、各試験飼料の窒素含量(17.06%)が同様となるようカゼインで調整した。また、各試験飼料については、100g当たり、カルシウム 300mg、リン 230mg及びマグネシウム 50mgを配合した。
Claims (6)
- 次の(1)〜(4)の特性を有する乳タンパク質画分。
(1) 乳由来であること。
(2) ソディウムドデシルサルフェート−ポリアクリルアミドゲル(SDS-PAGE)電気泳動で、分子量80,000〜85,000ダルトンの範囲のタンパク質成分を含有する画分であること。
(3) 構成アミノ酸組成中に塩基性アミノ酸を16〜18重量%含有し、かつ、塩基性アミノ酸/酸性アミノ酸比が0.65〜0.85の範囲であること。
(4) 破骨細胞形成抑制作用を有すること。 - 請求項1に記載の乳タンパク質画分をタンパク質分解酵素で分解して得られる乳タンパク質画分分解物。
- 請求項1又は2に記載の乳タンパク質画分又は乳タンパク質画分分解物を含有する破骨細胞形成抑制剤。
- 請求項1又は2に記載の乳タンパク質画分又は乳タンパク質画分分解物を配合した破骨細胞形成抑制用医薬品。
- 請求項1又は2に記載の乳タンパク質画分又は乳タンパク質画分分解物を配合した破骨細胞形成抑制用飲食品。
- 請求項1又は2に記載の乳タンパク質画分又は乳タンパク質画分分解物を配合した破骨細胞形成抑制用飼料。
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US (1) | US20100234296A1 (ja) |
EP (1) | EP2208734B1 (ja) |
JP (1) | JPWO2009057287A1 (ja) |
KR (1) | KR20100100845A (ja) |
CN (1) | CN101842385B (ja) |
AT (1) | ATE542832T1 (ja) |
AU (1) | AU2008320277A1 (ja) |
CA (1) | CA2704092C (ja) |
DK (1) | DK2208734T3 (ja) |
ES (1) | ES2378213T3 (ja) |
NZ (1) | NZ585092A (ja) |
PT (1) | PT2208734E (ja) |
WO (1) | WO2009057287A1 (ja) |
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US8580740B2 (en) * | 2007-11-01 | 2013-11-12 | Megmilk Snow Brand Co., Ltd. | Bone resorption inhibitory food material for inhibiting bone resorption |
US20100298204A1 (en) * | 2007-11-01 | 2010-11-25 | Snow Brand Milk Products Co., Ltd. | Food material for promoting the differentiation of osteoblast and inhibiting the differentiation of osteoclast |
NZ584888A (en) * | 2007-11-01 | 2012-08-31 | Megmilk Snow Brand Co Ltd | Bone-strengthening food material |
JP2012077015A (ja) * | 2010-09-30 | 2012-04-19 | Snow Brand Milk Products Co Ltd | 皮膚コラーゲン産生促進剤 |
JP5993308B2 (ja) | 2011-01-26 | 2016-09-14 | 雪印メグミルク株式会社 | 感覚改善剤 |
JP5969225B2 (ja) * | 2012-03-09 | 2016-08-17 | 雪印メグミルク株式会社 | 骨強化剤 |
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Publication number | Publication date |
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EP2208734B1 (en) | 2012-01-25 |
EP2208734A1 (en) | 2010-07-21 |
NZ585092A (en) | 2013-03-28 |
AU2008320277A1 (en) | 2009-05-07 |
DK2208734T3 (da) | 2012-05-14 |
CN101842385A (zh) | 2010-09-22 |
PT2208734E (pt) | 2012-03-29 |
CA2704092C (en) | 2017-06-20 |
ATE542832T1 (de) | 2012-02-15 |
CN101842385B (zh) | 2013-11-27 |
WO2009057287A1 (ja) | 2009-05-07 |
US20100234296A1 (en) | 2010-09-16 |
ES2378213T3 (es) | 2012-04-10 |
CA2704092A1 (en) | 2009-05-07 |
EP2208734A4 (en) | 2010-11-03 |
KR20100100845A (ko) | 2010-09-15 |
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