JPS61277626A - Placenta extract containing superoxide dismutase in high concentration and production thereof - Google Patents
Placenta extract containing superoxide dismutase in high concentration and production thereofInfo
- Publication number
- JPS61277626A JPS61277626A JP60117919A JP11791985A JPS61277626A JP S61277626 A JPS61277626 A JP S61277626A JP 60117919 A JP60117919 A JP 60117919A JP 11791985 A JP11791985 A JP 11791985A JP S61277626 A JPS61277626 A JP S61277626A
- Authority
- JP
- Japan
- Prior art keywords
- placenta
- placenta extract
- extract
- superoxide dismutase
- high concentration
- 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.)
- Pending
Links
Abstract
Description
【発明の詳細な説明】
〈産業上の利用分野〉
本発明はスーパーオキシドジスムターゼ（Ｓｕｐｅｒ−
ｏｘｉｄｅ ｄｌａｍｕｔａｓｅ Ｅ Ｃ１，１５，１
，１，以下ＳＯＤと称す〕を高濃度に含有することを特
徴とする胎盤エキスおよびその製法にかんする。さらに
詳しくは、本発明はＳＯＤ濃度が１００単位以上であり
、アルカリ性ホスホ７アターゼ濃度が１００キング・ア
ームストロング単位以上である胎盤エキスおよびその製
法にかんする。DETAILED DESCRIPTION OF THE INVENTION <Industrial Application Field> The present invention relates to superoxide dismutase (Super-
oxide dlamutase E C1,15,1
, 1, hereinafter referred to as SOD] and a method for producing the same. More specifically, the present invention relates to a placenta extract having an SOD concentration of 100 units or more and an alkaline phospho-7 atase concentration of 100 King-Armstrong units or more, and a method for producing the same.
〈従来の技術〉
胎盤エキス中には、非常に多くの生理活性物質が含まれ
ておシ、アミノ酸、ホルモン、ビタミン、酵素などを挙
げることができる。し之がって、これらの有用物質を有
効に利用しようという考えにより、胎盤エキスの製造が
樵々検討され、ある程度実用化されている、しかしなが
ら、現在市販されている胎盤エキス中のＳＯＤ活性全測
定したところ、１００単位／Ｍ以上のＳＯＤを含む胎盤
エキスは皆無であった。このことは、従来の胎盤エキス
の製造にあたり、ＳＯＤは考慮されてｈな−ことになる
。<Prior Art> Placenta extract contains a large number of physiologically active substances, such as milk, amino acids, hormones, vitamins, and enzymes. Therefore, with the idea of making effective use of these useful substances, the production of placenta extracts has been extensively studied and has been put into practical use to some extent. As a result of measurement, there was no placenta extract containing SOD of 100 units/M or more. This means that SOD must be taken into account when manufacturing conventional placenta extracts.
〈発明“が解決しようとする問題点〉
本発明は前記従来技術において考慮されてｈなかったＳ
ＯＤを高濃度に含有する胎盤エキスおよびその製造法を
提供することを目的とする、〈問題点を解決する乏めの
手段〉
ＳＯＤはスーパーオキシドアニオンラジカル（０□；）
の不均化反応Ｃ２０，； ＋ ２Ｈ＋→０□＋Ｈ２０，
）を触媒する酵素であって、Ｑ、 ｅの毒性から生物を
保傾するものと考えられている。<Problems to be solved by the invention> The present invention solves S
The purpose is to provide a placenta extract containing a high concentration of OD and a method for producing the same. <A poor means to solve the problem> SOD is superoxide anion radical (0□;)
Disproportionation reaction C20,; + 2H+→0□+H20,
), and is thought to protect living organisms from the toxicity of Q and e.
本発明者らは、過酸化脂質産生の主原因となる０ヨ；を
消去し、活性酸素の毒性を最も強く抑制する酵素として
のＳＯＤに注目してきた。また本酵素が胎盤に比較的多
く存在しかつ安定なものであることを明らかにしてきた
。そこで、本発明者らは、５ＯＤ１に高濃度に含む胎盤
エキスの製造ＶＣついて研究を行ったところ、従来の胎
盤エキスの規格に適合し、しかもＳＯＤを高濃度（１９
０単位／ ｍ１以上）に含む胎盤エキスの製造法を確立
し、本方法によ、Ｄ、ＳＯＤを高濃度に含有する胎盤エ
キスｆ！：鯛造した。即ち本発明は、ＳＯＤを高娘度に
含有することをＷ徴とする胎盤エキスおよびその製造法
にかんする。The present inventors have focused on SOD as an enzyme that eliminates the main cause of lipid peroxide production and suppresses the toxicity of active oxygen most strongly. We have also revealed that this enzyme is present in relatively large amounts in the placenta and is stable. Therefore, the present inventors conducted research on the production VC of placenta extract containing a high concentration of 5OD1, and found that it complies with the standards for conventional placenta extracts and also contains a high concentration of SOD (19
We have established a method for producing a placenta extract that contains D and SOD at a high concentration f! :I made sea bream. That is, the present invention relates to a placenta extract whose W characteristic is that it contains SOD at a high concentration, and a method for producing the same.
本発明においては、まず新鮮なヒト、ウシ、ブタ等の哺
乳動物の胎盤が原料として使用されるが、これらの凍結
保存品も使用可能である。このような原料胎盤を冷生理
食塩水にて充分に洗浄した後、冷却下細かく切断するか
もしくは凍結粉砕を行い、これに４１１１Ｍ水およびｎ
−ブタノールを添加してホモジナイズする。精製水およ
びブタノールの使用ｊｌハ、夫々１〜２倍盪でよい。ホ
モジナイズが充分に行われたならば、このものを８０℃
の湯浴中で加温し、内容物が４５−６５℃になつ念時点
で氷冷する。水冷後、約８０００回転／分にて２０分間
、４℃にて遠心分離を行って上清液を集める。In the present invention, fresh placenta from mammals such as humans, cows, and pigs is first used as a raw material, but cryopreserved products thereof can also be used. After sufficiently washing such a raw placenta with cold physiological saline, it is cut into small pieces under cooling or freeze-pulverized, and then added with 4111M water and n
- Add butanol and homogenize. Purified water and butanol may be used in an amount of 1 to 2 times each. Once homogenization is complete, heat the mixture to 80°C.
Warm in a hot water bath, and cool on ice until the contents reach 45-65°C. After cooling with water, centrifugation is performed at about 8,000 rpm for 20 minutes at 4° C. to collect the supernatant.
この上清液は淡黄色で筐明であり一、ＳＯＤ活性は１０
０単位／ｍ１以上を含有して−る。この上清液ば、必要
により、Ｃへ１−セルロースなどｏカラム・クロマトグ
ラフィを行って、より高虜度［ＳＯＤを含有する胎盤エ
キスを得ること７つ；できる。このようにして得られた
１治盤エキスにはＳＯＤ活性が１００単位／づ以上含ま
れており、ものによっては１０００単位／ｍ）以上と高
羨度であった。また、有用な酵素の指標となっているア
ルカリ性ホスファターゼ活性は３００単位７ｍ１（キン
グ。アームストロング単位）から１５０００単位と高濃
度Ｖこ存在してｂた。本胎盤エキスに０．０１−０．０
５チの一度になるようにパラベンを添加することにより
、腐敗、白濁等の変化をおさえることができる。This supernatant liquid was pale yellow and clear, and the SOD activity was 10.
Contains 0 units/ml or more. If necessary, this supernatant liquid can be subjected to o-column chromatography such as 1-cellulose to obtain a placenta extract containing SOD with a higher concentration. The thus obtained Ichiban extract contained SOD activity of 100 units/m or more, and in some cases had a high level of SOD activity of 1000 units/m or more. In addition, alkaline phosphatase activity, which is an indicator of a useful enzyme, was present in high concentrations ranging from 300 units (King Armstrong units) to 15,000 units (7ml). 0.01-0.0 for this placenta extract
By adding parabens at once, it is possible to suppress changes such as spoilage and cloudiness.
以下に本発明の実施例を示すが、これによって本発明を
制限するものではない。Examples of the present invention are shown below, but the present invention is not limited thereto.
なお、実施例中のアルカリホスファターゼ活性測定法お
よびＳＯＤ活性測定法は以下に示す方法によつ比。In addition, the alkaline phosphatase activity measurement method and the SOD activity measurement method in the examples are based on the methods shown below.
注、アルカリホスファターゼ活性測定法Ｋｉｎｄ−Ｋｉ
ｎｇ変法（Ｊ、Ｃｌ１ｎ、Ｐａｔｈｏｌ、、７，３２２
゜（１９５４））に従す、検体１ ｄｌが、１５分間に
遊離するフェノールの１■をもって、■キング・アーム
ストロング（Ｋ −Ａ）単位とした。Note: Alkaline phosphatase activity assay Kind-Ki
ng modified method (J, Cl1n, Pathol, 7,322
(1954)), the amount of phenol released per 1 dl of sample in 15 minutes was defined as a King Armstrong (K-A) unit.
スーパーオキシドジスムターゼ活性測定法Ｍｃｃｏｒｄ
とＦｒ１ｄｏｖｉｃｈの方法ＣＪ、Ｂｉｏｌ、Ｃｈｅｍ
、。Superoxide dismutase activity assay McCord
and Fr1dovich's method CJ, Biol, Chem
,.
２４４．６０４９．（１９６９））に従論、４トクロー
ムＣの還元速、Ｖを５０％阻害する酸を１単位とし友。244.6049. (1969)), the reduction rate of 4-tochrome C is determined by using one unit of acid that inhibits V by 50%.
実施例 ｌ
ヒトの新鮮な胎盤３００ｇを１０倍量の冷生理食塩水Ｃ
でて十分洗滌したのち、４℃にて細切すし念。このもの
に２倍量の精製水と２倍量のｎ−ブタノールを加え、ホ
モジナイズした（約１．５４’）。Example l 300g of fresh human placenta was added to 10 times the volume of cold physiological saline C.
After washing thoroughly, cut into pieces at 4℃. To this was added twice the amount of purified water and twice the amount of n-butanol, and homogenized (approximately 1.54').
その後、８０℃の湯浴中で内容物を撹拌しながら加温し
、内容物が６５℃になった時点で、氷冷した。今後遠心
分離を行い（８０００回転／分、２０分間、４℃）上清
を集めｅ（０，８１）。このものは淡黄色でわずかに生
体臭を有する澄明な液体であり、アルカリホスファター
ゼの活性は１５０００単位（キング・アームストロング
単位）であり、ＳＯＤ活性は１２００単位／ｄであった
。本胎盤エキスに０．０５％になるように、パラベンを
添加することによシ、腐敗、白濁等の変化を１年以内に
は観察できなかった。Thereafter, the contents were heated while stirring in a water bath at 80°C, and when the contents reached 65°C, they were cooled on ice. Thereafter, centrifugation was performed (8000 rpm, 20 minutes, 4°C) and the supernatant was collected e(0,81). This was a pale yellow, clear liquid with a slight biological odor, and the alkaline phosphatase activity was 15,000 units (King Armstrong units), and the SOD activity was 1,200 units/d. By adding paraben to this placenta extract to a concentration of 0.05%, no changes such as rot, rot, or cloudiness were observed within one year.
実施例 ２
新鮮な牛脂盤２ Ｋｑを冷生理食塩水にて十分洗滌した
。その後凍結粉砕を行い、その粉砕物にＮ農水４１％ｎ
−ブタノール２１を加え、よく撹拌した。次に５ｏｏｏ
回転／分、４℃、２０分間遠心分離を行い、上清を集め
た。集めた上清を湯浴中で４５℃にまで加温した後、友
だちに氷冷し之。Example 2 Fresh beef tallow disc 2 Kq was thoroughly washed with cold physiological saline. After that, freeze pulverization is performed, and the pulverized product contains 41% N agricultural water.
-Butanol 21 was added and stirred well. Next 5ooo
Centrifugation was performed at 4° C. for 20 minutes at revolutions/minute, and the supernatant was collected. The collected supernatant was heated to 45°C in a hot water bath, and then cooled on ice by a friend.
その後、再度、上記条件にて遠心分離を行い、上溝’ｔ
３７℃にて減圧濃縮した。この濃縮物を精製水で平衡化
し７’ｈＣＭ−セルロースカラム（３ｃｐｓ Ｘ２０α
）に添加し、素通シ画分を集め胎盤エキスを製造し念。After that, centrifugation is performed again under the above conditions, and the upper groove't
It was concentrated under reduced pressure at 37°C. This concentrate was equilibrated with purified water and loaded onto a 7'hCM-cellulose column (3 cps
) and collect the fractions to produce placenta extract.
ここで得られ次胎盤エキスのアルカリホスファターゼ活
性は、３００単位（キング・アームス）ａング単位）で
あり、ＳＯＤ活性は８００単位／ｄであり之。ま之、本
品は、淡黄色から黄色の澄明な液体であり、わずかに生
体臭を有し、０．０３％のパラベンの添加によ４５１０
℃の保存下で１年以上安定であつ九。The alkaline phosphatase activity of the placental extract obtained here was 300 units (King Arms), and the SOD activity was 800 units/d. However, this product is a clear liquid with a pale yellow to yellow color and has a slight biological odor.
Stable for over 1 year when stored at ℃.
実施例 ３
実施例１にて得られた胎盤エキスを精製水で１０倍希釈
した。次に酸性処理を行い、中性にした活性炭カラム（
５ｍＸ ｌ Ｏｃｍ）を流下させることにより、澄明な
液体を得次。本品のアルカリホスファターゼ活性は、１
０５０単位（キング・アームス）ｏング単位）であり、
ＳＯＤ活性は１０５単位／ｄであり、淡黄色であった。Example 3 The placenta extract obtained in Example 1 was diluted 10 times with purified water. Next, an activated carbon column (
A clear liquid was then obtained by flowing down 5 mX l Ocm). The alkaline phosphatase activity of this product is 1
050 units (King Arms),
The SOD activity was 105 units/d, and the color was pale yellow.
また、無臭であツタ。パラベンを０．０３１になるよう
に添加することにより２年以上安定であった。It is also odorless and ivy. It was stable for more than 2 years by adding paraben to 0.031.
Claims (6)
ることを特徴とする胎盤エキス。(1) A placenta extract characterized by containing superoxide dismutase at a high concentration.
／ｍｌ以上である特許請求の範囲第１項記載の胎盤エキ
ス。(2) The placenta extract according to claim 1, which has a superoxide dismutase concentration of 100 units/ml or more.
／ｍｌ以上、およびアルカリ性ホスファターゼ濃度が１
００キング・アームストロング単位以上である特許請求
の範囲第１項記載の胎盤エキス。(3) Superoxide dismutase concentration is 100 units/ml or more and alkaline phosphatase concentration is 1
The placenta extract according to claim 1, which has a content of 00 King Armstrong units or more.
の範囲第１項記載の胎盤エキス。(4) The placenta extract according to claim 1, wherein the placenta extract is a mammalian placenta extract.
請求の範囲第４項記載の胎盤エキス。(5) The placenta extract according to claim 4, wherein the placenta is fresh human, bovine, or porcine placenta.
理食塩水にて充分に洗滌し、冷却下細切りしたものに、
精製水およびｎ−ブタノールを加えてホモジナイズし、
これを加温した後氷冷し、このものを８０００回転／分
で２０分間遠心分離した上清を分取することよりなるス
ーパーオキシドジスムターゼを高濃度に含有する胎盤エ
キスの製造法。(6) Fresh mammalian placenta such as human, bovine, porcine, etc., thoroughly washed with cold physiological saline and cut into thin pieces under cooling.
Add purified water and n-butanol and homogenize,
A method for producing a placenta extract containing a high concentration of superoxide dismutase, which comprises heating the product, cooling it on ice, centrifuging the product at 8,000 rpm for 20 minutes, and collecting the supernatant.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60117919A JPS61277626A (en) | 1985-05-31 | 1985-05-31 | Placenta extract containing superoxide dismutase in high concentration and production thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60117919A JPS61277626A (en) | 1985-05-31 | 1985-05-31 | Placenta extract containing superoxide dismutase in high concentration and production thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPS61277626A true JPS61277626A (en) | 1986-12-08 |
Family
ID=14723423
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP60117919A Pending JPS61277626A (en) | 1985-05-31 | 1985-05-31 | Placenta extract containing superoxide dismutase in high concentration and production thereof |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS61277626A (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63254181A (en) * | 1987-04-09 | 1988-10-20 | Kobayashi Kooc:Kk | Antioxidant composition |
| US7374754B2 (en) | 2003-09-02 | 2008-05-20 | Essential Skincare, Llc | Use of placental alkaline phosphatase to promote skin cell proliferation |
| WO2010138024A3 (en) * | 2009-05-29 | 2011-02-10 | Общество С Ограниченной Ответственностью "Медицинский Исследовательский Центр "Иммункулус" | Agent for protecting brain cells |
| US8101211B2 (en) | 2000-12-15 | 2012-01-24 | Kabushiki Kaisha Yakult Honsha | Compositions for retarding skin aging |
| US8460654B2 (en) | 2005-12-28 | 2013-06-11 | Zoltan Laboratories Llc | Use of alkaline phosphatase to maintain healthy tissue mass in mammals |
| CN103181553A (en) * | 2011-12-30 | 2013-07-03 | 广州拜迪生物医药有限公司 | Pig placentin production method |
| US8603464B2 (en) | 2002-12-12 | 2013-12-10 | Zoltan Laboratories Llc | Alkaline phosphatase to reduce weight gain or induce weight loss |
-
1985
- 1985-05-31 JP JP60117919A patent/JPS61277626A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63254181A (en) * | 1987-04-09 | 1988-10-20 | Kobayashi Kooc:Kk | Antioxidant composition |
| JPH0781140B2 (en) * | 1987-04-09 | 1995-08-30 | 株式会社コーセー | Antioxidant composition |
| US8101211B2 (en) | 2000-12-15 | 2012-01-24 | Kabushiki Kaisha Yakult Honsha | Compositions for retarding skin aging |
| US8603464B2 (en) | 2002-12-12 | 2013-12-10 | Zoltan Laboratories Llc | Alkaline phosphatase to reduce weight gain or induce weight loss |
| US7374754B2 (en) | 2003-09-02 | 2008-05-20 | Essential Skincare, Llc | Use of placental alkaline phosphatase to promote skin cell proliferation |
| US8460654B2 (en) | 2005-12-28 | 2013-06-11 | Zoltan Laboratories Llc | Use of alkaline phosphatase to maintain healthy tissue mass in mammals |
| WO2010138024A3 (en) * | 2009-05-29 | 2011-02-10 | Общество С Ограниченной Ответственностью "Медицинский Исследовательский Центр "Иммункулус" | Agent for protecting brain cells |
| CN103181553A (en) * | 2011-12-30 | 2013-07-03 | 广州拜迪生物医药有限公司 | Pig placentin production method |
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