JP4578821B2 - Pharmaceutical composition for the treatment of nephritis - Google Patents
Pharmaceutical composition for the treatment of nephritis Download PDFInfo
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- JP4578821B2 JP4578821B2 JP2004043523A JP2004043523A JP4578821B2 JP 4578821 B2 JP4578821 B2 JP 4578821B2 JP 2004043523 A JP2004043523 A JP 2004043523A JP 2004043523 A JP2004043523 A JP 2004043523A JP 4578821 B2 JP4578821 B2 JP 4578821B2
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- nephritis
- lactalbumin
- pharmaceutical composition
- treatment
- active oxygen
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- 201000008383 nephritis Diseases 0.000 title claims description 14
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- 108090000942 Lactalbumin Proteins 0.000 claims description 19
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 19
- 239000001301 oxygen Substances 0.000 claims description 19
- 229910052760 oxygen Inorganic materials 0.000 claims description 19
- 235000021241 α-lactalbumin Nutrition 0.000 claims description 18
- 230000000694 effects Effects 0.000 claims description 16
- 102000016938 Catalase Human genes 0.000 claims description 9
- 108010053835 Catalase Proteins 0.000 claims description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 7
- 201000010099 disease Diseases 0.000 claims description 6
- 239000004480 active ingredient Substances 0.000 claims description 5
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- 235000013336 milk Nutrition 0.000 description 8
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- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 230000009471 action Effects 0.000 description 4
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- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
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- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
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- 238000002835 absorbance Methods 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 239000005018 casein Substances 0.000 description 2
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 2
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- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 description 1
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Description
本発明は、α−ラクトアルブミンを有効成分とする腎炎の治療用医薬組成物に関する。 The present invention relates to a pharmaceutical composition for treating nephritis comprising α-lactalbumin as an active ingredient.
活性酸素とは、スーパーオキサイドや過酸化水素、ヒドロキシラジカル、一重項酸素等を指し、安定な通常の酸素と異なり反応性に富んでいる。マクロファージは自ら活性酸素を生成し、異物を攻撃し排除している。しかし、過剰に産生した活性酸素は、正常細胞にも作用するので、生体に対して種々の障害も引き起こすことが知られている。例えば、脂質の過酸化による膜の損傷、タンパク質の酸化修飾によるタンパク質の構造変化、ＤＮＡの切断等の結果、細胞に障害作用を示し、様々な疾病の原因ともなることが明らかにされている。 Active oxygen refers to superoxide, hydrogen peroxide, hydroxy radical, singlet oxygen, and the like, and is rich in reactivity unlike normal normal oxygen. Macrophages themselves generate active oxygen and attack and eliminate foreign substances. However, it is known that the excessively produced active oxygen also acts on normal cells and thus causes various damages to the living body. For example, as a result of damage to the membrane due to lipid peroxidation, structural change of the protein due to oxidative modification of the protein, DNA cleavage, etc., it has been clarified that it shows a damaging effect on cells and causes various diseases.
抗酸化作用を有する食品成分としては、β−ラクトグロブリンが知られているが、食品中の酸化防止用（特許文献１参照）であり、生体内の活性酸素を低減する作用を有しているかは不明である。
安全で且つ活性酸素低減作用（活性酸素濃度を減少させる作用のみならず、これに起因して生じると考えられる種々の病気の予防、改善の作用をも含む。）を有し、医薬に用いられるものがあれば、人の健康にとって非常に有用であり、その必要性は極めて大きい。近年のようにストレスが多く、それに起因する多くの病気が発生している状況においては特にその必要性が大きいことから、安全で且つ優れた活性酸素低減作用を有する製品の開発が強く望まれていた。
本発明は、上記現状に鑑みてなされたものであり、乳由来のα−ラクトアルブミンを原料として用い、生体内において安全で且つ優れた活性酸素低減効果を有する腎炎の治療用医薬組成物を提供することを目的とする。
Has a safe and active oxygen-reducing action (not only act to reduce the active oxygen concentration, prevention of various diseases which are thought to occur due to this, including the effects of improvement.), Used in the pharmaceutical if there is, is very useful to take to human health, the need is very large. The development of a product that is safe and has an excellent action for reducing active oxygen is strongly desired, especially in the situation where there is a lot of stress as in recent years and many diseases resulting from it have occurred. It was.
The present invention has been made in view of the above situation, and provides a pharmaceutical composition for treating nephritis that is safe in vivo and has an excellent effect of reducing active oxygen, using α-lactalbumin derived from milk as a raw material. The purpose is to do.
以上の観点より、本発明者等は、乳由来組成物に着目して種々検討したところ、α−ラクトアルブミンが、生体内において活性酸素低減作用を有し、腎炎の治療用医薬組成物の有効成分となりうることを明らかにし、本発明を完成するに至ったものである。 From the above viewpoint, the present inventors have made various study focusing on the milk-derived compositions, alpha-lactalbumin, have a reactive oxygen reduction effect in vivo, the effective of a pharmaceutical composition for the treatment of nephritis It has been clarified that it can be a component, and the present invention has been completed.
すなわち、本発明は、
（１）α−ラクトアルブミンを有効成分として５〜１０％含有してなる、腎炎疾患時のカタラーゼ活性増強により活性酸素を低減させるための腎炎の治療用医薬組成物、
（２）α−ラクトアルブミン含有量が９〜１０％である（１）記載の腎炎の治療用医薬組成物、
からなる。
That is, the present invention
(1) alpha-lactalbumin comprising 5-10% as an active ingredient, a pharmaceutical composition for the treatment of nephritis to reduce the active oxygen by catalase activity enhancement during nephritic diseases,
(2) The pharmaceutical composition for the treatment of nephritis according to (1), wherein the α-lactalbumin content is 9 to 10%,
Consists of.
本発明に係る組成物は、生体内における活性酸素低減作用に優れることから、活性酸素低減作用に起因する効果に有効である。従って、本発明に係る組成物は、活性酸素種が原因となる腎炎に優れた効果を発揮するものと考えられる。
また、本発明に係る組成物に含まれるα−ラクトアルブミンは乳由来であることから、吸収効果に優れると共に、経口投与により簡易に摂取でき、有害な成分及び重金属は認められず安全である。
Since the composition according to the present invention is excellent in the active oxygen reducing action in a living body, it is effective for the effect resulting from the active oxygen reducing action. Thus, the composition according to the present invention is believed to exhibit excellent effects on nephritis active oxygen species are responsible.
Moreover, since α-lactalbumin contained in the composition according to the present invention is derived from milk, it has an excellent absorption effect and can be easily ingested by oral administration, and no harmful components and heavy metals are recognized and is safe.
本発明は、α−ラクトアルブミンを有効成分とする、腎炎疾患時のカタラーゼ活性増強により活性酸素を低減させるための腎炎の治療用医薬組成物を提供する。本発明は、乳由来タンパク質の１つであるα−ラクトアルブミンにカタラーゼ活性増強作用を有することを見出したことに基づくものである。 The present invention provides a pharmaceutical composition for the treatment of nephritis comprising α-lactalbumin as an active ingredient for reducing active oxygen by enhancing catalase activity during nephritis disease . The present invention is based on the finding that α-lactalbumin, one of milk-derived proteins, has a catalase activity enhancing action.
したがって、本発明のα−ラクトアルブミンは、生体内において血清中の活性酸素低減作用を示す。該α−ラクトアルブミンは、医薬組成物の有効成分として添加することが可能である。 Therefore, α-lactalbumin of the present invention exhibits an action of reducing active oxygen in serum in vivo. The α-lactalbumin can be added as an active ingredient of a pharmaceutical composition .
α−ラクトアルブミンとは、乳の主要タンパク質で、乳汁中に少量（牛乳中に約0.5%）含まれるアルブミンである。本発明ではα−ラクトアルブミンを５〜１０％、さらに好ましくは９〜１０％用いる。α−ラクトアルブミンは精製物を用いてもよいが、乳またはホエイに含まれているのでこれらをそのまま用いてもよいし、粗精製したものを用いても構わない。 α-Lactalbumin is a major protein of milk and is an albumin contained in a small amount in milk (about 0.5% in milk). In the present invention, α-lactalbumin is used in an amount of 5 to 10%, more preferably 9 to 10% . α-Lactalbumin may be used as a purified product, but since it is contained in milk or whey, these may be used as they are, or a crude product may be used.
過酸化水素は、細胞内に浸透しやすく、そこで鉄や銅イオンと結びついてしまえば、たちまちハイドロキシラジカルという活性酸素に変身してしまう性格を持っている。また、スーパーオキサイドラジカルと反応することで、ハイドロキシラジカルや一重項酸素に変わったりもする。このように、毒性の高い化合物が過酸化水素である。
一方、カタラーゼは、過酸化水素を水と酸素に変換する生体内酵素で、酵素活性が低下すると、活性酸素が体内にたまり、疾病を誘発しやすくなる。
Hydrogen peroxide easily penetrates into cells, and if it is combined with iron or copper ions, it has the property of being transformed into active oxygen called a hydroxyl radical. Moreover, by reacting with a superoxide radical, it may be changed to a hydroxy radical or singlet oxygen. Thus, hydrogen peroxide is a highly toxic compound.
Catalase, on the other hand, is an in vivo enzyme that converts hydrogen peroxide into water and oxygen. When the enzyme activity decreases, active oxygen accumulates in the body, making it easier to induce disease.
α−ラクトアルブミンを製剤化して医薬品とする場合には、治療目的や投与経路等に応じて剤形を選択することができ、例えば、錠剤、丸剤、散剤、液剤、懸濁剤、乳剤、顆粒剤、カプセル剤、注射剤、坐剤、エリキシル剤、シロップ剤、浸剤、煎剤等が挙げられる。また製剤化のために、必要に応じて充填剤、増量剤、結合剤、保湿剤、崩壊剤、界面活性剤、滑沢剤等の希釈剤あるいは賦形剤を用いることができる。また、この医薬製剤中に着色剤、保存剤、香料、風味剤、甘味剤等や他の医薬品を医薬製剤中に含有させてもよい。 When α-lactalbumin is formulated into a pharmaceutical product, the dosage form can be selected according to the therapeutic purpose, administration route, etc., for example, tablets, pills, powders, solutions, suspensions, emulsions, Examples include granules, capsules, injections, suppositories, elixirs, syrups, soaking agents, and decoction. For formulation, diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and lubricants can be used as necessary. In addition, a colorant, a preservative, a fragrance, a flavoring agent, a sweetening agent, and other pharmaceuticals may be included in the pharmaceutical preparation.
以下、実施例により本発明を具体的に説明するが、本発明は、これら実施例に制限されるものではない。 EXAMPLES Hereinafter, although an Example demonstrates this invention concretely, this invention is not restrict | limited to these Examples.
[実施例１]
Wistar系雄ラット（３週齢）を１週間CE-2（日本クレア株式会社製、水分8.93%、粗タンパク質25.40%、粗脂肪4.43%、粗繊維4.13%、粗灰分6.93%、可溶性無窒素物50.18%）で飼育し、その後５日間は20%カゼイン標準食（20C、表１）を水道水と共に自由摂取させ予備飼育を行った。購入日から予備飼育９日目まで５室ケージで飼育し、10日目より代謝ケージに移した。
実験開始日（０日目）に各群の体重が等しくなるように３群（６頭／群）に分け、腎炎２群には、抗ラット糸球体基底膜ウサギ抗血清0.25mL/ratをWistar系雄ラットの尾静脈に注射し、糸球体腎炎を惹起させた。翌日（１日目）、ウサギγ−グロブリン(8mg/0.1mL生理食塩水/rat）：Freund's complete adjuvant=1:1のエマルジョンを後肢フットパッドに0.2mL/rat皮下注射した。実験０日目から各群共に実験食として、20Cおよび9%ラクトアルブミン食（9α-La、表１）をmiliQ水で固めたものを14日間自由摂取させた。20Cを摂取する正常群も設けた。実験開始14日目の午前９時に飼料を取り除き、水は与えたまま、午後１時に肝臓を採取した。
[Example 1]
Wistar male rats (3 weeks old) for 1 week CE-2 (manufactured by CLEA Japan, moisture 8.93%, crude protein 25.40%, crude fat 4.43%, crude fiber 4.13%, crude ash content 6.93%, soluble nitrogen-free substance 50.18%), and for the next 5 days, 20% casein standard diet (20C, Table 1) was freely ingested with tap water and preliminarily reared. The animals were reared in a five-chamber cage from the date of purchase to the 9th day of preliminary breeding, and transferred to the metabolic cage from the 10th day.
The group was divided into 3 groups (6 animals / group) so that the weight of each group would be equal on the start day of experiment (day 0), and anti-rat glomerular basement membrane rabbit antiserum 0.25 mL / rat was added to Wistar for 2 groups of nephritis. Injected into the tail vein of male male rats to induce glomerulonephritis. The next day (Day 1), rabbit γ-globulin (8 mg / 0.1 mL saline / rat): Freund's complete adjuvant = 1: 1 emulsion was subcutaneously injected into the hind leg footpad at 0.2 mL / rat. From day 0 of the experiment, each group was allowed to freely ingest a 20C and 9% lactalbumin diet (9α-La, Table 1) solidified with miliQ water as an experimental diet for 14 days. There was also a normal group taking 20C. At 9 am on the 14th day from the start of the experiment, the feed was removed, and the liver was collected at 1 pm with water.
[抗酸化酵素活性の測定]
ラット解剖時に、肝臓0．2gを精秤し、液体窒素で氷結させ、−84℃で保存した。試験管に0.1％TritonX-100-50mMリン酸緩衝液を5mL入れ、ポリトロンで氷冷下ホモジナイズした。続いて、超遠心用チューブに氷冷下0.1％TritonX-100-50mMリン酸緩衝液5mLで共洗いしながら移した後、19000×g、4℃、30分間超遠心した。上清をバイアルに分注し、測定まで冷凍保存した。
測定試料（酵素液）2mLに30mM H2O2-50mMリン酸緩衝液を1mLを加え、攪拌し240nmで吸光度の経時変化を測定した（15秒間）。対照は、酵素液2mLに50mMリン酸緩衝液を1mL加え、攪拌し240nmで吸光度を測定したものを用いた。
その結果、対照群に比べ、20C摂取群は腎炎によるカタラーゼ活性が低下するが、α−ラクトアルブミン摂取群は腎炎によるカタラーゼ活性低下を抑制するばかりでなく、対照群に比べても有意にカタラーゼ活性を上昇させることがわかった（図１）。
[Measurement of antioxidant enzyme activity]
At the time of rat dissection, 0.2 g of the liver was precisely weighed, frozen with liquid nitrogen, and stored at -84 ° C. 5 mL of 0.1% TritonX-100-50 mM phosphate buffer was added to the test tube, and homogenized with polytron under ice-cooling. Subsequently, the sample was transferred to an ultracentrifuge tube while being washed with 5 mL of 0.1% TritonX-100-50 mM phosphate buffer under ice cooling, and then ultracentrifuged at 19000 × g, 4 ° C. for 30 minutes. The supernatant was dispensed into vials and stored frozen until measurement.
1 mL of 30 mM H 2 O 2 -50 mM phosphate buffer was added to 2 mL of the measurement sample (enzyme solution), stirred, and the change with time in absorbance was measured at 240 nm (15 seconds). As a control, 1 mL of 50 mM phosphate buffer was added to 2 mL of enzyme solution, and the mixture was stirred and the absorbance was measured at 240 nm.
As a result, compared to the control group, the catalase activity due to nephritis decreased in the 20C intake group, but the α-lactalbumin intake group not only suppressed the decrease in catalase activity due to nephritis, but also significantly reduced the catalase activity compared to the control group. (Fig. 1).
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