JPS6237611B2 - - Google Patents
Info
- Publication number
- JPS6237611B2 JPS6237611B2 JP56018429A JP1842981A JPS6237611B2 JP S6237611 B2 JPS6237611 B2 JP S6237611B2 JP 56018429 A JP56018429 A JP 56018429A JP 1842981 A JP1842981 A JP 1842981A JP S6237611 B2 JPS6237611 B2 JP S6237611B2
- Authority
- JP
- Japan
- Prior art keywords
- immune complex
- immune
- protease
- acidic
- human igg
- 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.)
- Expired
Links
- 208000024781 Immune Complex disease Diseases 0.000 claims description 11
- 108091005508 Acid proteases Proteins 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 6
- 229940124597 therapeutic agent Drugs 0.000 claims description 5
- 230000002485 urinary effect Effects 0.000 claims description 4
- 239000004480 active ingredient Substances 0.000 claims description 3
- 102000035195 Peptidases Human genes 0.000 description 18
- 108091005804 Peptidases Proteins 0.000 description 18
- 239000004365 Protease Substances 0.000 description 18
- 230000002378 acidificating effect Effects 0.000 description 18
- 230000000694 effects Effects 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 238000012360 testing method Methods 0.000 description 6
- 230000007062 hydrolysis Effects 0.000 description 5
- 238000006460 hydrolysis reaction Methods 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- 241000283973 Oryctolagus cuniculus Species 0.000 description 4
- 230000000295 complement effect Effects 0.000 description 4
- 230000008021 deposition Effects 0.000 description 4
- 241000700159 Rattus Species 0.000 description 3
- 102000004142 Trypsin Human genes 0.000 description 3
- 108090000631 Trypsin Proteins 0.000 description 3
- 239000000427 antigen Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000005227 gel permeation chromatography Methods 0.000 description 3
- 210000000265 leukocyte Anatomy 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 206010043778 thyroiditis Diseases 0.000 description 3
- 239000012588 trypsin Substances 0.000 description 3
- 210000002700 urine Anatomy 0.000 description 3
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 229920005654 Sephadex Polymers 0.000 description 2
- 239000012507 Sephadex™ Substances 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229940124599 anti-inflammatory drug Drugs 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 201000008383 nephritis Diseases 0.000 description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 238000000159 protein binding assay Methods 0.000 description 2
- QZAYGJVTTNCVMB-UHFFFAOYSA-N serotonin Chemical compound C1=C(O)C=C2C(CCN)=CNC2=C1 QZAYGJVTTNCVMB-UHFFFAOYSA-N 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000000451 tissue damage Effects 0.000 description 2
- 231100000827 tissue damage Toxicity 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- LPXQRXLUHJKZIE-UHFFFAOYSA-N 8-azaguanine Chemical compound NC1=NC(O)=C2NN=NC2=N1 LPXQRXLUHJKZIE-UHFFFAOYSA-N 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 102000003908 Cathepsin D Human genes 0.000 description 1
- 108090000258 Cathepsin D Proteins 0.000 description 1
- 102000004178 Cathepsin E Human genes 0.000 description 1
- 108090000611 Cathepsin E Proteins 0.000 description 1
- 102000029816 Collagenase Human genes 0.000 description 1
- 108060005980 Collagenase Proteins 0.000 description 1
- GUBGYTABKSRVRQ-WFVLMXAXSA-N DEAE-cellulose Chemical compound OC1C(O)C(O)C(CO)O[C@H]1O[C@@H]1C(CO)OC(O)C(O)C1O GUBGYTABKSRVRQ-WFVLMXAXSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 102000001554 Hemoglobins Human genes 0.000 description 1
- 108010054147 Hemoglobins Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000016387 Pancreatic elastase Human genes 0.000 description 1
- 108010067372 Pancreatic elastase Proteins 0.000 description 1
- 241001092459 Rubus Species 0.000 description 1
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 1
- 102000009524 Vascular Endothelial Growth Factor A Human genes 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- 238000010306 acid treatment Methods 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 238000005377 adsorption chromatography Methods 0.000 description 1
- 108010027597 alpha-chymotrypsin Proteins 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 229940127218 antiplatelet drug Drugs 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 210000002469 basement membrane Anatomy 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 229960002424 collagenase Drugs 0.000 description 1
- 230000004154 complement system Effects 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000000875 corresponding effect Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 230000008394 fibrinolytic abnormality Effects 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 229960001340 histamine Drugs 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 208000024326 hypersensitivity reaction type III disease Diseases 0.000 description 1
- 230000000951 immunodiffusion Effects 0.000 description 1
- 229960003444 immunosuppressant agent Drugs 0.000 description 1
- 239000003018 immunosuppressive agent Substances 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 238000001155 isoelectric focusing Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 229950000964 pepstatin Drugs 0.000 description 1
- 108010091212 pepstatin Proteins 0.000 description 1
- FAXGPCHRFPCXOO-LXTPJMTPSA-N pepstatin A Chemical compound OC(=O)C[C@H](O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)C[C@H](O)[C@H](CC(C)C)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)NC(=O)CC(C)C FAXGPCHRFPCXOO-LXTPJMTPSA-N 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229940012957 plasmin Drugs 0.000 description 1
- 239000000106 platelet aggregation inhibitor Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 201000001474 proteinuria Diseases 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 229940076279 serotonin Drugs 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 238000003307 slaughter Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 229940126585 therapeutic drug Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000001541 thymus gland Anatomy 0.000 description 1
- 210000001685 thyroid gland Anatomy 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 208000025883 type III hypersensitivity disease Diseases 0.000 description 1
- 230000008728 vascular permeability Effects 0.000 description 1
- 230000002227 vasoactive effect Effects 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Description
本発明はヒト尿中酸性プロテアーゼを有効成分
とする免疫複合体病治療剤に関する。
慢性関節リウマチ、全身紅斑性狠瘡（SLE）あ
るいはルーブス腎炎に代表される免疫複合体病
（イミユーン コンプレツクス デイズイーズ、
immune complex disease）はその名の通り、
種々の抗原と抗体との複合体すなわち免疫複合体
により惹き起こされる疾患である。免疫複合体病
の発症機序は複雑であり、不明な点も多いが、お
およそ次のような経過をたどるものと考えられて
いる。すなわち、細菌やウイルス感染などによつ
て組織に障害が起こると、新たに出現した自己抗
原やウイルス感染細胞などに対する抗体が産生さ
れ、これらの抗体は対応する抗原と反応して免疫
複合体をを形成する。免疫複合体は補体系、血小
板などを活性化し、ヒスタミン、セロトニンなど
の血管作働物質を遊離するため血管透過性が亢進
する。次いで、流血中の免疫複合体が透過性の亢
進した血管壁に入り込み、基底膜に沿つて沈着す
る。沈着した免疫複合体に補体が作用して生成さ
れる白血球遊走因子によつて免疫複合体の沈着部
位に多核白血球が集簇する。多核白血球は免疫複
合体と反応すると、種々の組織障害物質、例えば
カテプシンＤ，Ｅ、コラゲナーゼ、エラスター
ゼ、血管透過性因子（パーミアビリテイーフアク
ター、permeability factor）などを放出し、これ
らの物質によつて遂に組織障害が惹き起こされ
る。SLEなどの免疫複合体病患者の血中補体価は
一般に低く、病状の悪化と補体価の低下は密接に
相関し、この補体価の低下は抗原−抗体反応部
位、例えば腎、血管などで多量の補体が消費され
るためであるとされている。また、免疫複合体は
血液凝固系とも関連し、障害組織へのフイブリノ
イド沈着を促進するなど、その多様な障害機構に
よつて重篤な病変へと進展させるものと考えられ
ている。
現在、これら免疫複合体病の治療には、亢進し
た免疫系を抑制し、局所の炎症、疼痛を鎮めるた
めのステロイド剤をはじめとする免疫抑制剤や抗
炎症剤、あるいは血管内で生じた凝固線溶系の異
常を改善するための抗凝固剤や抗血小板剤などが
使用されている。しかしながら、これらの薬剤は
作用が弱く、副作用が強いため必ずしも満足でき
る治療効果が得られていないことから、安全且つ
治療効果の高い薬剤の開発が切望されている。
このような背景に鑑み、本発明者らは免疫複合
体病のより有効な治療剤を開発すべく、長年にわ
たり研究を重ねた結果、ヒト尿中酸性プロテアー
ゼが免疫複合体を特異的に分解することを見出
し、さらに、この酸性プロテアーゼが各種免疫複
合体病を著明に抑制することを確認し、本発明を
完成するに至つた。
本発明の免疫複合体病治療剤の有効成分である
酸性プロテアーゼは公知の酵素であるが〔ミルス
キーら（Mirsky et al.），ジヤーナル オブ ク
リニカル インベステイゲーシヨン（J.Clin.
Invest.）27巻、818頁、1948年〕、従来、免疫複
合体病に対する治療剤としては用いられたことが
なかつた。この酸性プロテアーゼは蛋白質を精製
する場合に用いる一般的方法、例えば塩析法、無
機吸着体による吸着クロマトグラフイー、イオン
交換樹脂によるイオン交換クロマトグラフイー、
分子ふるい効果を有するゲルクロマトグラフイー
などを適宜組み合わせることによりヒト尿から採
取することができる。
例えば、ヒト尿をセイフアーら（Seijffers）の
方法〔アメリカン ジヤーナル オブ フイジオ
ロジー（Amer.J.Physiol.）206巻、1106頁、1964
年〕に準じ、0.1M酢酸緩衝液（PH5.3）にて平衡
化したDEAE−セルロースカラムを通過させるこ
とにより、酸性プロテアーゼを吸着させたのち、
0.3Mの塩化ナトリウムを含む同緩衝液にて溶出
する。溶出液を濃縮後、0.9％の生理食塩水に膨
潤せしめたセフアデツクスＧ−100によるゲルク
ロマトグラフイーにてさらに精製し、酸処理を行
うことにより本発明の酸性プロテアーゼを得るこ
とができる。
本法により得た酸性プロテアーゼは、セフアデ
ツクスＧ−100ゲルクロマトグラフイーによる分
析の結果、分子量32000−38000であり、アンフオ
ライン等電点電気泳動法による等電点は１〜３で
あり極大吸収278nm、ニンヒドリン反応陽性、水
に易溶、エーテル、クロロホルムに不溶である。
また、この酸性プロテアーゼは、PH7.0以下の酸
性領域にて、ヘモグロビンに対して高い水解活性
を示すが、ペプスタチンによつて著明に抑制され
る性質を有する。また、この酸性プロテアーゼは
PH7.0以下の酸性領域にて安定、PH8.0以上のアル
カリ性にて不安定である。以下、この酸性プロテ
アーゼの薬理作用および毒性を試験例にて説明す
る。
試験例１ 免疫複合体水解作用
可溶性免疫複合体〔ヒトIgG−抗ヒトIgG抗体
（家兎）〕15mgと酸性プロテアーゼ、トリプシン、
α−キモトリプシンあるいはプラスミン各３mgを
１mlのリン酸緩衝液（0.06M，PH6.0）に溶解
し、37℃で60分間インキユベーシヨンした。20％
過塩素酸溶液１mlを加え反応を停止せしめ、上清
の280nmの吸光度を測定した。また、免疫複合体
の代りに、ヒトIgGを用い、同様の操作を行い、
両者の水解比を比較した。なお、水解活性はヒト
IgGを酸性プロテアーゼで処理したときの値を
100として相対値で表わした。結果を第１表に示
す。
酸性プロテアーゼは他のプロテアーゼに比し、
正常ヒトIgGよりも可溶性免疫複合体を選択的に
水解した。
The present invention relates to a therapeutic agent for immune complex diseases containing human urinary acid protease as an active ingredient. Rheumatoid arthritis, systemic erythematosus erythematosus (SLE), and immune complex diseases represented by rubus nephritis.
As the name suggests, immune complex disease)
It is a disease caused by complexes of various antigens and antibodies, that is, immune complexes. Although the onset mechanism of immune complex diseases is complex and there are many unknown points, it is thought that the disease progresses roughly as follows. In other words, when tissue damage occurs due to bacterial or viral infection, antibodies against newly appeared self-antigens or virus-infected cells are produced, and these antibodies react with the corresponding antigens to form immune complexes. Form. Immune complexes activate the complement system, platelets, etc., and release vasoactive substances such as histamine and serotonin, leading to increased vascular permeability. Immune complexes in the bloodstream then enter the hyperpermeable vessel walls and are deposited along the basement membrane. Multinucleated leukocytes gather at the site of immune complex deposition by leukocyte migration factors produced by the action of complement on the deposited immune complexes. When polynuclear leukocytes react with immune complexes, they release various tissue-damaging substances, such as cathepsin D and E, collagenase, elastase, and vascular permeability factor. Eventually, tissue damage is caused. Blood complement levels in patients with immune complex diseases such as SLE are generally low, and the worsening of the condition is closely correlated with the decrease in complement levels. This is thought to be due to the consumption of large amounts of complement. Immune complexes are also associated with the blood coagulation system, and are thought to lead to the development of serious lesions through various damage mechanisms, such as promoting fibrinoid deposition in damaged tissues. Currently, treatments for these immune complex diseases include immunosuppressants and anti-inflammatory drugs such as steroids to suppress the overactive immune system and calm local inflammation and pain, as well as anti-inflammatory drugs that suppress the overactive immune system and suppress local inflammation and pain. Anticoagulants and antiplatelet agents are used to improve fibrinolytic abnormalities. However, these drugs have weak effects and strong side effects, and therefore do not necessarily provide satisfactory therapeutic effects.Therefore, there is a strong desire for the development of safe and highly therapeutic drugs. In view of this background, the present inventors have conducted research over many years to develop more effective therapeutic agents for immune complex diseases, and as a result, they have discovered that human urinary acid protease specifically degrades immune complexes. They discovered that this acidic protease significantly suppresses various immune complex diseases, and completed the present invention. Acidic protease, which is the active ingredient of the therapeutic agent for immune complex diseases of the present invention, is a known enzyme [Mirsky et al., Journal of Clinical Investigation (J.Clin.
Invest.) Vol. 27, p. 818, 1948], and had never been used as a therapeutic agent for immune complex diseases. This acidic protease can be purified using general methods used to purify proteins, such as salting-out method, adsorption chromatography using an inorganic adsorbent, ion-exchange chromatography using an ion-exchange resin,
It can be collected from human urine by appropriately combining gel chromatography, etc., which has a molecular sieving effect. For example, the method of Seijffers et al. [American Journal of Physiol., Vol. 206, p. 1106, 1964]
After adsorbing acidic protease by passing it through a DEAE-cellulose column equilibrated with 0.1M acetate buffer (PH5.3),
Elute with the same buffer containing 0.3M sodium chloride. After concentrating the eluate, it is further purified by gel chromatography using Sephadex G-100 swollen in 0.9% physiological saline, followed by acid treatment to obtain the acidic protease of the present invention. The acidic protease obtained by this method has a molecular weight of 32,000-38,000 as a result of analysis using Sephadex G-100 gel chromatography, an isoelectric point of 1 to 3 using ampholine isoelectric focusing, and a maximum absorption of 278 nm. Positive reaction, easily soluble in water, insoluble in ether and chloroform.
Furthermore, this acidic protease exhibits high hydrolysis activity for hemoglobin in an acidic region of pH 7.0 or lower, but has the property of being significantly inhibited by pepstatin. In addition, this acidic protease
Stable in acidic areas below PH7.0, unstable in alkaline areas above PH8.0. The pharmacological action and toxicity of this acidic protease will be explained below using test examples. Test Example 1 Immune complex hydrolysis 15 mg of soluble immune complex [human IgG - anti-human IgG antibody (rabbit)], acidic protease, trypsin,
3 mg each of α-chymotrypsin or plasmin was dissolved in 1 ml of phosphate buffer (0.06M, PH6.0) and incubated at 37°C for 60 minutes. 20%
The reaction was stopped by adding 1 ml of perchloric acid solution, and the absorbance of the supernatant at 280 nm was measured. Also, instead of the immune complex, human IgG was used and the same procedure was performed.
The water splitting ratios of both were compared. In addition, the hydrolytic activity is
The value when IgG is treated with acidic protease is
Expressed as a relative value as 100. The results are shown in Table 1. Compared to other proteases, acidic proteases
Soluble immune complexes were selectively hydrolyzed over normal human IgG.
【表】
試験例２ 免疫複合体水解作用
可溶性免疫複合体〔ヒトIgG−抗ヒトIgG抗体
（家兎）〕15mgと酸性プロテアーゼあるいはトリプ
シン0.3mgを１mlのリン酸緩衝液（0.06M、PH
6.0）に溶解し、ついで、ラツト血清あるいは同
緩衝液250μを添加し、37℃で60分間インキユ
ベーシヨンした。反応終了後、残存するヒトIgG
を抗ヒトIgG家兎血清を用い一次元免疫拡散法に
より測定し、水解されたヒトIgGの百分率を算出
した。結果を第２表に示す。
酸性プロテアーゼの免疫複合体水解活性は血清
添加によつて影響されなかつた。一方、トリプシ
ンの水解活性は血清添加により、著しく低下し
た。[Table] Test Example 2 Immune complex hydrolysis 15 mg of soluble immune complex [human IgG - anti-human IgG antibody (rabbit)] and 0.3 mg of acidic protease or trypsin were mixed in 1 ml of phosphate buffer (0.06M, PH
6.0), then 250μ of rat serum or the same buffer was added and incubated at 37°C for 60 minutes. Human IgG remaining after the reaction
was measured by one-dimensional immunodiffusion method using anti-human IgG rabbit serum, and the percentage of hydrolyzed human IgG was calculated. The results are shown in Table 2. The immune complex hydrolysis activity of acidic protease was not affected by serum addition. On the other hand, the hydrolysis activity of trypsin was significantly decreased by the addition of serum.
【表】
試験例３ 甲状腺炎抑制作用
小谷ら（臨床免疫 ９巻、８号、635頁、1977
年）の方法に準じて行なつた。すなわち、１群
100匹のBUF／HDK雄性ラツト（６週令）の胸腺
を摘除し、２週間毎に１回、200radのＸ線照射を
４回くり返し、胸腺摘除14週後に、全屠殺した。
甲状腺を摘出してパラフイン包埋後、ヘマトキシ
リン−エオジン染色およびアザン染色を施し、そ
の単核球浸潤、小胞破壊、線維化の程度を指標に
して甲状腺炎の重症度を０から＋４に分類した。
なお、酸性プロテアーゼは１日１回静脈内に投与
し、対照には、プロテアーゼ活性を失活させた標
品を同様に投与した。結果を第３表に示す。
対照群に比し、酸性プロテアーゼ投与群におい
ては、用量依存的に、甲状腺炎の発症率が減少
し、重症度も軽減された。[Table] Test Example 3 Thyroiditis suppression effect Kotani et al. (Clinical Immunology Vol. 9, No. 8, p. 635, 1977)
It was carried out according to the method of 2010). That is, group 1
The thymuses of 100 BUF/HDK male rats (6 weeks old) were removed, X-ray irradiation at 200 rad was repeated 4 times once every 2 weeks, and all rats were sacrificed 14 weeks after thymectomy.
After removing the thyroid gland and embedding it in paraffin, it was stained with hematoxylin-eosin and Azan staining, and the severity of thyroiditis was classified from 0 to +4 based on the degree of mononuclear cell infiltration, vesicular destruction, and fibrosis. .
Note that acidic protease was administered intravenously once a day, and as a control, a preparation in which protease activity had been inactivated was administered in the same manner. The results are shown in Table 3. Compared to the control group, the incidence and severity of thyroiditis was reduced in a dose-dependent manner in the acid protease administration group.
【表】
試験例４ 免疫複合体腎炎抑制作用
１群10匹のC57BL系雄性マウスに、ヒトIgG−
抗ヒトIgG抗体（家兎）複合体200mg／Kgを１日
３回８時間毎に３日間静注した。４日目に屠殺し
て、腎を摘出し、糸球体への免疫複合体（I.C.）
の沈着をフルオレツセン イソチオシアネート
（FITC）標識した抗家兎IgG抗体（ヤギ）を用い
た螢光抗体法により観察した。さらに血清中の免
疫複合体をClqバインデイングアツセイ法（Clq
binding assay法）を用い測定した。屠殺前に尿
を採取できた34例については、尿蛋白測定試験紙
を用い、尿蛋白量を測定した。なお酸性プロテア
ーゼは１日３回、免疫複合体注入直後に静注し、
対照には失活させた酸性プロテアーゼを同様に静
注した。結果を第４表に示す。
血中免疫複合体、蛋白尿、糸球体へのI.C.沈着
いずれもが酸性プロテアーゼ投与により抑制され
た。[Table] Test Example 4 Immune complex nephritis suppressive effect Human IgG-
Anti-human IgG antibody (rabbit) complex 200 mg/Kg was intravenously injected every 8 hours three times a day for 3 days. On the 4th day, sacrifice the kidney, remove the kidney, and transfer immune complexes (IC) to the glomerulus.
The deposition was observed by fluorescent antibody method using anti-rabbit IgG antibody (goat) labeled with fluorescein isothiocyanate (FITC). Furthermore, immune complexes in serum were analyzed using Clq binding assay (Clq
binding assay method). For the 34 animals for which urine could be collected before slaughter, the amount of urinary protein was measured using urine protein measurement test strips. Acid protease was injected intravenously three times a day immediately after injection of the immune complex.
As a control, inactivated acidic protease was similarly injected intravenously. The results are shown in Table 4. Blood immune complexes, proteinuria, and IC deposition in glomeruli were all suppressed by acid protease administration.
【表】
テアーゼ
0.3mg〓Kg 128±9 5〓7 8〓10
1.0mg〓Kg 88±12＊＊ 4〓10 5〓10＊
3.0mg〓Kg 53±7＊＊ 3〓9＊ 2〓10＊
＊
[Table] Tease
0.3mg〓Kg 128±9 5〓7 8〓10
1.0mg〓Kg 88±12 ＊＊ 4〓10 5〓10 ＊
3.0mg〓Kg 53±7 ＊＊ 3〓9 ＊ 2〓10 ＊
*
Claims (1)
免疫複合体病治療剤。1. A therapeutic agent for immune complex diseases containing human urinary acid protease as an active ingredient.
Priority Applications (13)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP56018429A JPS57131725A (en) | 1981-02-10 | 1981-02-10 | Remedy for immune complex disease |
| CA000395742A CA1181005A (en) | 1981-02-10 | 1982-02-08 | Therapeutic agent for treatment of allergic diseases, immune complex diseases and tumors |
| GB8203682A GB2095993B (en) | 1981-02-10 | 1982-02-09 | Compositions containing human urinary acid protease |
| AU80290/82A AU531314B2 (en) | 1981-02-10 | 1982-02-09 | Therapeutic agent containing human urinary acid protease |
| SE8200748A SE455163B (en) | 1981-02-10 | 1982-02-09 | UROPEPSIN FOR USE AS A THERAPEUTIC AGAINST ALLERGIC DISEASES, IMMUNE COMPLEX DISEASES AND TUMORS |
| CH782/82A CH653557A5 (en) | 1981-02-10 | 1982-02-09 | THERAPEUTIC AGENT SUITABLE FOR TREATING ALLERGIC CONDITIONS, IMMUNE COMPLEX DISEASES AND TUMORS. |
| IT47761/82A IT1154280B (en) | 1981-02-10 | 1982-02-10 | PROTEA-BASED THERAPEUTIC AGENT ACIDS AND ITS USE IN PATIENTS WITH ALLERGIC DISORDERS, IMMUNE-COMPLEX DISEASES AND CANCERS |
| DE8282100973T DE3273953D1 (en) | 1981-02-10 | 1982-02-10 | Therapeutic agent containing a human urinary pepsin |
| DE19823204631 DE3204631A1 (en) | 1981-02-10 | 1982-02-10 | THERAPEUTIC AGENT AND ITS USE |
| EP82100973A EP0059346B1 (en) | 1981-02-10 | 1982-02-10 | Therapeutic agent containing a human urinary pepsin |
| NL8200509A NL8200509A (en) | 1981-02-10 | 1982-02-10 | MEDICINAL PRODUCT FOR ALLERGY DISEASES, IMMUNE COMPLEX DISEASES AND TUMORS; METHOD FOR TREATING PATIENTS SUFFERING FROM THIS |
| FR8202145A FR2499409A1 (en) | 1981-02-10 | 1982-02-10 | THERAPEUTIC AGENT BASED ON ACIDIC PROTEASE FOR THE TREATMENT OF ALLERGIC DISORDERS, IMMUNOCOMPLEX DISEASES AND TUMORS |
| US06/365,465 US4540569A (en) | 1981-02-10 | 1982-04-05 | Method for treatment of allergic disorders and immune complex diseases |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP56018429A JPS57131725A (en) | 1981-02-10 | 1981-02-10 | Remedy for immune complex disease |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS57131725A JPS57131725A (en) | 1982-08-14 |
| JPS6237611B2 true JPS6237611B2 (en) | 1987-08-13 |
Family
ID=11971396
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP56018429A Granted JPS57131725A (en) | 1981-02-10 | 1981-02-10 | Remedy for immune complex disease |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS57131725A (en) |
-
1981
- 1981-02-10 JP JP56018429A patent/JPS57131725A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS57131725A (en) | 1982-08-14 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Olson | Role of heparin as a protective agent following reduction of renal mass | |
| Axelsson et al. | Renal damage after prolonged exposure to cadmium: an experimental study | |
| Morris et al. | Isolation of renin granules from rat kidney cortex and evidence for an inactive form of renin (prorenin) in granules and plasma | |
| Davies et al. | The effect of trypan blue, suramin and aurothiomalate on the breakdown of 125I-labelled albumin within rat liver lysosomes | |
| JPH0544478B2 (en) | ||
| JPS62174023A (en) | Anticoagulant substance, production thereof and anticoagulant agent containing said substance as active ingredient | |
| Bützow et al. | Purification and characterization of placental protein 5 | |
| US5262178A (en) | Therapeutic use of enkephalinase | |
| España et al. | Complexes of tissue kallikrein with protein C inhibitor in human semen and urine | |
| CA1198672A (en) | Fibrinolytically active agent and a method for the preparation thereof | |
| US4479937A (en) | Process of treating inflammation with human urinary thiol protease inhibitor | |
| JPS6237611B2 (en) | ||
| Rosenblum et al. | Hepatobiliary and pancreatic clearance of circulating pancreatic amylase | |
| US4540569A (en) | Method for treatment of allergic disorders and immune complex diseases | |
| Mackenzie et al. | Rapid renal failure in a case of multiple myeloma: the role of Bence Jones proteins | |
| US5403585A (en) | Therapeutic use of enkephalinase | |
| Bocci | Catabolism of plasma proteins | |
| Grant et al. | Identification of a defence mechanism in vivo against the leakage of enterokinase into the blood | |
| JPS6238329B1 (en) | ||
| CA1207262A (en) | Human leucocyte pepsin-like enzyme, method for preparation of the enzyme, and method and therapeutic agent for treating allergic disorders, immune complex diseases and tumors containing the same as an effective ingredient | |
| CA1222693A (en) | Membrane-associated proteins (mp.sub.2), a process for obtaining them, and their use | |
| Largman et al. | Clearance and metabolism of circulating pancreatic proelastase in the rat | |
| US3522229A (en) | Glycoprotein from saliva or salivary glands having gastric acid secretion inhibitory as well as anti-ulcerative activity and method of collecting the same | |
| JPS60188400A (en) | Tissue protein pp19 and obtaining method | |
| US6103691A (en) | Utilisation of angiogenin and/or α1 -microglobulin and/or complement factor D in order to inhibit the secretion of proteins |