• 약학회지
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• 제48권1호
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• pp.82-87
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• 2004

Mungbean trypsin inhibitor (MBTI) was isolated and purified from Mung bean which has been used as a galenic and traditional food. MBTI and poly(ethylene glycol) were conjugated by using water soluble carbodiimide. We evaluated the therapeutic value of the MBTI and MBTI-PEG conjugate using animal models, sublethal septic shock model in guinea pig caused by pseudomonal elastase, shock model in rat caused by lipopolysaccharide, and the vascular permeability test by using pseudomonal elastase. In two shock model in guinea p Is and in rat, hypotesion shock was inhibited by pretreatment of MBTI. And also the vascular permeability caused by pseudomonal elastase reduced by pretreatment of MBTI. Also, therapeutic value of the MBTI-PEG conjugate was evaluated by using the sublethal septic shock model caused by pseudomonal elastase. The MBTI-PEG conjugate was more effective than native MBTI against pseudomonal elastase induced septic shock in guinea pig model.

• BMB Reports
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• 제33권2호
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• pp.112-119
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• 2000

Three kinds of serine protease inhibitors, members of the Bowman-Birk trypsin inhibitor, were purified from Dolichos lablab seeds and named Dolichos protease inhibitor 1, 2 and 3 (DI-1, DI-2 and DI-3), respectively. Each inhibitor showed a single band with gel mobility at around 15.9, 12.1 and 14.6 kDa on 20% SDS-PAGE under reducing conditions. To characterize inhibitory specificity, the inhibition constant (Ki) for these inhibitors was measured against several known serine proteases. All three Dolichos protease inhibitors (DI-1, DI-2 and DI-3) inhibited the activity of trypsin and plasmin, but had no effect on thrombin and kallikrein (either for human plasma kallikrein or for porcine pancreas kallikrein). DI-1 inhibited chymotrypsin most effectively (Ki = $3.6{\times}10^{-9}\;M$), while DI-2 displayed inhibitory activity for porcine pancreatic elastase (Ki = $6.2{\times}10^{-8}\;M$). Pre-treatment of the 33 mg/kg of DI-mixture (active fractions from $C_{18}$ open column chromatography that included DI-1, DI-2 and DI-3) inhibited the induction of pseudomonal elastase-induced septic hypotension and prevented an increase in bradykinin generation in pseudomonal elastase-treated guinea pig plasma. Also, the increase of kallikrein activity, by injection of pseudomonal elastase, was inhibited by the pretreatment of the DI-mixture in a guinea pig. Since the DI-mixture had no inhibitory effect on kallikrein activity when Z-Phe-Arg-MCA was used as a substrate in vitro, its inhibitory activity in the pseudomonal elastase-induced septic hypotension model might not be due to a direct inhibition of plasma kallikrein in the activation cascade of the Hageman factor and prekallikrein system. These results suggest that the Dolichos DI-mixture might be used as an inhibitor in pathogenic bacterial protease-induced septic shock.

• Journal of Pharmaceutical Investigation
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• 제30권3호
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• pp.159-166
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• 2000

Kunitz-type soybean trypsin inhibitor (SBTI) and chondroitin sulfate (A, and C type) were conjugated using sodium periodate method. And the physicochemical, pharmacokinetic properties and immunogenecity of the conjugates (Chon-A-SBTI or Chon-C-SBTI) were characterized. We expected the conjugation using chondroitin sulfate to reduce the immunogenecity and to improve the pharmacological effect. As the results, the mean molecular weight of the conjugate highly increased. After I.V. injection of the radiolabeled conjugates or native SBTI into mice, it was found that native SBTI showed rapid elimination from plasma, whereas Chon-A-SBTI and Chon-C-SBTI were slowly eliminated. Organ distribution of the two agents at 30 min after I.V. injection was different : Chon-A-SBTI or Chon-C-SBTI accumulated to a large extent in the liver (13% in Chon-A-SBTI and 16% in Chon-C-SBTI), whereas native SBTI was taken up more rapidly by the kidney (107% dose/g of tissue) and excreated into the urine (26%). In addition we evaluated the therapeutic value of the conjugates by using the sublethal septic shock model caused by pseudomonal elastase and tested the immunogenecity by passive cutaneous anaphylaxis shock (PCA). The conjugates were more effective than native SBTI against pseudomonal elastase induced septic shock in guinea pig. In case of the conjugates, the pharmacological and therapeutic effect lasted over 3 hours long. In immunogenecity test, both of the conjugates showed the reduction of their immunogenecity, especially Chon-A-SBTI looked most effective.

• 생명과학회지
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• 제12권5호
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• pp.528-534
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• 2002

우리나라에서 식용으로 뿐만아니라 한방재료로 널리 사용되고 있는 녹두(vigna radiata L. wilczek) 로부터 trypsin inhibitor (Mung bean trypsin inhibitor, MBTI)를 분리정제하여 그 특성을 조사하였다 또한 병태동물모델 즉, septic shock induced guinea pig model을 이용하여 MBTI의 약물학적 효과를 평가하였다. MBTI의 분리 및 정제과정은 Sephadex C-50 chromatography, DEAE-celluloseion exchange chromatography 및 trypsin affinity column 을 차례로 이용하였다. 정제한 MBTI는 전기영동 및 아미노산 서열분석결과 분자량 약 8,000 Da 의 BBI-type (Bowman-birk inhibitor type)임을 알 수 있었으며 이들의 생화학적 특성을 구명하였다. 또한 pseudomonal elastase로 유도된 septic shock guinea pig model에서 MBTI 10 mg/kg를 전처치한 결과 hypotention shock 유발이 억제됨을 알 수 있었다.