초록집 ( 1993 . 4 . 15 ) : 1993 년도 신약개발 연구결과 / 미발표 결과 목차 ; 폐염균 다당류 정제 방법의 개발 및 한국형 폐염균의 수집

임관기,김수남,강연현,이동권 한국응용약물학회 1993 학술대회 Vol.1993 No.1

한국 독사독으로부터의 혈전 용해제 개발에 관한 연구;2. 살모사 (A. bromhoffi brevicaudus) 사독 Protease 의 특성과 혈전 용해능에 관한 연구

김종호,김병재,이문한,이혜숙,이항,임종섭,채창수 한국응용약물학회 1995 Biomolecules & Therapeutics(구 응용약물학회지) Vol.3 No.2

The biochemical properties of the fibrinolytic protease of 50,800 Da isolated from the venom of Agkistrodon blomhoffi brevicaudus were characterized. The enzyme hydrolyzed the carboxyl side of arginine in the synthetic chromogenic peptides, N-Benzoyl-Phe-Val-Arg-pNA and N p-Tosyl-Gly-Pro-Arg-pNA, and the enzyme activity was inhibited by phenylmethylsulfonylfluoride indicating that the enzyme belongs to the serine protease family. The protease showed maximum activity at pH 7.5 and inhibited by ZnCl₂, CuSO₄, but not by soybean trypsin inhibitor, pepstatin A, 2-mercaptoethanol and EDTA. The Km value determined with Np-Tosyl-Gly-Pro-Arg-pNA was 0.2 mM. The thrombolytic activity of the purified enzyme was evaluated by platelet aggregation test in rabbits. While the platelet count ratio in blood of the rabbits injected with thrombin alone declined from 1.0 to 0.6 within 7 min and maintained around 0.6 for 24 hours thereafter, the ratio rapidly recovered from around 0.6 to 0.8 in 1 hr, to 1.0 in 24 hrs when the rabbits were sequentially treated with thrombin and the purified enzyme. The result showed that the serine protease from A. blomhoffi brevicoudus of 50,800 Da had a thrombolytic activity in vivo and the enzyme might be developed as a therapuetic agent for the treatment of thrombic disease.

제 3 회 신약개발 연구발표회 초록집 ( 1994 . 4 . 15 ~ 16 ) : 포스터발표 제 2 분과 ( 생명공학 , 약효검색 , 약리대사 ) ; 리보솜 불활성화 단백질의 삼차원 구조 규명과 그 구조의 신약개발에의 응용

서세원 한국응용약물학회 1994 학술대회 Vol.1994 No.1

포스터 발표 초록집 ( 1997 . 4 . 18 ) : 제 2 분과 6 ( 생명공학 ) ; 한국형 유산균의 in vitro에서 장내유해효소의 억제효과

김동현,이승원,김숙영,한명주,박혜영,배은아 한국응용약물학회 1997 학술대회 Vol.1997 No.1

한국 독사독으로부터의 혈전 용해제 개발에 관한 연구 II. 살모사(A. bromhoffi brevicaudus) 사독 Protease의 특성과 혈전 용해능에 관한 연구

김병재,이문한,임종섭,이항,이혜숙,김종호,채창수,Kim,,Byoung-Jae,Lee,,Mun-Han,Rim,,Jong-Seop,Lee,,Hang,Lee,,Hye-Suk,Kim,,Jong-Ho,Chai,,Chang-Su 한국응용약물학회 1995 Biomolecules & Therapeutics(구 응용약물학회지) Vol.3 No.2

The biochemical properties of the fibrinolytic protease of 50,800 Da isolated from the venom of Kgdistrodon blomhoffi brevicaudus were characterized. The enzyme hydrolyzed the carboxyl side of arginine in the synthetic chromogenic peptides, N-Benzoyl-Phe-Val-Arg-pNA and N-p-Tosyl-Gly-Pro-Arg-pNA, and the enzyme activity was inhibited by phenylmethylsulfonylfluoride indicating that the enzyme belongs to the serine protease family. The pretense showed maximum activity at pH 7.5 and inhibited by ZnCl$_2$, CuSO$_4$, but not by soybean trypsin inhibitor, pepstatin A, 2-mercaptoethanol and EDTA. The fm value determined with N-p-Tosyl-Gly-Pro-Arg-pNA was 0.2 mM. The thrombolytic activity of the purified enzyme was evaluated by platelet aggregation test in rabbits. While the platelet count ratio in blood of the rabbits injected with thrombin alone declined from 1.0 to 0.6 within 7 min and maintained around 0.6 for 24 hours thereafter, the ratio rapidly recovered from around 0.6 to 0.8 in 1 hr, to 1.0 in 24 hrs when the rabbits were sequentially treated with thrombin and the purified enzyme. The result showed that the serine protease from A. blomhoffi brevicoudus of 50,800 Da had a thrombolytic activity in vivo and the enzyme might be developed as a therapuetic agent for the treatment of thrombic disease.

'94 신약개발의 실제와 문제 ( 1994 . 11. 4 - 5 ) : 심포지움 2 ; 신약창출 및 검색 ; 리포좀의 면역학적 응용

정흥석 한국응용약물학회 1994 학술대회 Vol.1994 No.1

제 3 회 신약개발 연구발표회 초록집 ( 1994 . 4 . 15 ~ 16 ) : 포스터발표 제 2 분과 ( 생명공학 , 약효검색 , 약리대사 ) ; 한국형 B 형 간염 바이러스 elongated X 단백질의 기능 및 간암 유발 기작에 관한 연구

노현모 한국응용약물학회 1994 학술대회 Vol.1994 No.1

한국 Streptomyces sp. 로 부터 분리한 방향족 화합물과 지질 화합물의 세포독성 연구

염곤,신석우 한국응용약물학회 1997 Biomolecules & Therapeutics(구 응용약물학회지) Vol.5 No.2

In an effort to screen new selective antitumor agents from the broth of soil microorganism, cytotoxicity oriented screening was performed against tumor cells and 3 compounds (Compound 1, 2 and 3) were isolated from Streptomyces parvullus ISP 5048 and their chemical structures were determined. Among these compounds, Compound 2 showed the highest cytotoxicity against P388D1 and L1210. While the IC_(50) values of compound 2 against P388D1 and L1210 were 0.073 ㎍/㎖ and 0.07 ㎍/㎖, respectively, and the IC_(50) value of Compound 3 was 0.17 ㎍/㎖ against human lung cancer cells, A549, the cytotoxicity of Compound 2 and 3 against normal cell line, Vero E6 cell was about 4- and 8-fold lower than that of adriamycin. Based on the chemical analysis data, Compound 3 was octacosamicine A, a known antibiotic, which was reported by Dobasih et al. (1988). Taken together the results demonstrated that Compound 2 and Compound 3 has the possibility to be developed as antitumor agent because of its potent cytotoxicity as well as high selectivity against various cancer cell lines.

한국 독사독으로부터의 혈전 용해제 개발에 관한 연구 : 1. 살모사 (A. bromhoffi brevicaudus) 사독 Protease 의 정제에 관한 연구 I. Purification of a Protease from the Venom of A. bromhoffi brevicaudus

김종호,김병재,이문한,이혜숙,이항,임종섭,채창수 한국응용약물학회 1995 Biomolecules & Therapeutics(구 응용약물학회지) Vol.3 No.2

Fibrinolytic and fibrinogenolytic activities of the venoms from the Korean snakes, Agkistrodon caliginosus, Agkistrodon saxatilis and Agkistrodon blomhoffi brevicaudus were compared by fibrin-plate method and polyacrylamide gel electrophoresis, respectively. The venom from A. blomhoffi brevicaudus showed the highest degree of fibrin(ogen)olytic activity, and a protease with the fibrin(ogen)olytic activity was purified by p-aminobenzamidine affinity chromatography and DEAF ion-exchange chromatography. The purified enzyme had a molecular weight of 50,800 and a capability to degrade the Bβ-chain of fibrinogen preferentially to the Aα-chain, but not the γ-chain. Fibrinolytic activity of the purified enzyme was approximately 3.8 plasmin unit/mg protein.

32P-Postlabelling 방법의 응용 : Azo 색소 및 Flavonoid 화합물에의해 유도되는 DNA Adduct 의 검출에 관한 연구 A Study on Detection of DNA Adduct Induced by Azo Dyes and Flavonoid Compounds

김재현,박창원,박정식,홍연탁 한국응용약물학회 1993 Biomolecules & Therapeutics(구 응용약물학회지) Vol.1 No.1

DNA adducts induced by putative chemical related to carcinogenesis were detected and determined by (32)^P-Postlabelling assay after exposure of 4 compounds comprising two azo dyes (amaranth, new coccine) and two flavonoid compounds (rutin, quercetin) to ICR mouse. DNA was isolated from mouse liver and digested enzymatically to deoxyribonucleoside 3'-monophosphate. The postincubation of DNA digests with nuclease P1 before (32)^P-labelling enhanced the technique's sensitivity. Nuclease P1 cleaves deoxyribonucleoside 3'-monophosphates of normal nucleotides to deoxyribonucleosides which do not serve as substrates for polynucleotide kinase, while most of adducts were found to be totally or partially resistant to the 3'-dephosphorylating action of nuclease Pl. The adducted deoxyribonucleoside 3'-monophosphate was converted to 5'-(32)^P-labelled deoxynucleoside 3',5'-bisphosphate by T4 polynucleotide kinase. The nucleotides were separated by anion-exchange thin layer chromatography(TLC) on polyethyleneimine cellulose by 4-dimensional or 2-dimensional TLC then detected by autoradiography. The results show that DNA adducts were detected in liver DNA of ICR mouse after administration of amaranth and quercetin by 2-dimensional and/or 4-dimensional TLC.