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재조합 인간 상피세포성장인자(DWP401)의 흰쥐에서의 in vivo와 in vitro 대사
고여욱(Yeo Wook Koh),남권호(Kouen Ho Nam),정주영(Ju Young Jung),박승국(Seung Kook Park),유영효(Young Hyo Yu),김재환(Jae Hwan Kim),한건(Kun Han),박명환(Myung Hwan Park),심창구(Chang Koo Shim) 대한약학회 1997 약학회지 Vol.41 No.3
Metabolism of DWP401, recombinant juman epidermal growth factor, was examined in vivo and in vitro in rats. When 125I-labeled DWP401 was administered at a dose of 50 mcg/kg by i.v. injection. 125I-labeled DWP401 was rapidly degraded within 30 minytes above 93%. Thin layer chromatography analysis of urine collected for 24 hr after i.v. administration of 125I-labeled DWP401 showed ohly one spot on a X-ray film which was considered as diiodo-tyrosine. This result suggests tha 125I-labeled DWP401 was completely digested into free amino acids without any specific intermediate polypeptides. About 42.1% of the administered iodine was recovered in 24 hr. For in vitro degradation study, 125I-labeled DWP401 was added to plama and tissue homogenates of rats and incubated at 37oC. Almost 98% of the added radioactivity recovered from the protein fraction of the liver, kidey, small intestine, stomach and spleen decreased rapidly. For examplem the recovery rates of 125I-labeled DWP401 were 58.6, 63.2, 39.9, 52.9 and 66.8% after 4hrs of incubation in respective organ homogenates.
재조합 인간 상피세포성장인자(DWP401)의 흰쥐에서의 약물동태
정주영(Joo Young Chung),고여욱(Yeo Wook Koh),남권호(Kwon Ho Nam),조재열(Jae Youl Cho),박승국(Seung Kook Park),유영효(Young Hyo Yu),김재환(Jae Hwan Kim),한건(Kun Han),박명환(Myung Hwan Park),심창구(Chang Koo Shim) 대한약학회 1997 약학회지 Vol.41 No.3
Pharmacokinetics of DWP401, a recombinant human epidermal growth factor (rhEGF), was studied using radioimmunoassay (RIA) and 125I-DWP401 in rats. When DWP401 was administered i.v. at doses of 50 and 500 mcg/kg, the plasma DWP401 disappeared biiexponentially with terminal half life of 4.7 and 92.8 min. The Cmax and Tmax after s.c. administration of ti at doses of 50 and 500 mcg/kg were determined to be 23.6 and 17.5 ng/ml at 50 mcg/kg, and 261.4 ng/ml and 36.8 min, respectively. Both the total urinary and biliary recoveries of intact DWP401 2343 very low (<0.4%), probably due to its extensive degradation in the body. the concentration ratio of DWP401 between the organ and plasma decreased especially in the liver and kidney as the dose and time after the dose increased. For example, the liver/plasma and kidney/plasma concentration ratio of DWP401 at 2.5 min after i.v. doses of 50 mcg/kg were comparable and much larger than unity. But, the ratio at 2.5 min after i.v. doses of 500mcg/kg was much larger in the kidney that in than in the liver. These results suggest that the systemic administration of DWP401 might be subject to rapid and extensive clearance from circulation within several hour after main distrbution to liver and kidney.
Escherichia coli 및 Bacillus subtilis 간의 새로운 shuttle vector 의 조성
고여욱,양재명,이희명,이청호 한국유전학회 1988 Genes & Genomics Vol.10 No.1
A new shuttle vector replicating stably both in Escherichia coli and Bacillus subtilis was constructed. pUBHR isolated from B. subtilis DB 104 and pBR322 isolated from E. coli HB101 were digested with BamHI, ligated, and transformed into E. coli. Plasmid DNAs from four transformants with Ap^rKm^rTc^s (ampiciilin-resistant, kanamycin-resistant, tetracycline-sensitive) marker genes were isolated, digested with BamHI, and analyzed on agarose gel. The size of the plasmid was 8.1kb long which corresponded to the sum of 3.7kb fragment from pUBHR and 4.4kb fragment from pBR322. EcoRI digestion patterns showed that it consisted of two recombinant plasmids ligated in opposite orientation. These recombinant plasmids were named pKR317 and pKR318. pKR317 was transformed into B. subtilis 168. Three kanamycin-resistant transformatns were selected and analyzed. The size and restriction pattern of the pKR317 isolated from B. subtilis were found to be identical to pKR 317 isolated from E. coli. These results suggested that the pKR317 was replicated, and the kanamycin gene on the plasmid was expressed both in E. coli and B. subtilis. These properties make pKR317 potentially useful as an appropritate shuttle vector for E. coli and B. subtilis.
상피세포 성장인자의 경피흡수 : 정상피부 , 각질제거피부 및 화상피부에 있어서
심창구,조애리,이정욱,안병락,정주영,고여욱 한국약제학회 1996 Journal of Pharmaceutical Investigation Vol.26 No.1
In vivo and in vitro skin permeation of recombinant ^(125)I-EGF through normal. stripped and the first degree burn skin were studied. The in vitro skin permeation rate through the first degree burn skin (296 cpm/㎠/hr) and the stripped skin (1131 cpm/㎠/hr) were 3.5 times and 13 times higher, respectively, as compared with the one through normal skin. In vivo absorption study with the first degree burn skin, the peak concentration of EGF in the skin was achieved at 1-3 hr and decreased afterward up to 8 hr with an elimination constant of 1.31 × 10³g/㎖/hr. To investigate the higher elimination rate of EGF in burn skin, binding and metabolism studies were conducted. No significant metabolism of EGF in burn skin (100℃, 5second burning) was observed. With the presence of unlabelled-EGF, ^(125)I-EGF permeation through the burn skin showed higher permeation rate than the one without unlabelled-EGF. The result may indicate that EGF-receptor binding play a role in determining the skin permeation rate.