재조합 인터페론 알파의 표준화에 관한 연구

손여원,신원,정자영,최영주,정지원,오일웅,진재호,박선영,박태성 식품의약품안전청 2000 식품의약품안전청 연보 Vol.4 No.-

우리나라에서 유통되고 있는 재조합 인터페론 알파의 역가시험을 표준화하기 위하여 국내제조 5개사,수입 1개사 및 식품의약품안전청이 참여하여 공동연구를 실시하였다. 공동연구 참여사의 역가시험법의 상대적인 감도와 상용표준품의 활성을 비교 ·보정하기 위하여 공통의 인터페론 알파 시험물질에 대하여 5회 이상의 독립적인 시험들 실시하고 시험결과의 정확도, 정밀포 및 재현성을 비교하였다. 보다 정확한 분석을 위하여 모든 참여사는 자사 제품의 품질관리에 적용되고 있는 역가시험법과 더불어 식품의약품안전청에서 제공한 참조씨험법을 함께 시험하였고 시험결과를 자사의 계산법과 동시에 평행선분석법으로 각각 계산하여 그 차이를 비교하였다. 그 결과 시험의 정밀도와 재현성은 각 참여사의 자사 뵉가시험법과 평행선분석법을 적용하였을 째 보다 우수하게 나타났고 대부븐의 참여사에서 1표시역가의 80%~l%', 신뢰구간 '표시울가의 64%~l56%' 내에 분포하여 상대적인 감도가 고르면서 정확포가 높았다. 또한 각 참여사의 자사 결과계산법과 평행선분석법 간의 차이를 't-Test'로 분석한 결과 유의한 차이를 나타내지 않았으며 모든 찹여사의 두 가지 시험법과 분석법으로부터 얻은 평균값들의 차이를 분석한 결과 유의한 차이를 나타내지 않아 모든 참여사의 인터페론 역가시헌이 '표준화'되어 있는 것으로 분석되었으며 평행선분석법 및 신뢰구간에 대한 적용가능성을 제시하였다. The specific activiw of recombinant interferons made by different manufacturers can vary and bioassay systems which are utilized to determine the biological potency ofinterferon may he affected by a number of factors, such as ceIB lines, viruses and the statisticalanalysis of the assay. Tllerefore the bioassay of interferon, like as other biological products, isessentially comparative and thus requires a fHxed reference standard and standardized assayconditions. A collaborative study was performed for standardization of interferon bloassay. Sixlaboratories of interferon Danufacturers and KFDA were participated in this study. Alllaboratories measured the potency of'the same inteferon samples'by their own routinemethods and the reference method which was offered by KFBA. The results were analysed byboth ways using their own data analysis methods and the usual statistical methods for aparallel line assay The relatiue sensitivities of each assay system and the potency of eachworking standard of the participants were compared by assessing the assay performance svchas accuracy, precision and reproducibility. The results showed best pr·ecision and reproducibiBitywhen the potency was measured by the manufacturer's routine methods and calculated byparallel line analysis. Tte estimated potency was from 80% to 125% and the confidence limitwas from 64% to 156% of the stated potency in most laboratofes, which showed goodaccuracy. Differences in data analysis between the manufacturer's routine analytical method andthe Parallel line assay were not significant by't-Test'and differences in all results fromroutine assay and reference assay also were not significant by'analysis of valiance'. Based onthe results of the collaboriltive study, all participants were'standardized'in the interferonbioassay and we may consider the change of the data analysis of Inteferon potency to thestatistical method for a parallel line assay.

생물의약품의 심사 방향 : 기준 및 시험방법 심사

손여원 한국에프디시규제과학회(구 한국에프디시법제학회) 2006 FDC법제연구 Vol.1 No.2

COVID19 회복기 환자의 폐 영상학적 변화 양상에 대한 고찰

손여원,김연재 대한내과학회 2022 대한내과학회 추계학술대회 Vol.2022 No.2

Guinea Pig간장에서 GTP결합단백질의 특성연구(Ⅳ) : Rat brain에서 small GTO-binding Protein Rho6의 표적단백질에 관한 연구

손여원,홍성화,신원,김호식,정지원,오호정,최영주,김병헌,이숙연,김용관,김선미,민홍기,전용성 식품의약품안전청 1997 식품의약품안전청 연보 Vol.1 No.-

이종간 원형질체 융합을 이용한 acetaminophen 생산균주 개량

손여원(Yeo Won Shon),정대영(Dae Young Jung),이상섭(Sang Sup Lee),민홍기(Hong Ki Min) 대한약학회 1994 약학회지 Vol.38 No.5

Acetaminophen, a widely used analgesic, can be formed by N-acetylation and p-hydroxylation of aniline. Interspecific protoplast fusion technique was used to get acetaminophen directly from aniline and to increase the productivity of acetaminophen. Three auxotrophic mutants were obtained from S. griseus(ATCC 13273) and S. hygroscopicus(KCTC 1089) by N-methyl-N''-nitro-N-nitrosoguanidine(NTG) treatment. Regeneration frequencies of S. griseus(his-), S. griseus(lys- ), S. hygroscopicus(arg-) were 42%, 45%, and 31%, respectively. Fusion of protoplasts carrying different auxotrophic markers was achieved by treatment with polyethylene glycol. When protoplasts were treated with 50% polyethylene glycol for 3 minutes, the fusion frequency between S. griseus(his- ) and S. hygroscopicus(arg-) was 3.8X10-5. The fusion frequency between S. griseus(lys-) and S. hygroscopicus(arg- ) was 5.6X10-4. When we checked the production of acetaminophen, thirty-four out of the fifty-six fusants produced larger amounts of acetaminophen than the parent strains did. Nine fusants produced twice more and twenty-five fusants produced one to two times more of acetaminophen than their parents.

TGF-β-induced Transcriptional Activation of MMP-2 is Mediated by Activating Transcription Factor (ATF)2 in Human Breast Epithelial Cells

김은숙,손여원,문애리 덕성여자대학교 대학원 2008 덕성여자대학교 대학원 논문집 Vol.10 No.-

We have previously shown that transforming growth factor (TGF)-βup-regulates matris metalloproteinase (MMP)-2 leading to the induction of oncogenic signaling in preneoplastic MCF10A human breast epithelial cells. The present study investigated the mechanism of transcriptional regulation of MMP-2 by TGF-β in MCF10A cells. By using 50 deletion constructs of MMP-2 promoter, we demonstrated that binding sites for p53, S1, AP-1and Sp1, and to a lesser extent CREB, GCN-His and PEA3, were potential cis-acting elements for TGF-β-induced transcriptional activation of MMP-2 in MCF10A cells. Since activating transcription factor (ATF)2 was shown to mediate the TGF-β induced cellular responses, we examined the involvement of ATF2 in TGF-β-activated MMP-2 gene transcription. TGF-β increased DNA binding activity of AP-1 in which ATF2 was involved as evidenced by electrophoretic mobility shift assay. TGF-βinduced phosphorylation of ATF2 through p38 MAPK signaling. A dominant-negative (DN) ATF2 significantly inhibited the TGF-β-induced up-regulation of MMP-2, but not that of MMP-9, suggesting that ATF2may be a transcription factor responsible for transcriptional activation of MMP-2 gene by TGF-β. Invasive and migratory phenotypes induced by TGF-β were significantly inhibited by DN ATF2, indicating a critical role of ATF2 in TGF-β-induced oncogenic progression of MCF10A cells. Taken together, this study demonstrates that ATF2 mediates the TGF-β-induced MMP-2 transcriptional activation, elucidating a molecular mechanism for the malignant progression of human breast epithelial cells exerted by TGF β.

지속성 진통작용 물질인 Capsaicinoid 의 흰쥐 척수내 Substance P 함량 및 생합성에 미치는 영향

김해영,손여원,이상섭,이승기 ( Hai Young Kim,Yeo Won Sohn,Sang Sup Lee,Seung Ki Lee,Seung Ki Lee ) 생화학분자생물학회 1990 BMB Reports Vol.23 No.2

We aimed in this study to evaluate the effects of capsaicinoids on the cellular concentration of substance P(SP) and substance P-mRNA in rat spinal cord. This study was based on the fact that longlasting analgesic, capsaicin caused depletion of SP from primary afferent neurons in dorsal root ganglia. After administration of 6-paradol (80, 500, 1000 ㎎/㎏) or capsaicin (80 ㎎/㎏) in rats, the cellular concentration of SP and its mRNA in spinal cord were analyzed by radioimmunoassay and slot blot hybridization analysis respectively. 6-Paradol, like capsaicin, reduced in dose-dependent manner the content of SP in rat spinal cord. Additionaliv, SP-mRNA concentration was also reduced to 61% of the control level at 3 h after capsaicin administration and was restored to the control level after 1 day. In 6-paradol treated group, substance P-mRNA concentration was not detectably reduced at 3 hours but significantly reduced to 49% of the control level at 1 day after 6-paradol treatment. These results support for the previous notion suggesting that the action mechanism of capsaicinoid analgesia may be primarily based on the depletion of SP from primary afferent neurons. These results also suggest that capsaicinoid analgesics may play a role at the transcription level of SP synthesis in rat spinal neurons.

지속성 진통작용 물질인 Capsaicinoids의 흰쥐 척수내 Substance P 함량 및 생합성에 미치는 영향

김해영,손여원,이상섭,이승기,Kim, Hai-Young,Sohn, Yeo-Won,Lee, Sang-Sup,Lee, Seung-Ki 생화학분자생물학회 1990 한국생화학회지 Vol.23 No.2

강력한 지속적 진통 효과를 나타내는 것으로 알려진 capsaicin은 작용기전은 밝혀져 있지 않으나 dorsal spinal cord와 dorsal root ganglia에 작용하여 신경전달물질이라 생각되는 substance P(SP)를 선택적으로 감소시키므로 지속적인 진통효과를 나타낼 것이라고 제시된 바 있다. 따라서, 본 연구에서는 capsaicin과, 그 analogue로 capsaicin과는 달리 신경 독성이 거의 없는 6-paradol이 척수내 신경조직에 존재하는 SP 농도변화에 미치는 효과 및 SP-mRNA 생합성에 미치는 영향을 분석하여, 이들 capsaicinoids의 진통 효과와 SP-생합성에 미치는 영향과의 상관관계를 규명코자 하였다. 생후 6주된 Sprague-Dawley male rat에 6-paradol(80, 500, 1000 mg/kg) 또는 capsaicin(80 mg/kg)을 ether 마취상태에서 피하주사하고 3시간, 1일, 5일 후에 척수내 SP 함량을 radioimmunoassay 방법으로 측정한 결과, 3시간부터 vehicle 투여군에 비해 capsaicin과 6-paradol 투여군에서 용량 의존적으로 유의성 있는 SP 함량 감소를 보였고, 그 효과가 5일까지도 유지되었다. 또한 이들 약물의 SP-유전자 전사활성에 미치는 영향을 분석하기 위해 slot blot hybridization 방법으로 SP-mRNA의 농도를 측정한 결과, capsaicin 투여군에서는 SP-mRNA 농도가 3시간에 유의적으로 61%로 감소하였으나 1일 후에 90%로 회복되었다. 그러나 6-paradol(1000 mg/kg) 투여군에서는 3시간에는 유의적 감소를 보이지 않았고 1일 후에 49%로 감소하였다. 본 연구 결과는 capsaicinoids 투여 후 초기의 척수내 SP 함량의 감소는 신경조직으로부터의 SP 유리에 따른 것이고, 그 다음의 지속적인 SP의 신경세포내 감소현상은 감각신경에서의 SP 생합성과정 중 SP-mRNA 합성단계에서의 억제효과 때문이라는 가설을 뒷받침하는 결과로 판단된다. We aimed in this study to evaluate the effects of capsaicinoids on the cellular concentration of substance P(SP) and substance P-mRNA in rat spinal cord. This study was based on the fact that long-lasting analgesic, capsaicin caused depletion of SP from primary afferent neurons in dorsal root ganglia. After administration of 6-paradol (80, 500, 1000 mg/kg) or capsaicin (80 mg/kg) in rats, the cellular concentration of SP and its mRNA in spinal cord were analyzed by radioimmunoassay and slot blot hybridization analysis respectively. 6-Paradol, like capsaicin, reduced in dose-dependent manner the content of SP in rat spinal cord. Additionallv, SP-mRNA concentration was also reduced to 61% of the control level at 3 h after capsaicin administration and was restored to the control level after 1 day. In 6-paradol treated group, substance P-mRNA concentration was not detectably reduced at 3 hours but significantly reduced to 49% of the control level at 1 day after 6-paradol treatment. These results support for the previous notion suggesting that the action mechanism of capsaicinoid analgesia may be primarily based on the depletion of SP from primary afferent neurons. These results also suggest that capsaicinoid analgesics may play a role at the transcription level of SP synthesis in rat spinal neurons.

배양 인형 림프구에 대한 Capsaicin의 유전독성 ( Genotoxicity of Capsaicin in Cultured Human Lymphocytes )

이상섭,박영호,손여원,류수정,서영준 한국환경성돌연변이발암원학회 1995 한국환경성돌연변이·발암원학회지 Vol.15 No.2

사람 폐암세포주에서의 bcl-2 안티센스 처리에 의한 효과

김선미,정자영,오호정,손여원 한국독성학회 2002 Toxicological Research Vol.18 No.4

Apoptosis, or programmed cell death, is a genetically regulated pathway that is altered in many cancers. Overexpression of bcl-2 leads to resistance to apoptosis and promotes tumorigenesis. To determine the effects of bcl-2 antisense treatment in human lung cancer cell lines, a 20 mer full phosphorothioate oligonucleotide (ODN) targeted at the coding region of the bcl-2 mRNA was synthesized. Western blot analyses were used to examine bcl-2 protein level in five human non-small cell lung cancer (NSCLC) cell lines (NCI-H226, SK-MES-1, NCI-H358, NCI-H522 and NCI-H1299) and four human small cell lung cancer (SCLC) cell lines (NCI-H69, NCI-H417, HCC-2108 and SW2). Three out of five NSCLC (NCI-H226, KS-MES-1 and NCI-H1299) and all of SCLC cell lines expressed Bcl-2 protein. Treatment of these cells with antisense ODN for 48 hours reduced their viability and Bcl-2 protein levels. As a conclusion, bcl-2 antisense treatment appears reduction of the Bcl-2 protein levels and cytotoxic effect including apoptosis in human lung cacer cell lines.