• 약학회지
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• 제45권2호
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• pp.190-204
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• 2001

This study was conducted to investigate for its effects on reproductive and developmental toxicity of recombinant human epidermal growth factor (rhEGF) in Sprague-Dawley rats. Male rats were administered rhEGF at doses of 1, 10, 100, and 1000$\mu$g/kg/day, respective1y, by subcutaneous injection from 63 days before and throughout to mating period until the day before sacrifice. Female rats were administered rhEGF at the same doses from 14 days before mating to day 20 of gestation or to day 21 of lactation. We examined the male and female fertility indices and maternal toxicity of F0 parental animals. Also, we examined the external, visceral, or skeletal malformation of fetuses, growth and development, behavior, and/or reproductive performance of F1 animals. At the highest dose (1,000 $\mu$g/kg), the mean body weights of F0 animals were significantly increased in males and females at 3 or 2 weeks after treatment, respective1y. No clinical signs and food intakes were observed at any time during the experimental period by rhEGF treatment. In autopsy examination, the relative and absolute liver weights significantly increased in both sexes of 1,000 $\mu$g/kg. At the highest dose (1,000 $\mu$g/kg), there was a statistically significant increase of pregnancy period and the number of dead fetuses. Moreover, significant increase of mean fetal body weight and decrease of number of live fetuses, which related to the difficult dilivery were observed in highest dose group. In Fl examination, no adverse effects on external, visceral, and skeletal malformation, physical and functional development, behavior or reproductive ability of Fl animals were observed in any group. Also, there was no significant difference between control and treated groups in copulation or fertility indices of Fl animals. These results indicate that rhEGF had no adverse effect on fertility and reproductive ability of Sprague-Dawley rats.

• Radiation Oncology Journal
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• 제24권3호
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• pp.179-184
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• 2006

목 적: 재조합 표피성장인자는(rhEGF) 다양한 표피와 상피 세포의 증식을 자극하는 것으로 알려져 있다. 본 연구에서는 방사선이 조사된 섬유아세포의 증식에 rhEGF의 효과를 알아보고자 하였다. 대상 및 방법: 인간에서 기원한 섬유아세포를 초대배양(primary culture)한 세포를 이용하였다. 대웅제약에서 유전자 재조합하여 대장균에서 발현하여 생산한 rhEGF를 제공 받아 사용하였다. 방사선 조사는 4 MV 선형가속기(CLINAC 600C, Varian, Palo Alto, CA, USA)를 이용하여 분당 2 Gy 내외의 선량률로 균일하게 조사하였다. 조사된 방사선량은 8 Gy이었다. 생존세포수는 trypan blue 염색법을 이용하였고, rhEGF에 의한 세포주기의 변화를 관찰하기 위하여 유세포 분석법을 시행하였다. 결 과: 4 Gy의 방사선을 조사한 후 7일째까지 생존 세포수를 trypan blue 염색법을 이용하여 측정한 결과 모든 rhEGF 농도(1.0 nM, 10 nM, 100 nM, 1,000 nM )에서 방사선 조사 단독군보다 생존세포 수가 많았다. 방사선을 조사하지 않은 섬유아세포에서 rhEGF를 10 nM처리한 후 FACS scan을 시행한 결과 세포주기 중에서 S기 비율이 증가하였다. 결 론: 방사선이 조사된 섬유아세포에서 rhEGF를 투여하면 rhEGF를 투여하지 않은 섬유아세포에 비해서 세포증식이 가속됨을 확인할 수 있었다.

• 한국미생물·생명공학회지
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• 제24권4호
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• pp.472-477
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• 1996

Natural human epidermal growth factor (nhEGF) was purified from pregnant human urine by benzoic acid adsorption, DEAE-Sepharose ion exchange, and immunoaffinity chromatography. The purified nhEGF was further separated into four fractions using Bondapak C$_{18}$ HPLC system. Following characterization by Western blot analysis and double immu- nodiffusion, we found that each fraction corresponds to four derivatives of the nhEGF. For biological analysis of nhEGF, we optimized the labeling time and serum concentration for the incorporatioin of 5-bromo-2'-deoxy uridine (BrdU), a non-radioactive alternative for [$^{3}$H]-thymidine uptake, into NIH 3T3 cells. The DNA synthesis of NIH 3T3 cells was gradually increased at the nhEGF concentrations between 0.1 - 10 ng/ml in the Dulbecco's Modified Eagles Medium (DMEM) containing 0.2% Fetal calf serum (FCS). When we assayed the biological activity of four fractions, the activity of the second fraction was superior to that of the others. Based on the results from the HPLC analysis spiked with recombinant human epidermal growth factor (rhEGF) and amino acid sequencing, we concluded that the second fraction was nhEGF and the other three fractions were the derivatives of nhEGF. In addition, the proportion of nhEGF was approximately 46% is compared with that of the other three derivatives.

• 한국미생물·생명공학회지
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• 제24권5호
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• pp.553-559
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• 1996

We have designed nucleotide sequences of hEGF structural gene to eliminate the N-terminal methionine residue incorporated during the translation initiation step, and constructed recombinant human epidermal growth factor (rhEGF) secretion plasmids pYHB101, and pYHB2 in which pelB signal sequence-hEGF gene was expressed under the control of the T7, and tac promoter, respectively. We also constructed pYHB1 vector which contains rhEGF gene controlled by T7 promoter. The transformant with pYHB101 showed relatively slow growth pattern compared to the transformant with pYHB1. However, we observed that the transformant with pYHB101 secreted rhEGF of 13 mg/l significantly after 5 hr induction with 1 mM IPTG and that the T7 promoter was more effective than tac promoter when connected to pelB signal sequence. The amount of rhEGF was 14 mg/l under the sub-optimized condition.

• Bulletin of the Korean Chemical Society
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• 제35권8호
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• pp.2290-2294
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• 2014

For the present study, rhEGF was encapsulated into solid lipid nanoparticles (SLNs). The SLNs were prepared by the $W_1/O/W_2$ double emulsification method combined with the high pressure homogenization method and the physical properties such as particle size, zeta-potential and encapsulation efficiency were measured. The overall particle morphology of SLNs was investigated using a transmission electron microscopy (TEM). The percutaneous skin permeation and accumulation property of rhEGF was evaluated using Franz diffusion cell system along with confocal laser scanning microscopy (CLSM). The mean particle size of rhEGF-loaded SLNs was $104.00{\pm}3.99nm$ and the zeta-potential value was in the range of -$36.99{\pm}0.54mV$, providing a good colloidal stability. The TEM image revealed a spherical shape of SLNs about 100 nm and the encapsulation efficiency was $18.47{\pm}0.22%$. The skin accumulation of rhEGF was enhanced by SLNs. CLSM image analysis provided that the rhEGF rat skin accumulation is facilitated by an entry of SLNs through the pores of skin.

• 한국산학기술학회논문지
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• 제22권6호
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• pp.542-549
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• 2021

본 연구의 목적은 재조합 인간 상피세포 성장인자(hEGF)의 발현 확인 및 정제의 용이성을 위해 단백질의 N-말단에 융합된 부분을 제거하기위하여 기존의 고가의 효소를 사용하지 않고 간단한 화학처리로 융합 태그를 절단하면서도 여전히 친화성 크로마토그래피로 정제가 가능한 재조합 hEGF의 경제적이며 공정이 단순화된 제조법을 제공하는 것이다. 인간 상피세포 성장 인자는 인간 세포 성장 및 증식에 매우 중요한 호르몬이며 이 단백질에 대한 발현 및 정제에 관한 많은 연구가 보고 되었다. 본 연구에서는 hEGF 유전자를 대장균 코돈에 최적화 하였으며 N-말단에 Hydroxylamine에 의한 절단이 가능한 Asparagine과 Glycine이 발현되도록 포함하여 설계하였다. 제조한 DNA를 대장균 발현 벡터인 pRSET_A에 삽입하여 발현용 균주 BL21 (DE3)에 형질전환 시켰으며 재조합 융합 단백질은 대장균에서 샤페론 벡터 pG-Tf2와 성공적으로 공발현 되었다. 발현된 융합 단백질은 Ni-NTA 컬럼 크로마토그래피로 정제한 후 Hydroxylamine으로 처리해 N-말단 융합부분을 제거하였으며 SDS-PAGE를 통해 확인하였다. ELISA 분석 결과 재조합 EGF의 활성이 상업용 hEGF와 92% 이상 유사한 것으로 나타났으며 피부 섬유아세포의 세포증식을 촉진하는 것으로 확인 되었다.

• Archives of Plastic Surgery
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• 제41권5호
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• pp.466-471
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• 2014

Background Bleeding can be a problem in wound debridement. In search for an effective hemostatic agent, we experimented with a chitosan film combined with the recombinant human epidermal growth factor (rh-EGF), hypothesizing that it would achieve effective hemostasis and simultaneously enhance arterial healing. Methods Forty-eight Sprague-Dawley rats were used, and 96 puncture wounds were made. The wounds were divided into the following four groups: treated with sterile gauze, treated with gelatin sponge, treated with chitosan, and treated with chitosan combined with rh-EGF. Immediate hemostasis was evaluated, and arterial healing was observed histologically. Results Groups B, C, and D showed a significant rate of immediate hemostasis as compared to group A (P<0.05), but there were no significant differences among groups B, C, and D. Histologically, only group D showed good continuity of the vessel wall after 1 week. It was the only group to show smooth muscle cell nuclei of the vessel wall. Conclusions We observed that chitosan has an effective hemostatic potential and the mix of rh-EGF and chitosan does not interfere with chitosan's hemostatic capabilities. We also identified enhanced healing of vessel walls when rh-EGF was added to chitosan. Further research based on these positive findings is needed to evaluate the potential use of this combination on difficult wounds like chronic diabetic ulcerations.

• Radiation Oncology Journal
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• 제25권4호
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• pp.242-248
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• 2007

목적: 마우스 모델에서 cisplatin과 방사선조사로 구내염을 유발시킨 후 재조합 표피성장인자 (recombinant human epidermal growth factor, rhEGF)을 처치하여 그 효과를 알아보고자 하였다. 대상 및 방법: 마우스 24마리를 대상으로 정상대조군 8마리와 실험군은 각각 rhEGF 처치군과 미처치군으로 8마리씩 분류하였으며 실험군은 10 mg/kg의 cisplatin을 방사선조사 첫날 동시에 1회 복강 내 투여하였고, 방사선조사는 5일간 1회 5 Gy씩, 5일간 25 Gy를 조사하였다. RhEGF 처치군은 방사선조사 2일 전부터 2일간 1 mg/kg의 rhEGF를 피하주사하였으며 방사선조사3일째부터 3일간 1 mg/kg를 피하주사하여 총 5 mg/kg의 rhEGF를 투여하였다. 마우스의 체중변화, 먹이섭취 및 조직학적 변화를 평가하였다. 결 과: 마우스 체중변화는 rhEG F처치군이 실험 3일째부터 5일간 rhEGF 미처치군과 비교하여 통계적으로 유의한 체중의 차이를 관찰할 수 있었다. 먹이섭취는 실험군(rhEGF 처치군과 미처치군)에서 실험 5일째까지 감소하였다가 13일째부터 먹이섭취의 증가를 보여주었다. Cisplatin과 방사선조사 후 7일째의 조직학적 검사에서는 rhEGF 처치군에서 마우스의 점막 표피층의 변성이 관찰되었으나 rhEGF 미처치군에서는 점막층의 염증 반응이 관찰되었다. 결 론: Cisplatin과 방사선 조사로 마우스에서 발생한 구내염에서 rhEGF를 투여한 경우 체중 변화와 먹이섭취의 유의한 개선을 관찰하였으며 조직학적 검사에서 점막 손상의 회복을 확인하였다. 향후 임상적으로 rhEGF가 항암화학요법과 방사선치료로 인한 구내염을 줄일 수 있는 가능성을 확인하였다.값은 11개월이었고 2년, 3년 생존율은 31.5%, 15.8%였다. 결 론: 2기 췌장암 환자들은 국소 재발 및 원격 전이의 가능성이 높은 고위험군으로 국소 제어율 및 전체 생존율의 향상을 위해서 수술 후 효과적인 방사선치료의 적극적인 시행 및 이후의 보조적인 전신 항암화학요법을 권고하여 시행하는 것이 바람직하다. 평가 검사는 모두 제조사의 시험지침서에 부합하였다. 결론적으로, 이는 본 GE $Advance^{TM}$ PET 시스템이 임상에의 적용에 적합함을 보여주었다.mens사의 상용 분석 프로그램에는 해당 기능이 없어 계산값의 비교를 통한 검증은 수행하지 못하였고, 수화(hydration)와 탈수(dehydration) 각각의 조건에 따른 신장기능 분석에 적용하였을 때, 판별 기준이 되는 유의미한 값을 산출하여 타당성 검증을 대신하였다. 결론: 이 연구에서 개발한 핵의학 영상의 정량적 분석을 통한 신장기능 분석 프로그램을 사용하여 신장 기능을 분석한 결과, 타당성 있는 결과를 도출하여 그 유용성을 입증하였다. 이 개발 프로그램은 좀 더 다양한 임상응용 목적의 분석기능을 사용자가 직접 개발, 추가하기가 용이하여 기존 상용 프로그램보다 연구적 활용범위가 크다고 사료된다.2.2{\pm}0.4\;(1.6{\sim}3.2){\mu}g/ml$와 $1.4{\pm}0.2\;(0.8{\sim}1.6){\mu}g/ml$로서(p=0.16), 표준균주 3종 모두에서 Infecton의 MBC 또한 ciprofloxacin에 비해 $2{\sim}4$배가 높았다. 결론: Tc-99m Infecton은 ciprofloxacin 보다는 약하였지만 표준균주에 대해 생체외 항균력을 보였다.를

• 약학회지
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• 제42권5호
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• pp.534-539
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• 1998

Effect of recombinant human epidermal growth factor (rhEGF, DWP401) on fetal external, visceral and skeletal malformation during organogenesis was examined. Pregnant Sprauge-Daw ley rats were administered with 0.2, 1 and 5mg/kg/day subcutaneously on gestation day 6 through 16. Dams were sacrified at 20th day of gestation. Materal body weight, food consumption and clinical observation were not changed. Significant dose-dependent increase of relative and absolute liver weight were observed in the treatment group, whereas other organ weights were not changed. Placental weight of 1 and 5mg/kg/day group and number of resorption in 5mg/kg/day treatment group were significantly increased. External and visceral malformation of fetuses were not observed with treatment. However, skeletal variations(increase of asymmetry sternebrae, decrease of dumb-bell and asymmetry sternbrae at 5mg/kg/day, and fused stemebrae at 5mg/kg/day) were observed. These results showed that rhEGF (DWP401) may not have embryo and/or fetal developmental toxicity effect in rats.

• Journal of Pharmaceutical Investigation
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• 제28권2호
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• pp.99-107
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• 1998

The objective of this study was to develop effective oral formulations of rhEGF for gastric ulcer healing using polycarbophil. hydroxypropylcellulose(HPC) and sucralfate as its bioadhesive bases. Cytoprotective effects of rhEGF, cell proliferation and differentiation. on the ulcers induced by ethanol or acetic acid in rats were studied. rhEGF release from HPC formulation was much faster than that from polycarbophil formulation. HPC formulation combined with small amount of sucralfate showed much slower release of rhEGF than only HPC base only. rhEGF preparations with bioadhesive polymers showed better effects on the healing of gastric ulcers than EGF solution when administered orally. When rhEGF preparations were administered at once and the animals were under starvation, polycarbophil formulation showed better effect on gastric ulcers than HPC formulation. Otherwise, when rhEGF preparations were given more than three times and the rats were fed normally, HPC formulation showed good healing efficacy of ulcers compared to polycarbophil formulation. rhEGF showed dose-dependent effect on the healing of both chronic and acute ulcers.