• 생명과학회지
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• 제20권11호
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• pp.1577-1581
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• 2010

조혈촉진 cytokine인 Granulocyte colony stimulating factor (G-CSF)는 골수세포를 자극하여 granulocyte로 증식, 분화시키는 기능을 가지며, 현재 아주 고가의 치료제로 사용되고 있다. 인간 G-CSF (hG-CSF)를 아직 시도되지 않은 누에 유래 세포주인 BM5 세포에서 발현시키고 생산 효율을 높이기 위해 hG-CSF cDNA를 변형하였다. hG-CSF의 cDNA의 endoplasmic reticulum (ER) signal sequence 부분을 누에의 소포체에서 분비되는 단백질인 prophenoloxidase (PPAE), protein disulfide isomerase (PDI)와 bombyxin (BX)에서 유래한 누에특이 ER signal sequence로 대체한 hG-CSF의 cDNA 함유 벡터를 구축하였다. 이들 벡터를 사용하여 형질전환한 BM5 세포의 배양액에 분비된 G-CSF 단백질을 western blot으로 분석하여 발현을 확인하였다. 누에특이 ER signal sequence들로 대체된 hG-CSF cDNA를 포함하는 벡터에 의한 hG-CSF 단백질 생산이 인간 G-CSF cDNA가 든 벡터에 의한 hG-CSF의 생산보다 월등히 효율적이었다. 또한, PPAE-signal sequence를 포함하는 hG-CSF 단백질은 배양배지에서 형질전환 4일 후에 최고에 달하였고, 7 일째까지 비슷한 양이 배지 내에서 검출되었다. 이상의 결과는 인간유래 유전자가 곤충세포 내에서 발현 될 때 인간유래 유전자 보다는 곤충 유전자발현 시스템에 맞게 변형했을 경우 더 효율적인 단백질 발현을 얻을 수 있음을 보여 준다.

• IMMUNE NETWORK
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• 제1권1호
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• pp.32-35
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• 2001

Background: rhGM-CSF has been shown to enhance the migration and proliferation of endothelial cells and to promote keratinocyte growth. This study was tried to evaluate the effect of rhGM-CSF dressing on the uninfected wounds. Methods: Thirty Sprague-dawley white mice(250-300g) were selected in this study. The number of wound with the diameter of 5 mm, was 3 in left and 3 in right at the symmetric sites, respectively. The site of rhGM-CSF dressing was decided by a randomization. rhGM-CSF($Leucogen^{(R)}$) was diluted in the distilled water($5{\mu}g/mL$). The experimental wound group was dressed by l mL of distilled water mixed with rhGM-CSF and control wound group was dressed by l mL of distilled water. The dressing was done, every 24 hours. The criteria of comparison were the duration of wound healing duration, histologic findings and the bacterial culture of wound sites. Results: The duration of wound healing was $10.3{\pm}1.7days$ in experimental group and $10.2{\pm}2.8days$ in control group, without significant difference. There was no specific difference of histologic findings between both groups. The pathogen was not found, at all. Conclusion: It seems to be that rhGM-CSF has no prominent effect on the uninfected wound healing in the mice without immune suppression.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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• pp.314-314
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• 1994

대상환자는 15예로, 14예에서 평가 가능하였으며 남녀비는 8:6, 중앙연령 32세(10-70세) 이었고, 대상질환은 악성골옥종 7예, 악성임파종 2예, 위암 2예, 폐암 2예, 그리고 자궁평활근육종 1예였다. rhGM-CSF 50 $\mu\textrm{g}$/$m^2$ 3예, 100 $\mu\textrm{g}$/$m^2$ 3예, 150 $\mu\textrm{g}$/$m^2$ 3예, 250 $\mu\textrm{g}$/$m^2$ 3예, 350 $\mu\textrm{g}$/$m^2$ 3예, 500 $\mu\textrm{g}$/$m^2$ 6예, 700 $\mu\textrm{g}$/$m^2$ 용량 3예에서 시행되었다. 1주기 시행한 환자는 7예, 2주기 5예, 3주기, 4주기 각각 1예씩 있다. 부작용은 50-150 $\mu\textrm{g}$/$m^2$에서 WHO grade I의 발열, 전신쇠약, 식용부진등이 관찰되었으나 grade II이상의 부작용은 없었다. 250 $\mu\textrm{g}$/$m^2$이상의 용량에서도 grade II의 발열이 관찰되었을 뿐 다른 중증의 부작용은 관찰되지 않았다. 최고용량인 700 $\mu\textrm{g}$/$m^2$ 에서도 grade II의 발열외의 중한 부작용은 관찰되지 않았다. 각 용량에 따른 백혈구 증가율(%투여제2일/투여제1일)은 130-500% 이었다. rhGM-CSF는 투여 2-4 시간후 혈중최고치 (0.42-15.4 ng/ml)가 관찰되었으며 투여 12시간까지 0.2-2 ng/ml 의 농도가 지속되었다. 소변내 rh GM-CSF 배설량은 총투여량의 1% 미만이었다.

• Biomolecules & Therapeutics
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• 제2권3호
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• pp.281-285
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• 1994

Neutropenia is a major dose-limiting factor in cancer chemotherapy diminishing its usefulness and increase patient's susceptibility to infectious disease. Some recombinant human granulocyte colony stimulating factors(rhG-CSFs) are in use to reduce the risk of this serious side effect. In this study, we examined the pharmacological properties of DA-3030, a rhG-CSF expressed in E. coli. DA-3030 100 and $\mu\textrm{g}$/kg, i. v., had no significant effect on the central nervous, gastrointestinal system in mice and cardiovascular system in rabbits, but it slightly inhibited the spontaneous motility of isolated nonpregnant uterus in rats. It also had no influence on excretion of urinary electrolytes. DA-3030 administered for successive 3 days increased the blood WBC count in zymosan air pouch inflammed rats and in normal rats. These results indicate that DA-3030 has little side effects in animals.

• Toxicological Research
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• 제23권4호
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• pp.383-389
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• 2007

Recombinant human granulocyte-macrophage colony stimulating factor (hGM-CSF) regulates proliferation and differentiation of hematopoietic progenitor cells and modulates function of the mature hematopoietic cells. In the previous study, we reported that hGM-CSF could be produced in transgenic rice cell suspension culture, termed rhGM-CSF. In the present study we examined the single and repeated dose toxicity of rice cells-derived hGM-CSF in SD rats. During single dose toxicity study for 7 days, there were no any toxic effects at any dose of from 10 to $1000{\mu}g/kg$. The lethal dose ($LD_{50}$) was not found in this range. Moreover, repeated dose toxicity study of 14-days period and at the doses of 50 and $200{\mu}g/kg$ (i. v.) of rhGM-CSF did not show any changes in food and water intake. There were also no significant changes in both body and organ weights between the control and the test groups. The hematological and blood biochemical parameters were statistically not different in all the groups. These results suggest that rhGM-CSF has no toxicity in SD rats.

• 대한수의학회지
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• 제57권3호
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• pp.155-158
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• 2017

Surgical castration performed to reduce male-associated problems results in pain and microbial infections in male animals. Therefore, immunocontraception, which is mediated by the animal's own antibodies against reproductive hormones, has been recommended as an alternative to surgical castration when considering the animal's welfare. In this study, a new immunocontraceptive vaccine composed of six tandem copies of gonadotropin-releasing hormone (GnRH) fused to rat granulocyte-macrophage colony-stimulating factor (GM-CSF) was developed, and its efficacy was evaluated in male rats. Three different doses (10, 50, and $100{\mu}g$) of recombinant GM-CSF-GnRH protein were injected three times at intervals of two weeks into male rats. The rats vaccinated with three doses of GM-CSF-GnRH produced a significantly higher level of antibodies against GnRH than that in the negative control rats. Severe atrophy of gonads was observed in rats vaccinated with three doses of GM-CSF-GnRH but not in the negative control rats. The results reveal that the new GnRH vaccine conjugated with rat GM-CSF induces efficient immunocontraception in male rats. This formulation of the immunocontraceptive vaccine would be applicable to both domestic and pet male animals.

• Toxicological Research
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• 제24권4호
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• pp.315-320
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• 2008

Recombinant human granulocyte-macrophage colony stimulating factor (hGM-CSF) is a glycoprotein and hematopoietic growth factors that regulates the proliferation of myeloid precursor cells and activates mature granulocytes and macrophages. In a previous study, we reported that hGM-CSF could be produced in transgenic rice cell suspension culture, termed rhGM-CSF. In the present study, we examined the repeated dose toxicity of rhGM-CSF in SD rats. The repeated dose toxicity study was performed at each dose of 50 and 200 ${\mu}g/kg$ subcutaneous administration of rhGM-CSF everyday for 28-days period. The results did not show any changes in food and water intake. There were also no significant changes in both body and organ weights between the control and the tested groups. The hematological and blood biochemical parameters were statistically not different in all groups. These results suggest that rhGM-CSF may show no repeated dose toxicity in SD rats under the conditions.

• Biomolecules & Therapeutics
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• 제2권3호
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• pp.247-255
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• 1994

As a series of safety studies of DA-3030, a new rhO-CSF, its local irritancy was examined in the rabbits after the following treatment; application into the conjunctival sac of the eye(single), subcutaneous injection(single), intramuscular injection(single), and intravenous injection(8-day repeated). In addition, paravenous irritation of DA-3030 was investigated in mice. The results obtained were as follows. 1. In the result of ocular irritation test, 0.03% solution of DA-3030 could be considered as a non-irritating material. 2. The local irritation of DA-3030 by an injection of 0.5mι of its solution subcutaneously or intramuscularly was negligible and not so much different from that of saline. 3. In the vascular irritancy test, macro- and microscopic observations revealed that the irritating activity of DA-3030 in blood vessels was not different from that of saline when they were injected once a day into vein retroauricularis of rabbits for 8 days.4. The paravenous administration of DA-3030 did not induce any abnormal changes at injection sites except mild swelling in 1 mouse at 3 hours after injection which was thought to be due to slow absorption. The above-mentioned results suggest that DA-3030 has no irritating activity when injected through intravenous or subcutaneous route for clinical practice as 0.03% solution.

• Biomolecules & Therapeutics
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• 제2권4호
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• pp.310-315
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• 1994

The pharmacokinetics and tissue distribution of DA-3030 (recombinant human granulocyte colony-stimulating factor, rhG-CSF, recently manufactured by Dong-A research laboratory of Dong-A Pharmaceutical Company) were compared with reported data in the literature. After intravenous(i.v.) administration of DA-3030, at dose of 5, 10 and 100 $\mu\textrm{g}$/kg to rats, some pharmacokinetic parameters, such as terminal half-lives(1.05, 1.19 and 1.83 hr, respectively) and clearance (84.0, 54.8 and 45.5 mι/hr/kg, repectively), were dose-dependent. This could be due to the saturable metabolism of DA-3030 in rats. Similar results were also reported. After subcutaneous(s.c.) and intramuscular(i.m.) administrations of DA-3030, 10 $\mu\textrm{g}$/kg to rats, the extent of bioavailability(absolute bioavailability) were incomplete; the values were 23.3 and 18.2% after s.c. and i.m. injections, respectively, due to the degradation of DA-3030 by protease. After 7-consecutive day i.v. administrations of DA-3030, 10 $\mu\textrm{g}$/kg/day, to rats, the plasma concentrations and pharmacokinetic parameters of DA-3030 were not significantly different from those in single administration. In mice and dogs at DA-3030 dose of 10 $\mu\textrm{g}$/kg, the plasma concentrations of DA-3030 were also declined rapidly with terminal half-lives of 1.31 and 1.15 hr, respectively. DA-3030 was highly concentrated in the kidney after i.v. administration of DA-3030, 10 $\mu\textrm{g}$/kg, to rats, and the results were similar to those obtained using radiolabelled rhG-CSF in the literature. Above data indicate that DA-3030 has similar properties to rhG-CSF manufactured by other companies in view of pharmacokinetics.

• Biomolecules & Therapeutics
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• 제6권2호
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• pp.145-150
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• 1998

Administration of 3 type KGCs [recombinant human granulocyte colony-stimulating factor (rhG-CSF)] to mice with cyclophosphamide (CPA)-induced neutropenia for 4 consecutive days from the day after the CPA dosing (100 mg/kg) resulted in a dose dependent increase in the peripheral blood neutrophil count 6 hours after the final KGC injection. Within the KGC dose range of 0.1 to 40$\mu$g Per mouse Per day, there was a sigmoidal relationship between the logarithm of the dose and the peripheral blood neutrophil count (relative value for neutrophil count of the basal dose) in the treated mice. The sigmoidal relationship of test KGC preparations shows that there is a saturation point in terms of efficacy. Compared with e(fact of KGC-Orange, Green, and Blue, KGC-Orange recovers neutrophils more effectively than the others do.