• Biomolecules & Therapeutics
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• 제6권1호
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• pp.50-55
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• 1998

Antigenic potential of a recombinant human erythropoietin (rhEPO) produced by Dong-A charm. Co. Ltd. was examined by active systemic anaphylaxis (ASA) test in guinea pigs, mouse-rat passive cutaneous anaphylaxis (PCA) reaction and passive hemagglutination (PHA) test. In ASA test, rhEPO induced the signs of restlessness, rubbing or licking nose, sneezing and coughing in the animals immunized with rhEPO 1000 lU/kg alone or rhEPO 1000 lU/kg incorporated into Freund\\\\`s complete adjuvant. In the mouse-rat PCA test, only one of six sera from the animals immunized with rhEPO 1000 lUng incorporated into Alum showed positive result. In the PHA test, rhEPO revealed negative results in all of the rhEPO-immunized groups. From these results, rhEPO was considered to produce IgE in guinea pigs and mice, but not IgG and/or IsM in mice. The results of this study were similar to those of the other recombinant human erythropoietin and these positive results were thought to be caused due to the fact that rhEPO were heterogeneous proteins to guinea pigs and mice. Considering the fact that rhErO has an identical structure with indigenous human erythropoietin, rhEPO is not thought to cause immunological problems in clinical use.

• KSBB Journal
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• 제32권2호
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• pp.90-95
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• 2017

Mammalian cell cultures have been used extensively to produce proteins for therapeutic agent because of their ability to perform post-translational modification including glycosylation. To produce recombinant protein, many factors and parameter are considered such as media composition, host cell type, and culture process. In this study, recombinant human erythropoietin (rhEPO) producing cell line was established by using glutamine synthetase system. To reduce serum concentration in media, we compared direct adaptation with step adaptation. Cell growth was faster in step adaptation. In low-level serum media, there were insufficient glucose for cell growth. Thus, we added glucose in low-level serum media from 2 g/L to 4.5 g/L. Titer of rhEPO was higher than other conditions at 4.5 g/L of glucose. Additionally, N-methyl-D-aspartate (NMDA), 13-cis-retinal, and pluronic F-68 (PF-68) were added to enhance productivity in CHO cell cultures. In conclusion, we applied CHO cell producing rhEPO to low-level of serum in media using step-adaptation. Also, we confirmed positive effect of NMDA, 13-cis-retinal, and PF-68.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제37권1호
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• pp.9-14
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• 2011

Recently, the population of patients who refuse transfusion has increased for both religious and non-religious reasons, even in life threatening emergency situations. Their refusal has highlighted the need to develop nonblood transfusion surgery techniques to decrease the risk from blood transfusions. A 57-year woman with an ulcerative lesion on the gingiva of the right upper molar area visited the department of oral and maxillofacial surgery in Dankook University Dental Hospital. After a preliminary evaluation, the patient was diagnosed with squamous cell carcinoma. As she refused blood transfusion during surgery for religious reasons, surgery was planned using recombinant human erythropoietin (rHuEPO) without a blood transfusion. The patient underwent a partial maxillectomy, supraomohyoid neck dissection, free radial forearm flap and split thickness skin graft under general anesthesia. rHuEPO and iron were used before and after surgery. The hemoglobin/hematocrit (Hb/Hct) level, iron (Fe) and total iron-binding capacity (TIBC) were assessed. The patient recovered completely without any blood transfusions. rHuEPO is a viable alternative for patients with religious objections to receiving blood transfusions.

• Archives of Plastic Surgery
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• 제33권5호
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• pp.621-626
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• 2006

Purpose: Erythropoietin is traditionally known to regulate erythropoiesis, but recently its protective effect against ischemia-reperfusion injury has been studied mainly in cardiovascular and neuronal systems. This study was planned to investigate the effects of recombinant human erythropoietin on ischemia-reperfusion injury in rat TRAM flap model. Methods: Superiorly based TRAM flap was elevated and ischemic insult was given for four hours. Thirty minutes before reperfusion, single dose recombinant human Erythropoietin(5000IU/kg) was injected via intraperitoneal route in the treatment group. At 24 hours postoperatively, systemic neutrophil count, tissue myeloperoxidase activity, malonyldialdehyde amount, nitric oxide content, tissue water content and histologic finding of inflammation was evaluated. On 10 days postoperatively, flap survival rate, angiogenesis and change in hematocrit level was evaluated. Results: Tissue nitric oxide level was significantly higher and myeloperoxidase activity was significantly lower in the treatment group 24 hours after reperfusion. Tissue water content was significantly lower in the treatment group. Perivascular neutrophil infiltration and intravascular adhesion was marked in the control group. Mean flap survival after ten days was 69% in the treatment group, and 47% in the control group, demonstrating a significant difference. Neovascularization in the treatment group also outnumbered the control group. No significant hematocrit rise was noted ten days after erythropoietin administration. Conclusion: Recombinant human Erythropoietin improved flap survival in ischemia-reperfusion injured rat TRAM flaps, at least partially owing to suppressed inflammation, increased nitric oxide, and enhanced angiogenesis.

• Biomolecules & Therapeutics
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• 제6권3호
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• pp.317-327
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• 1998

The subchronic toxicity study of rHuEPO, a newly developed recombinant erythropoietin, was investigated for 13 weeks in Beagle dogs intravenously treated with doses of 100, 500 and 2,500 lU/kg/day. There were no significant changes in body weight, food intake, physical and opthalmic examination, urine analysis, etc. Any toxic response was not observed except for enlarged spleen and extramedullary hematopoiesis. These results indicate that the no-observed adverse effect level (NOAEL) of rHuEPO is 100 lU/kg in Beagle dogs.

• Toxicological Research
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• 제12권1호
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• pp.101-111
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• 1996

The local irritation studies of DA-3285, recombinant human erytropoietin(rHu-EPO), were carried out in rabbits after the following treatment; single application into the conjunctival sac of the eye, single subcutaneous injection, 7-day repeated subcutaneous injection and 8-day repeated infusion into the ear vein. Also, the local irritancy of DA-3285 leaked around vein was studied in mice by single perivascular injection. The results obtained were as follows. In the result of ocular irritation test, DA-3285 could be considered as a non-irritating material. In single and 7-day repeated subcutaneous irritation test, the irritancy of DA-3285 was not so much different from that of saline. The vascular irritancy of DA3285 by 8-day repeated infusion was negligible and similar to that of saline. And the irritancy of DA3285 by perivascular injection was comparable to that of saline. These results indicate that DA-3285 has no irritating activity when injected through subcutaneous or intravenous route for clinical practice as 3.5% solution.

• Biomolecules & Therapeutics
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• 제4권4호
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• pp.334-338
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• 1996

Antigenicity of DA-3285, human recombinant erythropoietin which was produced from mammalian cells, was examined in guinea pigs and mice. In active systemic anaphylactic test, mild anaphylactic signs were observed in guinea pigs sensitized subcutaneously with DA-3285 or DA-3285 incorporated with complete Freund's adjuvant(CFA) when challenged with high dose(1000 IU/Kg) of DA-3285. Other groups showed negative responses. In mouse-rat passive cutaneous anaphylaxis test, 20% sera of mice immunized with DA-3285 or DA-3285 mixed with aluminum hydroxide(alum) showed mild positive responses. In the case of indirect haemagglutination reaction(IHA) test, when sheep red blood cells coated with DA-3285 was incubated with mouse serum, all the serum samples were showed negative responses. These results suggest that DA-3285 has a very weak antigenic potential and probably would not induce systemic allergic reactions in clinical uses.

• Biomolecules & Therapeutics
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• 제11권2호
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• pp.126-132
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• 2003

Efficacy and in vivo bioassay of recombinant human erythropoietin (rh-EPO, DWP413) was investigated. Efficacy studies on erythropoiesis were conducted in normal, cisplatin-induced anemic rats and acute hemorrhage - induced anemic rats. Animals were treated intravenously with DWP413 for 5 days, the changes in the number of red blood cells (RBC), hematocrit value (Hct), hemoglobin concentration (Hb) and reticulocyte were examined. In normal rats, at the doses of 50, 250, and 1250 IU/kg/day, in cisplatin-induced anemic rats, at the doses of 50, 100 and 200 IU/kg/day, RBC, Hb, Hct and reticulocyte were increased dose-depen-dently. And in acute hemorrhage-induced anemic rats, DWP413 (150, 450 and 1350 IU/kg/day) significantly increased RBC, Hb, Hct and reticulocytes. In histopathological findings of kidney, cisplatin alone treated rats expressed severe glomerulus and tubular damage. But in the DWP413 treated rats after cisplatin treatment, these were not remarkable compared to cisplatin alone treated rats. In vivo bioassay, DWP413 had 102.43% of bioactivity compared to erythropoietin BRP(Biological Reference Product, European Directorate for the Quality of Medicines). These results suggest that DWP413 might be useful for the therapy of anemia induce by renal failure and acute blood loss.

• Biomolecules & Therapeutics
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• 제4권2호
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• pp.154-161
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• 1996

General pharmacological properties of LB-00014, an erythropoietin which was produced by recombinant DNA technique in Biotech Research Institute, LG Chemical Ltd. were examined. The administration of LB-00014 (60, 600, 6000 IU/kg, iv) in mice had no effect in general behavior and central nervous system, and no influences on normal body temperature, writhing syndromes induced by 0.7% acetic acid solution and chemoshock produced by strychnine and pentetrazole solution. LB-00014 (60, 600, 6000 IU/kg, iv) given to anesthetized rabbits showed no effect on blood pressure of carotid artery and respiration rates, and it did not influence the responses produced by injection of acetylcholine or epinephme. The administration of LB-00014 (601, 600, 6000 IU/kg, iv) in rats had no effect on the plasma prothrombin time, activated partial thromboplastin time and hemolytic action. The platelet aggregation induced by collagen in human plasma was not influenced by LB-00014 (10, 100, 1000 lU/kg, iv). It showed no direct effect at 100 and 1000IU/m1 in isolated stomach fundus and uterus of rats and ileum of guinea-pig, and it also had no inhibition of contraction produced by acetylcholine, oxytocin, serotonin and histamine in the above-mentioned preparations. It did not influence gastric secretion, pH and acid output at 6000 IU/kg, iv in rats, but showed a significant increase in intestinal propulsion of mice at 6000 IU/kg, iv. Its administration (60, 600, 6000 lU/kg, iv) caused no effect on urine and electrolyte excretion in rats. These results indicate that LB-00014 does not exsert any of serious pharmacological effects.

• Journal of Yeungnam Medical Science
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• 제11권1호
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• pp.115-126
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• 1994

1991년 1월부터 1993년 2월까지 영남대학교 의과대학 부속병원에서 출생하여 신생아 중환자실에서 치료 받았던 12명의 미숙아를 대상으로 무작위로 대조군과 rHuEPO투여군으로 나누어 체중 kg당 100unit의 rHuEPO을 1주간 3회씩 4주동안 피하주사하고 rHuEPO투여 시작과 같이 체중 kg당 3mg의 elemental iron을 매일 경구투여 하였고 출생 후 수유가 가능할 때 부터 25unit씩의 비타민 E를 매일 경구투여 하였다. 망상적혈구, 혈색소치, 백혈구수, 혈소판수, 혈청 철, ferritin, 총 철결합능, erythropoietin농도, 활력징후, 체중, 간기능, 신장기능 및 피하주사부 위의 국소 부작용을 비교해 본 결과 치료 시작전 erythropoietin의 농도는 대조군과 rHuEPO투여군 사이에 차이는 없었으며 모두 정상 범위에 속했고 rHuEPO투여군에서는 치료 1주에서 3주사이에 급격한 망상적혈구의 경향이 관찰되었고 전반적으로 혈색소치, 백혈구수, 혈소판수, 혈청 철, ferritin, TIBC등에서는 차이가 없었다. 그리고 체중증가 및 활력증상, 간기능, 신장기능에서도 대조군과 rHuEPO투여군 사이에서 차이가 없었고 rHuEPO 투여 기간 동안 계속 정상적으로 유지되었다. 결론적으로 우리의 연구에서는 rHuEPO치료로 뚜렷한 혈색소치 및 헤마토크리의 증가를 볼 수 없었지만 망상적혈구가 치료 초기에 증가되는 경향이 관찰되었고 치료로 인한 특이할 만한 부작용이 없었으므로 첫째, 치료 기간 중의 예견할 수 없이 발생하는 패혈증, 무시할 수 없는 채혈량과 spontaneous endogenous erythropoietin생산이 rHuEPO의 반응에 미치는 영향, 둘째. rHuEPO의 가장 높은 효과를 얻기 위한 보다 적절한 투여용량, 기간 및 rHuEPO의 투여를 출생 후부터 바로 예방 목적으로 할 것인지 또는 출생 몇 주 후부터 rescue therapy로 할 것인지 투여 시작 시기, 셋째, 용량의 증가 및 감소에 동반되는 부작용의 정도등을 고려해서 앞으로 더 많은 미숙아를 대상으로 지속적인 연구를 한다면 rHuEPO을 이용해 더욱 더 활발하고 효과적으로 미숙아 빈혈을 치료할 수 있으리라 기대한다.