• Toxicological Research
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• v.13 no.1_2
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• pp.149-152
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• 1997

This test was performed to evaluate the ocular and skin irritation of EPO (Erythropoietin). The results as follows: 1. Ocular irritation test There were no observed clinical signs, body weght changes by EPO during experimental period. The acute ocular irritation index(A.O.I.), mean ocular irritation index(M.O.I.) and Day-7 individual ocular irritation index(I.O.I.) of EPO at dose of 1000U and 10, 000U were 0, respectively. Therefore we evaluated that EPO was non-toxic to eyes. 2. Skin irritation test There were no observed clinical signs, body weght changes and gross pathologic findings by EPO during experimental period. There were no observed erythema, eschar formation and edema formation on intact and abraded skin treated by EPO. The primary irritation index(P.I.I.) of EPO at dose of 1000U and 10, 000U were 0, respectively and were evaluated none irritating product about skin irritation.

• Biomolecules & Therapeutics
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• v.6 no.1
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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.

• Toxicological Research
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• v.13 no.1_2
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• pp.131-138
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• 1997

Group of 40 male and 40 female Sprague-Dawley rats were given daily intravenous injections of different dosage of Erythropoietin (EPO), 80 IU/ kg/day (low dosage group), 400 IU/ kg/day (middle dosage group), or 2000 IU /kg/day (high dosage group)for 4 weeks by tail vein according to Established Regulation of Korean National Institute of Safety Research (1994. 4. 14). Appearance, behavior, mortality, and food consumption of rats of treated groups were not affected during the experimental periods. No significant EPO (erythropoietin)-related changes were found in urinalysts, eye examination, hematology, serum chemistry, and organ weight. No histopathological lesions were observed in both control and treatment groups. Our results strongly suggest that no toxic changes were found in rat treated intravenously with EPO (erythropoietin)for 4 weeks.

• Biomolecules & Therapeutics
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• v.8 no.2
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• pp.171-178
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• 2000

To evaluate the pharmacokinetic properties and tissue distribution of a newly developed recombinant human erythropoietin (GC-rhEPO), we analyzed the plasma and tissue levels of erythropoietin by an ELISA after intravenous (IV) and subcutaneous (SC) adminstration to the male rats at the doses of 20, 100, 500 or 2,500 unit/kg. After single IV bolus injection of GC-rhEPO, the plasma concentration was rapidly increased and decreased with two phases with half-lives of 13.4 min and 2.94 hours. AUC was increased dose- dependently but plasma half-lives remained constant regardless of GC-rhEPO doses. Following SC administration, the plasma concentration increased slowly with half-life of 9.2 hours and reached peak at 8 hours. Mean residence time and bioavailability were 18.2 hours and 44%, respectively. After single IV dose of 100 unit/kg, tissue GC-rhEPO level was higher in bone marrow and spleen, while the depletion rate was slower in liver and bone marrow, indicating the higher affinity of GC-rhEPO to bone marrow. Taken together, the experimental results indicate that GC-rhEPO contained the typical pharmacokinetic properties and the tissue distribution patterns inherent to human erythropoietin.

• Toxicological Research
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• v.11 no.1
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• pp.77-80
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• 1995

Antigenic potential of genetically-engineered human erythropoietin (EPO) was assessed in guinea pigs (active systemic anaphylaxis [ASA] ; passive cutaneous anaphylaxis [PCA]) and in vitro (hemagglutination test [PHA]). In ASA, EPO at 70~700 U/kg elicited a weak anaphylactic response tvhereas the positive control ovalbumin (OVA) did cause intensive responses leading to death in 40% animals. However, the extract of CHO cells, to which EPO gene was introduced, did not cause any symptom. In PCA and PHA tests, neither EPO nor CHO cell extract induced positive responses. OVA, in contrast, produced high titers in both PCA and PHA tests. It was concluded that, in light of the fact that EPO was slightly antigenic only in ASA but not in PCA or PHA and also that human EPO is a foreign protein to guinea pigs, the present EPO may not be antigenic in humans.

• Biomolecules & Therapeutics
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• v.2 no.4
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• pp.343-346
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• 1994

In vivo activity of recombinant human erythropoietin (rh-EPO) has been examined using polycythemic model in mice and acute hemorrhage model in rats. The number of reticulocytes in blood stream was increased after a single injection of rh-EPO depending on the dosage of rh-EPO in polycythemy model. It seemed that optimal dose of rh-EPO for polycythemic mice was around 1-10 U/kg. Rh-EPO also showed the effectiveness for increase of reticulocyte numbers both in male and female rats after bleeding. The number of reticulocytes and the change of hemoglobin concentration in the blood stream of normal rats has been examined after injection of rh-EPO. The maximum value of reticulocyte was observed on the 6th day of the injection in these normal rats. In addition, the increase of reticulocyte and the concentration of hemoglobin were dependent on the dosage of rh-EPO. The increase of hemoglobin concentration was continued to the 9th day after injection. In this study, the efficacy of rh-EPO was confirmed in both mice and rats.

• Toxicological Research
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• v.13 no.1_2
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• pp.139-147
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• 1997

Four-week toxicity of EPO(erythropoietin) was investigated using New Zealand White rabbits according to the established regulations of Korean National Institute of Safety Research. Rabbits were administered intravenously seven days per week for 28 days with dosage of 0, 80, 400 and 2000IU/kg B. W./day. Animals administered with EPO showed no significant changes of body weight, water consumption and feed consumption, and no clinical signs and death. They were not significantly different from the control group in hematological and serum biochemical analysis, urinalysis, prothrombin time, and partial thromboplastin time. In this study, we concluded that EPO had no toxic effect in the New Zealand White rabbits when they were administered intravenously below 2000IU/ kg B.W./ day for 28 days.

• Toxicological Research
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• v.13 no.1_2
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• pp.127-130
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• 1997

Acute toxicity of EPO(Erythropoietin) was investigated using rats and beagle dogs according to Established Regulation of Korean National Institute of Safety Research (1994. 4. 14). Rats and beagle dogs were injected intravenously with dosages of 20000 IU/kg, 2000 IU/kg, 200 IU/kg, 20 IU/kg and 2 IU/kg. In animals injected with EPO, there were neither dead animals nor significant changes of body weights. In addition, no differences were found between control and treated groups in clinical signs and autopsy findings. Therefore $LD_50$ of EPO was considered to be higher than 20000 IU/ kg B. W. in rats and beagle dogs.

• BMB Reports
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• v.30 no.5
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• pp.315-319
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• 1997

It was discovered that monoclonal anti-erythropoietin (EPO) antibody CFC-6 can neutralize EPO-induced cell activation. To know the binding site of CFC-6, recombinant human erythropoietin (rhEPO) was digested with Glu-C, followed by a separation using high performance liquid chromato graphy (HPLC). Each HPLC fraction was blotted on the nitrocellulose membrane and the membrane was treated with anti-EPO antibody CFC-6 and anti-mouse antibody which is modified with peroxidase. Only one spot showed the color and the fraction was sequenced by Edman degradation. The results suggest that CFC-6 recognizes amino acid sequence V63-W-Q-G-L-A-L-L-S-E72 which is a part of helix II of the EPO molecule. Binding of CFC-6 to EPO may inhibit EPO binding to its receptor, which implies that the antibody binding site and the receptor binding site are close or overlapping.

• Journal of Yeungnam Medical Science
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• v.11 no.1
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• pp.115-126
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• 1994

The recent availability of recombinant human erythropoietin has opened new perspectives in the management of a variety of anemias. Clinical trials have been initiated in several countries using different approaches and methodology. We randomly assigned twelve premature infants(gestational age < 32 week) at high risk of requiring erythrocyte transfusion for anemia of prematurity with either subcutaneous recombinant human erythropoietin or a placebo. Treatment with rHuEPO was initiated at a dose of 100 units/kg day for 3 days a week. All patients were given supplemental oral iron therapy at a dose of 3 mg/kg per day, as tolerated and oral vitamin E at a dose of 25 units per day. Treated and control babies did not differ with respect to weight, hematocrit, overall mean reticulocyte count or rate of growth respectively. However, reticulocyte counts increased earlier in patients given rHuEPO. We conclude that rHuEPO administration is safe and feasible at the dose studied.