Guinea pig 및 mouse 에 있어서 인형 과립구 콜로니 자극인자 DA-3030 의 항원성

백남기(Nam Gi Baik),강경구(Kyung Koo Kang),이순복(Soon Bok Lee),김원배(Won Bae Kim),양중익(Jung Ick Yang) 한국응용약물학회 1994 Biomolecules & Therapeutics(구 응용약물학회지) Vol.2 No.3

This study was conducted to investigate antigenic potential of DA-3030, a recombinant human granulocyte-colony stimulating factor, in guinea pigs and mice. In the active systemic anaphylaxis test, the guinea pigs sensitized with 1.25 or 12.5 ㎍/head of DA-3030 alone did not show any anaphylactic reaction. In the homologous passive cutaneous anaphylaxis reaction, anti-DA-3030 antibody was not detected in guinea pigs sensitized with 1.25 or 12.5 ㎍/head of DA-3030 alone. On the other hand, the guinea pigs sensitized with 12.5 ㎍/heed of DA-3030 incorporated in Freund`s complete adjuvant(FCA) or 1 mg/head of ovalbumin incorporated in FCA showed anaphylactic reaction. Anti-DA-3030 antibody was also detected in those guinea pigs. In immunodiffusion test using the sera sensitized with DA-3030 incorporated in FCA, precipitating antibodies were detected only in the sera sensitized with DA-3030 or DA-3030 incorporated in FCA showed. In 24-hour heterologous PCA reaction with sera of C57BL/6 mice immunized with 1.25 or 12.5 ㎍/head of DA-3030 alone, none of the sera showed positive reaction. But sera of the animals immunized with 12.5 ㎍/head of DA-3030 incorporated in aluminum hydroxide gel(Alum) or 5 ㎍/head of ovalbumin incorporated in alum showed positive PCA reaction. DA-3030 did not cause anaphylactic shock or passive cutaneous anaphylaxis in guinea pigs and mice when given alone although DA-3030 incorporated in FCA or Alum induced anaphylactic shock and passive cutaneous anaphylaxis. From these results, it may be concluded the DA-3030 does not induce systemic allergic reaction when administered alone in its clinical use.

비 마약성 진통제 DA-5018 의 랫드에 대한 4 주 경구투여 아급성독성

백남기(Nam Gi Baik),안병옥(Byoung Ok Ahn),김원배(Won Bae Kim),양중익(Junn Ick Yang),김옥진(Ok Jin Kim),강경구(Kyung Koo Kang) 한국응용약물학회 1997 Biomolecules & Therapeutics(구 응용약물학회지) Vol.5 No.2

4-week repeated dose toxicity of DA-5O18, a new capsaicin analogue analgesic agent, was examined in SD rats at dosage levels of 0, 0.4, 2, 10 and 50 mg/kg/day. DA-5018 was administered orally to 17 males and 17 females per group at doses of 0, 10 and 50 mg/kg and to 12 males and 12 females per group at doses of 0.4 and 2 mg/kg. After the administration period, 5 males and 5 females at the 0, 10 and 50 mg/kg were placed on withdrawal for 2 weeks. Treatment-related clinical signs were observed at 10 and 50 mg/kg. Clinical signs observed immediately after the administration of DA-5018 were grooming, sedation or depression, lacrimation, ataxia, reddening of extremities and ears, ventral or lateral recumbency, respiratory distress, cyanosis and convulsion. Delayed-type clinical signs including focal scabbing and depilation around nose were also observed 1 or 2 weeks after the start of administrarion of DA-5018. Only at the 50 mg/kg group, corneal opacities, reduced body weight gain (male) and death (male 6/17, female 3/17) were noted. In blood biochemical analysis, serum levels of glucose and triglyceride decreased at 10 and 50 mg/kg. In hematological examination, there were increases in the number of red blood cell, hemoglobin content and percent of hematocrit at 10 and 50 mg/kg. Pulmonary enlargement and hemorrhagic spot, focal scabbing and depilation around nose and corneal opacities were seen at the necropsy of the animals died during the dosing of DA-5018 50 mg/kg. Focal scabbing and depilation arowtd nose were observed in the animals terminally necropsied at doses of 10 and 50 mg/kg. Histopathological examination revealed pulmonary hemorrhage, focal necrosis in the scabbed area, corneal necrosis, fibrosis and neovasculization in the stroma. At 0.4 and 2 mg/kg, there were no significant toxic changes attributable to the administration of DA-5018. In conclusion, target organs following to 4-week repeated dose of DA-5018 in the rat were determined to be lung, skin and eyes. Definite toxic dose and no-observed-adverse-effect-level (NOAEL) were estimated to be 50 and 2 mg/kg/day, respectively.

새로운 Authracycline 계 항암성 항생물질 DA-125 의 랫드에 대한 4주 용량설정시험 ( DRF ) 과 13 주 아급성 독성시험

백남기(Nam Gi Baik),안병옥(Byoung Ok Ahn),이순복(soon Bok Lee),이상득(Sang Deuk Lee),김원배(Won Bae Kim),양중익(Jung Ick Yang),(Eric J . F . Spicer),(Susan Novitsky),(Lee Bernal),(Pamela Ball),(Mary Ellen Mckenna),(K . 한국응용약물학회 1994 Biomolecules & Therapeutics(구 응용약물학회지) Vol.2 No.2

This study was conducted to investigate the repeated dose toxicity of DA-125, a new anthracycline antitumor antibiotic, in rats. Before the 13-week main study, a 4-week dose-range finding (DRF) study was carried out. The administration of DA-125 intravenously at dosage levels of 0, 0.125, 0.5, 2.0, and 8.0 ㎎/㎏/day to rats for 4 weeks resulted in premature deaths of all animals in the 8.0 ㎎/㎏/day group and in the deaths of 4 males and 4 females at 2.0 ㎎/㎏/day. Body weights were markedly reduced in the 8.0 ㎎/㎏/day group and showed dose-related decreases in all treatment groups when compared with the control group. Reductions in weight gain were slight and not significantly different at 0.125 ㎎/㎏/day but animals receiving 0.5 ㎎/㎏/day showed more marked decreases in gain in a clear dose-related manner. Based on the results of the above DRF study, a 13-week repeated dose intravenous toxicity study in rats with DA-125 was performed at a dose level of 0, 0.012, 0.08 and 0.3 ㎎/㎏/day. No treatment related effects were noted in behavior or body weight in all treatment groups. One male at the highest dose level died on study day 26, but the death could not be related to test article toxicity. Swelling and scabbing of the ears was present in all of the groups, including the control group. There were no treatment related changes in the hematological, biochemical or urinalysis values in all treatment groups. Thymus weights were significantly reduced in males receiving 0.3 ㎎/㎏/day and they were sligltly, and not significantly, reduced in females of the same group. While there were no associated histological changes. Treatment related necrosis was found in the tail vein (injection site) at 0.08 and 0.3 ㎎/㎏/day. On the basis of these results, the no observed effect level (NOEL) was 0.012 ㎎/㎏/day and the maximum tolerated dose (MTD) was estimated to be more than 0.3 ㎎/㎏/day under the conditions tested.

비 마약성 진통제 DA-5018 의 변이원성 연구

강경구(Kyung Koo Kang),백남기(Nam Gi Baik),김원배(Won Bae Kim),양중익(Junn Ick Yang) 한국응용약물학회 1996 Biomolecules & Therapeutics(구 응용약물학회지) Vol.4 No.3

DA-5018, a non-narcotic analgesic agent, was examined for mutagenicity in the reverse mutation test on bacteria, chromosomal aberration test on cultured mammalian cells and micronucleus test on mice. The reverse mutation test was performed by a plate incorporation method with or without a metabolic activation system(S9 mix) using Salmonella typhimurium strain TA100, TA1535, TA98 and TA1537. DA-5018 did not significantly increase revenant colonies in any of the test strains under any conditions at concentrations ranging from 0.0049 to 1.25 ㎎/plate, compared with the vehicle control. In the chromosomal aberration test using cultured Chinese Hamster Lung(CHL) cells, DA-5018 did not increase the number of aberrant cells in the presence or absence of S9 mix at concentrations of 0.016 mM/plate to 0.25 mM/plate, compared with the vehicle control. In the micronucleus test, male ICR mice were given DA-5018 intraperitoneally at a dose level of 0.55, 1.10 and 2.20 ㎎/kg. The incidence of bone marrow micronucleated polychromatic erythrocytes in the DA-5018 treated mice was not significantly different from that of the vehicle control. These results indicate that DA-5018 does not have mutagenic potential under the present test conditions.

천연형 인성장호르몬 DA-3002 의 항원성

강경구(Kyung Koo Kang),백남기(Nam Gi Baik),김원배(Won Bae Kim),양중익(Jung Ick Yang) 한국응용약물학회 1995 Biomolecules & Therapeutics(구 응용약물학회지) Vol.3 No.4

DA-3002 is a genuine human growth hormone produced by Dong-A Pharm. Co. Ltd. research laboratory using recombinant DNA technic. In this study, antigenic potential of DA-3002 was examined by active systemic anaphyaxis(ASA) in guinea pigs, mouse-rat passive cutaneous anaphylaxis(PCA) and passive hemagglutination(PHA) test as a part of safety research. DA-3002 induced anaphylactic shock in ASA test using guinea pigs immunized with DA-3002 alone or DA-3002 incoporated into Freund`s complete adjuvant(FCA) when challenged with 10 times higher dose of anticipated clinical dose of DA-3002, In the mouse-rat PCA and PHA test, DA-3002 also showed positive results. DA-3002, therfore, was considered to produce IgE, IgG and/or IgM in mice. The results of this study were similar to those of the other human growth hormones and these positive results were thought to be caused due to the fact that both DA-3002 and the other human growth hormones were heterogenous proteins to guinea pigs and mice. Considering the fact that DA-3002 is a genuine human growth hormone of which structure is identical with indigenous human growth hormone, DA-3002 is thought not to cause immunological problems in clinical use.

천연형 사람 적혈구 조혈인자의 항원성시험

강경구(Kyung Koo Kang),조현(Hyeon Cho),백남기(Nam Gi Baik),김원배(Won Bae Kim) 한국응용약물학회 1998 Biomolecules & Therapeutics(구 응용약물학회지) Vol.6 No.1

Antigenic potential of a recombinant human erythropoietin (rhEPO) produced by Dong-A Pharm. 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 IU/kg alone or rhEPO 1000 IU/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 IU/kg 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 IgM 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 rhEPO has an identical structure with indigenous human erythropoietin, rhEPO is not thought to cause immunological problems in clinical use.

애엽추출물 항궤양제 DA-9601 의 랫드에 대한 4 주 경구 반복투여 독성연구

김옥진(Ok Jin Kim),강경구(Kyung Koo Kang),김동환(Dong Hwan Kim),백남기(Nam Gi Baik),안병옥(Byoung Ok Ahn),김원배(Won Bae Kim),양중익(Junn Ick Yang) 한국응용약물학회 1996 Biomolecules & Therapeutics(구 응용약물학회지) Vol.4 No.4

This study was conducted to investigate the repeated dose toxicity of DA-9601, an antiulcer agent of Artemisia spp. extract, in rats. DA-9601 was administered orally once a day for 4 weeks to 10 males and 10 females per group at doses of 0(vehicle control), 125, 500 or 2000 mg/kg/day. Throughout the study, no treatment-related deaths and clinical signs were observed. In female rats receiving 125 mg/kg of DA-9601, water consumption increased slightly on day 4, 11 and 25. Hematological examination showed a decrease of MCV and an increase of PLT in male rats at the doses of 500 and 2000 mg/kg groups. Blood biochemistry revealed slight decreases of cholesterol, BUN and Na in male rats and decreases of total bilirubin and creatinine and slight increases of globulin and Cl in female rats. The organ weights at the end of 4 weeks showed slight changes in some organs of treated groups. But, all these changes were not considered to be of toxicological importance, because they did not show dose-response relationship and relevance to gross and microscopic findings. Histopathologically, abnormal treatment-related changes were not observed in any organ and target organs were not detected. On the basis of these results, the NOAEL(no-observed-adverse-effect level) of DA-9601 was estimated to be more than 2000 mg/kg/day under the conditions tested.

재조합 사람 적혈구 조혈인자 DA-3585 의 랫드에 대한 단회 및 4주 반복 정맥투여 독성시험

김동환(Dong Hwan Kim),조현(Hyeon Cho),강경구(Kyung Koo Kang),백남기(Nam Gi Baik),김원배(Won Bae Kim) 한국응용약물학회 1998 Biomolecules & Therapeutics(구 응용약물학회지) Vol.6 No.2

DA-3585 is a recombinant human erythropoietin produced by Dang-A pharmaceutical Co. Ltd. using recombinant DNA technique: Recently, recombinant human erythropoietin (rHu-EPO) has been used to treat various types of anemia. In this study, we examined acute and subacute toxicity of DA-3585 in rats. DA3585 was intravenously administered to rats at dose levels of 0, 6,250, 12,500 and 25,000 IU/kg for single dose toxicity study and at dose levels of 0, 100, 500 and 2,500 IU/kg daily for 4 week-repeated dose toxicity study. In the single dose toxicity study, there were no death, clinical signs and changes in body weight gain related to the treatment. Necropsy revealed no evidence of toxicity related to DA-3585. In the repeated dose toxicity study, all the rats survived throughout the study. There were no treatment-related changes in clinical signs, food and water intake, and body weight. Hematological examination showed increases in the number of erythrocytes, hemoglobin concentration, hematocrit value and mean corpuscular volume, and decrease in the number of platelet in 500 and 2,500 IU/kg dosed groups. Extramedullary hematopoiesis in the spleen and erythroid hyperplasia in the bone marrow were noted as treatment-related histological changes. Toxicologically significant changes were not observed in blood biochemistry, urinalysis, organ weights and in any other examinations. The treatment-related changes observed in this study were hematological or histological changes associated with pharmacological effects of DA-3585. On the basis of the results of this study, LDP value of DA-3585 was above 25,000 IU/kg and the no-observed-adverse-effect-level was estimated to be 100 IU/kg.

사람 적혈구 조혈인자 DA-3585 의 토끼에 대한 단회 및 4주 반복투여 독성시험

조현(Hyeon Cho),김동환(Dong Hwan Kim),강경구(Kyung Koo Kang),백남기(Nam Gi Baik),김원배(Won Bae Kim) 한국응용약물학회 1998 Biomolecules & Therapeutics(구 응용약물학회지) Vol.6 No.2

DA-3585, a biosynthetic recombinant human erythropoietin has been developed as a treatment for anemia associated with chronic renal failure in Dong-A pharmaceutical Co. Ltd. This study was carried out to assess its acute and subacute toxicities in rabbits. DA-3585 was intravenously administered to rabbits at dose levels of 6250, 12500 or 25000 IU/kg for single dose toxicity study and at dose levels of 100, 500 or 2500 IU/kg/day for 4-week repeated dose toxicity study. In the acute toxicity study, dose up to 25000 IU/kg had no adverse effect on the behavior or body weight gain. Pathological examinations revealed no abnormal gross lesions related to DA-3585. In the subacute toxicity study, all animals survived until termination of treatment. DA-3585 had no influence on clinical signs, food and water intake or on body weight changes. Hematological examination showed increases in the number of RBC, hemoglobin contents and hematocrit values with a dose dependent manner in the animals treated with DA-3585. Histopathological examination revealed erythroid hyperplasia in the bone marrow and extramedullary hematopoiesis in the liver. The changes detected in the hematological and histopathological examination presumably represent exaggerated pharmacological effects of erythropoietin. The NOAEL (no-observed-adverse-effect-level) of DA-3585 was estimated to be 100 IU/kg/ day under this study condition.

천연형 인성장호르몬 DA-3002 의 변이원성 연구

강경구(Kyung Koo Kang),김옥진(Ok Jin Kim),신동환(Dong Hwan Shin),백남기(Nam Gi Baik),안병옥(Byoung Ok Ahn),김원배(Won Bae Kim),양중익(Junn Ick Yang) 한국응용약물학회 1995 Biomolecules & Therapeutics(구 응용약물학회지) Vol.3 No.4

DA-3002, an authentic recombinant human growth hormane(rhGH), was examined for mutagenicity in the reverse mutation test on bacteria, in the chromosomal aberration test on cultured mammalian cells and in the micronucleous test on mice. The reverse mutation test was performed by a plate incorporation method with or without a metabolic activation system(S9 Mix) using Salmonella typhimurium strain TA100, TA1535, TA98 and TA1537. DA-3002 did not significantly increase revertant colonies in any of the test strains under any conditions at dose levels ranging from 0.0125 to 0.4 IU/plate, compared with the vehicle control. In the chromosomal aberration test using cultured Chinese hamster lung(CHL) cells, DA-3002 did not increase the number of aberrant cells in the presence or absence of S9 mix at concentrations of 0.0125 IU/ml to 0.4 IU/ml, compared with the vehicle control. In the micronucleus test, male ICR mice were given DA-3002 intraperitoneally at a dose level of 20, 40 and 80 IU/kg. The incidence of bone marrow micronucleated polychromatic erythrocytes in the DA-3002 treated mice did not differ from that of the vehicle control. These results indicate that DA-3002 doesn`t have mutagenic potential under the present test conditions.