CAJ | 학술논문

开发了用家蚕蛹表达的人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)的口服药物.试验评价了猕猴、大鼠和小鼠口服rhGM-CSF毒性情况,包括遗传毒性、单剂量和重复剂量的全身毒性以及生殖毒性.毒理学试验显示:静脉给药在NIH小鼠中最大耐受剂量为3 300 μg/kg,LD50为2 020 μg/kg ,在SD大鼠中LD50为3 660 μg/kg.在猕猴和大鼠的重复剂量毒性试验结果中显示:口服rhGM-CSF后,除了雌性SD大鼠的血糖含量外,其余各项指标包括白细胞、红细胞、血红蛋白含量、血小板数量以及血细胞凝集等都正常.雌性大鼠每天口服剂量为1 250 μg/kg时,给药后血糖浓度显著增加(P＜0.001),但在恢复期间可恢复到正常水平.微核实验、CHL染色体畸变试验和埃姆斯测验法都表明无论是在体内还是体外服用rhGM-CSF均未见遗传毒性,普通的生殖毒性试验和围产期及致畸敏感期的毒性试验均未见异常现象,表明口服rhGM-CSF无明显生殖毒性.临床前毒性试验表明口服超过临床剂量10倍的rhGM-CSF与严重的慢性中毒没有联系,其在药理学有效剂量的范围内是很安全的.
개발료용가잠용표체적인립세포-거서세포집락자격인자(rhGM-CSF)적구복약물.시험평개료미후、대서화소서구복rhGM-CSF독성정황,포괄유전독성、단제량화중복제량적전신독성이급생식독성.독이학시험현시:정맥급약재NIH소서중최대내수제량위3 300 μg/kg,LD50위2 020 μg/kg ,재SD대서중LD50위3 660 μg/kg.재미후화대서적중복제량독성시험결과중현시:구복rhGM-CSF후,제료자성SD대서적혈당함량외,기여각항지표포괄백세포、홍세포、혈홍단백함량、혈소판수량이급혈세포응집등도정상.자성대서매천구복제량위1 250 μg/kg시,급약후혈당농도현저증가(P＜0.001),단재회복기간가회복도정상수평.미핵실험、CHL염색체기변시험화애모사측험법도표명무론시재체내환시체외복용rhGM-CSF균미견유전독성,보통적생식독성시험화위산기급치기민감기적독성시험균미견이상현상,표명구복rhGM-CSF무명현생식독성.림상전독성시험표명구복초과림상제량10배적rhGM-CSF여엄중적만성중독몰유련계,기재약이학유효제량적범위내시흔안전적.
We developed an oral drug of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) expressed in silkworm pupae. Our goal in this study was to determine the toxicity of oral drug of rhGM-CSF in monkeys (Macaca mulatto), rats and mice, including genetic toxicology, single-and repeat-dose systemic toxicity and reproductive toxicity. Toxicology tests showed that the maximum tolerated dose (I.v) was 3 300 μg/kg, and that the LD50 (I.v) was 2 020 μg/kg in NIH mice, and 3.660 μg/kg in SD rats respectively. Repeat-dose toxicity test of oral drug of rhGM-CSF in monkeys and rats showed that all indices (including white blood cell, red blood cell, haematoglobin content, platelet number, and hemagglutination) were normal, with the exception of glucose content in female rats, which increased significantly (P＜0.001) after oral administration of the high-dose 1 250 μg·kg-1·d-1 rhGM-CSF while it reversibly was normal during recovery. It was found that rhGM-CSF oral drug showed no genotoxicity either in vitro or in vivo through the micronucleus test, CHL chromosome aberration test and Ames test. No abnormality was found in general reproductive toxicity test, and toxicity test in perinatal stage and sensitive period to teratogenic agent, which suggests little reproductive toxicity. Preclinical toxicity indicated administration of the rhGM-CSF oral drug in excess of ten times as many as clinical dosages was not associated with severe chronic toxicities and it is very safe at the range of pharmacologically functional dose. This study represents the first report of detailed toxicological evaluation of human protein drug expressed in silkworm using baculovirus expression system.