11-Hydroxyaclacinomycin X의 종간 스케일링에 있어서 단백결합의 역할 = Role of Protein Binding in the Interspecies Scaling of 11-Hydroxyaclacinomycin X

다국어 초록 (Multilingual Abstract)

11-Hydroxyaclacinomycin X (HAMX) is isolated from S. galiaeus 3113 (pMC213) with main anti-cancer effects. The pharmacokinetics of HAMX, which has the inhibitory effects on DNA synthesis as well as topoisomerase like other anthracyclines, was investigated in mice, rat, rabbits and beagle dogs after i.v. bolus administration. To support its pharmacokinetic study which was essential for the pre-clinical research for new drug development, the total body clearance (CLt), apparent volume of distribution at steady state (Vdss) and terminal half-life (t1/2) were calculated. CLt, Vdss and t1/2 were 73.7 ml/mim/kg, 18.1 L/kg and 219 min in mice, 64.0 ml/min/kg, 10.9 L/kg and 227 min in rats, 47.2 ml/min/kg, 21.3 L/kg and 206 min in rabbits, and 27.4 ml/min/kg, 2.24 L/kg and 118 min in dogs, respectively. Based on animal data, CLt, Vdss and t1/2 of HAMX was predicted in humans using simple allometry and invariant time methods. The corresponding allometric equations were CLt = 45.896W0.8452, CLt × MLP = 31.175W1.1405, Vdss = 10140W0.8653 and t1/2 = 178.43W-0.0883. These allometric equations were extrapolated to predict CLt, Vdss and t1/2 in human based on 70 kg body weight. Plasma concentration of HAMX was also predicted by several species-invariant time transformations using equivalent time, Kallynochron, Apolysichron and Dienerichron. The human pharmacokinetic parameters of CLt, Vdss and t1/2 predicted by the simple allometry and various species-invariant time methods were 17∼42 ml/min/kg, 6∼19 L/kg and 112.5∼399 min, respectively. Plasma and cytosol protein binding of drug is an important factor that influences its pharmacokinetic parameters. To predict human CLt based on the unbound drug concentration in plasma and cytosol (fu), protein binding of the drug was assessed in vitro at five concentration levels (1, 5, 10, 50, 100 μg/ml) in plasma and cytosol using equilibrium dialysis and followed by HPLC analysis. The method was optimized against some influencing factors during both experimental procedures. And it was validated to be specific, sensitive, accurate, precise and of satisfactory recovery. The free fraction in plasma (fu) was 0.104 in mouse, 0.081 in rat, 0.049 in rabbit, 0.012 in dog, 0.011 in human, respectively. It exhibited a difference from mouse to human for the plasma and cytosol protein binding of 11-Hydroxyaclacinomycin X. Unbound clearance (CLt, u) was 707.68 ml/min/kg in mouse, 790.12 ml/min/kg in rat, 963.26 ml/min/kg in rabbit, 2258.33 ml/min/kg in dog, respectively. Based on animal data, the unbound clearance of HAMX was predicted in humans. The results indicated that HAMX data obtained from animals could be utilized to generate preliminary estimates of the pharmacokinetic parameters in humans. These parameter can serve as guidelines for better planning of clinical studise.

목차 (Table of Contents)

• Ⅰ. 서론 1
• Ⅱ. 연구 방법 6
• 1. 실험 재료 6
• 1.1. 시약 6
• 1.2. 실험 기기 6
• 1.3. 실험 동물 7
• 2. 11-Hydroxyaclacinomycin X의 HPLC 분석조건 8
• 2.1. 형광 흡수파장의 검토 8
• 2.2. 이동상 용매 선택 8
• 2.3. 컬럼의 선택 8
• 2.4. 추출 용매의 선택 8
• 3. 동물 실험 9
• 3.1. 마우스에서 의 정맥투여 후 혈장 채취 9
• 3.2. 흰 쥐에서 HAMX의 정맥투여 후 혈장 채취 9
• 3.3. 토끼에서 HAMX의 정맥투여 후 혈장 채취 9
• 3.4. 비글견에서 HAMX의 정맥투여후 혈장 채취 9
• 3.5. 혈장 중 HAMX 농도 정량 12
• 3.6. 혈장 중 HAMX 농도의 약물동태학적 해석 12
• 3.6.1. 정맥주사 후 속도론적 파라미터 산출 12
• 3.6.2. 모델 비의존적 해석 13
• 4. In-vitro 에서의 단백결합 실험 15
• 4.1. 혈장 단백결합 실험 15
• 4.2. 간세포질(Cytosol)의 분리 15
• 4.3. 간세포질내 단백결합 실험 15
• 4.4. 혈장 및 간세포질 시료 중 11-Hydroxyaclacinomycin X 농도정량 16
• 5. 단백결합 실험 데이터 해석 18
• 5.1. Langmuir type plot 18
• 5.2. Langmuir type plot 의 변형 18
• 6. Interspecies scaling 20
• 6.1. Allometric equation 20
• 6.1.1. 사람에서의 클리어런스 예측 20
• 6.1.2. 사람에서의 분포용적 예측 21
• 6.1.3. 사람에서의 소실 반감기 예측 21
• 6.2. Invariant time methods 21
• 6.2.1. Equivalent time 22
• 6.2.2. Kallynochrons or Elementary dedrick plot 22
• 6.2.3. Apolysichrons or Complex dedrick plot 22
• 6.2.4. Dienetichrons 23
• Ⅲ. 결과 및 고찰 24
• 1. 11-Hydroxyaclacinomycin X의 정맥주사 후 체내 동태 24
• 1.1. 마우스에서의 체내 동태 24
• 1.2. 흰 쥐에서의 체내 동태 24
• 1.3. 토끼에서의 체내 동태 25
• 1.4. 비글견에서의 체내 동태 25
• 2. In-vitro에서 단백결합 실험 30
• 2.1. 혈장 30
• 2.2. 간세포질(Cytosol) 30
• 3. Interspecies scaling 38
• 3.1. Allometric equation 38
• 3.1.1. 사람에서의 클리어런스 예측 38
• 3.1.2. 사람에서의 분포용적 예측 38
• 3.1.3. 사람에서의 소실 반감기 예측 38
• 3.2. Invariant time methods 39
• 3.2.1. Equivalent time 39
• 3.2.2. Kallynochrons or Elementary dedrick plot 39
• 3.2.3. Apolysichrons or Complex dedrick plot 40
• 3.2.4. Dienetichrons 40
• Ⅳ. 결론 52
• Ⅴ. 참고문헌 53