評估插管小鼠經靜脈及口服多次劑量投與質體

評估插管小鼠經靜脈及口服多次劑量投與質體 DN A (pMBP-Lac Z) 聚合微膠體溶液之藥物動力學

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(40?g DNA/150 ?L) 與 pMBP-Lac Z 聚合微膠體溶液到插管小鼠，其體內

pMBP-Lac Z 之藥物動力學參數及體內之分佈情形。血液及組織檢品利用即時聚 合酶連鎖反應法 (Q-PCR) 分析檢品中 pMBP-Lac Z 含量。實驗初步結果顯示，

在頸靜脈投與 (IVJ) 三次總劑量後之血液檢品中發現， pMBP-Lac Z (P) 之曲線下 面積 (area under the curve, AUC) 為 1.56 x 1012 (min x copies/?L) ，投與 pMBP-Lac Z 聚合微膠體溶液 (P/PM) 之 AUC 為 5.29 x 1011 (min x copies/?L) 。而排除半衰 期的部份，投與 P 為 436 分鐘，投與 P/PM 為 344 分鐘。經由尾靜脈投與 (IVT) P/PM 後的血中濃度曲線圖發現， AUC 為 5.12 x 1011 (min x copies/?L) ，其半衰 期為 97 分鐘。口服投與三個劑量的 P/PM ，其 AUC 為 1.73 x 107 (min x

copies/?L) ，半衰期為 58 分鐘。利用 IVJ 和 IVT 投與單一劑量 P 和 P/PM 的 5, 30 和 60 分鐘後，觀察到 DNA 主要分佈於心臟、肺臟、腦、脊髓和腎臟等高血流 的組織，若經由口服投與 P 和 P/PM 後，以十二指腸及胃部組織為主要分佈組織。

此外，體外 DNase 安定性試驗發現， PM 存在下確實能對 DNA 有保護效果，而 pMBP-Lac Z 在體外的血漿安定性試驗發現，不論是 P 或 P/PM 之安定性，在 20 分鐘都已觀察不到完整之 pMBP-Lac Z 存在，若是胃沖洗液的安定性試驗，則是 P 或 P/PM 皆在 60 分鐘才開始出現 DNA 片段。綜合以上結果，體內實驗發現，

不同投與途徑下確實會影響其體內的組織分佈情形。

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A clear understanding of the tissue distribution of plasmid DNA and its complexes with nonviral vectors i s a prerequisite for a strategy for developing novel in vivo gene transfer methods. Therefore the aim of this study was to evaluate the pMBP-Lac Z plasmid DNA, which is driven by specific, myelin basic protein

（ MBP ） gene promoter into oligodendrocytes, with or without polymeric micelles

(PM) in the systemic distribution of pharmacokinetics (PK) profiles via 3 dose (40 ug DNA/150 uL) of jug ular vein injection (IVJ), tail vein injection (IVT) or oral

administration in cannulated mice. The distribution of pharmacokinetics profiles were quantitated using rea l-time quantitation polymerase chain reaction (Q-PCR). After 3

dose of multiple jugular vein injection (IVJ) , the area under the curve (AUC) of pMBP-Lac Z (P) and with polymeric micelles (P/PM) in plasma were 1.56 x 10^12 (min x copies/μL) and 5.29 x 10^11 (min x copies /uL), respectively. And the elimination half-lives of pMBP-Lac Z (P) and with PM (P/PM) in plasma were 436 min and 344 min, respectively. For 3 dose of multiple tail vein injection (IVT), the

AUC of P/PM in plasma was 5.12 x 10^11 (min x copies/uL), and the elimination half-lives was 97 min. A fter 3 dose of multiple oral administrations, the AUC was 1.73 x 10^7 (min x copies/uL), and the eliminati on half-lives was 58 min. At 5, 30 and 60 min after single IVJ or IVT of P or P/PM, the higher tissue distri bution was found in heart, lung, brain, spinal cord and kidney tissues which were high blood flow tissues.

Then, after single oral administration, the duodenum and stomach were the

major distributed tissues. The in vitro DNAse stability test of pMBP-Lac Z was confirmed that the PM can protect the DNA, and the in vitro serum stability of pMBP-Lac Z were indicated that both of with or witho ut formulated were not detectable within 20 min. And the in vitro gastric washing fluid stability of pMBP- Lac Z were indicated that both of with or without PM had degradation at 60 min. The different administrati on routes suggest influence the biodistribution of plasmid.

Evaluation of pharmacokinetics of pMBP-Lac Z with polymeric micelles via multiple-dose intravenous

and oral administrations in cannulated mice