Artemisinin bioavailability via an orally consumed dry leaf herbal therapeutic.

描述（由申请人提供）：青蒿素（AN）由植物黄花蒿（Artemisia annua L.）生产，作为ACT（青蒿素联合疗法）递送，是一种经证实的倍半萜内酯治疗疟疾的药物，并有望治疗许多其他疾病。这种重要的草药也引起了北美市场的兴趣，特别是通过其抗炎作用促进一般健康和愈合。最近，我们报道了整株A. annua （pACT）对寄生虫病的有效性至少是纯AN的5倍，生物利用度是纯AN的40倍，并且对耐药性的抗性更强。总之，这些结果表明，pACT的天然化学复杂性促进了这些反应。植物产生的一些内源性小分子具有抗疟原虫活性；有些还显示出与AN对抗恶性疟疾的治疗协同作用。然而，这些辅助化合物（ACs），主要是类黄酮和萜烯，对AN的生物利用度的影响知之甚少。它们通过肠壁进入血清的途径也不清楚。我们的基础研究将有助于解释植物基质及其内源小分子如何影响AN的生物利用度，这对提高对pACT功效的整体理解具有重要意义。在我们的动物研究中发现，在pACT中使用的A. annua栽培（SAM）中证实存在目标ac，可能会提高AN的生物利用度。为了确定AC是否增强了通过肠壁的运输，我们将使用体外消化模型Caco-2，与纯试验AC的纯AN +组合来测量AN和AC的通透性。这将告诉我们纯ac如何影响a肠道通道，并表明是否有任何ac也通过肠壁，但没有背景植物基质。为了确定pACT植物基质如何影响AN的生物利用度，我们将通过体外肠道消化SAM产生的pACT植物基质（含ACs）来测量AN的Caco-2通透性。我们还将测量通过的ac的水平。这告诉我们有多少AN和ac从Caco-2系统的顶端移动到基底外侧，并成为我们的比较对照，将根据Aim 3的结果进行测量。目的3：利用GLS植物基质的肠道消化液，我们将系统地将AN与GLS中已知不存在（或几乎不存在）但存在于SAM （pACT植物）中的单个ac一起添加，以确定哪种能有效地改善AN的通过。这将告诉我们哪个ac在帮助AN通过时是有效的，但是在一个零植物矩阵下。目的4：利用已有的pact喂养小鼠血清，我们还将提取并测量血液中ACs的存在。我们已经从最近的PK实验中获得了现成的冷冻血清提取物（见第3.3.3节）。虽然我们现在已经测量了这些提取物中的AN，但我们一直在等待，直到我们积累了相关的AC标准，才能量化这些血清样本中的AN水平。这能告诉我们血清中是否有ac。

DESCRIPTION (provided by applicant): Artemisinin (AN), produced by the plant Artemisia annua L. and delivered as ACT (Artemisinin Combination Therapy), is a proven sesquiterpene lactone therapeutic for treating malaria and shows promise against many other diseases. This important medicinal herb is also becoming of interest to the N. American market, particularly to promote general health and healing via its anti-inflammatory effects. Recently we reported that whole plant A. annua (pACT) is at least fivefold more effective against parasitemia, >40-fold more bioavailable, and much more resilient to development of drug resistance than pure AN. Together these results suggest the native chemical complexity of pACT is facilitating these responses. Some endogenous small molecules produced by the plant have shown antiplasmodial activity; some also show therapeutic synergism with AN vs. falciparum malaria. However, little is known about the effect of these assistive compounds (ACs), mainly flavonoids and terpenes, on bioavailability of AN. Their passage through the intestinal wall into the serum is also not clear. Our fundamental studies will help explain how the plant matrix and its endogenous small molecules affect bioavailability of AN, important to improving overall understanding of pACT efficacy. Aim 1 The target ACs, verified to be present in the A. annua cultivar (SAM) used in pACT, may enhance bioavailability of AN as observed in our animal studies. To determine if the ACs enhance transport through the intestinal wall, we will use the in vitro digestion model, Caco-2, with pure AN + combinations of the pure test ACs to measure AN and AC permeability. This will tell us how the pure ACs affect AN intestinal passage and indicate if any of the ACs also pass through the intestinal wall, but absent the background plant matrix. Aim 2 To determine how the plant matrix of pACT affects bioavailability of AN, we will measure the Caco-2 permeability of AN from ex vitro intestinal digestion of the pACT plant matrix produced from SAM (contains ACs). We will also measure the level of passaged ACs. This tells us how much AN and ACs move from the apical to basolateral side of the Caco-2 systems and becomes our comparative control against which results of Aim 3 will be measured. Aim 3 Using intestinal digestate of the GLS plant matrix we will systematically add AN along with individual ACs known to be absent (or nearly so) in GLS, but present in SAM (the pACT plant), to determine which is effective in improving the passage of AN. This will tell us which of the ACs is effective in assisting the passage of AN, but with a null plant matrix. Aim 4 Using serum already on hand from pACT-fed mice, we will also extract and measure ACs for their presence in the bloodstream. We already have on hand frozen serum extracts from our recent PK experiment (see Sec. 3.3.3). While we have now measured AN in those extracts, we have waited until we have accumulated relavant AC standards to quantify their levels in these serum samples. This will tell us if any ACs actually appears in the serum.