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.）并作为ACT（青蒿素联合疗法）递送，是经证实的用于治疗疟疾的倍半萜内酯治疗剂，并显示出对抗许多其他疾病的前景。这种重要的药草也引起了N.美国市场，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A. annua（pACT）对抗寄生虫血症的有效性比纯AN高至少5倍，生物可利用性高> 40倍，并且对耐药性的发展具有更大的弹性。总之，这些结果表明pACT的天然化学复杂性促进了这些反应。由植物产生的一些内源性小分子已显示出抗疟原虫活性;一些还显示出与AN对恶性疟疾的治疗协同作用。然而，很少有人知道这些辅助化合物（AC），主要是黄酮类化合物和萜烯，对AN的生物利用度的影响。它们通过肠壁进入血清的通道也不清楚。我们的基础研究将有助于解释植物基质及其内源性小分子如何影响AN的生物利用度，这对于提高对pACT疗效的整体理解至关重要。目的1目的活性炭，经鉴定存在于A.在我们的动物研究中观察到，在pACT中使用的annua栽培品种（SAM）可以提高AN的生物利用度。为了确定AC是否增强通过肠壁的转运，我们将使用体外消化模型Caco-2，其具有纯测试AC的纯AN+组合来测量AN和AC渗透性。这将告诉我们纯AC如何影响AN肠道通道，并指示是否有任何AC也通过肠壁，但不存在背景植物基质。目的2为了确定pACT植物基质如何影响AN的生物利用度，我们将从离体肠消化的pACT植物基质（含有AC）测量AN的Caco-2渗透性。我们还将测量传代AC的水平。这告诉我们有多少AN和AC从Caco-2系统的顶侧移动到基底侧，并成为我们的比较对照，将根据目标3的结果进行测量。目的3使用GLS植物基质的肠内消化状态，我们将系统地添加AN沿着已知在GLS中不存在（或几乎不存在）但存在于SAM（pACT植物）中的单个AC，以确定哪种在改善AN的传代中是有效的。这将告诉我们哪些AC在辅助AN通过方面是有效的，但是具有零植物矩阵。目的4使用来自pACT喂养小鼠的血清，我们还将提取和测量AC在血流中的存在。我们手头已经有了最近PK实验的冷冻血清提取物（见第二节）。3.3.3）。虽然我们现在已经测量了这些提取物中的AN，但我们一直等到我们积累了相关的AC标准品才能量化这些血清样品中的AN水平。这能告诉我们血清中是否真的出现了进化人

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.