Artemisinin bioavailability via an orally consumed dry leaf herbal therapeutic.

描述(申请人提供)：青蒿素(AN)，由植物青蒿素(Artemisia annua L.)生产，以ACT(青蒿素联合疗法)的形式提供，是一种已被证实的倍半萜内酯疗法，用于治疗疟疾，并显示出对许多其他疾病的治疗前景。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，我们报道了全植物青蒿素(PACT)对寄生虫血症的疗效至少是纯植物青蒿素的五倍，生物利用度是纯植物青蒿素的40倍，而且对耐药性的产生有更强的适应能力。综上所述，这些结果表明PACT的天然化学复杂性促进了这些反应。一些由该植物产生的内源小分子显示出抗疟活性；一些还显示出对抗恶性疟疾的治疗协同作用。然而，人们对这些辅助性化合物(主要是黄酮类和萜类)对AN生物利用度的影响知之甚少。它们穿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，这对于提高对PACT疗效的整体理解非常重要。目的1在PACT中证实存在于黄花蒿品种(SAM)中的靶向乙酰胆碱能提高AN的生物利用度，正如我们的动物研究所观察到的那样。为了确定ACS是否增强了通过肠壁的转运，我们将使用体外消化模型Caco-2，与纯测试ACS的纯AN+组合来测量AN和AC的通透性。这将告诉我们纯粹的ACS是如何影响肠道的，并表明是否有任何ACS也穿过肠壁，但没有背景植物基质。目的为了确定PACT的植物基质对AN生物利用度的影响，我们测定了SAM(含ACS)产生的PACT植物基质的体外肠道消化对AN的Caco-2通透性。我们还将测量传代的急性冠脉综合征的水平。这告诉我们AN和ACS从CACO-2系统的顶端移动到基底外侧的程度，并成为我们对照目标3的结果进行测量的比较对照。目的3利用GLS植物基质的肠道消化作用，我们将系统地将AN与已知在GLS中缺失(或几乎如此)但存在于SAM(PACT植物)中的单个ACS一起添加，以确定哪一种在促进AN通过方面是有效的。这将告诉我们，哪些ACS在帮助AN通过方面是有效的，但植物矩阵为空。目的4利用PACT喂养的小鼠已有的血清，我们还将提取并测量其在血流中的存在。我们手头已经有了我们最近PK实验的冷冻血清提取物(见SEC。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标准才能量化它们在这些血清样本中的水平。这会告诉我们血清中是否真的出现了急性冠脉综合征。

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.