【正文】 初步研究表明，MOMIPP誘發(fā)空泡的能力并不僅限于癌細(xì)胞（圖5EH的）。一個(gè)可能的例外在一份報(bào)告中被指出，報(bào)告稱一個(gè)被稱為“C2的”查爾酮的衍生物可誘導(dǎo)膠質(zhì)瘤細(xì)胞以非凋亡的死亡機(jī)制死亡，其中涉及自噬泡的積累。因此，MOMIPP不大可能通過蛋白質(zhì)的親電改造誘導(dǎo)細(xì)胞死亡。在為SAR研究服務(wù)的化合物庫形成過程中，我們進(jìn)行了各種indole3carboxaldehydes和芳香酮在哌啶催化下的ClaisenSchmidt縮合反應(yīng)，從而能有效提供吲哚查耳酮。C, 53%. Compound 40: POCl3, DMF, 0 176。二苯甲酮類似物（化合物，苯甲酰MIPP30）用兩個(gè)步驟制備，首先從2 甲基吲哚的Vilsmeier–Haack中間體（圖3A）酰化開始。這些觀察表明，雖然MOMIPP會對正常細(xì)胞產(chǎn)生毒性，但與癌癥細(xì)胞系，如U251和MCF7相比，正常細(xì)胞對這類化合物相對不太敏感。當(dāng)用集落形成實(shí)驗(yàn)來評估細(xì)胞增殖能力和長期活力時(shí)，MOMIPP和MIPP的區(qū)別就表現(xiàn)非常明顯（圖4）。Scheme 2. Analogues with 5′ Modifications of the Indole Ring Generated by Functionalizing the Indole Prior to Introduction of the Pyridine MoietyaaConditions and reagents: Compound 22: BBr3, CH2Cl2, ?78 176。在生理?xiàng)l件下使用MIPP的局限性之一是其在水溶液中始終有一定溶解度。C, 61 and 95% for 15 and 21, respectively. (C) POCl3, DMF, 0 176。（ 10）Figure 1. Structures of pounds 1 and 2, initially found to be active inducers of methuosis(10) as pared with mercially available analogues 3–8, which failed to induce the hallmarks of methuosis in culture U251 GBM cells.因?yàn)樾枰狹IPP的濃度≥10μM才能有效誘導(dǎo)methuosis，所以我們開始組裝MIPP相關(guān)化合物庫，并與經(jīng)效能提高的確定的目標(biāo)類似物進(jìn)行初步特區(qū)的比較。在此，我們報(bào)告了相關(guān)化合物庫合成及構(gòu)效關(guān)系（SAR）的研究從而導(dǎo)出（1）定義的主要特點(diǎn)，為誘導(dǎo)methuosis活動，（2）確定改進(jìn)生物活性的衍生物以及發(fā)展一個(gè)可能適合在未來目標(biāo)識別工作中用作光親探測器的疊狀類似物做準(zhǔn)備。因此，它可能可以作為一種能夠在癌癥中通過methuosis形式來促進(jìn)細(xì)胞凋零的藥物原型，這種原理與傳統(tǒng)細(xì)胞死亡（如細(xì)胞凋亡）的機(jī)理不同。（3，4）此外，膠質(zhì)母細(xì)胞瘤細(xì)胞通過提高他們修復(fù)DNA損傷的能力而獲得對烷化劑的抗藥性。查耳酮由一個(gè)1,3 二苯基2 丙烯1框架組成，這種框架是黃酮類天然產(chǎn)物的前體組成。因此，一種類似物的初始構(gòu)成被合成，以用來研究調(diào)查吡啶氮的位置對生物活性的影響。如表1所示，盡管他們在短期的增長/可行性實(shí)驗(yàn)中細(xì)胞毒性不一致。我們在不同pKa值的多個(gè)基地進(jìn)行了篩選，從K2CO3到Cs2CO3，以及乙胺，tetramethylguanidine，1,8 二氮雜雙環(huán)[]螺7烯（DBU），氫化鈉。 (2) 5 N NaOH, 99%. Compound 26: 4acetylpyridine, piperidine, MeOH, reflux, 34%. Compound 27: 4acetylpyridine, piperidine, MeOH, reflux, 21%.化合物19（MOMIPP）與化合物2（MIPP）生物活性比較上述研究確定化合物19是最有力誘導(dǎo)methuosis的化合物。 SD from three separate cultures.Figure 3. Comparison of the abilities of MOMIPP and MIPP to induce the morphological hallmarks of methuosis. One day after plating, U251 GBM cells were treated with MOMIPP or MIPP at final concentrations of (A) or 10 μM (B). Controls received an equivalent volume of vehicle (DMSO). Cells were observed by phase contrast microscopy on three sequential days after addition of the pounds, without changing the medium or replenishing the pounds. Methuosis is characterized by extensive accumulation of phaselucent cytoplasmic vacuoles, with eventual cell rounding and detachment from the substratum as viability Figure 4. Comparison of the abilities of MOMIPP and MIPP to inhibit survival of U251 GBM cells in colonyforming assays. (A) Cells were plated for colonyforming assays as described in the Experimental Section. One day after plating the cells, MOMIPP (●) or MIPP (■) was added to the medium at the indicated concentrations, and cells were maintained in the presence of the pounds for 48 h. Thereafter, the pounds were removed, and colonies 50 cells were counted after 2 weeks. Each point represents the mean (177。SD) from four separate wells. (G) Normal human skin fibroblasts were treated with 10 μM MOMIPP or an equivalent volume of DMSO (control) and examined by phasecontrast microscopy after 24 h. (H) Normal human fibroblasts were seeded in 96well plates at subconfluent density (1000 cells/well) or confluent density (10000 cells/well). After 24 h, fresh medium containing 10 μM MOMIPP or an equal volume of vehicle (DMSO) was added. MTT viability assays were performed at a 48 h end point. Values are the means (177。隨后的形態(tài)和可行性研究顯示，與芐衍生物（化合物14）不同的是，當(dāng)在U251細(xì)胞（數(shù)據(jù)未顯示）測試時(shí)，沒有一個(gè)苯甲酮類似物（30日和31日）能誘導(dǎo)methuosis。溫和重氮化條件下，氨基吲哚轉(zhuǎn)化為芳香疊氮化合物34。首先，關(guān)于吲哚環(huán)，氮的甲基化（化合物20）或去除2 甲基（化合物13）降低但并未消除活性。后者的化合物被設(shè)計(jì)成許多秋水仙堿的和能結(jié)合β微管蛋白的天然產(chǎn)品bretastatins的類似物。在這方面， MIPP吲哚環(huán)的5位引入光活性疊氮化合物后產(chǎn)生了一種保留了良好的methuosis誘導(dǎo)活性（圖6）的衍生物，我們的這項(xiàng)發(fā)現(xiàn)提供了幾種按照確定的步驟鑒定蛋白質(zhì)受體的途徑。后者可能