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<title>Antagonists of IAP-family anti-apoptotic proteins - Probe 1 - Probe Reports from the NIH Molecular Libraries Program - NCBI Bookshelf</title>
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<meta name="robots" content="INDEX,FOLLOW,NOARCHIVE" /><meta name="citation_inbook_title" content="Probe Reports from the NIH Molecular Libraries Program [Internet]" /><meta name="citation_title" content="Antagonists of IAP-family anti-apoptotic proteins - Probe 1" /><meta name="citation_publisher" content="National Center for Biotechnology Information (US)" /><meta name="citation_date" content="2010/09/02" /><meta name="citation_author" content="Kate Welsh" /><meta name="citation_author" content="Hongbin Yuan" /><meta name="citation_author" content="Derek Stonich" /><meta name="citation_author" content="Ying Su" /><meta name="citation_author" content="Xocella Garcia" /><meta name="citation_author" content="Michael Cuddy" /><meta name="citation_author" content="Richard Houghten" /><meta name="citation_author" content="Eduard Sergienko" /><meta name="citation_author" content="John C Reed" /><meta name="citation_author" content="Robert Ardecky" /><meta name="citation_author" content="Santhi Reddy Ganji" /><meta name="citation_author" content="Marcos Lopez" /><meta name="citation_author" content="Shakeela Dad" /><meta name="citation_author" content="Thomas DY Chung" /><meta name="citation_author" content="Nicholas Cosford" /><meta name="citation_pmid" content="21433357" /><meta name="citation_fulltext_html_url" content="https://www.ncbi.nlm.nih.gov/books/NBK47341/" /><link rel="schema.DC" href="http://purl.org/DC/elements/1.0/" /><meta name="DC.Title" content="Antagonists of IAP-family anti-apoptotic proteins - Probe 1" /><meta name="DC.Type" content="Text" /><meta name="DC.Publisher" content="National Center for Biotechnology Information (US)" /><meta name="DC.Contributor" content="Kate Welsh" /><meta name="DC.Contributor" content="Hongbin Yuan" /><meta name="DC.Contributor" content="Derek Stonich" /><meta name="DC.Contributor" content="Ying Su" /><meta name="DC.Contributor" content="Xocella Garcia" /><meta name="DC.Contributor" content="Michael Cuddy" /><meta name="DC.Contributor" content="Richard Houghten" /><meta name="DC.Contributor" content="Eduard Sergienko" /><meta name="DC.Contributor" content="John C Reed" /><meta name="DC.Contributor" content="Robert Ardecky" /><meta name="DC.Contributor" content="Santhi Reddy Ganji" /><meta name="DC.Contributor" content="Marcos Lopez" /><meta name="DC.Contributor" content="Shakeela Dad" /><meta name="DC.Contributor" content="Thomas DY Chung" /><meta name="DC.Contributor" content="Nicholas Cosford" /><meta name="DC.Date" content="2010/09/02" /><meta name="DC.Identifier" content="https://www.ncbi.nlm.nih.gov/books/NBK47341/" /><meta name="description" content="Multiple member of Inhibitor of Apoptosis Proteins (IAPs) family are simultaneously overexpressed in many cancers. As a result, antisense or RNA interference (RNAi) methods for assessing function of these proteins are lacking. Selective probing the binding of small molecule compounds against either the BIR2 or BIR3 SMAC (second mitochondria-derived activator of caspase)-binding sites on a number of IAPs, (e.g. XIAP, cIAP1, cIAP2) may generate useful tools for elucidating their role in cancer biology. The goals of this project were: (1) to generate small molecule compounds that mimic the effects of SMAC peptides to inhibit the function of IAPs; (2) to find non-peptidyl small molecule chemical inhibitors that have advantages over SMAC peptides, in terms of cell permeability, stability, in vivo pharmacology and potency, ideally 1 uM or better; and (3) to identify compounds that inhibit the BIR2 domain of XIAP, either selectively or in addition to inhibiting the BIR3 domain. The selective probe ML101 (CID-25241665) mimics the binding of a single AVPI binding motif in demonstrating potent binding affinity and selectivity for BIR1/2 domain of XIAP." /><meta name="og:title" content="Antagonists of IAP-family anti-apoptotic proteins - Probe 1" /><meta name="og:type" content="book" /><meta name="og:description" content="Multiple member of Inhibitor of Apoptosis Proteins (IAPs) family are simultaneously overexpressed in many cancers. As a result, antisense or RNA interference (RNAi) methods for assessing function of these proteins are lacking. Selective probing the binding of small molecule compounds against either the BIR2 or BIR3 SMAC (second mitochondria-derived activator of caspase)-binding sites on a number of IAPs, (e.g. XIAP, cIAP1, cIAP2) may generate useful tools for elucidating their role in cancer biology. The goals of this project were: (1) to generate small molecule compounds that mimic the effects of SMAC peptides to inhibit the function of IAPs; (2) to find non-peptidyl small molecule chemical inhibitors that have advantages over SMAC peptides, in terms of cell permeability, stability, in vivo pharmacology and potency, ideally 1 uM or better; and (3) to identify compounds that inhibit the BIR2 domain of XIAP, either selectively or in addition to inhibiting the BIR3 domain. 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<div class="pre-content"><div><div class="bk_prnt"><p class="small">NCBI Bookshelf. A service of the National Library of Medicine, National Institutes of Health.</p><p>Probe Reports from the NIH Molecular Libraries Program [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2010-. </p></div><div class="iconblock clearfix whole_rhythm no_top_margin bk_noprnt"><a class="img_link icnblk_img" title="Table of Contents Page" href="/books/n/mlprobe/"><img class="source-thumb" src="/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-mlprobe-lrg.png" alt="Cover of Probe Reports from the NIH Molecular Libraries Program" height="100px" width="80px" /></a><div class="icnblk_cntnt eight_col"><h2>Probe Reports from the NIH Molecular Libraries Program [Internet].</h2><a data-jig="ncbitoggler" href="#__NBK47341_dtls__">Show details</a><div style="display:none" class="ui-widget" id="__NBK47341_dtls__"><div>Bethesda (MD): National Center for Biotechnology Information (US); 2010-.</div></div><div class="half_rhythm"><ul class="inline_list"><li style="margin-right:1em"><a class="bk_cntns" href="/books/n/mlprobe/">Contents</a></li></ul></div><div class="bk_noprnt"><form method="get" action="/books/n/mlprobe/" id="bk_srch"><div class="bk_search"><label for="bk_term" class="offscreen_noflow">Search term</label><input type="text" title="Search this book" id="bk_term" name="term" value="" data-jig="ncbiclearbutton" /> <input type="submit" class="jig-ncbibutton" value="Search this book" submit="false" style="padding: 0.1em 0.4em;" /></div></form></div></div><div class="icnblk_cntnt two_col"><div class="pagination bk_noprnt"><a class="active page_link prev" href="/books/n/mlprobe/ml102/" title="Previous page in this title">< Prev</a><a class="active page_link next" href="/books/n/mlprobe/ml100/" title="Next page in this title">Next ></a></div></div></div></div></div>
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<div class="main-content lit-style" itemscope="itemscope" itemtype="http://schema.org/CreativeWork"><div class="meta-content fm-sec"><h1 id="_NBK47341_"><span class="title" itemprop="name">Antagonists of IAP-family anti-apoptotic proteins - Probe 1</span></h1><p class="contrib-group"><span itemprop="author">Kate Welsh</span>, <span itemprop="author">Hongbin Yuan</span>, <span itemprop="author">Derek Stonich</span>, <span itemprop="author">Ying Su</span>, <span itemprop="author">Xocella Garcia</span>, <span itemprop="author">Michael Cuddy</span>, <span itemprop="author">Richard Houghten</span>, <span itemprop="author">Eduard Sergienko</span>, <span itemprop="author">John C Reed</span>, <span itemprop="author">Robert Ardecky</span>, <span itemprop="author">Santhi Reddy Ganji</span>, <span itemprop="author">Marcos Lopez</span>, <span itemprop="author">Shakeela Dad</span>, <span itemprop="author">Thomas DY Chung</span>, and <span itemprop="author">Nicholas Cosford</span>.</p><p class="small">Received: <span itemprop="datePublished">May 18, 2009</span>; Last Update: <span itemprop="dateModified">September 2, 2010</span>.</p></div><div class="jig-ncbiinpagenav body-content whole_rhythm" data-jigconfig="allHeadingLevels: ['h2'],smoothScroll: false" itemprop="text"><div id="_abs_rndgid_" itemprop="description"><p>Multiple member of Inhibitor of Apoptosis Proteins (IAPs) family are simultaneously overexpressed in many cancers. As a result, antisense or RNA interference (RNAi) methods for assessing function of these proteins are lacking. Selective probing the binding of small molecule compounds against either the BIR2 or BIR3 SMAC (second mitochondria-derived activator of caspase)-binding sites on a number of IAPs, (e.g. XIAP, cIAP1, cIAP2) may generate useful tools for elucidating their role in cancer biology. The goals of this project were: (1) to generate small molecule compounds that mimic the effects of SMAC peptides to inhibit the function of IAPs; (2) to find non-peptidyl small molecule chemical inhibitors that have advantages over SMAC peptides, in terms of cell permeability, stability, in vivo pharmacology and potency, ideally 1 uM or better; and (3) to identify compounds that inhibit the BIR2 domain of XIAP, either selectively or in addition to inhibiting the BIR3 domain. The selective probe ML101 (CID-25241665) mimics the binding of a single AVPI binding motif in demonstrating potent binding affinity and selectivity for BIR1/2 domain of XIAP.</p></div><div class="h2"></div><p><b>Assigned Assay Grant #:</b> R03 MH081277-01</p><p><b>Screening Center Name & PI:</b>
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<i>Conrad Prebys</i> Center for Chemical Genomics (<i>formerly Burnham Center for Chemical Genomics</i>) & Dr. John C. Reed</p><p><b>Chemistry Center Name & PI:</b>
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<i>Conrad Prebys</i> Center for Chemical Genomics (<i>formerly Burnham Center for Chemical Genomics)</i> & Dr. John C. Reed</p><p><b>Assay Submitter & Institution:</b> Dr. John C Reed & Sanford-Burnham Medical Research Institute (<i>formerly Burnham Institute for Medical Research</i>)</p><p><b>PubChem Summary Bioassay Identifier (AID):</b>
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<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1638" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1638</a></p><div id="ml101.s2"><h2 id="_ml101_s2_">Probe Structure & Characteristics</h2><div id="ml101.fu1" class="figure"><div class="graphic"><img src="/books/NBK47341/bin/ml101fu1.jpg" alt="Image ml101fu1" /></div></div><div id="ml101.tu1" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK47341/table/ml101.tu1/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml101.tu1_lrgtbl__"><table><thead><tr><th id="hd_h_ml101.tu1_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">CID/ML</th><th id="hd_h_ml101.tu1_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Target Name</th><th id="hd_h_ml101.tu1_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">IC50/EC50 (nM) [SID, AID]</th><th id="hd_h_ml101.tu1_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Anti-target Name(s)</th><th id="hd_h_ml101.tu1_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">IC50/EC50 (μM) [SID, AID]</th><th id="hd_h_ml101.tu1_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Selectivity</th><th id="hd_h_ml101.tu1_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary Assay(s) Name: IC50/EC50 (nM) [SID, AID]</th></tr></thead><tbody><tr><td headers="hd_h_ml101.tu1_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">CID-25241665<br /><a href="/pcsubstance/?term=ML101[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML101</a></td><td headers="hd_h_ml101.tu1_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BIR1/2<br /><br />XAIP</td><td headers="hd_h_ml101.tu1_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">IC50 4000 (nM)<br /><a href="https://pubchem.ncbi.nlm.nih.gov/substance/57643995" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-57643995</a><br /><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1749" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1749</a></td><td headers="hd_h_ml101.tu1_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BIR3</td><td headers="hd_h_ml101.tu1_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">IC50 32.6 (µM)<br /><a href="https://pubchem.ncbi.nlm.nih.gov/substance/57643995" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-57643995</a><br /><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1750" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1750</a></td><td headers="hd_h_ml101.tu1_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">8.2X</td><td headers="hd_h_ml101.tu1_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">N/A</td></tr></tbody></table></div></div></div><div id="ml101.s3"><h2 id="_ml101_s3_">Recommendations for the scientific use of this probe</h2><p>Various IAP-family proteins are over-expressed in specific types of cancer and some tumors over-express more than one member of the family simultaneously. Given that multiple IAPs are simultaneously over-expressed in many cancers, antisense or RNA interference (RNAi) methods for assessing function of these proteins are often lacking. Therefore, probing the competition of small molecule compounds against either the BIR2 or BIR3 SMAC-binding sites on Inhibitor of Apoptosis Protein (IAP) such as XIAP, cIAP1, cIAP2 or other IAPs from human and other organisms will serve as useful tools for elucidating their roles in cancer biology.</p></div><div id="ml101.s4"><h2 id="_ml101_s4_">1. Scientific Rationale for Project</h2><div id="ml101.s5"><h3>Specific aims</h3><p>The goal of this project was to generate small molecule compounds that mimic the effects of SMAC peptides, inhibiting the function of Inhibitor of Apoptosis Proteins (IAPs).</p></div><div id="ml101.s6"><h3>Background and Significance</h3><p>Apoptosis, or programmed cell death, is a critical cellular process in normal development and homeostasis of multicellular organisms (<a class="bk_pop" href="#ml101.r1">1</a>). Caspases are the executioners in both intrinsic and extrinsic pathways of apoptosis by cleaving a plethora of cellular components (<a class="bk_pop" href="#ml101.r2">2</a>). These intracellular proteases are suppressed by Inhibitor of Apoptosis Proteins (IAPs), a family of evolutionarily conserved anti-apoptotic proteins (<a class="bk_pop" href="#ml101.r3">3</a>). The X-linked inhibitor of apoptosis, XIAP, is a key member of the family of intrinsic inhibitors of apoptosis proteins (IAP), which block cell death both <i>in vitro</i> and <i>in vivo</i> by virtue of inhibition of two effector caspases (caspase-3 and -7) and an initiator caspase (caspase-9). XIAP contains three baculoviral IAP repeat (BIR) domains, defined by a novel ~80 amino acid motif. The third domain (BIR3) of XIAP selectively binds to the amino terminus of the caspase-9 linker peptide while the BIR2 domain inhibits both caspase-3 and caspase-7. This inhibiting activity of XIAP is negatively regulated by at least two XIAP interacting proteins, XAF1 and SMAC/DIABLO (second mitochondria-derived activator of caspase/direct IAP binding protein with low pl). As a dimer, SMAC targets both the BIR2 and BIR3 domains of XIAP. Proteins released from mitochondria (SMAC and HtrA2) (<a class="bk_pop" href="#ml101.r4" data-bk-pop-others="ml101.r5 ml101.r6">4–6</a>) can competitively displace IAPs from the Caspases, thus helping to drive apoptosis. It has been shown that only a few residues at the N-terminus of activated SMAC protein (4′mer) are sufficient to affect the release of IAPs from Caspases (<a class="bk_pop" href="#ml101.r7">7</a>).</p><p>In this study we investigated small molecule compounds that mimicked the effect of SMAC in antagonizing IAPs by causing them to release Caspases. Non-peptidyl chemical inhibitors would have advantages over SMAC peptides, in terms of cell permeability, stability, and <i>in vivo</i> pharmacology. Several strategies were employed for the design of small-molecular inhibitors of XIAP, including the structure-based rational design and synthesis of small molecules that mimic the binding interactions between this protein and SMAC.</p></div></div><div id="ml101.s7"><h2 id="_ml101_s7_">2. Project Description</h2><div id="ml101.s8"><h3>a. Describe the original goal for probe</h3><p>All current probes are polypeptides derived from natural IAP inhibitors (e.g. SMAC or HtrA2), with the 4-mer being the smallest that can displace IAPs from caspases. This project looked for non-peptidyl small molecule chemical inhibitors that would have advantages over SMAC peptides, in terms of cell permeability, stability, <i>in vivo</i> pharmacology and ideally having potency of 1 µM or better. In particular, the goal was to identify compounds that inhibit the BIR2 domain of XIAP, either selectively or in addition to inhibiting the BIR3 domain.</p></div><div id="ml101.s9"><h3>b. Assay implementation and screening</h3><div id="ml101.s10"><h4>i. PubChem Bioassay Name(s), AID(s), Assay-Type (Primary, DR, Counterscreen, Secondary)</h4><div id="ml101.tu2" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK47341/table/ml101.tu2/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml101.tu2_lrgtbl__"><table><thead><tr><th id="hd_h_ml101.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">PubChemBioAssay Name</th><th id="hd_h_ml101.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">AIDs</th><th id="hd_h_ml101.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Probe Type</th><th id="hd_h_ml101.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Assay Type</th><th id="hd_h_ml101.tu2_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Assay Format</th><th id="hd_h_ml101.tu2_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Assay Detection & well format</th></tr></thead><tbody><tr><td headers="hd_h_ml101.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Chemical Antagonists IAP-family anti-apoptotic proteins—primary screen and HTS/DPI liquid hit confirms</td><td headers="hd_h_ml101.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1018" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1018</a></td><td headers="hd_h_ml101.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Inhibitor</td><td headers="hd_h_ml101.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Primary</td><td headers="hd_h_ml101.tu2_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Biochemical</td><td headers="hd_h_ml101.tu2_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Fluorescence Polarization (384)</td></tr><tr><td headers="hd_h_ml101.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">XIAP-Bir3 for Chemical Antagonists of Bir1/2 domains of IAP-family anti-apoptotic proteins—liquid hit confirmation from DPI</td><td headers="hd_h_ml101.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1513" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1513</a></td><td headers="hd_h_ml101.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Inhibitor</td><td headers="hd_h_ml101.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Counterscreen</td><td headers="hd_h_ml101.tu2_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Biochemical</td><td headers="hd_h_ml101.tu2_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Fluorescence Polarization (384)</td></tr><tr><td headers="hd_h_ml101.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">SAR analysis of Antagonists of IAP-family anti-apoptotic proteins –Dry powder [Confirmatory]</td><td headers="hd_h_ml101.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1749" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1749</a></td><td headers="hd_h_ml101.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Inhibitor</td><td headers="hd_h_ml101.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Counterscreen</td><td headers="hd_h_ml101.tu2_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Biochemical</td><td headers="hd_h_ml101.tu2_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Fluorescence Polarization (96)</td></tr><tr><td headers="hd_h_ml101.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">SAR analysis of Antagonists of XIAP-Bir3 domain of IAP-family anti-apoptotic-dry powder.</td><td headers="hd_h_ml101.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1750" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1750</a></td><td headers="hd_h_ml101.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Inhibitor</td><td headers="hd_h_ml101.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Counterscreen</td><td headers="hd_h_ml101.tu2_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Biochemical</td><td headers="hd_h_ml101.tu2_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Fluorescence Polarization (96)</td></tr></tbody></table></div></div></div><div id="ml101.s11"><h4>ii. Assay Rationale & Description</h4><p>This XIAP dose response assay was developed and performed to confirm hits originally identified in the XIAP HTS binding assay (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1018" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1018</a>) and to study the structure-activity relationship on analogues of the confirmed hits. Compounds were acquired from commercial sources or synthesized in-house.</p><p>The rationale for the assay was based on the disruption of fluorescence polarization resulting from binding of a his-tagged-BIR1-BIR2 (bacoloviral IAP repeat, “Bir1/2”) domain protein derived from two of the three conserved caspase binding “BIR” domains of XIAP to a rhodamine tagged 7-mer N-terminal SMAC peptide.</p><div id="ml101.s12"><h5>Assay materials</h5><div id="ml101.t1" class="table"><h3><span class="label">Table 1</span><span class="title">Reagents used for the uHTS experiments</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK47341/table/ml101.t1/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml101.t1_lrgtbl__"><table class="no_top_margin"><thead><tr><th id="hd_h_ml101.t1_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Reagent</th><th id="hd_h_ml101.t1_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Vendor</th></tr></thead><tbody><tr><td headers="hd_h_ml101.t1_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Expression plasmid for BIR12 of XIAP cloned</td><td headers="hd_h_ml101.t1_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Dr. Reed’s laboratory</td></tr><tr><td headers="hd_h_ml101.t1_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">BIR1/2 protein</td><td headers="hd_h_ml101.t1_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Purified in Dr. Reed’s laboratory</td></tr><tr><td headers="hd_h_ml101.t1_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">1 M Hepes buffer</td><td headers="hd_h_ml101.t1_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Qmega Scientific, Inc</td></tr><tr><td headers="hd_h_ml101.t1_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">TCEP</td><td headers="hd_h_ml101.t1_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Sigma</td></tr><tr><td headers="hd_h_ml101.t1_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">0.5 M Tween 20</td><td headers="hd_h_ml101.t1_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">BIO-RAD</td></tr><tr><td headers="hd_h_ml101.t1_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">SMAC-rhodamine</td><td headers="hd_h_ml101.t1_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Synthesized in Dr. Houghten’s laboratory at The Torrey Pines Institute for Molecular Studies (TPIMS)</td></tr><tr><td headers="hd_h_ml101.t1_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">SMAC-7mer</td><td headers="hd_h_ml101.t1_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Synthesized in Dr. Houghten’s laboratory at The Torrey Pines Institute for Molecular Studies (TPIMS)</td></tr></tbody></table></div></div><div id="ml101.s13"><h5>Bir1/2 assay materials</h5><ol><li class="half_rhythm"><div>Bir1/2 protein and rhodamine-SMAC peptide (AVPIAQK-rhodamine) were provided by Prof. John Reed (Burnham Institute for Medical Research, San Diego, CA)</div></li><li class="half_rhythm"><div>Assay buffer: 31.25 mM HEPES-NaOH, pH 7.5, 1.25 mM TCEP, 0.00625% Tween 20.</div></li><li class="half_rhythm"><div>Bir1/2 working solution contained 2.5 µM Bir1/2 in the assay buffer.</div></li><li class="half_rhythm"><div>Rhodamine-SMAC working solution contained 50 nM FITC-Tr3-R8 peptide in the assay buffer.</div></li></ol></div><div id="ml101.s14"><h5>Bir1/2 protocol</h5><ol><li class="half_rhythm"><div>4 uL of serially diluted compounds in DMSO were dispensed in columns 3–22 of Greiner 384-well black small-volume plates (784076). Compounds were serially diluted in DMSO to have duplicate 10-point curves with 2-fold dilution between concentrations. Columns 1–2 and 23–24 were added with 4 uL of 10% DMSO.</div></li><li class="half_rhythm"><div>Positive control wells, that contained no Bir1/2, were assigned to columns 1–2 and were added 8 uL of assay buffer using WellMate bulk dispenser (Matrix).</div></li><li class="half_rhythm"><div>8 uL of Bir1/2 working solution was added to columns 3–24 using WellMate bulk dispenser (Matrix). Negative control wells that contained DMSO were assigned to columns 23–24.</div></li><li class="half_rhythm"><div>Plates were briefly spinned down and incubated for 1h at room temperature.</div></li><li class="half_rhythm"><div>8 uL of Rhodamine-SMAC working solution was added to the whole plate using WellMate bulk dispenser (Matrix).</div></li><li class="half_rhythm"><div>Final concentrations of the components in the assay were as follows:</div><ol class="lower-alpha"><li class="half_rhythm"><div>25 mM HEPES-NaOH, pH 7.5, 1 mM TCEP, 0.005% Tween 20.</div></li><li class="half_rhythm"><div>20 nM Rhodamine-SMAC (columns 1–24)</div></li><li class="half_rhythm"><div>1 µM Bir1/2 (columns 2–24)</div></li><li class="half_rhythm"><div>2 % DMSO (columns 1–24</div></li><li class="half_rhythm"><div>20 µM compounds (columns 3–24)</div></li></ol></li><li class="half_rhythm"><div>Plates were incubated for 30 min at room temperature protected from direct light.</div></li><li class="half_rhythm"><div>Fluorescence polarization was measured on an EnVision plate reader (Perkin Elemer) using rhodamine filters: excitation filter − 540 nm, emission filter 590 nm, dichroic mirror − 560 nm. The signal for each well was acquired for 100 ms.</div></li><li class="half_rhythm"><div>Data analysis was performed using CBIS software (ChemInnovations, Inc).</div></li><li class="half_rhythm"><div>Fluorescence intensity of each sample was normalized to the average fluorescence intensity value of the plate negative control wells to calculate F_ratio parameter.</div></li></ol><p>195576 compounds were tested in this assay. The average Z′ for this assay was 0.85, the signal to background ratio was 3.8, the signal to noise ratio was 46.6 and the signal window was 34.7.</p></div><div id="ml101.s15"><h5>Rationale for confirmatory, counter and selectivity assays</h5><p>SMAC is synthesized and enters the inner membrane space of mitochondria (<a class="bk_pop" href="#ml101.r4">4</a>,<a class="bk_pop" href="#ml101.r5">5</a>). The N-terminal of SMAC is cleaved off generating a new N-terminal AVPI sequence (<a class="bk_pop" href="#ml101.r4">4</a>,<a class="bk_pop" href="#ml101.r5">5</a>). On release of the activated SMAC from the mitochondria, it binds to a SMAC binding pocket on BIR2 and BIR3 competing off Caspase-3/7 and Caspase-9, respectively, and thus driving the activation of the executioner and initiator caspases by releasing the caspase from the BIR2 or BIR3 SMAC binding domains (<a class="bk_pop" href="#ml101.r7">7</a>,<a class="bk_pop" href="#ml101.r8">8</a>). It is established that the binding site of BIR3 for SMAC requires only the first 4 amino acids of activated SMAC (<a class="bk_pop" href="#ml101.r7">7</a>). The first 7 amino acids of activated SMAC are AVPIAQK (<a class="bk_pop" href="#ml101.r7">7</a>). We utilized the ɛ-amino group of lysine to couple the rhodamine to the peptide at the C-terminal. This spaced the rhodamine away from the SMAC binding pocket allowing the AVPI portion of the fluorescent probe to bind without hinderance to the BIR2 pocket yet still having the rhodamine probe tumbling with the rate of the protein.</p><p>We chose to target Caspase-3 and -7 and looked at the competition of compounds against SMAC-rhodamine in the presence of BIR1/2 where only the BIR2 domain was capable of binding SMAC-rhodamine. Development of the fluorescence polarization assay used the SMAC-7mer as a competitor for the SMAC-rhodamine (<a class="bk_pop" href="#ml101.r10">10</a>). A similar assay was developed for BIR3 and was utilized as a counterscreen.</p></div></div></div><div id="ml101.s16"><h4>iii. Summary of Results</h4><p>We developed a binding assay based upon fluorescence polarization, using a short peptide representing residues from the N-terminus of activated SMAC with an attached fluorochrome. This fluorescence polarization assay (FPA) formed the basis for a high-throughput competitive displacement assay that was optimized for screening. We screened the NIH compound library using this FPA to identify chemical compounds that compete with the SMAC peptide for binding to IAPs. A secondary assay was also devised, so the hits would be independently confirmed. However, the single hit from the conventional HTS screen was very weak (> 25 μM IC<sub>50</sub>), we therefore decided to take a structure-based rational design approach to develop novel chemical probes of XIAP. Chemistry has made significant progress towards this goal and the results are described in section 3c below.</p></div></div><div id="ml101.s17"><h3>b. Probe Optimization</h3><div id="ml101.s18"><h4>i. Description of SAR & chemistry strategy that led to the probe</h4><p>The design and synthesis of novel antagonists of XIAP that mimic the biological properties of the SMAC peptide has attracted considerable attention in the last few years. Short peptides (3–7 amino acids) typically demonstrate superior affinity for the BIR3 domain of XIAP to the detriment of its binding affinity for the BIR2 domain. With this in mind, most research to date has focused on the search for small molecules that disengage XIAP from caspase-9 by inhibiting the BIR3 interaction. <a class="figpopup" href="/books/NBK47341/figure/ml101.f1/?report=objectonly" target="object" rid-figpopup="figml101f1" rid-ob="figobml101f1">Figure 1</a> represents the evolution of the SAR studies that we undertook, <i>via</i> chemical synthesis assisted with a structure-based approach, to develop new BIR2-selective SMAC peptidomimetics. X-ray crystal structures of the BIR2 (PDB code <b>1i3o</b>) and BIR3 (PDB code <b>1g73</b>) domains of XIAP were used to guide the structure-based molecular design. One key difference between the BIR2 and BIR3 protein structures at the SMAC binding site is that BIR3 has a sub-pocket on the right (<a class="figpopup" href="/books/NBK47341/figure/ml101.f2/?report=objectonly" target="object" rid-figpopup="figml101f2" rid-ob="figobml101f2">Fig.2B</a>) occupied by the isoleucine (Ile) residue of the SMAC N-terminal AVPI sequence, whereas in Bir2 this region is a shallow surface (<a class="figpopup" href="/books/NBK47341/figure/ml101.f2/?report=objectonly" target="object" rid-figpopup="figml101f2" rid-ob="figobml101f2">Fig.2A</a>). This difference in the SMAC binding site therefore forms a rational basis for designing selective Bir2 antagonists. As a result, various substitutions at R4 group in <a class="figpopup" href="/books/NBK47341/figure/ml101.f1/?report=objectonly" target="object" rid-figpopup="figml101f1" rid-ob="figobml101f1">Figure 1</a> were designed to specifically target this non-conserved binding regions in BIR2 and BIR3. In addition, two complementary approaches drove our synthetic efforts: (<a class="bk_pop" href="#ml101.r1">1</a>) studies to determine the effect of different substituents on the potency and selectivity of inhibitors of the BIR1/2 vs. BIR3 domains of XIAP; and (<a class="bk_pop" href="#ml101.r2">2</a>) studies to determine the effect of the stereochemistry of amino acids in P1–3 on the potency and selectivity of inhibitors of the BIR1/2 vs. BIR3 domains.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml101f1" co-legend-rid="figlgndml101f1"><a href="/books/NBK47341/figure/ml101.f1/?report=objectonly" target="object" title="Figure 1" class="img_link icnblk_img figpopup" rid-figpopup="figml101f1" rid-ob="figobml101f1"><img class="small-thumb" src="/books/NBK47341/bin/ml101f1.gif" src-large="/books/NBK47341/bin/ml101f1.jpg" alt="Figure 1. SAR strategy for tripeptide 1." /></a><div class="icnblk_cntnt" id="figlgndml101f1"><h4 id="ml101.f1"><a href="/books/NBK47341/figure/ml101.f1/?report=objectonly" target="object" rid-ob="figobml101f1">Figure 1</a></h4><p class="float-caption no_bottom_margin">SAR strategy for tripeptide <i>1</i>. </p></div></div><div id="ml101.f2" class="figure bk_fig"><div class="graphic"><img src="/books/NBK47341/bin/ml101f2.jpg" alt="Figure 2. Protein surfaces of BIR2 (A) and BIR3 (B) at the SMAC binding site; docked poses of CID25241665 (MLS-0391005) in BIR2 (C) and CID25241639 (MLS-0390982) in BIR3 (D)." /></div><h3><span class="label">Figure 2</span><span class="title">Protein surfaces of BIR2 (A) and BIR3 (B) at the SMAC binding site; docked poses of CID25241665 (MLS-0391005) in BIR2 (C) and CID25241639 (MLS-0390982) in BIR3 (D)</span></h3></div><p>We synthesized a library of 47 compounds which were evaluated for binding to the BIR1/2 or BIR3 domains of XIAP by employing fluorescence polarization (FP) assays. The IC<sub>50</sub> values for selected inhibitors are summarized in <a class="figpopup" href="/books/NBK47341/table/ml101.t2/?report=objectonly" target="object" rid-figpopup="figml101t2" rid-ob="figobml101t2">Table 2</a>. Although all the compounds shown here were synthesized employing natural L-amino acids, several derivatives were synthesized using unnatural D-amino acids. The use of these unnatural building blocks in general resulted in inactive compounds (IC<sub>50</sub>>100 μM) for both the BIR1/2 and BIR3 domains. Compounds CID25241665 (MLS-0391005) and CID25241679 (MLS-0391014), represented by the sequence Ala1-Val2-Pro3, a methyl substituted N-terminus and a quinoline scaffold in the C-terminal capping group, showed high BIR1/2 affinity and selectivity (eight and nine-fold respectively relative to BIR3). This is consistent with our molecular modeling studies. Our docking results suggested that this quinoline group sat close to the side chain of the residue Lys206 in BIR2 and may form favorable H-bonding (<a class="figpopup" href="/books/NBK47341/figure/ml101.f2/?report=objectonly" target="object" rid-figpopup="figml101f2" rid-ob="figobml101f2">Fig. 2C</a>). The substitution of the quinoline fragment for a phenyl hydrazine (CID25241673 (MLS-0391011)) showed an almost three-fold improved binding affinity for BIR1/2, albeit losing most of the selectivity that characterizes the two aforementioned compounds. Interestingly, the (<i>R</i>)-tetrahydronaphthyl amide derivative CID25241639 (MLS-0390982) exhibited an extraordinary affinity for BIR3 with an IC<sub>50</sub>= 0.28 μM. The substitution of the Val2 residue for the amino acids Ile2 or Abu2 afforded (<i>R</i>)-tetrahydronaphthyl analogues CID25241667 (MLS-0391006) and CID25241659 (MLS-0391002) respectively. Although this modification translated into a small erosion of their binding affinities, both compounds still exhibited very high affinity and selectivity for the BIR3 domain of XIAP. Our modeling results showed that the (R)-tetrahydronaphthyl group of MLS-0391002 was likely bound to the sub-pocket of Bir3 in a similar fashion as the ILE residue of the SMAC tetrapeptide AVPI (<a class="figpopup" href="/books/NBK47341/figure/ml101.f2/?report=objectonly" target="object" rid-figpopup="figml101f2" rid-ob="figobml101f2">Fig.2B, D</a>). Compound CID25241687 (MLS-0390979), which contains the sequence Ala1-Val2-Pro3, a non-substituted N-terminus and a diphenylmethane fragment in the C-terminal capping group, follows the same trend as the other three highly BIR3 selective analogues.</p><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml101t2"><a href="/books/NBK47341/table/ml101.t2/?report=objectonly" target="object" title="Table 2" class="img_link icnblk_img figpopup" rid-figpopup="figml101t2" rid-ob="figobml101t2"><img class="small-thumb" src="/books/NBK47341/table/ml101.t2/?report=thumb" src-large="/books/NBK47341/table/ml101.t2/?report=previmg" alt="Table 2. Binding affinity of representative analogues for the XIAP BIR1/2 and BIR3 domains." /></a><div class="icnblk_cntnt"><h4 id="ml101.t2"><a href="/books/NBK47341/table/ml101.t2/?report=objectonly" target="object" rid-ob="figobml101t2">Table 2</a></h4><p class="float-caption no_bottom_margin">Binding affinity of representative analogues for the XIAP BIR1/2 and BIR3 domains. </p></div></div><p>To help validate our synthetic approach, we decided to undertake the challenging synthesis of a previously reported bivalent Smac peptidomimetic, which represents the current state-of-the-art for inhibitors that bind to the BIR2 domain of XIAP. To this end we completed the synthesis of the bivalent inhibitor CID25181204 (MLS-0390866) and this was tested in the FP assays (<a class="figpopup" href="/books/NBK47341/figure/ml101.f3/?report=objectonly" target="object" rid-figpopup="figml101f3" rid-ob="figobml101f3">Figure 3</a>). As shown in <a class="figpopup" href="/books/NBK47341/figure/ml101.f3/?report=objectonly" target="object" rid-figpopup="figml101f3" rid-ob="figobml101f3">Figure 3</a>, the affinity values obtained were not as potent (or selective) as the results for the best new inhibitors summarized in <a class="figpopup" href="/books/NBK47341/table/ml101.t2/?report=objectonly" target="object" rid-figpopup="figml101t2" rid-ob="figobml101t2">Table 2</a>, suggesting that our new inhibitors are an improvement over the state-of-the-art compound CID25181204 (MLS-0390866).</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml101f3" co-legend-rid="figlgndml101f3"><a href="/books/NBK47341/figure/ml101.f3/?report=objectonly" target="object" title="Figure 3" class="img_link icnblk_img figpopup" rid-figpopup="figml101f3" rid-ob="figobml101f3"><img class="small-thumb" src="/books/NBK47341/bin/ml101f3.gif" src-large="/books/NBK47341/bin/ml101f3.jpg" alt="Figure 3. Bivalent Smac peptidomimetic CID-25181204 (MLS-0390866)." /></a><div class="icnblk_cntnt" id="figlgndml101f3"><h4 id="ml101.f3"><a href="/books/NBK47341/figure/ml101.f3/?report=objectonly" target="object" rid-ob="figobml101f3">Figure 3</a></h4><p class="float-caption no_bottom_margin">Bivalent Smac peptidomimetic CID-25181204 (MLS-0390866). </p></div></div><p>In summary, the SAR studies carried out around the tripeptide <b>1</b> have established that the presence of a quinoline substituent in the C-terminal capping group is important for selective binding of compounds to the BIR1/2 domain of XIAP. To our knowledge there is no precedent for this observation in the literature.</p></div></div></div><div id="ml101.s19"><h2 id="_ml101_s19_">3. Probe</h2><div id="ml101.s20"><h3>a. Chemical name</h3><p>formic acid; (2S)-1-[(2S)-3-methyl-2-[[(2S)-2 (methylamino)propanoyl]- amino]butanoyl]-N-quinolin-5-ylpyrrolidine-2-carboxamide <b>[<a href="/pcsubstance/?term=ML101[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML101</a>]</b></p></div><div id="ml101.s21"><h3>b. Probe chemical structure</h3><div id="ml101.fu2" class="figure"><div class="graphic"><img src="/books/NBK47341/bin/ml101fu9.jpg" alt="Image ml101fu9" /></div></div></div><div id="ml101.s22"><h3>c. Structural verification information of probe SID</h3><p><a href="https://pubchem.ncbi.nlm.nih.gov/substance/57643995" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-57643995</a></p><div id="ml101.fu3" class="figure"><div class="graphic"><img src="/books/NBK47341/bin/ml101fu10.jpg" alt="Image ml101fu10" /></div></div><div id="ml101.fu4" class="figure"><div class="graphic"><img src="/books/NBK47341/bin/ml101fu11.jpg" alt="Image ml101fu11" /></div></div></div><div id="ml101.s23"><h3>d. PubChem CID (corresponding to the SID)</h3><p>CID-25241665</p></div><div id="ml101.s24"><h3>e. Availability from a vendor</h3><p>This probe is not commercially available from vendors</p></div><div id="ml101.s25"><h3>f. MLS#'s of probe molecule and related samples that were submitted to the SMR collection</h3><div id="ml101.t3" class="table"><h3><span class="label">Table</span><span class="title">XIAP Probe and analogs submitted</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK47341/table/ml101.t3/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml101.t3_lrgtbl__"><table class="no_top_margin"><thead><tr><th id="hd_h_ml101.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Probe/analog</th><th id="hd_h_ml101.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS-# (BCCG#)</th><th id="hd_h_ml101.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">CID</th><th id="hd_h_ml101.t3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">SID</th><th id="hd_h_ml101.t3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Source (vendor or BCCG syn)</th><th id="hd_h_ml101.t3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Amt (mg)</th><th id="hd_h_ml101.t3_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Date ordered/submitted</th></tr></thead><tbody><tr><td headers="hd_h_ml101.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Probe</td><td headers="hd_h_ml101.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">0391005</td><td headers="hd_h_ml101.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">25241665</td><td headers="hd_h_ml101.t3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/57643995" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">57643995</a></td><td headers="hd_h_ml101.t3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BCCG syn</td><td headers="hd_h_ml101.t3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">15</td><td headers="hd_h_ml101.t3_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">5/18/09</td></tr><tr><td headers="hd_h_ml101.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Analog1</td><td headers="hd_h_ml101.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">0391014</td><td headers="hd_h_ml101.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">25241679</td><td headers="hd_h_ml101.t3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/57644002" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">57644002</a></td><td headers="hd_h_ml101.t3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BCCG syn</td><td headers="hd_h_ml101.t3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">15</td><td headers="hd_h_ml101.t3_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">5/18/09</td></tr><tr><td headers="hd_h_ml101.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Analog2</td><td headers="hd_h_ml101.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">0391011</td><td headers="hd_h_ml101.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">25241673</td><td headers="hd_h_ml101.t3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/57643999" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">57643999</a></td><td headers="hd_h_ml101.t3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BCCG syn</td><td headers="hd_h_ml101.t3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">15</td><td headers="hd_h_ml101.t3_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">5/18/09</td></tr><tr><td headers="hd_h_ml101.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Analog3</td><td headers="hd_h_ml101.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">0390982</td><td headers="hd_h_ml101.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">25241639</td><td headers="hd_h_ml101.t3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/57643981" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">57643981</a></td><td headers="hd_h_ml101.t3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BCCG syn</td><td headers="hd_h_ml101.t3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">15</td><td headers="hd_h_ml101.t3_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">5/18/09</td></tr><tr><td headers="hd_h_ml101.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Analog4</td><td headers="hd_h_ml101.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">0391006</td><td headers="hd_h_ml101.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">25241667</td><td headers="hd_h_ml101.t3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/57643996" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">57643996</a></td><td headers="hd_h_ml101.t3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BCCG syn</td><td headers="hd_h_ml101.t3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">15</td><td headers="hd_h_ml101.t3_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">5/18/09</td></tr><tr><td headers="hd_h_ml101.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Analog5</td><td headers="hd_h_ml101.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">0391002</td><td headers="hd_h_ml101.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">25241659</td><td headers="hd_h_ml101.t3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/57643992" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">57643992</a></td><td headers="hd_h_ml101.t3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BCCG syn</td><td headers="hd_h_ml101.t3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">15</td><td headers="hd_h_ml101.t3_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">5/18/09</td></tr><tr><td headers="hd_h_ml101.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Analog6</td><td headers="hd_h_ml101.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">0390979</td><td headers="hd_h_ml101.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">25241687</td><td headers="hd_h_ml101.t3_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/57644006" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">57644006</a></td><td headers="hd_h_ml101.t3_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BCCG syn</td><td headers="hd_h_ml101.t3_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">15</td><td headers="hd_h_ml101.t3_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">5/18/09</td></tr></tbody></table></div></div></div><div id="ml101.s26"><h3>g. Mode of action for biological activity of probe</h3><p>The probe reported mimics the binding of a single AVPI binding motif to the BIR1/2 domain of XIAP.</p></div><div id="ml101.s27"><h3>h. Detailed synthetic pathway for making probe</h3><div id="ml101.f4" class="figure bk_fig"><div class="graphic"><img src="/books/NBK47341/bin/ml101f4.jpg" alt="Scheme 1. Synthesis of probe." /></div><h3><span class="label">Scheme 1</span><span class="title">Synthesis of probe</span></h3></div></div><div id="ml101.s28"><h3>i. Probe properties (solubility, absorbance/fluorescence, reactivity, toxicity, etc.)</h3><p>Our probe has demonstrated potent binding affinity and selectivity for BIR1/2 domain of XIAP.</p></div><div id="ml101.s29"><h3>j. Properties Computed from Structure</h3><div id="ml101.tu3" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK47341/table/ml101.tu3/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml101.tu3_lrgtbl__"><table><tbody><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Molecular Weight</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">471.54932</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Molecular formula</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">C<sub>24</sub>H<sub>33</sub>N<sub>5</sub>O<sub>5</sub></td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">H-Bond Donor</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">4</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">H-Bond Acceptor</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">7</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Rotatable Bond Count</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">7</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Tautomer Count</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">8</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Exact Mass</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">471.248169</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">MonoIsotopic Mass</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">471.248169</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Topological Polar Surface Area</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">141</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Heavy Atom Count</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">34</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Formal Charge</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">0</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Complexity</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">665</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Isotope Atom Count</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">0</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Defined Atom StereoCenter Count</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">3</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Undefined Atom StereoCenter Count</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">0</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Defined Bond StereoCenter Count</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">0</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Undefined Bond StereoCenter Count</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">0</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Covalently-Bonded Unit Count</td><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">2</td></tr></tbody></table></div></div></div></div><div id="ml101.s30"><h2 id="_ml101_s30_">4. Appendices</h2><div id="ml101.s31"><h3>a. Comparative data showing probe specificity for target</h3><p>A similar assay has been described in the literature (<a class="bk_pop" href="#ml101.r9">9</a>,<a class="bk_pop" href="#ml101.r10">10</a>,<a class="bk_pop" href="#ml101.r11">11</a>). In this case, a fluorescein molecule is coupled to the SMAC peptide. In this study rhodamine was preferred since it was further red shifted than flourescein allowing for less sensitivity in the assay to compounds which are auto-fluorescent.</p></div></div><div id="ml101.s32"><h2 id="_ml101_s32_">5. Bibliography</h2><dl class="temp-labeled-list"><dt>1.</dt><dd><div class="bk_ref" id="ml101.r1">Thompson CB. Apoptosis in the pathogenesis and treatment of disease. <span><span class="ref-journal">Science. </span>1995;<span class="ref-vol">267</span>:1456.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/7878464" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 7878464</span></a>]</div></dd><dt>2.</dt><dd><div class="bk_ref" id="ml101.r2">Riedl SJ, Shi Y. Molecular mechanisms of caspase regulation during apoptosis. <span><span class="ref-journal">Nature Reviews Moleculart Cell Biology. </span>2004;<span class="ref-vol">5</span>:897.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/15520809" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15520809</span></a>]</div></dd><dt>3.</dt><dd><div class="bk_ref" id="ml101.r3">Deveraux QL, Reed JC. IAP family proteins: Suppressors of apoptosis. <span><span class="ref-journal">Genes Dev. </span>1999;<span class="ref-vol">13</span>:239.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/9990849" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 9990849</span></a>]</div></dd><dt>4.</dt><dd><div class="bk_ref" id="ml101.r4">Du C, Fang M, Li Y, Li L, Wang X. SMAC, a mitochondrial protein that promotes cytochrome c-dependent caspase activation by eliminating IAP inhibition. <span><span class="ref-journal">Cell. </span>2000;<span class="ref-vol">102</span>:33.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/10929711" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 10929711</span></a>]</div></dd><dt>5.</dt><dd><div class="bk_ref" id="ml101.r5">Verhagen AM, Ekert PG, Pakusch M, Silke J, Connolly LM, Reid GE, Moritz RL, Simpson RJ, Vaux DL. Identification of DIABLO, a Mammalian Protein that Promotes Apoptosis by Binding to and Antagonizing IAP Proteins. <span><span class="ref-journal">Cell. </span>2000;<span class="ref-vol">102</span>:43.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/10929712" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 10929712</span></a>]</div></dd><dt>6.</dt><dd><div class="bk_ref" id="ml101.r6">Suzuki Y, Imai Y, Nakayama H, Takahashi K, Takio K, Takahashi R. A serine protease, HtrA2, is released from the mitochondria and interacts with XIAP, inducing cell death. <span><span class="ref-journal">Molecular Cell. </span>2001;<span class="ref-vol">8</span>:613.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/11583623" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11583623</span></a>]</div></dd><dt>7.</dt><dd><div class="bk_ref" id="ml101.r7">Liu Z, Sun C, Olejniczak ET, Meadows RP, Betz SF, Oost T, Hermann J, Wu JC, Fesik SW. Structural basis for binding of SMAC/DIABLO to the BIR3 domain. <span><span class="ref-journal">Nature. </span>2000;<span class="ref-vol">408</span>:1004.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/11140637" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11140637</span></a>]</div></dd><dt>8.</dt><dd><div class="bk_ref" id="ml101.r8">Huang Y, Rich RL, Myszka DG, Wu H. Requirement of both the second and third domains for the relief of X-linked inhibitor of apoptosis protein (XIAP)-mediated caspase inhibition by SMAC. <span><span class="ref-journal">J. Biol. Chem. </span>2003;<span class="ref-vol">278</span>:49517.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/14512414" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 14512414</span></a>]</div></dd><dt>9.</dt><dd><div class="bk_ref" id="ml101.r9">Glover CJ, Hite K, DeLosh R, Scudiero DA, Fivash MJ, Smith LR, Fisher RJ, Wu J-W, Shi Y, Kipp RA, McLendon GL, Sausville EA, Shoemaker RH. A high-throughput screen for identification of molecular mimics of SMAC-DIABLO utilizing a fluorescence polarization assay. <span><span class="ref-journal">Anal. Biochem. </span>2003;<span class="ref-vol">320</span>:157.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/12927820" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 12927820</span></a>]</div></dd><dt>10.</dt><dd><div class="bk_ref" id="ml101.r10">Nikolovska-Coleska Z, Wang R, Fang X, Pan H, Tomita Y, Li P, Roller PP, Krajewski K, Saito NG, Stuckey JA, Wang S. Development
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and optimization of a binding assay for XIAP BIR3 domain using fluorescence
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polarization. <span><span class="ref-journal">Anal.
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Biochem. </span>2004;<span class="ref-vol">332</span>:261.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/15325294" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15325294</span></a>]</div></dd><dt>11.</dt><dd><div class="bk_ref" id="ml101.r11">Oost TK, Sun C, Armstrong RC, Al-Assaad A-S, Betz SF, Deckwerth TL, Ding H, Elmore SW, Meadows RP, Olejniczak ET, Oleksijew A, Olterdorf T, Rosenberg SH, Shoemaker AR, Tomaselli KJ, Zou H, Fesik SW. Discovery of potent antagonists of the antiapoptotic protein XIAP for the treatment of cancer. <span><span class="ref-journal">J. Med. Chem. </span>2004;<span class="ref-vol">47</span>:4417.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/15317454" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15317454</span></a>]</div></dd></dl></div><div style="display:none"><div style="display:none" id="figml101f2"><img alt="Image ml101f2" src-large="/books/NBK47341/bin/ml101f2.jpg" /></div></div><div id="bk_toc_contnr"></div></div></div>
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<div class="post-content"><div><div class="half_rhythm"><a href="/books/about/copyright/">Copyright Notice</a></div><div class="small"><span class="label">Bookshelf ID: NBK47341</span><span class="label">PMID: <a href="https://pubmed.ncbi.nlm.nih.gov/21433357" title="PubMed record of this page" ref="pagearea=meta&targetsite=entrez&targetcat=link&targettype=pubmed">21433357</a></span></div><div style="margin-top:2em" class="bk_noprnt"><a class="bk_cntns" href="/books/n/mlprobe/">Contents</a><div class="pagination bk_noprnt"><a class="active page_link prev" href="/books/n/mlprobe/ml102/" title="Previous page in this title">< Prev</a><a class="active page_link next" href="/books/n/mlprobe/ml100/" title="Next page in this title">Next ></a></div></div></div></div>
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<div xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance"></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Views</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="PDF_download" id="Shutter"></a></div><div class="portlet_content"><ul xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="simple-list"><li><a href="/books/NBK47341/?report=reader">PubReader</a></li><li><a href="/books/NBK47341/?report=printable">Print View</a></li><li><a data-jig="ncbidialog" href="#_ncbi_dlg_citbx_NBK47341" data-jigconfig="width:400,modal:true">Cite this Page</a><div id="_ncbi_dlg_citbx_NBK47341" style="display:none" title="Cite this Page"><div class="bk_tt">Welsh K, Yuan H, Stonich D, et al. Antagonists of IAP-family anti-apoptotic proteins - Probe 1. 2009 May 18 [Updated 2010 Sep 2]. In: Probe Reports from the NIH Molecular Libraries Program [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2010-. <span class="bk_cite_avail"></span></div></div></li><li><a href="/books/NBK47341/pdf/Bookshelf_NBK47341.pdf">PDF version of this page</a> (316K)</li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>In this Page</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="page-toc" id="Shutter"></a></div><div class="portlet_content"><ul xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="simple-list"><li><a href="#ml101.s2" ref="log$=inpage&link_id=inpage">Probe Structure & Characteristics</a></li><li><a href="#ml101.s3" ref="log$=inpage&link_id=inpage">Recommendations for the scientific use of this probe</a></li><li><a href="#ml101.s4" ref="log$=inpage&link_id=inpage">Scientific Rationale for Project</a></li><li><a href="#ml101.s7" ref="log$=inpage&link_id=inpage">Project Description</a></li><li><a href="#ml101.s19" ref="log$=inpage&link_id=inpage">Probe</a></li><li><a href="#ml101.s30" ref="log$=inpage&link_id=inpage">Appendices</a></li><li><a href="#ml101.s32" ref="log$=inpage&link_id=inpage">Bibliography</a></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Related information</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="discovery_db_links" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pcassay&DbFrom=books&Cmd=Link&LinkName=books_pcassay_probe&IdsFromResult=2358572" ref="log$=recordlinks">PubChem BioAssay for Chemical Probe</a><div class="brieflinkpop offscreen_noflow">PubChem BioAssay records reporting screening data for the development of the chemical probe(s) described in this book chapter</div></li><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pcsubstance&DbFrom=books&Cmd=Link&LinkName=books_pcsubstance&IdsFromResult=2358572" ref="log$=recordlinks">PubChem Substance</a><div class="brieflinkpop offscreen_noflow">Related PubChem Substances</div></li><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pubmed&DbFrom=books&Cmd=Link&LinkName=books_pubmed_refs&IdsFromResult=2358572" ref="log$=recordlinks">PubMed</a><div class="brieflinkpop offscreen_noflow">Links to PubMed</div></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Similar articles in PubMed</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="PBooksDiscovery_RA" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/21634080" ref="ordinalpos=1&linkpos=1&log$=relatedreviews&logdbfrom=pubmed"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> Antagonists of IAP-family anti-apoptotic proteins - Probe 2.</a><span class="source">[Probe Reports from the NIH Mol...]</span><div class="brieflinkpop offscreen_noflow"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> Antagonists of IAP-family anti-apoptotic proteins - Probe 2.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Lopez M, Welsh K, Yuan H, Stonich D, Su Y, Garcia X, Cuddy M, Houghten R, Sergienko E, Reed JC, et al. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Probe Reports from the NIH Molecular Libraries Program. 2010</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/18023397" ref="ordinalpos=1&linkpos=2&log$=relatedarticles&logdbfrom=pubmed">Design and characterization of bivalent Smac-based peptides as antagonists of XIAP and development and validation of a fluorescence polarization assay for XIAP containing both BIR2 and BIR3 domains.</a><span class="source">[Anal Biochem. 2008]</span><div class="brieflinkpop offscreen_noflow">Design and characterization of bivalent Smac-based peptides as antagonists of XIAP and development and validation of a fluorescence polarization assay for XIAP containing both BIR2 and BIR3 domains.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Nikolovska-Coleska Z, Meagher JL, Jiang S, Kawamoto SA, Gao W, Yi H, Qin D, Roller PP, Stuckey JA, Wang S. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Anal Biochem. 2008 Mar 1; 374(1):87-98. Epub 2007 Nov 20.</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/15485396" ref="ordinalpos=1&linkpos=3&log$=relatedarticles&logdbfrom=pubmed">Engineering ML-IAP to produce an extraordinarily potent caspase 9 inhibitor: implications for Smac-dependent anti-apoptotic activity of ML-IAP.</a><span class="source">[Biochem J. 2005]</span><div class="brieflinkpop offscreen_noflow">Engineering ML-IAP to produce an extraordinarily potent caspase 9 inhibitor: implications for Smac-dependent anti-apoptotic activity of ML-IAP.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Vucic D, Franklin MC, Wallweber HJ, Das K, Eckelman BP, Shin H, Elliott LO, Kadkhodayan S, Deshayes K, Salvesen GS, et al. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Biochem J. 2005 Jan 1; 385(Pt 1):11-20. </em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/17168540" ref="ordinalpos=1&linkpos=4&log$=relatedarticles&logdbfrom=pubmed">Design, synthesis, and biological activity of a potent Smac mimetic that sensitizes cancer cells to apoptosis by antagonizing IAPs.</a><span class="source">[ACS Chem Biol. 2006]</span><div class="brieflinkpop offscreen_noflow">Design, synthesis, and biological activity of a potent Smac mimetic that sensitizes cancer cells to apoptosis by antagonizing IAPs.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Zobel K, Wang L, Varfolomeev E, Franklin MC, Elliott LO, Wallweber HJ, Okawa DC, Flygare JA, Vucic D, Fairbrother WJ, et al. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">ACS Chem Biol. 2006 Sep 19; 1(8):525-33. </em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/25120954" ref="ordinalpos=1&linkpos=5&log$=relatedreviews&logdbfrom=pubmed"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> X-linked inhibitor of apoptosis protein - a critical death resistance regulator and therapeutic target for personalized cancer therapy.</a><span class="source">[Front Oncol. 2014]</span><div class="brieflinkpop offscreen_noflow"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> X-linked inhibitor of apoptosis protein - a critical death resistance regulator and therapeutic target for personalized cancer therapy.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Obexer P, Ausserlechner MJ. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Front Oncol. 2014; 4:197. 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