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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="Identification of Potent and Selective Thyroid Stimulating Hormone Receptor Agonists" /><meta name="citation_publisher" content="National Center for Biotechnology Information (US)" /><meta name="citation_date" content="2011/03/25" /><meta name="citation_author" content="Steve Titus" /><meta name="citation_author" content="Wenwei Huang" /><meta name="citation_author" content="Juan Marugan" /><meta name="citation_author" content="Noel Southall" /><meta name="citation_author" content="Jim Inglese" /><meta name="citation_author" content="Christopher Austin" /><meta name="citation_author" content="Wei Zheng" /><meta name="citation_author" content="Erika Englund" /><meta name="citation_author" content="Susanne Neumann" /><meta name="citation_author" content="Marvin Gershengorn" /><meta name="citation_pmid" content="21735603" /><meta name="citation_fulltext_html_url" content="https://www.ncbi.nlm.nih.gov/books/NBK56234/" /><link rel="schema.DC" href="http://purl.org/DC/elements/1.0/" /><meta name="DC.Title" content="Identification of Potent and Selective Thyroid Stimulating Hormone Receptor Agonists" /><meta name="DC.Type" content="Text" /><meta name="DC.Publisher" content="National Center for Biotechnology Information (US)" /><meta name="DC.Contributor" content="Steve Titus" /><meta name="DC.Contributor" content="Wenwei Huang" /><meta name="DC.Contributor" content="Juan Marugan" /><meta name="DC.Contributor" content="Noel Southall" /><meta name="DC.Contributor" content="Jim Inglese" /><meta name="DC.Contributor" content="Christopher Austin" /><meta name="DC.Contributor" content="Wei Zheng" /><meta name="DC.Contributor" content="Erika Englund" /><meta name="DC.Contributor" content="Susanne Neumann" /><meta name="DC.Contributor" content="Marvin Gershengorn" /><meta name="DC.Date" content="2011/03/25" /><meta name="DC.Identifier" content="https://www.ncbi.nlm.nih.gov/books/NBK56234/" /><meta name="description" content="Thyroid Stimulating Hormone (TSH) is a heterodimeric glycoprotein hormone that regulates thyroid homeostasis upon interaction with the TSH receptor (TSHR). TSH binds to the TSH receptor, which couples preferentially to the G-alpha (s) (Gs) protein, resulting in activation of adenylate cyclase and increase in cyclic adenosine 3′, 5′ monophosphate (cAMP). In this report, we describe the discovery and SAR studies of a series of dihydroquinazolin-4-ones as TSHR agonists. ML109 (CID-25246343) is the first selective and orally available small-molecule TSHR agonist, and the probe will be a useful pharmacological tool to study TSHR biology in thyroidal and extrathyroidal tissues." /><meta name="og:title" content="Identification of Potent and Selective Thyroid Stimulating Hormone Receptor Agonists" /><meta name="og:type" content="book" /><meta name="og:description" content="Thyroid Stimulating Hormone (TSH) is a heterodimeric glycoprotein hormone that regulates thyroid homeostasis upon interaction with the TSH receptor (TSHR). TSH binds to the TSH receptor, which couples preferentially to the G-alpha (s) (Gs) protein, resulting in activation of adenylate cyclase and increase in cyclic adenosine 3′, 5′ monophosphate (cAMP). In this report, we describe the discovery and SAR studies of a series of dihydroquinazolin-4-ones as TSHR agonists. ML109 (CID-25246343) is the first selective and orally available small-molecule TSHR agonist, and the probe will be a useful pharmacological tool to study TSHR biology in thyroidal and extrathyroidal tissues." /><meta name="og:url" content="https://www.ncbi.nlm.nih.gov/books/NBK56234/" /><meta name="og:site_name" content="NCBI Bookshelf" /><meta name="og:image" content="https://www.ncbi.nlm.nih.gov/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-mlprobe-lrg.png" /><meta name="twitter:card" content="summary" /><meta name="twitter:site" content="@ncbibooks" /><meta name="bk-non-canon-loc" content="/books/n/mlprobe/ml109/" /><link rel="canonical" href="https://www.ncbi.nlm.nih.gov/books/NBK56234/" /><link rel="stylesheet" href="/corehtml/pmc/css/figpopup.css" type="text/css" media="screen" /><link rel="stylesheet" href="/corehtml/pmc/css/bookshelf/2.26/css/books.min.css" type="text/css" /><link rel="stylesheet" href="/corehtml/pmc/css/bookshelf/2.26/css/books_print.min.css" type="text/css" media="print" /><style type="text/css">p a.figpopup{display:inline !important} .bk_tt {font-family: monospace} .first-line-outdent .bk_ref {display: inline} .body-content h2, .body-content .h2 {border-bottom: 1px solid #97B0C8} .body-content h2.inline {border-bottom: none} a.page-toc-label , .jig-ncbismoothscroll a {text-decoration:none;border:0 !important} .temp-labeled-list .graphic {display:inline-block !important} .temp-labeled-list img{width:100%}</style><script type="text/javascript" src="/corehtml/pmc/js/jquery.hoverIntent.min.js"> </script><script type="text/javascript" src="/corehtml/pmc/js/common.min.js?_=3.18"> </script><script type="text/javascript" src="/corehtml/pmc/js/large-obj-scrollbars.min.js"> </script><script type="text/javascript">window.name="mainwindow";</script><script type="text/javascript" src="/corehtml/pmc/js/bookshelf/2.26/book-toc.min.js"> </script><script type="text/javascript" src="/corehtml/pmc/js/bookshelf/2.26/books.min.js"> </script><meta name="book-collection" content="NONE" />
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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="#__NBK56234_dtls__">Show details</a><div style="display:none" class="ui-widget" id="__NBK56234_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/ml111/" title="Previous page in this title">< Prev</a><a class="active page_link next" href="/books/n/mlprobe/ml108/" 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="_NBK56234_"><span class="title" itemprop="name">Identification of Potent and Selective Thyroid Stimulating Hormone Receptor Agonists</span></h1><p class="contrib-group"><span itemprop="author">Steve Titus</span>, <span itemprop="author">Wenwei Huang</span>, <span itemprop="author">Juan Marugan</span>, <span itemprop="author">Noel Southall</span>, <span itemprop="author">Jim Inglese</span>, <span itemprop="author">Christopher Austin</span>, <span itemprop="author">Wei Zheng</span>, <span itemprop="author">Erika Englund</span>, <span itemprop="author">Susanne Neumann</span>, and <span itemprop="author">Marvin Gershengorn</span>.</p><a data-jig="ncbitoggler" href="#__NBK56234_ai__" style="border:0;text-decoration:none">Author Information and Affiliations</a><div style="display:none" class="ui-widget" id="__NBK56234_ai__"><p class="contrib-group"><h4>Authors</h4><span itemprop="author">Steve Titus</span>,<sup>*</sup> <span itemprop="author">Wenwei Huang</span>,<sup>*</sup> <span itemprop="author">Juan Marugan</span>,<sup>*</sup> <span itemprop="author">Noel Southall</span>,<sup>*</sup> <span itemprop="author">Jim Inglese</span>,<sup>*</sup> <span itemprop="author">Christopher Austin</span>,<sup>*</sup> <span itemprop="author">Wei Zheng</span>,<sup>*</sup> <span itemprop="author">Erika Englund</span>,<sup>*</sup> <span itemprop="author">Susanne Neumann</span>,<sup>A</sup> and <span itemprop="author">Marvin Gershengorn</span><sup>A</sup>.</p><h4>Affiliations</h4><div class="affiliation"><sup>*</sup>
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NIH Chemical Genomics Center, 9800 Medical Center Dr., Building B, Bethesda, MD, 20892-3370</div><div class="affiliation"><sup>A</sup>
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National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), NIH</div></div><p class="small">Received: <span itemprop="datePublished">September 1, 2009</span>; Last Update: <span itemprop="dateModified">March 25, 2011</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>Thyroid Stimulating Hormone (TSH) is a heterodimeric glycoprotein hormone that regulates thyroid homeostasis upon interaction with the TSH receptor (TSHR). TSH binds to the TSH receptor, which couples preferentially to the G-alpha (s) (Gs) protein, resulting in activation of adenylate cyclase and increase in cyclic adenosine 3′, 5′ monophosphate (cAMP). In this report, we describe the discovery and SAR studies of a series of dihydroquinazolin-4-ones as TSHR agonists. <a href="/pcsubstance/?term=ML109[synonym]" ref="pagearea=abstract&targetsite=entrez&targetcat=term&targettype=pubchem">ML109</a> (CID-25246343) is the first selective and orally available small-molecule TSHR agonist, and the probe will be a useful pharmacological tool to study TSHR biology in thyroidal and extrathyroidal tissues.</p></div><div class="h2"></div><p><b>Assigned Assay Grant #:</b> X01 <a href="/nuccore/1365463024" class="bk_tag" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=nuccore">MH080680</a></p><p><b>Screening Center Name & PI</b>: NIH Chemical Genomics Center, Christopher Austin</p><p><b>Chemistry Center Name & PI</b>: NIH Chemical Genomics Center, Christopher Austin</p><p><b>Assay Submitter & Institution</b>: Marvin Gershengorn, NIDDK</p><p><b>PubChem Summary Bioassay Identifier (AID)</b>: <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1401" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1401</a></p><div id="ml109.s1"><h2 id="_ml109_s1_">Probe Structure & Characteristics</h2><p>PubChem CID: 25246343</p><p>Probe MLSMR ID: MLS002576689</p><p>Internal ID: NCGC00161870</p><p>Chemical Formula: C<sub>31</sub>H<sub>29</sub>N<sub>3</sub>O<sub>5</sub></p><p>Exact Mass: 523.21</p><div id="ml109.fu1" class="figure bk_fig"><div class="graphic"><img src="/books/NBK56234/bin/ml109fu1.jpg" alt="ML109." /></div><h3><span class="title"><a href="/pcsubstance/?term=ML109[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML109</a></span></h3></div><div id="ml109.tu1" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK56234/table/ml109.tu1/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml109.tu1_lrgtbl__"><table><thead><tr><th id="hd_h_ml109.tu1_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">CID/ML#</th><th id="hd_h_ml109.tu1_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Target Name</th><th id="hd_h_ml109.tu1_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">IC<sub>50</sub>/EC<sub>50</sub> (nM) [SID, AID]</th><th id="hd_h_ml109.tu1_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Anti-target Name(s)</th><th id="hd_h_ml109.tu1_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">IC<sub>50</sub>/EC<sub>50</sub> (μM) [SID, AID]</th><th id="hd_h_ml109.tu1_1_1_1_6" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Fold Selective</th><th id="hd_h_ml109.tu1_1_1_1_7" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Secondary Assay(s) Name: IC<sub>50</sub>/EC<sub>50</sub> (nM) [SID, AID]</th></tr></thead><tbody><tr><td headers="hd_h_ml109.tu1_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">25246343/<a href="/pcsubstance/?term=ML109[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML109</a></td><td headers="hd_h_ml109.tu1_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">TSHR</td><td headers="hd_h_ml109.tu1_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">7900nM [<a href="https://pubchem.ncbi.nlm.nih.gov/substance/26755506" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">26755506</a>,<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1401" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1401</a>]</td><td headers="hd_h_ml109.tu1_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">FSHR</td><td headers="hd_h_ml109.tu1_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">>100μM [<a href="https://pubchem.ncbi.nlm.nih.gov/substance/26755506" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">26755506</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1403" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1403</a>]</td><td headers="hd_h_ml109.tu1_1_1_1_6" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">>10 fold</td><td headers="hd_h_ml109.tu1_1_1_1_7" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">TSHR cAMP ELISA 90nM [<a href="https://pubchem.ncbi.nlm.nih.gov/substance/26755506" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">26755506</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2104" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2104</a>]</td></tr><tr><td headers="hd_h_ml109.tu1_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_ml109.tu1_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_ml109.tu1_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td><td headers="hd_h_ml109.tu1_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">LHR</td><td headers="hd_h_ml109.tu1_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">>100μM [<a href="https://pubchem.ncbi.nlm.nih.gov/substance/26755506" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">26755506</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1402" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1402</a>]</td><td headers="hd_h_ml109.tu1_1_1_1_6" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">>10 fold</td><td headers="hd_h_ml109.tu1_1_1_1_7" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr></tbody></table></div></div></div><div id="ml109.s2"><h2 id="_ml109_s2_">Recommendations for scientific use of the probe</h2><p>This probe is a member of a series of TSHR agonists. The current state of the art is lacking any small molecule TSHR agonists. This probe can be used to study TSHR functions <i>in vitro</i> and <i>in vivo</i>, and could be used as a lead for development of drugs to replace recombinant human TSH in patients with thyroid cancer. It is highly selective for human TSHR over other glycoprotein hormone receptors, such as LHCGR and FSHR, and interacts with the receptor’s serpentine domain. In primary cultures of human thyrocytes, this compound increases mRNA levels for thyroglobulin, thyroperoxidase, sodium iodide symporter, and deiodinase type 2, as well as deiodinase type2 enzyme activity. Moreover, oral administration of the agonist stimulated thyroid function in mice, resulting in increased serum thyroxine and thyroidal radioiodide uptake<a class="bk_pop" href="#ml109.r1">1</a>. This report supplants the previous report of activity with the [2-(furan-2-ylmethylamino)-2-oxoethyl] adamantane-1-carboxylate series.</p></div><div id="ml109.s3"><h2 id="_ml109_s3_">1. Introduction</h2><p>Thyroid Stimulating Hormone (TSH) is an α/β heterodimeric glycoprotein hormone secreted from the anterior pituitary gland. It belongs to the glycoprotein hormone family, including Chorionic Gonadotropin (CG), Luteinizing Hormone (LH), and Follicle Stimulating Hormone (FSH)<a class="bk_pop" href="#ml109.r2">2</a>,<a class="bk_pop" href="#ml109.r3">3</a>. TSH binds to the TSH receptor (TSHR), which couples preferentially to the G-alpha (s) (Gs) protein, resulting in activation of adenylate cyclase and increase in cyclic adenosine 3′, 5′ monophosphate (cAMP). THSR is mainly expressed in thyroid follicular cells<a class="bk_pop" href="#ml109.r4">4</a> and regulates their growth and function<a class="bk_pop" href="#ml109.r5">5</a>. Recombinant TSH is currently used for follow-up in patients with thyroid cancer who are receiving thyroid hormone suppression therapy, specifically for screening of residual tumor after surgery<a class="bk_pop" href="#ml109.r6">6</a>, but it is difficult to produce and must be administered intramuscularly. An orally active small molecule TSHR agonist would serve as a valuable research tool for studying TSHR pharmacology and physiology, especially the extrathyroidal role of TSHR, and would have multiple advantages for therapeutic applications.</p><p>There are few literature reports of TSHR agonists (all from the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) group), and most are confined to a class of thienopyrimidines with little structural diversity and no selectivity for TSHR over LHR or FSHR. The thienopyrimidine <b>Org 41841</b> (<a class="figpopup" href="/books/NBK56234/figure/ml109.f1/?report=objectonly" target="object" rid-figpopup="figml109f1" rid-ob="figobml109f1">Fig. 1</a>) was first described in 2002 as an orally active LH agonist with <i>in vivo</i> efficacy in mice following oral administration<a class="bk_pop" href="#ml109.r7">7</a>. It was later found to have modest TSHR activity, with an EC<sub>50</sub> of 7700nM and 220nM for TSHR and LHCGR, respectively<a class="bk_pop" href="#ml109.r8">8</a>. This finding was followed with a series of synthesized analogs for SAR studies. Some of the analogs achieved LHR selectivity, but none were TSHR selective<a class="bk_pop" href="#ml109.r9">9</a>. Another TSHR agonist is from a high throughput screen at NCGC. <b>MLS000098157</b> was identified as a 2μM agonist of hTSHR from the 2-(furan-2-ylmethylamino)-2-oxoethyl] adamantane-1-carboxylate series (<a class="figpopup" href="/books/NBK56234/figure/ml109.f1/?report=objectonly" target="object" rid-figpopup="figml109f1" rid-ob="figobml109f1">Fig. 1</a>). That series is supplanted by this report.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml109f1" co-legend-rid="figlgndml109f1"><a href="/books/NBK56234/figure/ml109.f1/?report=objectonly" target="object" title="Figure 1" class="img_link icnblk_img figpopup" rid-figpopup="figml109f1" rid-ob="figobml109f1"><img class="small-thumb" src="/books/NBK56234/bin/ml109f1.gif" src-large="/books/NBK56234/bin/ml109f1.jpg" alt="Figure 1. TSHR agonists." /></a><div class="icnblk_cntnt" id="figlgndml109f1"><h4 id="ml109.f1"><a href="/books/NBK56234/figure/ml109.f1/?report=objectonly" target="object" rid-ob="figobml109f1">Figure 1</a></h4><p class="float-caption no_bottom_margin">TSHR agonists. </p></div></div></div><div id="ml109.s4"><h2 id="_ml109_s4_">2. Materials and Methods</h2><p>All commercially available reagents and solvents were purchased and used without further purification. All microwave reactions were carried out in a sealed microwave vial equipped with a magnetic stir bar and heated in a Biotage Initiator Microwave Synthesizer. All compounds for biological testing were purified using a Waters semi-preparative HPLC equipped with a Phenomenex Luna<sup>®</sup> C18 reverse phase (5 micron, 30 × 75 mm) column having a flow rate of 45 ml/min. The mobile phase was a mixture of acetonitrile and H<sub>2</sub>O, each containing 0.1% trifluoroacetic acid. During purification, a gradient of 30% to 80% acetonitrile over 8 minutes was used with fraction collection triggered by UV detection (220 nM). Pure fractions passed through PL-HCO<sub>3</sub> MP SPE (Varian) to remove trifluoroacetic acid and concentrated under vacuum on a lyophilizer. <sup>1</sup>H spectra were recorded using an Inova 400 (100) MHz spectrometer (Varian).</p><div id="ml109.s5"><h3>2.1. Assays</h3><div id="ml109.s6"><h4>i. qHTS Assay for agonists of the TSH receptor [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/926" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-926</a>; Primary DR]</h4><div id="ml109.s7"><h5>Assay Description</h5><p>TSHR is a seven-transmembrane receptor that couples to the Gs protein, resulting in activation of adenylate cyclase and increase in intracellular cAMP level. This cell-based assay utilized a cyclic nucleotide gated ion channel (CNG) as a biosensor for measurement of increase in intracellular cAMP level stimulated by TSHR agonists. The HEK293 cell line expressing human TSHR and a modified CNG was used as the primary screen assay<a class="bk_pop" href="#ml109.r9">9</a>,<a class="bk_pop" href="#ml109.r10">10</a>,<a class="bk_pop" href="#ml109.r11">11</a>. The membrane potential dye was used to detect the membrane depolarization upon the activation of CNG by increased cAMP level. This assay was optimized in a homogenous 1536-well plate format.</p></div><div id="ml109.s8"><h5>TSHR cell line and Parental cell lines used in membrane potential assay</h5><p>A HEK293 cell line stably expressing the TSHR as well as a modified CNG was purchased from BD biosciences (TSHR ACTOne cell line, Rockville, MD). When the TSHR is stimulated, intracellular cAMP level increases, which activates CNG channels to cause membrane depolarization. A fluorescent membrane potential dye (BD biosciences) was utilized to detect the membrane depolarization in this assay (<a class="figpopup" href="/books/NBK56234/figure/ml109.f2/?report=objectonly" target="object" rid-figpopup="figml109f2" rid-ob="figobml109f2">Fig. 2</a>). The cells were maintained in DMEM medium (Invitrogen) containing 10% FBS (Hyclone), 100 units/ml Penicillin, 100 ug/ml Streptomycin (Invitrogen), and 250 ug/ml Geneticin (Invitrogen) and 1 μg/ml Puromycin (Invitrogen) at 37°C in 5% CO2. A parental HEK293 cell line expressing CNG without TSHR was used in a parallel screen to eliminate compounds that activate targets other than TSHR.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml109f2" co-legend-rid="figlgndml109f2"><a href="/books/NBK56234/figure/ml109.f2/?report=objectonly" target="object" title="Figure 2" class="img_link icnblk_img figpopup" rid-figpopup="figml109f2" rid-ob="figobml109f2"><img class="small-thumb" src="/books/NBK56234/bin/ml109f2.gif" src-large="/books/NBK56234/bin/ml109f2.jpg" alt="Figure 2. Schematic illustration of the assay principle of the ACT: One cAMP assay for TSHR." /></a><div class="icnblk_cntnt" id="figlgndml109f2"><h4 id="ml109.f2"><a href="/books/NBK56234/figure/ml109.f2/?report=objectonly" target="object" rid-ob="figobml109f2">Figure 2</a></h4><p class="float-caption no_bottom_margin">Schematic illustration of the assay principle of the ACT: One cAMP assay for TSHR. A TSHR was co-expressed in a HEK293 cell line with a modified CNG channel. When TSH binds to TSHR, it increases the intracellular cAMP level. The CNG channels are activated <a href="/books/NBK56234/figure/ml109.f2/?report=objectonly" target="object" rid-ob="figobml109f2">(more...)</a></p></div></div></div><div id="ml109.s9"><h5>Assay Protocol</h5><p>Reagents used are listed in <a class="figpopup" href="/books/NBK56234/table/ml109.t1/?report=objectonly" target="object" rid-figpopup="figml109t1" rid-ob="figobml109t1">Table 1</a>. Cells from 4 confluent T175 flasks were seeded into 25 to 30 T175 flasks at a density of 3 to 4 million cells per flask. After 3 days of growth, the flasks were harvested, and the fresh cells were used for dispensing into 1536-well plates. This process was repeated on a daily basis for screening purposes. The cells were resuspended in 2% FCS DMEM medium at 500,000 cells/ml, and 4μl of resuspended cells were dispensed into each well of black, clear bottom, 1536-well tissue culture–treated plates using a Multidrop Combi dispenser. After overnight culture at 37°C with 5% CO2, the cells were generally 60% to 70% confluent in the 1536-well plates. After overnight incubation, 4μl of membrane potential dye containing 50μM of the phosphodiesterase inhibitor RO 20- 1724 was added to each well, and the plates were incubated for 60 min at room temperature. A total of 23nl of compound or positive control (30nM TSH or 10μM forskolin for TSHR- expressing or parental cells, respectively) in DMSO was added to each well using a pintool (Kalypsys, San Diego, CA), and the plates were further incubated for 30 min at room temperature. Plates were measured with an excitation of 535/20 nm, an emission of 590/20 nm, a gain of 150, and 5 flashes per well on an Envision plate reader (PerkinElmer, Boston, MA) (<a class="figpopup" href="/books/NBK56234/table/ml109.t2/?report=objectonly" target="object" rid-figpopup="figml109t2" rid-ob="figobml109t2">Table 2</a>).</p><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml109t1"><a href="/books/NBK56234/table/ml109.t1/?report=objectonly" target="object" title="Table 1" class="img_link icnblk_img figpopup" rid-figpopup="figml109t1" rid-ob="figobml109t1"><img class="small-thumb" src="/books/NBK56234/table/ml109.t1/?report=thumb" src-large="/books/NBK56234/table/ml109.t1/?report=previmg" alt="Table 1. Reagents and resources for the TSH receptor screen." /></a><div class="icnblk_cntnt"><h4 id="ml109.t1"><a href="/books/NBK56234/table/ml109.t1/?report=objectonly" target="object" rid-ob="figobml109t1">Table 1</a></h4><p class="float-caption no_bottom_margin">Reagents and resources for the TSH receptor screen. </p></div></div><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml109t2"><a href="/books/NBK56234/table/ml109.t2/?report=objectonly" target="object" title="Table 2" class="img_link icnblk_img figpopup" rid-figpopup="figml109t2" rid-ob="figobml109t2"><img class="small-thumb" src="/books/NBK56234/table/ml109.t2/?report=thumb" src-large="/books/NBK56234/table/ml109.t2/?report=previmg" alt="Table 2. Assay protocol for the TSHR and parental cell lines in 1536 well format." /></a><div class="icnblk_cntnt"><h4 id="ml109.t2"><a href="/books/NBK56234/table/ml109.t2/?report=objectonly" target="object" rid-ob="figobml109t2">Table 2</a></h4><p class="float-caption no_bottom_margin">Assay protocol for the TSHR and parental cell lines in 1536 well format. </p></div></div></div><div id="ml109.s10"><h5>Center Summary of Results</h5><p>Both TSHR and parental (lacking TSHR) lines were screened across 463 compound plates (for a total of 926 plates). The average signal-to-basal ratio for the entire screen was 2.01, the CV was 12.1%, and the Ẓ′ value was 0.4. Plates with a Ẓ′ factor less than 0.2 (approximately 5% of the total plates) were rescreened. The relatively low Ẓ′ is likely due to the use of HEK293 cells, which typically do not adhere well after treatment with membrane potential dyes.</p></div></div><div id="ml109.s11"><h4>ii. Counterscreen in ACTOne parental cell line for TSHR agonists. [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/398" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID:398</a>; Primary DR]</h4><div id="ml109.s12"><h5>Assay Description</h5><p>All 72,030 compounds were tested in qHTS format in the parental cell line lacking the TSH receptor.</p></div><div id="ml109.s13"><h5>Assay Protocol</h5><p>The assay protocol and reagents are identical to: qHTS Assay for TSHR agonists Primary screen [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/926" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID: 926</a>; Primary] but screened in the parental HEK 293 cell line, which lacks the TSH receptor.</p></div><div id="ml109.s14"><h5>Center Summary of Results</h5><p>Prioritized compounds had no activity in the parental cell line.</p></div></div><div id="ml109.s15"><h4>iii. Confirmation Concentration-Response Assay for TSHR agonists in ACTOne TSHR cells [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/939" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID:939</a>; Confirmatory DR]</h4><div id="ml109.s16"><h5>Assay Description</h5><p>To confirm activity in the original assay, select samples active in the primary screen were obtained in DMSO solution from the MLSMR and/or as powders from compound vendors.</p></div><div id="ml109.s17"><h5>Assay Protocol</h5><p>The assay protocol and reagents are identical to: qHTS Assay for TSHR agonists Primary screen [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/926" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID: 926</a>; Primary]</p></div><div id="ml109.s18"><h5>Center Summary of Results</h5><p>Prioritized compounds had a high confirmation rate.</p></div></div><div id="ml109.s19"><h4>iv. Counterscreen in ACTOne parental cell line for TSHR agonists. [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/953" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID:953</a>; Confirmatory DR]</h4><div id="ml109.s20"><h5>Assay Description</h5><p>To confirm activity in the original assay, select samples active in the primary screen were obtained in DMSO solution from the MLSMR and/or as powders from compound vendors.</p></div><div id="ml109.s21"><h5>Assay Protocol</h5><p>The assay protocol and reagents are identical to: qHTS Assay for TSHR agonists Primary screen [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/926" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID: 926</a>; Primary], but screened in the parental HEK 293 cell line, which lacks the TSH receptor.</p></div><div id="ml109.s22"><h5>Center Summary of Results</h5><p>Prioritized compounds were inactive in the parental cell line.</p></div></div><div id="ml109.s23"><h4>v. Counterscreen in TSHR cell line using Homogeneous Time Resolved Fluorescence (HTRF) cAMP assay in TSHR cells [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/933" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID:933</a>; Confirmatory DR]</h4><div id="ml109.s24"><h5>Assay Description</h5><p>Recently, a time resolved fluorescence resonance energy transfer (TR-FRET) based cAMP assay has been available for screens from Cisbio (12,13). It uses an anti-cAMP antibody labeled with Eu<sup>3+</sup>, which binds to a d2-dye labeled cAMP tracer, resulting in TR-FRET. The cAMP from the cell lysate can displace the d2 labeled cAMP tracer that interrupts the TR-FRET (<a class="figpopup" href="/books/NBK56234/figure/ml109.f3/?report=objectonly" target="object" rid-figpopup="figml109f3" rid-ob="figobml109f3">Fig. 3</a>). Because this cAMP assay uses a different detection system, it can help to further eliminate the false positive compounds.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml109f3" co-legend-rid="figlgndml109f3"><a href="/books/NBK56234/figure/ml109.f3/?report=objectonly" target="object" title="Figure 3" class="img_link icnblk_img figpopup" rid-figpopup="figml109f3" rid-ob="figobml109f3"><img class="small-thumb" src="/books/NBK56234/bin/ml109f3.gif" src-large="/books/NBK56234/bin/ml109f3.jpg" alt="Figure 3. Schematic of cAMP competitive immunoassay." /></a><div class="icnblk_cntnt" id="figlgndml109f3"><h4 id="ml109.f3"><a href="/books/NBK56234/figure/ml109.f3/?report=objectonly" target="object" rid-ob="figobml109f3">Figure 3</a></h4><p class="float-caption no_bottom_margin">Schematic of cAMP competitive immunoassay. </p></div></div></div><div id="ml109.s25"><h5>Assay Protocol</h5><p>The 95 selected compounds were tested for TSHR agonism in the TR-FRET based cAMP assay, which measures cAMP concentrations in cell lysates. The readout in the HTRF cAMP assay is distinct from that in the membrane potential dye measurement assay used in the qHTS, allowing us to further eliminate assay-related false positive compounds. Of the 95 compounds, 49 showed TSH agonist activity in the TSHR cell line without the activities in the parental cell line. Taken together, these results indicate that these 49 compounds are true small molecule TSHR agonists.</p></div><div id="ml109.s26"><h5>Assay Protocol</h5><p>HEK 293 cell lines stably expressing the TSH, FSH, or LH receptors were used as the counter-screens for these compounds in the TR-FRET cAMP assay format. These lines did not contain the CNG and were therefore incapable of being used in the primary assay format. An assay protocol was summarized in <a class="figpopup" href="/books/NBK56234/table/ml109.t2/?report=objectonly" target="object" rid-figpopup="figml109t2" rid-ob="figobml109t2">Table 2</a>. Cells were plated in white, solid bottom 1536 well tissue culture treated plates and assayed as described in <a class="figpopup" href="/books/NBK56234/table/ml109.t3/?report=objectonly" target="object" rid-figpopup="figml109t3" rid-ob="figobml109t3">Table 3</a>.</p><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml109t3"><a href="/books/NBK56234/table/ml109.t3/?report=objectonly" target="object" title="Table 3" class="img_link icnblk_img figpopup" rid-figpopup="figml109t3" rid-ob="figobml109t3"><img class="small-thumb" src="/books/NBK56234/table/ml109.t3/?report=thumb" src-large="/books/NBK56234/table/ml109.t3/?report=previmg" alt="Table 3. HTRF assay in TSH, LH, and FSH cell lines." /></a><div class="icnblk_cntnt"><h4 id="ml109.t3"><a href="/books/NBK56234/table/ml109.t3/?report=objectonly" target="object" rid-ob="figobml109t3">Table 3</a></h4><p class="float-caption no_bottom_margin">HTRF assay in TSH, LH, and FSH cell lines. </p></div></div><div id="ml109.t4" class="table"><h3><span class="label">Table 4</span><span class="title">Reagents for HTRF counterscreen</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK56234/table/ml109.t4/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml109.t4_lrgtbl__"><table class="no_top_margin"><thead><tr><th id="hd_h_ml109.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Reagents</th><th id="hd_h_ml109.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Resources</th><th id="hd_h_ml109.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Catalog Number</th></tr></thead><tbody><tr><td headers="hd_h_ml109.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">HTRF cAMP <i>dynamic</i> 2 kit</td><td headers="hd_h_ml109.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Cisbio</td><td headers="hd_h_ml109.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">62AM4PEJ</td></tr><tr><td headers="hd_h_ml109.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">DMEM Media</td><td headers="hd_h_ml109.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Invitrogen</td><td headers="hd_h_ml109.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">11765</td></tr><tr><td headers="hd_h_ml109.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Geneticin</td><td headers="hd_h_ml109.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Invitrogen</td><td headers="hd_h_ml109.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">10131027</td></tr><tr><td headers="hd_h_ml109.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Fetal Bovine Serum</td><td headers="hd_h_ml109.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Hyclone</td><td headers="hd_h_ml109.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">SH30071.03</td></tr><tr><td headers="hd_h_ml109.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Penicillin-Streptomycin</td><td headers="hd_h_ml109.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Invitrogen</td><td headers="hd_h_ml109.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">15140</td></tr><tr><td headers="hd_h_ml109.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">RO 20-1724</td><td headers="hd_h_ml109.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Sigma</td><td headers="hd_h_ml109.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">B8279</td></tr></tbody></table></div></div></div></div><div id="ml109.s27"><h4>vi. Counterscreen in TSHR cell line using Homogeneous Time Resolved Fluorescence (HTRF) cAMP assay in LH cells [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1402" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID:1402</a>; Confirmatory DR]</h4><div id="ml109.s28"><h5>Assay Description</h5><p>To confirm activity in the original assay, select samples active in the primary screen were obtained in DMSO solution from the MLSMR and/or as powders from compound vendors.</p></div><div id="ml109.s29"><h5>Assay Protocol</h5><p>The assay protocol and reagents are identical to the HTRF assay run in TSH cells qHTS Assay [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/933" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID: 933</a>; Primary].</p></div><div id="ml109.s30"><h5>Center Summary of Results</h5><p>Prioritized compounds had a high confirmation rate.</p></div></div><div id="ml109.s31"><h4>vii. Counterscreen in TSHR cell line using Homogeneous Time Resolved Fluorescence (HTRF) cAMP assay in FSH cells [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1403" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID:1403</a>; Confirmatory DR]</h4><div id="ml109.s32"><h5>Assay Description</h5><p>To confirm activity in the original assay, select samples active in the primary screen were obtained in DMSO solution from the MLSMR and/or as powders from compound vendors.</p></div><div id="ml109.s33"><h5>Assay Protocol</h5><p>The assay protocol and reagents are identical to the HTRF assay run in TSH cells qHTS Assay [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/933" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID: 933</a>; Primary].</p></div><div id="ml109.s34"><h5>Center Summary of Results</h5><p>Prioritized compounds had a high confirmation rate.</p></div></div><div id="ml109.s35"><h4>viii. Secondary Concentration-Response Assay for Agonists of the Thyroid Stimulating Hormone Receptor: ELISA Activity Detection [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2104" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID:2104</a>; Confirmatory DR]</h4><div id="ml109.s36"><h5>Assay Description</h5><p>To confirm activity in the original assay, select samples active in the confirmatory assays were resynthesized and assayed by the low-throughput ELISA assay format.</p></div><div id="ml109.s37"><h5>Assay Protocol</h5><p>The assay protocol and reagents are described in detail in reference <a class="bk_pop" href="#ml109.r11">11</a> (1) [<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2104" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID: 2104</a>; Secondary]. Transiently transfected HEK-EM293 cells or cells stably expressing TSHR, LHCGR, or FSHR were seeded into 96-well plates at a density of 50,000 cells/well in DMEM containing 10% FBS. Cells were cultured for 24 h before incubation for 1 h in serum-free DMEM containing 1mM 3-isobutyl-1-methylxanthine (Sigma) and TSH, LH, FSH, or small-molecule ligands in a humidified 5% CO2 incubator at 37°C. Following aspiration of the media, cells were lysed using lysis buffer of the cAMP-Screen Direct system (Applied Biosystems). The cAMP content of the cell lysate was determined using the method described in the manufacturer’s protocol. Reagents are listed in <a class="figpopup" href="/books/NBK56234/table/ml109.t5/?report=objectonly" target="object" rid-figpopup="figml109t5" rid-ob="figobml109t5">Table 5</a>.</p><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml109t5"><a href="/books/NBK56234/table/ml109.t5/?report=objectonly" target="object" title="Table 5" class="img_link icnblk_img figpopup" rid-figpopup="figml109t5" rid-ob="figobml109t5"><img class="small-thumb" src="/books/NBK56234/table/ml109.t5/?report=thumb" src-large="/books/NBK56234/table/ml109.t5/?report=previmg" alt="Table 5. Reagents for ELISA secondary screen." /></a><div class="icnblk_cntnt"><h4 id="ml109.t5"><a href="/books/NBK56234/table/ml109.t5/?report=objectonly" target="object" rid-ob="figobml109t5">Table 5</a></h4><p class="float-caption no_bottom_margin">Reagents for ELISA secondary screen. </p></div></div></div><div id="ml109.s38"><h5>Center Summary of Results</h5><p>Compounds were much more potent using ELISA assay than HTRF format assay.</p></div></div></div><div id="ml109.s39"><h3>2.2. Probe Chemical Characterization</h3><div id="ml109.s40"><h4>Synthesis of probe molecule</h4><p>The synthesis of the probe molecule commenced with amide formation between commercially available acid <b>1a</b> and benzyl amine to afford <b>3a</b> and subsequent demethylation to <b>3<sup>1</sup>a</b> (<a class="figpopup" href="/books/NBK56234/figure/ml109.f8/?report=objectonly" target="object" rid-figpopup="figml109f8" rid-ob="figobml109f8">Scheme 1</a>). The second component of the aminal was synthesized via ether formation between benzyl chloride <b>4a</b> and p-acetamide phenol to afford <b>5a</b>. The ytterbium catalyzed condensation between <b>3<sup>1</sup>a</b> and <b>5a</b> afforded the probe molecule <b>6a</b>.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml109f8" co-legend-rid="figlgndml109f8"><a href="/books/NBK56234/figure/ml109.f8/?report=objectonly" target="object" title="Scheme 1" class="img_link icnblk_img figpopup" rid-figpopup="figml109f8" rid-ob="figobml109f8"><img class="small-thumb" src="/books/NBK56234/bin/ml109f8.gif" src-large="/books/NBK56234/bin/ml109f8.jpg" alt="Scheme 1. Synthesis of probe." /></a><div class="icnblk_cntnt" id="figlgndml109f8"><h4 id="ml109.f8"><a href="/books/NBK56234/figure/ml109.f8/?report=objectonly" target="object" rid-ob="figobml109f8">Scheme 1</a></h4><p class="float-caption no_bottom_margin">Synthesis of probe. </p></div></div><p>MLS002576689 (CID: 25246343/<a href="/pcsubstance/?term=ML109[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML109</a>):</p><p><sup>1</sup>H NMR (400 MHz, DMSO-<i>d</i><sub>6</sub>) δppm 2.00 (s, 3 H), 3.78 (d, J=15.5Hz, 1H), 3.80 (s, 3H), 4.96 (s, 2 H), 5.15 (d, <i>J</i>=15.3 Hz, 1 H), 5.73 (d, <i>J</i>=1.96 Hz, 1 H), 6.05–6.08 (m, 2 H), 6.86–6.90 (m, 2 H), 7.01 – 7.11 (m, 2 H), 7.19 – 7.36 (m, 6 H), 7.41–7.48 (m, 4 H) 9.77 (s, 1 H), 12.26 (s, 1 H); HPLC: t<sub>R</sub> = 6.19 min, UV<sub>254</sub> = 96%; HRMS (ESI): <i>m/z</i> calcd for C<sub>31</sub>H<sub>29</sub>N<sub>3</sub>O<sub>5</sub> [M+1]+ 524.2185, found 524.2184. Solubility (PBS, pH = 7.4, 23 °C): 2μM.</p><div id="ml109.t6" class="table"><h3><span class="label">Table 6</span><span class="title">Compounds submitted to the MLSMR</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK56234/table/ml109.t6/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml109.t6_lrgtbl__"><table class="no_top_margin"><tbody><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:bottom;">MLS002576689</td><td rowspan="1" colspan="1" style="text-align:center;vertical-align:bottom;">Probe</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:bottom;">MLS003221407</td><td rowspan="1" colspan="1" style="text-align:center;vertical-align:bottom;">Analogue</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:bottom;">MLS003221408</td><td rowspan="1" colspan="1" style="text-align:center;vertical-align:bottom;">Analogue</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:bottom;">MLS003221409</td><td rowspan="1" colspan="1" style="text-align:center;vertical-align:bottom;">Analogue</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:bottom;">MLS003221410</td><td rowspan="1" colspan="1" style="text-align:center;vertical-align:bottom;">Analogue</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:bottom;">MLS003221411</td><td rowspan="1" colspan="1" style="text-align:center;vertical-align:bottom;">Analogue</td></tr></tbody></table></div></div></div></div><div id="ml109.s41"><h3>2.3. Probe Preparation</h3><div id="ml109.s42"><h4>Synthesis of 2,3-dihydroquinazolin-4(1H)-one (<b>6</b>) and quinazolin-4(1H)one (<b>7</b>) analogs</h4><p>As depicted in <a class="figpopup" href="/books/NBK56234/figure/ml109.f9/?report=objectonly" target="object" rid-figpopup="figml109f9" rid-ob="figobml109f9">scheme <b>2</b></a>, 2-aminobenzamides <b>3</b> were prepared by either amide couplings of 2-aminobenzoic acids <b>1</b> with different amines or reactions of isatoic anhydrides <b>2</b> with amines. Reactions of benzyl chlorides <b>4</b> with different phenols under microwave irradiation generated aldehydes <b>5</b>. Condensations of aldehydes <b>5</b> with 2-aminobenzamides <b>3</b> yielded 2,3-dihydroquinazolin-4-ones <b>6</b>. The 2,3-dihydroquinazolin-4-ones <b>6</b> were rapidly oxidized by DDQ at room temperature to produce quinazolin-4-ones <b>7</b>.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml109f9" co-legend-rid="figlgndml109f9"><a href="/books/NBK56234/figure/ml109.f9/?report=objectonly" target="object" title="Scheme 2" class="img_link icnblk_img figpopup" rid-figpopup="figml109f9" rid-ob="figobml109f9"><img class="small-thumb" src="/books/NBK56234/bin/ml109f9.gif" src-large="/books/NBK56234/bin/ml109f9.jpg" alt="Scheme 2. Analog synthesis." /></a><div class="icnblk_cntnt" id="figlgndml109f9"><h4 id="ml109.f9"><a href="/books/NBK56234/figure/ml109.f9/?report=objectonly" target="object" rid-ob="figobml109f9">Scheme 2</a></h4><p class="float-caption no_bottom_margin">Analog synthesis. Reagents and conditions: (i) DMC, DIPEA, r.t. 12 h; (ii) ACN, r.t.-50 °C. (iii) K<sub>2</sub>CO<sub>3,</sub> DMA, microwave heating, 150 °C, 10 min; (iv) Yb(OTf)<sub>3</sub>, DMA, microwave heating, 200 °C, 10 min; (v) DDQ, r.t. 1 h </p></div></div></div><div id="ml109.s43"><h4>General procedure for the synthesis of 2-aminobenzamides from isatoic anhydride</h4><p>Amines (1.05 mmol, 1.05 equiv) at room temperature were added to a solution of isatoic anhydride (0.163 g, 1.0 mmol, 1.0 equiv) in 10 mL of anhydrous acetonitrile. The resulting mixture was stirred at room temperature for 2 hours and heated at 50°C for 4 hours. Then, it was concentrated <i>in vacuo</i> to yield the products as solids in 90–99% yields.</p></div><div id="ml109.s44"><h4>2-Amino-N-benzylbenzamide</h4><div id="ml109.fu2" class="figure"><div class="graphic"><img src="/books/NBK56234/bin/ml109fu2.jpg" alt="Image ml109fu2" /></div></div><p><sup>1</sup>H NMR (400 MHz, CHLOROFORM-<i>d</i>) δ 4.61 (s, 1 H), 4.63 (s, 1 H), 5.58 (br. s., 2 H), 6.33 (br. s., 1 H), 6.62–6.66 (m, 1 H), 6.69–6.71 (m, 1 H), 7.19–7.25 (m, 1 H), 7.28 – 7.43 (m, 6 H); LCMS: (electrospray +ve), <i>m</i>/<i>z</i> 227.1 (MH)<sup>+</sup>; HPLC: t<sub>R</sub> = 4.38 min, UV<sub>254</sub> = 96%.</p></div><div id="ml109.s45"><h4>2-Amino-N-(furan-2-ylmethyl)benzamide</h4><div id="ml109.fu3" class="figure"><div class="graphic"><img src="/books/NBK56234/bin/ml109fu3.jpg" alt="Image ml109fu3" /></div></div><p><sup>1</sup>H NMR (400 MHz, CHLOROFORM-<i>d</i>) δ 4.60 (s, 1 H), 4.61 (s, 1 H), 5.57 (br. s., 2 H), 6.24 – 6.42 (m, 3 H), 6.59 – 6.74 (m, 2 H), 7.16 – 7.25 (m, 1 H), 7.33–7.39 (m, 2 H); LCMS: (electrospray +ve), <i>m</i>/<i>z</i> 217.1 (MH)<sup>+</sup>; HPLC: t<sub>R</sub> = 3.77 min, UV<sub>254</sub> = 98%.</p></div><div id="ml109.s46"><h4>2-Amino-N-benzyl-6-methoxybenzamide</h4><div id="ml109.fu4" class="figure"><div class="graphic"><img src="/books/NBK56234/bin/ml109fu4.jpg" alt="Image ml109fu4" /></div></div><p>2-chloro-1,3-dimethylimidazolinium chloride (1.099g, 6.5mmol, 1.3equiv) at room temperature was added to a solution of 2-amino-6-methoxybenzoic acid (0.841g, 5.0mmol, 1.0equiv), benzylamine (0.643g, 6.0mmol, 1.2equiv), and diisopropylethylamine (1.935g, 15.0mmol, 3.0equiv) in 50ml of dichloromethane. The mixture was stirred at room temperature for 6 hours, poured into water, and extracted with dichloromethane. The organic solution was successively washed with aqueous saturated NaHCO<sub>3</sub> and water. The organic layer was dried over MgSO<sub>4</sub> and the solvent was removed by rotary evaporator. The residue was purified by column chromatography (silica gel, 2% 2.0 M ammonia MeOH solution in CH<sub>2</sub>Cl<sub>2</sub>) to give 2-Amino-N-benzyl-6-methoxybenzamide (0.593g, 46%) as a solid. <sup>1</sup>H NMR (400 MHz, CHLOROFORM-<i>d</i>) δ 3.81 (s, 3 H), 4.62 (s, 1 H), 4.63 (s, 1 H), 6.07 (vb.s, 2 H), 6.19 (d, <i>J</i>=8.2 Hz, 1 H), 6.32 (d, <i>J</i>=8.2 Hz, 1 H), 7.07 (t, <i>J</i>=8.2 Hz, 1 H), 7.14 – 7.54 (m, 5 H), 8.05 (br. s., 1 H); HPLC: t<sub>R</sub> = 4.72 min, UV<sub>254</sub> = 99%; HRMS (ESI): <i>m/z</i> calcd for C<sub>15</sub>H<sub>16</sub>N<sub>2</sub>O<sub>2</sub> [M+1]+ 257.1296, found 257.1294.</p></div><div id="ml109.s47"><h4>N-(4-(5-(3-(Furan-2-ylmethyl)-4-oxo-1,2,3,4-tetrahydroquinazolin-2-yl)-2-methoxybenzyloxy)phenyl)acetamide</h4><div id="ml109.fu5" class="figure"><div class="graphic"><img src="/books/NBK56234/bin/ml109fu5.jpg" alt="Image ml109fu5" /></div></div><p>K<sub>2</sub>CO<sub>3</sub> (207mg, 1.5mmol, 5equiv) was added to a solution of 3-(chloromethyl)-4-methoxybenzaldehyde (55.2mg, 300μmol, 1.0equiv) and 4-acetamidophenol (54.4mg, 360μmol, 1.2equiv) in 1.5ml of anhydrous DMA. The mixture was heated in a microwave for 10 min at 150°C. After filtering off the solid, 2-amino-<i>N</i>-(furan-2-ylmethyl)benzamide (77.8mg, 360μmol, 1.2equiv) and Ytterbium trifluoromethanesulfonate (93mg, 150μmol, 0.5equiv) were added to the clear solution. The resulting mixture was heated in a microwave for 10 min at 200 °C. The product was isolated via preparative HPLC purification and solvent was removed by a GeneVac to give <i>N</i>-(4-(5-(3-(furan-2-ylmethyl)-4-oxo-1,2,3,4-tetrahydroquinazolin-2-yl)-2-methoxybenzyloxy)phenyl)acetamide (47.8mg, 32%) as a solid. <sup>1</sup>H NMR (400 MHz, CDCl<sub>3</sub>) δ 2.11 (s, 3 H), 3.72 (d, <i>J</i>=15.6 Hz, 1 H), 3.83 (s, 3 H), 4.25 (v.b.s, 1H), 5.01 (s, 2 H), 5.26 (d, <i>J</i>=15.6 Hz, 1 H), 5.71 (s, 1 H), 6.03 – 6.29 (m, 2 H), 6.48 (d, <i>J</i>=7.8 Hz, 1 H), 6.83 (dd, <i>J</i>=18.2, 8.8 Hz, 3 H), 7.10 – 7.61 (m, 7 H), 7.92 (d, <i>J</i>=7.0 Hz, 1 H); HPLC: t<sub>R</sub> = 5.40 min, UV<sub>254</sub> = 91%; HRMS (ESI): <i>m/z</i> calcd for C<sub>29</sub>H<sub>27</sub>N<sub>3</sub>O<sub>5</sub> [M+1]+ 498.2029, found 498.2025.</p></div><div id="ml109.s48"><h4>3-(Furan-2-ylmethyl)-2-(4-methoxy-3-(phenoxymethyl)phenyl)-2,3-dihydroquinazolin-4(1H)-one</h4><div id="ml109.fu6" class="figure"><div class="graphic"><img src="/books/NBK56234/bin/ml109fu6.jpg" alt="Image ml109fu6" /></div></div><p>K<sub>2</sub>CO<sub>3</sub> (319mg, 2.3mmol, 5equiv) was added to a solution of 3-(chloromethyl)-4-methoxybenzaldehyde (85mg, 462μmol, 1.0equiv) and phenol (52mg, 554μmol, 1.2equiv) in 2.0ml of anhydrous DMA. The mixture was heated in a microwave for 10 min at 150°C. After filtering off the solid, 2-amino-<i>N</i>-(furan-2-ylmethyl)benzamide (100mg, 462μmol, 1.0equiv) and Ytterbium trifluoromethanesulfonate (286 mg, 231μmol, 0.5equiv) were added to the clear solution. The resulting mixture was heated in a microwave for 10 min at 200°C. The product was isolated via preparative HPLC purification and solvent was removed via reduced pressure lyophilization to give 3-(furan-2-ylmethyl)-2-(4-methoxy-3-(phenoxymethyl)phenyl)-2,3-dihydroquinazolin-4(1H)-one (41.5mg, 20%) as a white solid after triturating with diethyl ether. <sup>1</sup>H NMR (400 MHz, CDCl<sub>3</sub>) δ 3.74 (d, <i>J</i>=15.6 Hz, 1 H), 3.86 (s, 3 H), 4.40 (v.b.s, 1H), 5.08 (s, 2 H), 5.31 (d, <i>J</i>=16.0 Hz, 1 H), 5.74 (s, 1 H), 6.13 (dd, <i>J</i>=3.3, 0.6 Hz, 1 H), 6.25 (dd, <i>J</i>=3.1, 1.9 Hz, 1 H), 6.48 (dd, <i>J</i>=8.0, 0.4 Hz, 1 H), 6.81–6.85 (m, 2H), 6.94–6.98 (m, 3H), 7.21 – 7.32 (m, 5 H), 7.48 (d, <i>J</i>=2.4 Hz, 1 H), 7.98 (dd, J=7.8, 1.4Hz, 1H); HPLC: t<sub>R</sub> = 6.42 min, UV<sub>254</sub> = 92%; HRMS (ESI): <i>m/z</i> calcd for C<sub>27</sub>H<sub>24</sub>N<sub>2</sub>O<sub>4</sub> [M+1]+ 441.1820, found 441.1821.</p></div><div id="ml109.s49"><h4>N-(4-(5-(3-(Furan-2-ylmethyl)-4-oxo-3,4-dihydroquinazolin-2-yl)-2-methoxybenzyloxy)phenyl)acetamide</h4><div id="ml109.fu7" class="figure"><div class="graphic"><img src="/books/NBK56234/bin/ml109fu7.jpg" alt="Image ml109fu7" /></div></div><p><sup>1</sup>H NMR (400 MHz, CHLOROFORM-<i>d</i>) δ 2.15 (s, 3 H), 3.95 (s, 3 H), 5.12 (s, 2 H), 5.19 (s, 2 H), 6.12 (d, <i>J</i>=2.7 Hz, 1 H), 6.22 – 6.28 (m, 1 H), 6.93 (d, <i>J</i>=9.0 Hz, 2 H), 7.00 (d, <i>J</i>=8.6 Hz, 1 H), 7.12 (br. s., 1 H), 7.24 (s, 1 H), 7.39 (d, <i>J</i>=9.0 Hz, 2 H), 7.46 – 7.56 (m, 2 H), 7.64 – 7.82 (m, 3 H), 8.33 (d, <i>J</i>=8.2 Hz, 1 H); HPLC: t<sub>R</sub> = 5.49 min, UV<sub>254</sub> = 95%; HRMS (ESI): <i>m/z</i> calcd for C<sub>29</sub>H<sub>25</sub>N<sub>3</sub>O<sub>5</sub> [M+1]+ 496.1872, found 496.1873.</p></div><div id="ml109.s50"><h4>N-(4-(5-(3-Benzyl-4-oxo-3,4-dihydroquinazolin-2-yl)-2-methoxybenzyloxy)phenyl)acetamide</h4><div id="ml109.fu8" class="figure"><div class="graphic"><img src="/books/NBK56234/bin/ml109fu8.jpg" alt="Image ml109fu8" /></div></div><p><sup>1</sup>H NMR (400 MHz, CHLOROFORM-<i>d</i>) δ 2.13 (s, 3 H), 3.91 (s, 3 H), 5.04 (s, 2 H), 5.26 (s, 2 H), 6.81 – 7.04 (m, 5 H), 7.11 – 7.26 (m, 4 H), 7.38 (d, <i>J</i>=9.0 Hz, 2 H), 7.46 – 7.58 (m, 2 H), 7.71 – 7.83 (m, 2 H), 8.36 (d, <i>J</i>=7.8 Hz, 1 H); HPLC: t<sub>R</sub> = 5.73 min, UV<sub>254</sub> = 98%; HRMS (ESI): <i>m/z</i> calcd for C<sub>31</sub>H<sub>27</sub>N<sub>3</sub>O<sub>4</sub> [M+1]<sup>+</sup> 506.2086, found 506.2082.</p></div><div id="ml109.s51"><h4>Synthesis of the probe</h4><div id="ml109.s52"><h5>2-Amino-N-benzyl-6-hydroxybenzamide (<b>3a</b>)</h5><div id="ml109.fu9" class="figure"><div class="graphic"><img src="/books/NBK56234/bin/ml109fu9.jpg" alt="Image ml109fu9" /></div></div><p>1-dodecanethiol (0.360g, 1.78 mmol, 2.0equiv) was added to a solution of 2-amino-N-benzyl-6-methoxybenzamide (0.228g, 0.89mmol, 1.0equiv) in 3 mL of anhydrous DMF, followed by adding NaOMe (0.385g of 25% solution in MeOH, 1.78, 2.0equiv). Five copies of this reaction were prepared, and the reaction mixtures were heated in a microwave at 150 °C for 10 min. The reaction mixtures were combined and passed through a silica gel plug, which was washed with an ethyl acetate-methanol mixture (1:1). The solvents were removed and the residue was purified by column chromatography (silica gel, eluent gradient 2:98 ethyl acetate-CH<sub>2</sub>Cl<sub>2</sub> to 1:1 ethyl acetate-CH<sub>2</sub>Cl<sub>2</sub>). The desired 2-Amino-N-benzyl-6-hydroxybenzamide (0.780 g, 72%) was isolated as an off-white solid after triturating with diethyl ether/Hexanes. <sup>1</sup>H NMR (400 MHz, CHLOROFORM-<i>d</i>) δ 3.55 – 4.20 (vb.s., 2 H), 4.60 (s, 1 H), 4.61 (s, 1 H), 6.25 (d, <i>J</i>=7.8 Hz, 1 H), 6.50 (d, <i>J</i>=8.2 Hz, 1 H), 7.09 (t, <i>J</i>=8.0 Hz, 1 H), 7.16 – 7.52 (m, 5 H), 8.58 (br. s., 1 H),12.05 (br. s., 1 H); HPLC: t<sub>R</sub> = 3.97 min, UV<sub>254</sub> = 98%; HRMS (ESI): <i>m/z</i> calcd for C<sub>14</sub>H<sub>14</sub>N<sub>2</sub>O<sub>2</sub> [M+1]+ 243.1140, found 243.1134.</p></div><div id="ml109.s53"><h5>N-(4-(5-(3-Benzyl-5-hydroxy-4-oxo-1,2,3,4-tetrahydroquinazolin-2-yl)-2-methoxybenzyloxy)phenyl)acetamide (<b>6a</b>)</h5><div id="ml109.fu10" class="figure"><div class="graphic"><img src="/books/NBK56234/bin/ml109fu10.jpg" alt="Image ml109fu10" /></div></div><p>K<sub>2</sub>CO<sub>3</sub> (690 mg, 5.0mmol, 5equiv) was added to a solution of 3-(chloromethyl)-4-methoxybenzaldehyde (184.6 mg, 1.00 mmol, 1.0equiv) and N-(4-hydroxyphenyl)acetamide (181mg, 1.20mmol, 1.2equiv) in 5.0ml of anhydrous DMA. The mixture was heated in a microwave at 150°C for 10 min. The solid was filtered and the clear solution was added to a microwave tube containing 2-amino-N-benzyl-6-hydroxybenzamide (264mg, 1.09mmol, 1.1equiv). After adding Ytterbium trifluoromethanesulfonate (310mg, 0.5mmol, 0.5equiv), the mixture was heated in a microwave at 200°C for 10 min. The reaction mixture was purified by HPLC. Pure fractions were dried down under vacuum on a lyophilizer. The desired product (132.3mg, 28%) was obtained as a solid after triturating with diethyl ether. <sup>1</sup>H NMR (400 MHz, DMSO-<i>d</i><sub>6</sub>) δppm 2.00 (s, 3 H), 3.78 (d, J=15.5Hz, 1H), 3.80 (s, 3H), 4.96 (s, 2 H), 5.15 (d, <i>J</i>=15.3 Hz, 1 H), 5.73 (d, <i>J</i>=1.96 Hz, 1 H), 6.05–6.08 (m, 2 H), 6.86–6.90 (m, 2 H), 7.01 – 7.11 (m, 2 H), 7.19 – 7.36 (m, 6 H), 7.41–7.48 (m, 4 H) 9.77 (s, 1 H), 12.26 (s, 1 H); HPLC: t<sub>R</sub> = 6.19 min, UV<sub>254</sub> = 96%; HRMS (ESI): <i>m/z</i> calcd for C<sub>31</sub>H<sub>29</sub>N<sub>3</sub>O<sub>5</sub> [M+1]+ 524.2185, found 524.2184.</p></div></div></div></div><div id="ml109.s54"><h2 id="_ml109_s54_">3. Results</h2><div id="ml109.s55"><h3>3.1. Summary of Screening Results</h3><p>72,030 compounds were screened in the primary assay<a class="bk_pop" href="#ml109.r14">14</a>. Some active compounds were chosen for confirmation according to the scheme shown in <a class="figpopup" href="/books/NBK56234/figure/ml109.f4/?report=objectonly" target="object" rid-figpopup="figml109f4" rid-ob="figobml109f4">Fig. 4</a>.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml109f4" co-legend-rid="figlgndml109f4"><a href="/books/NBK56234/figure/ml109.f4/?report=objectonly" target="object" title="Figure 4" class="img_link icnblk_img figpopup" rid-figpopup="figml109f4" rid-ob="figobml109f4"><img class="small-thumb" src="/books/NBK56234/bin/ml109f4.gif" src-large="/books/NBK56234/bin/ml109f4.jpg" alt="Figure 4. High-throughput screen." /></a><div class="icnblk_cntnt" id="figlgndml109f4"><h4 id="ml109.f4"><a href="/books/NBK56234/figure/ml109.f4/?report=objectonly" target="object" rid-ob="figobml109f4">Figure 4</a></h4><p class="float-caption no_bottom_margin">High-throughput screen. </p></div></div></div><div id="ml109.s56"><h3>3.2. Dose Response Curves for Probe</h3><div id="ml109.f5" class="figure bk_fig"><div class="graphic"><img src="/books/NBK56234/bin/ml109f5.jpg" alt="Figure 5. Activity of MLS002576689 in the ELISA cAMP assay." /></div><h3><span class="label">Figure 5</span><span class="title">Activity of MLS002576689 in the ELISA cAMP assay</span></h3></div></div><div id="ml109.s57"><h3>3.3. Scaffold/Moiety Chemical Liabilities</h3><p>A potential site for undesired chemical reactivity in the probe would be the aminal, which could undergo oxidation or hydrolysis to the amine and aldehyde. We found experimentally that the rate of oxidation is dependent on the aryl ring substitution. The 5-hydroxy substituted probe molecule is particularly resistant to oxidation. It was dissolved in acetonitrile and oxygen was bubbled through the solution, but no oxidation was witnessed, even after several hours. In fact, the probe compound only underwent oxidation when treated with MnO<sub>2</sub>. The other chemical concern for this scaffold would be hydrolysis. The hydrolytic stability of this scaffold was monitored at neutral and basic pH and the t<sub>1/2</sub> was 16 h. The stability, not surprisingly, suffered at low pH and the t<sub>1/2</sub> was 3 h<a class="bk_pop" href="#ml109.r1">1</a>.</p></div><div id="ml109.s58"><h3>3.4. SAR Tables</h3><div id="ml109.t7" class="table"><h3><span class="label">Table 7</span><span class="title">Structure-Activity relationship study<sup><a class="bk_pop" href="#ml109.tfn1">*</a></sup></span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK56234/table/ml109.t7/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml109.t7_lrgtbl__"><table class="no_margin"><thead><tr><th id="hd_h_ml109.t7_1_1_1_1" colspan="13" rowspan="1" style="text-align:center;vertical-align:middle;">
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<div class="graphic"><img src="/books/NBK56234/bin/ml109fu11.jpg" alt="Image ml109fu11.jpg" /></div></th></tr><tr><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">No</th><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">SID</th><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">CID</th><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">R1</th><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">R2</th><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">R3</th><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">R4</th><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">R5</th><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">R6</th><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">X</th><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Y</th><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">EC50</th><th headers="hd_h_ml109.t7_1_1_1_1" id="hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Emax</th></tr></thead><tbody><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">1</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/26755506" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">26755506</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">661788</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">
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<div class="graphic"><img src="/books/NBK56234/bin/ml109fu12.jpg" alt="Image ml109fu12.jpg" /></div></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">1.16</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">101</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">2</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/29216393" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">29216393</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">16759579</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.29</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">99.8</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">3</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/29216998" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">29216998</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">17757110</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">n-Bu</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.83</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">112</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">4</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/29218134" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">29218134</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">17757294</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">
|
||
<div class="graphic"><img src="/books/NBK56234/bin/ml109fu13.jpg" alt="Image ml109fu13.jpg" /></div></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">3.98</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">70.3</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">5</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/29216997" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">29216997</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">17757109</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">n.d.</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">39.5</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">6</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/29218154" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">29218154</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">17757314</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.96</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">97.4</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">7</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/57655046" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">57655046</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">25246367</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">4.48</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">49.1</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">8</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/26755506" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">26755506</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">25246343</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.09</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">100</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">9</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/29218135" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">29218135</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">17757295</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">
|
||
<div class="graphic"><img src="/books/NBK56234/bin/ml109fu14.jpg" alt="Image ml109fu14.jpg" /></div></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">n.d.</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">10.1</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">10</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/29216999" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">29216999</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">17757111</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">
|
||
<div class="graphic"><img src="/books/NBK56234/bin/ml109fu15.jpg" alt="Image ml109fu15.jpg" /></div></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">n.d.</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">19.7</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">11</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/29218136" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">29218136</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">17757296</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">
|
||
<div class="graphic"><img src="/books/NBK56234/bin/ml109fu16.jpg" alt="Image ml109fu16.jpg" /></div></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.43</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">102.5</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">12</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/103073331" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">103073331</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">49789101</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.33</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">93.3</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">13</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/103073332" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">103073332</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">49789102</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">1.86</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">36.8</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">14</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/103073333" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">103073333</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">49789103</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">F</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.28</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">94.7</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">15</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/29216969" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">29216969</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">2887852</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">
|
||
<div class="graphic"><img src="/books/NBK56234/bin/ml109fu17.jpg" alt="Image ml109fu17.jpg" /></div></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">5.09</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">75.1</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">16</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/29218141" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">29218141</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">17757301</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">2.16</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">46.3</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">17</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/103073337" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">103073337</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">49789106</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">2.57</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">25.6</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">18</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/103073338" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">103073338</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">49789107</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">4.34</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">24.8</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">19</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/103073339" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">103073339</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">46190126</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">S</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.08</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">95.7</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">20</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/29216989" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">29216989</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">17757102</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">
|
||
<div class="graphic"><img src="/books/NBK56234/bin/ml109fu18.jpg" alt="Image ml109fu18.jpg" /></div></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.93</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">59</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">21</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/103073340" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">103073340</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">49789108</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">_</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">_</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">_</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">n.d.</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">17.7</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">22</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/103073341" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">103073341</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">49789109</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH2</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">3.78</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">72.6</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">23</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/103073342" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">103073342</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">49789110</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHMs</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.98</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">90.8</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">24</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/29218140" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">29218140</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">17757300</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">CN</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">n.d.</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">11.7</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">25</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/103073343" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">103073343</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">49789111</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">F</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">n.d.</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">7.4</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">26</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/103073344" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">103073344</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">49789112</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">S</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.018</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">69.6</td></tr><tr><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">27</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/103073345" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">103073345</a></td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">49789113</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OH</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Bn</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">OMe</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">H</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NHAc</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">N</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_11" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">O</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_12" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">0.55</td><td headers="hd_h_ml109.t7_1_1_1_1 hd_h_ml109.t7_1_1_2_13" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">98.6</td></tr></tbody></table></div><div><div><dl class="temp-labeled-list small"><dt>*</dt><dd><div id="ml109.tfn1"><p class="no_margin">All analogs were synthesized. Activities (EC50) in μM were obtained from the Elisa assay (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2104" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2104</a>). E<sub>max</sub> is expressed as % of the maximal response of CID-25246343/<a href="/pcsubstance/?term=ML109[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML109</a>, set at 100%.</p></div></dd></dl></div></div></div></div><div id="ml109.s59"><h3>3.5. Cellular Activity</h3><p>Both primary and secondary assays are cell-based assays.</p></div><div id="ml109.s60"><h3>3.6. Profiling Assays</h3><p><i>In vitro</i> ADME studies were performed on the probe molecule (<a class="figpopup" href="/books/NBK56234/table/ml109.t8/?report=objectonly" target="object" rid-figpopup="figml109t8" rid-ob="figobml109t8">Table 8</a>). This data indicates that the probe precipitates out of solution at 2 μM, and exhibits a high rate of clearance and good absorption. Additionally, the probe does not inhibit CYP, is modestly affected by the Pgp inhibitor.</p><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml109t8"><a href="/books/NBK56234/table/ml109.t8/?report=objectonly" target="object" title="Table 8" class="img_link icnblk_img figpopup" rid-figpopup="figml109t8" rid-ob="figobml109t8"><img class="small-thumb" src="/books/NBK56234/table/ml109.t8/?report=thumb" src-large="/books/NBK56234/table/ml109.t8/?report=previmg" alt="Table 8. Probe (CID-25246343/ML109) ADME properties." /></a><div class="icnblk_cntnt"><h4 id="ml109.t8"><a href="/books/NBK56234/table/ml109.t8/?report=objectonly" target="object" rid-ob="figobml109t8">Table 8</a></h4><p class="float-caption no_bottom_margin">Probe (CID-25246343/ML109) ADME properties. </p></div></div><div id="ml109.t9" class="table"><h3><span class="label">Table 9</span><span class="title">Summary of data in PubChem</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK56234/table/ml109.t9/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml109.t9_lrgtbl__"><table class="no_top_margin"><thead><tr><th id="hd_h_ml109.t9_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">PubChem AID</th><th id="hd_h_ml109.t9_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Type</th><th id="hd_h_ml109.t9_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Target</th><th id="hd_h_ml109.t9_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Conc. Range</th><th id="hd_h_ml109.t9_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Samples Tested</th></tr></thead><tbody><tr><td headers="hd_h_ml109.t9_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/926" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">926</a></td><td headers="hd_h_ml109.t9_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Primary DR</td><td headers="hd_h_ml109.t9_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">TSHR cell line</td><td headers="hd_h_ml109.t9_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">0.5nM-46μM</td><td headers="hd_h_ml109.t9_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">72030</td></tr><tr><td headers="hd_h_ml109.t9_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/938" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">938</a></td><td headers="hd_h_ml109.t9_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Primary DR counterscreen</td><td headers="hd_h_ml109.t9_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Parental cell line</td><td headers="hd_h_ml109.t9_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">0.5nM-46μM</td><td headers="hd_h_ml109.t9_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">72030</td></tr><tr><td headers="hd_h_ml109.t9_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/939" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">939</a></td><td headers="hd_h_ml109.t9_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Counterscreen</td><td headers="hd_h_ml109.t9_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">TSHR cell line</td><td headers="hd_h_ml109.t9_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">0.5nM-46μM</td><td headers="hd_h_ml109.t9_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">151</td></tr><tr><td headers="hd_h_ml109.t9_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/933" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">933</a></td><td headers="hd_h_ml109.t9_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Counterscreen</td><td headers="hd_h_ml109.t9_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">TSHR cell line HTRF</td><td headers="hd_h_ml109.t9_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">0.5nM-46μM</td><td headers="hd_h_ml109.t9_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">346</td></tr><tr><td headers="hd_h_ml109.t9_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/953" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">953</a></td><td headers="hd_h_ml109.t9_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Counterscreen</td><td headers="hd_h_ml109.t9_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Parental cell line</td><td headers="hd_h_ml109.t9_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">0.5nM-46μM</td><td headers="hd_h_ml109.t9_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">151</td></tr><tr><td headers="hd_h_ml109.t9_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1403" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1403</a></td><td headers="hd_h_ml109.t9_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Counterscreen</td><td headers="hd_h_ml109.t9_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">FSHR cell line HTRF</td><td headers="hd_h_ml109.t9_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">0.5nM-46μM</td><td headers="hd_h_ml109.t9_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">29</td></tr><tr><td headers="hd_h_ml109.t9_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1402" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1402</a></td><td headers="hd_h_ml109.t9_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Counterscreen</td><td headers="hd_h_ml109.t9_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">LHR cell line HTRF</td><td headers="hd_h_ml109.t9_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">0.5nM-46μM</td><td headers="hd_h_ml109.t9_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">31</td></tr><tr><td headers="hd_h_ml109.t9_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2104" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2104</a></td><td headers="hd_h_ml109.t9_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Secondary</td><td headers="hd_h_ml109.t9_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">TSHR cell line</td><td headers="hd_h_ml109.t9_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">0.5nM-46μM</td><td headers="hd_h_ml109.t9_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">35</td></tr><tr><td headers="hd_h_ml109.t9_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1401" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1401</a></td><td headers="hd_h_ml109.t9_1_1_1_2" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Summary</td><td headers="hd_h_ml109.t9_1_1_1_3" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">N/A</td><td headers="hd_h_ml109.t9_1_1_1_4" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">N/A</td><td headers="hd_h_ml109.t9_1_1_1_5" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">N/A</td></tr></tbody></table></div></div></div></div><div id="ml109.s61"><h2 id="_ml109_s61_">4. Discussion</h2><p>Among the confirmed hits, NCGC00168126 (CID-661788) was the most selective for TSH receptor, with no detectable agonist activity at the closely related LHCGR or FSHR, and was therefore selected for chemistry optimization. We explored substitution patterns throughout the scaffold. The analogs were first evaluated using the HTRF cAMP assay (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/933" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-933</a>). However, due to increased satisfaction with the consistency of results from the ELISA assay (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2104" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-2104</a>), the ELISA assay was used to guide our SAR study. As shown in <a class="figpopup" href="/books/NBK56234/table/ml109.t7/?report=objectonly" target="object" rid-figpopup="figml109t7" rid-ob="figobml109t7">Table 7</a>, when R1 was either an OH or H (entries 12 and 2), there was no substantial change in activity. However, the R2 substitution influences activity. OH improved activity (entry 8), but NH<sub>2</sub>, NHMs and OMe (entries, 22, 23, and 7) all resulted in a loss of activity. Furyl, benzyl, butyl and phenethyl groups were tolerated at R3 (entries 1, 2, 3 and 11), but pyridyl, tertiary amine, or an alcohol (entries 4, 9 and 10) resulted in a loss of activity. This indicates that lipophilic groups are well tolerated at R3, but polar groups that could behave as either hydrogen bond donors or acceptors are detrimental to activity. When R3 was an H (entry 5), activity was completely lost. The removal of the OMe moiety at R4 resulted in an inactive compound (entry13). Activity was lost when R5 was a functionality other than H (entries 18 and 25), or when R6 was a functionality other than NHAc (entries 16, 17, 18, 24 and 25). The R5 and R6 containing aryl ring is crucial for activity and very sensitive to substitution patterns. This was further demonstrated in the inactive analog where this entire ring was removed (entry 21). Y was oxygen in most analogs tested, but the activity improved when Y was sulfur (entries 19 and 26). Most analogs tested were with X as NH, but several oxidized compounds (X = N) were tested (entries 6, 20, and 27). The oxidized compounds were less active than the un-oxidized counterparts, but remain interesting analogs due to the improved air and acid stability. <i>In vivo</i> studies (<a class="figpopup" href="/books/NBK56234/figure/ml109.f6/?report=objectonly" target="object" rid-figpopup="figml109f6" rid-ob="figobml109f6">Figure 6</a>) showed that CID-25246343/<a href="/pcsubstance/?term=ML109[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML109</a> could increase secretion of T4 and thyroidal iodide uptake in mice after administration by esophageal gavage, suggesting CID-25246343/<a href="/pcsubstance/?term=ML109[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML109</a> is an orally available small molecule that can stimulate thyroid gland function<a class="bk_pop" href="#ml109.r1">1</a>.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml109f6" co-legend-rid="figlgndml109f6"><a href="/books/NBK56234/figure/ml109.f6/?report=objectonly" target="object" title="Figure 6" class="img_link icnblk_img figpopup" rid-figpopup="figml109f6" rid-ob="figobml109f6"><img class="small-thumb" src="/books/NBK56234/bin/ml109f6.gif" src-large="/books/NBK56234/bin/ml109f6.jpg" alt="Figure 6. Stimulation by CID-25246343/ML109 of thyroxine (T4) secretion and thyroidal radioiodide uptake in mice." /></a><div class="icnblk_cntnt" id="figlgndml109f6"><h4 id="ml109.f6"><a href="/books/NBK56234/figure/ml109.f6/?report=objectonly" target="object" rid-ob="figobml109f6">Figure 6</a></h4><p class="float-caption no_bottom_margin">Stimulation by CID-25246343/ML109 of thyroxine (T4) secretion and thyroidal radioiodide uptake in mice. <i>A:</i> Vehicle (PEG 300, control) or CID-25246343/ML109 (2.5 mg) was given by esophageal gavage and serum T4 was measured 2 hr later. <i>B:</i> Vehicle (control) <a href="/books/NBK56234/figure/ml109.f6/?report=objectonly" target="object" rid-ob="figobml109f6">(more...)</a></p></div></div><div id="ml109.s62"><h3>4.1. Comparison to existing art and how the new probe is an improvement</h3><p>The probe is the first orally active TSHR agonist. The probe series represents the most potent and selective small molecule TSHR agonists reported to date.</p></div><div id="ml109.s63"><h3>4.2. Mechanism of Action Studies</h3><p>There have been several studies to validate the probe’s exact binding site on TSHR. It was predicted to bind to the serpentine domain of TSHR, which is different from the amino terminal ectodomain binding site of endogenous TSH. This was validated by testing its binding to a TSHR mutant with the amino-terminal ectodomain deleted (KFLR). KFLR is not activated by TSH, but the probe molecule did activate KFLR, albeit with 12% lower efficacy and lower potency (EC50 = 1.7 μM). The binding was then modeled and N5.47 was predicted to be critical for activity, and a site specific mutant validated this prediction<a class="bk_pop" href="#ml109.r1">1</a> (<a class="figpopup" href="/books/NBK56234/figure/ml109.f7/?report=objectonly" target="object" rid-figpopup="figml109f7" rid-ob="figobml109f7">Figure 7</a>).</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml109f7" co-legend-rid="figlgndml109f7"><a href="/books/NBK56234/figure/ml109.f7/?report=objectonly" target="object" title="Figure 7" class="img_link icnblk_img figpopup" rid-figpopup="figml109f7" rid-ob="figobml109f7"><img class="small-thumb" src="/books/NBK56234/bin/ml109f7.gif" src-large="/books/NBK56234/bin/ml109f7.jpg" alt="Figure 7. CID-25246343/ML109 activates TSHR by binding to the transmembrane helical bundle." /></a><div class="icnblk_cntnt" id="figlgndml109f7"><h4 id="ml109.f7"><a href="/books/NBK56234/figure/ml109.f7/?report=objectonly" target="object" rid-ob="figobml109f7">Figure 7</a></h4><p class="float-caption no_bottom_margin">CID-25246343/ML109 activates TSHR by binding to the transmembrane helical bundle. <i>A</i> The effects of CID-25246343/ML109 or TSH on cAMP accumulation in cells expressing TSHR, TSHR-KFLR in which the large amino-terminal ectodomain to which TSH binds, is deleted, <a href="/books/NBK56234/figure/ml109.f7/?report=objectonly" target="object" rid-ob="figobml109f7">(more...)</a></p></div></div></div><div id="ml109.s64"><h3>4.3. Planned Future Studies</h3><p>The oxidized compounds present an attractive scaffold for further optimization due to the improved air and acid stability over the probe molecule.</p></div></div><div id="ml109.s65"><h2 id="_ml109_s65_">5. References</h2><dl class="temp-labeled-list"><dt>1.</dt><dd><div class="bk_ref" id="ml109.r1">Neumann S, Huang W, Titus S, Krause G, Kleinau G, Alberobello AT, Zheng W, Southall NT, Inglese J, Austin CP, Celi FS, Gavrilova O, Thomas CJ, Raaka BM, Gershengorn MC. Small-molecule agonists for the thyrotropin receptor stimulate thyroid function in human thyrocytes and mice. <span><span class="ref-journal">Proc Natl Acad Sci U S A. </span>2009 Jul 28;<span class="ref-vol">106</span>(30):12471–6.</span> [<a href="/pmc/articles/PMC2708975/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC2708975</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/19592511" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 19592511</span></a>]</div></dd><dt>2.</dt><dd><div class="bk_ref" id="ml109.r2">Vassart G, Pardo L, Costagliola S. A molecular dissection of the glycoprotein hormone receptors. <span><span class="ref-journal">Trends in Biochemical Sciences. </span>2004;<span class="ref-vol">29</span>(3):119–126.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/15003269" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15003269</span></a>]</div></dd><dt>3.</dt><dd><div class="bk_ref" id="ml109.r3">Demeure MJ, Doffek KM, Wilson SD. Defective thyrotropin receptor G-protein cyclic adenosine monophosphate signaling mechanism in the FTC human follicular thyroid cancer cell line. <span><span class="ref-journal">Surgery. </span>1997;<span class="ref-vol">122</span>(6):1195–1201.</span> discussion 1201–1192. [<a href="https://pubmed.ncbi.nlm.nih.gov/9426438" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 9426438</span></a>]</div></dd><dt>4.</dt><dd><div class="bk_ref" id="ml109.r4">Latif R, Ando T, Daniel S, Davies TF. Localization and regulation of thyrotropin receptors within lipid rafts. <span><span class="ref-journal">Endocrinology. </span>2003;<span class="ref-vol">144</span>(11):4725–4728.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/12960014" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 12960014</span></a>]</div></dd><dt>5.</dt><dd><div class="bk_ref" id="ml109.r5">Davies TF, Ando T, Lin RY, Tomer Y, Latif R. Thyrotropin receptor-associated diseases: from adenomata to Graves disease. <span><span class="ref-journal">J Clin Invest. </span>2005;<span class="ref-vol">115</span>(8):1972–1983.</span> [<a href="/pmc/articles/PMC1180562/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC1180562</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/16075037" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 16075037</span></a>]</div></dd><dt>6.</dt><dd><div class="bk_ref" id="ml109.r6">Fernandes JK, Day TA, Richardson MS, Sharma AK. Overview of the management of differentiated thyroid cancer. <span><span class="ref-journal">Current Treatment Options in Oncology. </span>2005;<span class="ref-vol">6</span>(1):47–57.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/15610714" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15610714</span></a>]</div></dd><dt>7.</dt><dd><div class="bk_ref" id="ml109.r7">van Straten NC, Schoonus-Gerritsma GG, van Someren RG, Draaijer J, Adang AE, Timmers CM, Hanssen RG, van Boeckel CA. The first orally active low molecular weight agonists for the LH receptor: thienopyr(im)idines with therapeutic potential for ovulation induction. <span><span class="ref-journal">Chembiochem. </span>2002;<span class="ref-vol">3</span>(10):1023–1026.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/12362369" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 12362369</span></a>]</div></dd><dt>8.</dt><dd><div class="bk_ref" id="ml109.r8">Jäschke H, Neumann S, Moore S, Thomas CJ, Colson A-O, Costanzi S, Kleinau G, Jiang J-K, Paschke Raaka BM, Krause G, Gershengorn MC. A low molecular weight agonist signals by binding to the transmembrane domain of Thyroid-Stimulating Hormone Receptor (TSHR) and Luteinizing Hormone/Chorionic Gonadotropin Receptor (LHCGR). <span><span class="ref-journal">J. Biol. Chem. </span>2006;<span class="ref-vol">281</span>:9841–9844.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/16488885" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 16488885</span></a>]</div></dd><dt>9.</dt><dd><div class="bk_ref" id="ml109.r9">Moore S, Jaeschke H, Kleinau G, Neumann S, Costanzi S, Jiang J, Childress J, Raaka B, Colson A, Paschke R, Krause G, Thomas C, Gershengorn M. Evaluation of small-molecule modulators of the luteinizing hormone/choriogonadotropin and thyroid stimulating hormone receptors: structure-activity relationships and selective binding patterns. <span><span class="ref-journal">J. Med. Chem. </span>2006;<span class="ref-vol">49</span>(13):3888–3896.</span> [<a href="/pmc/articles/PMC2543117/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC2543117</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/16789744" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 16789744</span></a>]</div></dd><dt>10.</dt><dd><div class="bk_ref" id="ml109.r10">Titus S, Neumann S, Zheng W, et al. Quantitative high-throughput screening using a live-cell cAMP assay identifies small-molecule agonists of the TSH receptor. <span><span class="ref-journal">J Biomol Screen. </span>2008;<span class="ref-vol">13</span>(2):120–127.</span> [<a href="/pmc/articles/PMC2653065/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC2653065</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/18216391" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 18216391</span></a>]</div></dd><dt>11.</dt><dd><div class="bk_ref" id="ml109.r11">Reinscheid RK, Kim J, Zeng J, Civelli O. High-throughput real-time monitoring of Gs-coupled receptor activation in intact cells using cyclic nucleotide-gated channels. <span><span class="ref-journal">Eur J Pharmacol. </span>2003;<span class="ref-vol">478</span>(1):27–34.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/14555181" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 14555181</span></a>]</div></dd><dt>12.</dt><dd><div class="bk_ref" id="ml109.r12">Yao YLJ, Llorente I, DeBernardi M, Cao L. ACT: One technology: a live cAMP assay. Poster presentation in the 10th Anniversary Conference of Society of Biomolecular Sciences; 2004. </div></dd><dt>13.</dt><dd><div class="bk_ref" id="ml109.r13">Gabriel D, Vernier M, Pfeifer MJ, Dasen B, Tenaillon L, Bouhelal R. High throughput screening technologies for direct cyclic AMP measurement. <span><span class="ref-journal">Assay Drug Dev Technol. </span>2003;<span class="ref-vol">1</span>(2):291–303.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/15090194" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15090194</span></a>]</div></dd><dt>14.</dt><dd><div class="bk_ref" id="ml109.r14">Prystay L, Gagne A, Kasila P, Yeh LA, Banks P. Homogeneous cell-based fluorescence polarization assay for the direct detection of cAMP. <span><span class="ref-journal">J Biomol Screen. </span>2001;<span class="ref-vol">6</span>(2):75–82.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/11689101" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11689101</span></a>]</div></dd><dt>15.</dt><dd><div class="bk_ref" id="ml109.r15">Inglese J, Auld DS, Jadhav A, et al. Quantitative high-throughput screening: A titration-based approach that efficiently identifies biological activities in large chemical libraries. <span><span class="ref-journal">Proc Natl Acad Sci U S A. </span>2006;<span class="ref-vol">103</span>(31):11473–11478.</span> [<a href="/pmc/articles/PMC1518803/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC1518803</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/16864780" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 16864780</span></a>]</div></dd></dl></div><div id="bk_toc_contnr"></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/NBK56234/?report=reader">PubReader</a></li><li><a href="/books/NBK56234/?report=printable">Print View</a></li><li><a data-jig="ncbidialog" href="#_ncbi_dlg_citbx_NBK56234" data-jigconfig="width:400,modal:true">Cite this Page</a><div id="_ncbi_dlg_citbx_NBK56234" style="display:none" title="Cite this Page"><div class="bk_tt">Titus S, Huang W, Marugan J, et al. Identification of Potent and Selective Thyroid Stimulating Hormone Receptor Agonists. 2009 Sep 1 [Updated 2011 Mar 25]. 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/NBK56234/pdf/Bookshelf_NBK56234.pdf">PDF version of this page</a> (402K)</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="#ml109.s1" ref="log$=inpage&link_id=inpage">Probe Structure & Characteristics</a></li><li><a href="#ml109.s2" ref="log$=inpage&link_id=inpage">Recommendations for scientific use of the probe</a></li><li><a href="#ml109.s3" ref="log$=inpage&link_id=inpage">Introduction</a></li><li><a href="#ml109.s4" ref="log$=inpage&link_id=inpage">Materials and Methods</a></li><li><a href="#ml109.s54" ref="log$=inpage&link_id=inpage">Results</a></li><li><a href="#ml109.s61" ref="log$=inpage&link_id=inpage">Discussion</a></li><li><a href="#ml109.s65" ref="log$=inpage&link_id=inpage">References</a></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Related information</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="discovery_db_links" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pmc&DbFrom=books&Cmd=Link&LinkName=books_pmc_refs&IdsFromResult=2509788" ref="log$=recordlinks">PMC</a><div class="brieflinkpop offscreen_noflow">PubMed Central citations</div></li><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pcassay&DbFrom=books&Cmd=Link&LinkName=books_pcassay_probe&IdsFromResult=2509788" ref="log$=recordlinks">PubChem BioAssay for Chemical Probe</a><div class="brieflinkpop offscreen_noflow">PubChem BioAssay records reporting screening data for the development of the chemical probe(s) described in this book chapter</div></li><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pcsubstance&DbFrom=books&Cmd=Link&LinkName=books_pcsubstance&IdsFromResult=2509788" ref="log$=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=2509788" ref="log$=recordlinks">PubMed</a><div class="brieflinkpop offscreen_noflow">Links to PubMed</div></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Similar articles in PubMed</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="PBooksDiscovery_RA" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/23658967" ref="ordinalpos=1&linkpos=1&log$=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> Identification of Thyroid Stimulating Hormone Receptor Inverse Agonists.</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> Identification of Thyroid Stimulating Hormone Receptor Inverse Agonists.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" 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xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">J Biomol Screen. 2008 Feb; 13(2):120-7. Epub 2008 Jan 23.</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/7901757" ref="ordinalpos=1&linkpos=4&log$=relatedarticles&logdbfrom=pubmed">Substitutions of different regions of the third cytoplasmic loop of the thyrotropin (TSH) receptor have selective effects on constitutive, TSH-, and TSH receptor autoantibody-stimulated phosphoinositide and 3',5'-cyclic adenosine monophosphate signal generation.</a><span class="source">[Mol Endocrinol. 1993]</span><div class="brieflinkpop offscreen_noflow">Substitutions of different regions of the third cytoplasmic loop of the thyrotropin (TSH) receptor have selective effects on constitutive, TSH-, and TSH receptor autoantibody-stimulated phosphoinositide and 3',5'-cyclic adenosine monophosphate signal generation.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Kosugi S, Okajima F, Ban T, Hidaka A, Shenker A, Kohn LD. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Mol Endocrinol. 1993 Aug; 7(8):1009-20. </em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/28484426" ref="ordinalpos=1&linkpos=5&log$=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> Structural-Functional Features of the Thyrotropin Receptor: A Class A G-Protein-Coupled Receptor at Work.</a><span class="source">[Front Endocrinol (Lausanne). 2...]</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> Structural-Functional Features of the Thyrotropin Receptor: A Class A G-Protein-Coupled Receptor at Work.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Kleinau G, Worth CL, Kreuchwig A, Biebermann H, Marcinkowski P, Scheerer P, Krause G. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Front Endocrinol (Lausanne). 2017; 8:86. 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