714 lines
No EOL
98 KiB
HTML
714 lines
No EOL
98 KiB
HTML
<?xml version="1.0" encoding="utf-8"?>
|
||
<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
|
||
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">
|
||
|
||
<head><meta http-equiv="Content-Type" content="text/html; charset=utf-8" />
|
||
<!-- AppResources meta begin -->
|
||
<meta name="paf-app-resources" content="" />
|
||
<script type="text/javascript">var ncbi_startTime = new Date();</script>
|
||
|
||
<!-- AppResources meta end -->
|
||
|
||
<!-- TemplateResources meta begin -->
|
||
<meta name="paf_template" content="" />
|
||
|
||
<!-- TemplateResources meta end -->
|
||
|
||
<!-- Logger begin -->
|
||
<meta name="ncbi_db" content="books" /><meta name="ncbi_pdid" content="book-part" /><meta name="ncbi_acc" content="NBK47340" /><meta name="ncbi_domain" content="mlprobe" /><meta name="ncbi_report" content="record" /><meta name="ncbi_type" content="fulltext" /><meta name="ncbi_objectid" content="" /><meta name="ncbi_pcid" content="/NBK47340/" /><meta name="ncbi_pagename" content="Discovery of a Highly Selective in vitro and in vivo M1 Allosteric Agonist Probe - Probe Reports from the NIH Molecular Libraries Program - NCBI Bookshelf" /><meta name="ncbi_bookparttype" content="chapter" /><meta name="ncbi_app" content="bookshelf" />
|
||
<!-- Logger end -->
|
||
|
||
<title>Discovery of a Highly Selective in vitro and in vivo M1 Allosteric Agonist Probe - Probe Reports from the NIH Molecular Libraries Program - NCBI Bookshelf</title>
|
||
|
||
<!-- AppResources external_resources begin -->
|
||
<link rel="stylesheet" href="/core/jig/1.15.2/css/jig.min.css" /><script type="text/javascript" src="/core/jig/1.15.2/js/jig.min.js"></script>
|
||
|
||
<!-- AppResources external_resources end -->
|
||
|
||
<!-- Page meta begin -->
|
||
<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="Discovery of a Highly Selective in vitro and in vivo M1 Allosteric Agonist Probe" /><meta name="citation_publisher" content="National Center for Biotechnology Information (US)" /><meta name="citation_date" content="2010/10/04" /><meta name="citation_author" content="Craig Lindsley" /><meta name="citation_author" content="Michelle Lewis" /><meta name="citation_author" content="Dave Weaver" /><meta name="citation_pmid" content="21433356" /><meta name="citation_fulltext_html_url" content="https://www.ncbi.nlm.nih.gov/books/NBK47340/" /><link rel="schema.DC" href="http://purl.org/DC/elements/1.0/" /><meta name="DC.Title" content="Discovery of a Highly Selective in vitro and in vivo M1 Allosteric Agonist Probe" /><meta name="DC.Type" content="Text" /><meta name="DC.Publisher" content="National Center for Biotechnology Information (US)" /><meta name="DC.Contributor" content="Craig Lindsley" /><meta name="DC.Contributor" content="Michelle Lewis" /><meta name="DC.Contributor" content="Dave Weaver" /><meta name="DC.Date" content="2010/10/04" /><meta name="DC.Identifier" content="https://www.ncbi.nlm.nih.gov/books/NBK47340/" /><meta name="description" content="Previous attempts to develop compounds that are highly selective for M1 or other specific muscarinic acetylcholine receptor (mAChR) subtypes have failed because of the high conservation of the ACh binding site and difficulty in developing truly specific compounds. Basic and clinical studies suggest that mAChR activation has the potential to be disease modifying, as well as to provide palliative cognitive therapy. Probes developed from these efforts will greatly advance the current state of the art by aiding in the understanding of M1's role in cell-based physiology and may extend the clinical understanding of psychotic and cognitive symptoms associated with neurodegenerative disorders such as Alzheimer's Disease and schizophrenia. This probe report describes ML071 (CID-25010775), a highly selective, potent M1 agonist that activates M1 by virtue of binding at an allosteric site in the third extracellular loop of the M1 receptor. The compound represents a "best in class" probe that provides unprecedented mAChR selectivity, clean ancillary pharmacology, saline solubility, and central penetrance. Furthermore, it can be employed for in vitro and in vivo studies to examine the role of selective M1 receptor activation." /><meta name="og:title" content="Discovery of a Highly Selective in vitro and in vivo M1 Allosteric Agonist Probe" /><meta name="og:type" content="book" /><meta name="og:description" content="Previous attempts to develop compounds that are highly selective for M1 or other specific muscarinic acetylcholine receptor (mAChR) subtypes have failed because of the high conservation of the ACh binding site and difficulty in developing truly specific compounds. Basic and clinical studies suggest that mAChR activation has the potential to be disease modifying, as well as to provide palliative cognitive therapy. Probes developed from these efforts will greatly advance the current state of the art by aiding in the understanding of M1's role in cell-based physiology and may extend the clinical understanding of psychotic and cognitive symptoms associated with neurodegenerative disorders such as Alzheimer's Disease and schizophrenia. This probe report describes ML071 (CID-25010775), a highly selective, potent M1 agonist that activates M1 by virtue of binding at an allosteric site in the third extracellular loop of the M1 receptor. The compound represents a "best in class" probe that provides unprecedented mAChR selectivity, clean ancillary pharmacology, saline solubility, and central penetrance. Furthermore, it can be employed for in vitro and in vivo studies to examine the role of selective M1 receptor activation." /><meta name="og:url" content="https://www.ncbi.nlm.nih.gov/books/NBK47340/" /><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/ml071/" /><link rel="canonical" href="https://www.ncbi.nlm.nih.gov/books/NBK47340/" /><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" />
|
||
|
||
<!-- Page meta end -->
|
||
<link rel="shortcut icon" href="//www.ncbi.nlm.nih.gov/favicon.ico" /><meta name="ncbi_phid" content="CE8E13397D65E8410000000000E000C5.m_13" />
|
||
<meta name='referrer' content='origin-when-cross-origin'/><link type="text/css" rel="stylesheet" href="//static.pubmed.gov/portal/portal3rc.fcgi/4216699/css/3852956/3985586/3808861/4121862/3974050/3917732/251717/4216701/14534/45193/4113719/3849091/3984811/3751656/4033350/3840896/3577051/3852958/4008682/4207974/4206132/4062871/12930/3964959/3854974/36029/4128070/9685/3549676/3609192/3609193/3609213/3395586.css" /><link type="text/css" rel="stylesheet" href="//static.pubmed.gov/portal/portal3rc.fcgi/4216699/css/3411343/3882866.css" media="print" /></head>
|
||
<body class="book-part">
|
||
<div class="grid">
|
||
<div class="col twelve_col nomargin shadow">
|
||
<!-- System messages like service outage or JS required; this is handled by the TemplateResources portlet -->
|
||
<div class="sysmessages">
|
||
<noscript>
|
||
<p class="nojs">
|
||
<strong>Warning:</strong>
|
||
The NCBI web site requires JavaScript to function.
|
||
<a href="/guide/browsers/#enablejs" title="Learn how to enable JavaScript" target="_blank">more...</a>
|
||
</p>
|
||
</noscript>
|
||
</div>
|
||
<!--/.sysmessage-->
|
||
<div class="wrap">
|
||
<div class="page">
|
||
<div class="top">
|
||
<div id="universal_header">
|
||
<section class="usa-banner">
|
||
<div class="usa-accordion">
|
||
<header class="usa-banner-header">
|
||
<div class="usa-grid usa-banner-inner">
|
||
<img src="https://www.ncbi.nlm.nih.gov/coreutils/uswds/img/favicons/favicon-57.png" alt="U.S. flag" />
|
||
<p>An official website of the United States government</p>
|
||
<button class="non-usa-accordion-button usa-banner-button" aria-expanded="false" aria-controls="gov-banner-top" type="button">
|
||
<span class="usa-banner-button-text">Here's how you know</span>
|
||
</button>
|
||
</div>
|
||
</header>
|
||
<div class="usa-banner-content usa-grid usa-accordion-content" id="gov-banner-top" aria-hidden="true">
|
||
<div class="usa-banner-guidance-gov usa-width-one-half">
|
||
<img class="usa-banner-icon usa-media_block-img" src="https://www.ncbi.nlm.nih.gov/coreutils/uswds/img/icon-dot-gov.svg" alt="Dot gov" />
|
||
<div class="usa-media_block-body">
|
||
<p>
|
||
<strong>The .gov means it's official.</strong>
|
||
<br />
|
||
Federal government websites often end in .gov or .mil. Before
|
||
sharing sensitive information, make sure you're on a federal
|
||
government site.
|
||
</p>
|
||
</div>
|
||
</div>
|
||
<div class="usa-banner-guidance-ssl usa-width-one-half">
|
||
<img class="usa-banner-icon usa-media_block-img" src="https://www.ncbi.nlm.nih.gov/coreutils/uswds/img/icon-https.svg" alt="Https" />
|
||
<div class="usa-media_block-body">
|
||
<p>
|
||
<strong>The site is secure.</strong>
|
||
<br />
|
||
The <strong>https://</strong> ensures that you are connecting to the
|
||
official website and that any information you provide is encrypted
|
||
and transmitted securely.
|
||
</p>
|
||
</div>
|
||
</div>
|
||
</div>
|
||
</div>
|
||
</section>
|
||
<div class="usa-overlay"></div>
|
||
<header class="ncbi-header" role="banner" data-section="Header">
|
||
|
||
<div class="usa-grid">
|
||
<div class="usa-width-one-whole">
|
||
|
||
<div class="ncbi-header__logo">
|
||
<a href="/" class="logo" aria-label="NCBI Logo" data-ga-action="click_image" data-ga-label="NIH NLM Logo">
|
||
<img src="https://www.ncbi.nlm.nih.gov/coreutils/nwds/img/logos/AgencyLogo.svg" alt="NIH NLM Logo" />
|
||
</a>
|
||
</div>
|
||
|
||
<div class="ncbi-header__account">
|
||
<a id="account_login" href="https://account.ncbi.nlm.nih.gov" class="usa-button header-button" style="display:none" data-ga-action="open_menu" data-ga-label="account_menu">Log in</a>
|
||
<button id="account_info" class="header-button" style="display:none" aria-controls="account_popup" type="button">
|
||
<span class="fa fa-user" aria-hidden="true">
|
||
<svg xmlns="http://www.w3.org/2000/svg" viewBox="0 0 24 24" width="20px" height="20px">
|
||
<g style="fill: #fff">
|
||
<ellipse cx="12" cy="8" rx="5" ry="6"></ellipse>
|
||
<path d="M21.8,19.1c-0.9-1.8-2.6-3.3-4.8-4.2c-0.6-0.2-1.3-0.2-1.8,0.1c-1,0.6-2,0.9-3.2,0.9s-2.2-0.3-3.2-0.9 C8.3,14.8,7.6,14.7,7,15c-2.2,0.9-3.9,2.4-4.8,4.2C1.5,20.5,2.6,22,4.1,22h15.8C21.4,22,22.5,20.5,21.8,19.1z"></path>
|
||
</g>
|
||
</svg>
|
||
</span>
|
||
<span class="username desktop-only" aria-hidden="true" id="uname_short"></span>
|
||
<span class="sr-only">Show account info</span>
|
||
</button>
|
||
</div>
|
||
|
||
<div class="ncbi-popup-anchor">
|
||
<div class="ncbi-popup account-popup" id="account_popup" aria-hidden="true">
|
||
<div class="ncbi-popup-head">
|
||
<button class="ncbi-close-button" data-ga-action="close_menu" data-ga-label="account_menu" type="button">
|
||
<span class="fa fa-times">
|
||
<svg xmlns="http://www.w3.org/2000/svg" viewBox="0 0 48 48" width="24px" height="24px">
|
||
<path d="M38 12.83l-2.83-2.83-11.17 11.17-11.17-11.17-2.83 2.83 11.17 11.17-11.17 11.17 2.83 2.83 11.17-11.17 11.17 11.17 2.83-2.83-11.17-11.17z"></path>
|
||
</svg>
|
||
</span>
|
||
<span class="usa-sr-only">Close</span></button>
|
||
<h4>Account</h4>
|
||
</div>
|
||
<div class="account-user-info">
|
||
Logged in as:<br />
|
||
<b><span class="username" id="uname_long">username</span></b>
|
||
</div>
|
||
<div class="account-links">
|
||
<ul class="usa-unstyled-list">
|
||
<li><a id="account_myncbi" href="/myncbi/" class="set-base-url" data-ga-action="click_menu_item" data-ga-label="account_myncbi">Dashboard</a></li>
|
||
<li><a id="account_pubs" href="/myncbi/collections/bibliography/" class="set-base-url" data-ga-action="click_menu_item" data-ga-label="account_pubs">Publications</a></li>
|
||
<li><a id="account_settings" href="/account/settings/" class="set-base-url" data-ga-action="click_menu_item" data-ga-label="account_settings">Account settings</a></li>
|
||
<li><a id="account_logout" href="/account/signout/" class="set-base-url" data-ga-action="click_menu_item" data-ga-label="account_logout">Log out</a></li>
|
||
</ul>
|
||
</div>
|
||
</div>
|
||
</div>
|
||
|
||
</div>
|
||
</div>
|
||
</header>
|
||
<div role="navigation" aria-label="access keys">
|
||
<a id="nws_header_accesskey_0" href="https://www.ncbi.nlm.nih.gov/guide/browsers/#ncbi_accesskeys" class="usa-sr-only" accesskey="0" tabindex="-1">Access keys</a>
|
||
<a id="nws_header_accesskey_1" href="https://www.ncbi.nlm.nih.gov" class="usa-sr-only" accesskey="1" tabindex="-1">NCBI Homepage</a>
|
||
<a id="nws_header_accesskey_2" href="/myncbi/" class="set-base-url usa-sr-only" accesskey="2" tabindex="-1">MyNCBI Homepage</a>
|
||
<a id="nws_header_accesskey_3" href="#maincontent" class="usa-sr-only" accesskey="3" tabindex="-1">Main Content</a>
|
||
<a id="nws_header_accesskey_4" href="#" class="usa-sr-only" accesskey="4" tabindex="-1">Main Navigation</a>
|
||
</div>
|
||
<section data-section="Alerts">
|
||
<div class="ncbi-alerts-placeholder"></div>
|
||
</section>
|
||
</div>
|
||
<div class="header">
|
||
<div class="res_logo"><h1 class="res_name"><a href="/books/" title="Bookshelf home">Bookshelf</a></h1><h2 class="res_tagline"></h2></div>
|
||
<div class="search"><form method="get" action="/books/"><div class="search_form"><label for="database" class="offscreen_noflow">Search database</label><select id="database"><optgroup label="Recent"><option value="books" selected="selected" data-ac_dict="bookshelf-search">Books</option><option value="pubmed">PubMed</option><option value="clinvar">ClinVar</option><option value="refseq" class="last">RefSeq</option></optgroup><optgroup label="All"><option value="gquery">All Databases</option><option value="assembly">Assembly</option><option value="biocollections">Biocollections</option><option value="bioproject">BioProject</option><option value="biosample">BioSample</option><option value="books" data-ac_dict="bookshelf-search">Books</option><option value="clinvar">ClinVar</option><option value="cdd">Conserved Domains</option><option value="gap">dbGaP</option><option value="dbvar">dbVar</option><option value="gene">Gene</option><option value="genome">Genome</option><option value="gds">GEO DataSets</option><option value="geoprofiles">GEO Profiles</option><option value="gtr">GTR</option><option value="ipg">Identical Protein Groups</option><option value="medgen">MedGen</option><option value="mesh">MeSH</option><option value="nlmcatalog">NLM Catalog</option><option value="nuccore">Nucleotide</option><option value="omim">OMIM</option><option value="pmc">PMC</option><option value="protein">Protein</option><option value="proteinclusters">Protein Clusters</option><option value="protfam">Protein Family Models</option><option value="pcassay">PubChem BioAssay</option><option value="pccompound">PubChem Compound</option><option value="pcsubstance">PubChem Substance</option><option value="pubmed">PubMed</option><option value="snp">SNP</option><option value="sra">SRA</option><option value="structure">Structure</option><option value="taxonomy">Taxonomy</option><option value="toolkit">ToolKit</option><option value="toolkitall">ToolKitAll</option><option value="toolkitbookgh">ToolKitBookgh</option></optgroup></select><div class="nowrap"><label for="term" class="offscreen_noflow" accesskey="/">Search term</label><div class="nowrap"><input type="text" name="term" id="term" title="Search Books. Use up and down arrows to choose an item from the autocomplete." value="" class="jig-ncbiclearbutton jig-ncbiautocomplete" data-jigconfig="dictionary:'bookshelf-search',disableUrl:'NcbiSearchBarAutoComplCtrl'" autocomplete="off" data-sbconfig="ds:'no',pjs:'no',afs:'no'" /></div><button id="search" type="submit" class="button_search nowrap" cmd="go">Search</button></div></div></form><ul class="searchlinks inline_list"><li>
|
||
<a href="/books/browse/">Browse Titles</a>
|
||
</li><li>
|
||
<a href="/books/advanced/">Advanced</a>
|
||
</li><li class="help">
|
||
<a href="/books/NBK3833/">Help</a>
|
||
</li><li class="disclaimer">
|
||
<a target="_blank" data-ga-category="literature_resources" data-ga-action="link_click" data-ga-label="disclaimer_link" href="https://www.ncbi.nlm.nih.gov/books/about/disclaimer/">Disclaimer</a>
|
||
</li></ul></div>
|
||
</div>
|
||
|
||
|
||
|
||
<!--<component id="Page" label="headcontent"/>-->
|
||
|
||
</div>
|
||
<div class="content">
|
||
<!-- site messages -->
|
||
<!-- Custom content 1 -->
|
||
<div class="col1">
|
||
|
||
</div>
|
||
|
||
<div class="container">
|
||
<div id="maincontent" class="content eight_col col">
|
||
<!-- Custom content in the left column above book nav -->
|
||
<div class="col2">
|
||
|
||
</div>
|
||
|
||
<!-- Book content -->
|
||
|
||
|
||
<!-- Custom content between navigation and content -->
|
||
<div class="col3">
|
||
|
||
</div>
|
||
|
||
<div class="document">
|
||
<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="#__NBK47340_dtls__">Show details</a><div style="display:none" class="ui-widget" id="__NBK47340_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/ml075/" title="Previous page in this title">< Prev</a><span class="inactive page_link next">Next ></span></div></div></div></div></div>
|
||
<div class="main-content lit-style" itemscope="itemscope" itemtype="http://schema.org/CreativeWork"><div class="meta-content fm-sec"><h1 id="_NBK47340_"><span class="title" itemprop="name">Discovery of a Highly Selective <i>in vitro</i> and <i>in
|
||
vivo</i> M1 Allosteric Agonist Probe</span></h1><p class="contrib-group"><span itemprop="author">Craig Lindsley</span>, <span itemprop="author">Michelle Lewis</span>, and <span itemprop="author">Dave Weaver</span>.</p><a data-jig="ncbitoggler" href="#__NBK47340_ai__" style="border:0;text-decoration:none">Author Information and Affiliations</a><div style="display:none" class="ui-widget" id="__NBK47340_ai__"><p class="contrib-group"><h4>Authors</h4><span itemprop="author">Craig Lindsley</span>,<sup>1</sup> <span itemprop="author">Michelle Lewis</span>,<sup>2</sup> and <span itemprop="author">Dave Weaver</span><sup>3</sup>.</p><h4>Contact</h4><div class="affiliation"><sup>1</sup> <span class="email-label">Email: </span><a href="mailto:dev@null" data-email="ude.tlibrednav@yelsdnil.giarc" class="oemail">ude.tlibrednav@yelsdnil.giarc</a></div><div class="affiliation"><sup>2</sup> <span class="email-label">Email: </span><a href="mailto:dev@null" data-email="ude.tlibrednav@siwel.ellehcim" class="oemail">ude.tlibrednav@siwel.ellehcim</a></div><div class="affiliation"><sup>3</sup> <span class="email-label">Email: </span><a href="mailto:dev@null" data-email="ude.tlibrednav@revaew.divad" class="oemail">ude.tlibrednav@revaew.divad</a></div></div><p class="small">Received: <span itemprop="datePublished">December 3, 2008</span>; Last Update: <span itemprop="dateModified">October 4, 2010</span>.</p></div><div class="jig-ncbiinpagenav body-content whole_rhythm" data-jigconfig="allHeadingLevels: ['h2'],smoothScroll: false" itemprop="text"><div id="_abs_rndgid_" itemprop="description"><p>Previous attempts to develop compounds that are highly selective for M1 or other specific muscarinic acetylcholine receptor (mAChR) subtypes have failed because of the high conservation of the ACh binding site and difficulty in developing truly specific compounds. Basic and clinical studies suggest that mAChR activation has the potential to be disease modifying, as well as to provide palliative cognitive therapy. Probes developed from these efforts will greatly advance the current state of the art by aiding in the understanding of M1's role in cell-based physiology and may extend the clinical understanding of psychotic and cognitive symptoms associated with neurodegenerative disorders such as Alzheimer's Disease and schizophrenia. This probe report describes ML071 (CID-25010775), a highly selective, potent M1 agonist that activates M1 by virtue of binding at an allosteric site in the third extracellular loop of the M1 receptor. The compound represents a "best in class" probe that provides unprecedented mAChR selectivity, clean ancillary pharmacology, saline solubility, and central penetrance. Furthermore, it can be employed for in vitro and in vivo studies to examine the role of selective M1 receptor activation.</p></div><div class="h2"></div><p><b>Assigned Assay Grant #:</b> X01 MH077606-01</p><p><b>Screening Center Name & PI:</b> Vanderbilt Screening Center for GPCRs,
|
||
Ion Channels, and Transporters, David Weaver</p><p><b>Chemistry Center Name & PI:</b> Vanderbilt Specialized Chemistry Center
|
||
for Accelerated Probe Development, Craig Lindsley</p><p><b>Assay Submitter & Institution:</b> P. Jeffrey Conn, Vanderbilt
|
||
University</p><p><b>PubChem Summary Bioassay Identifier (AID):</b>
|
||
<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1798" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1798</a></p><div id="ml071.s2"><h2 id="_ml071_s2_">Probe Structure & Characteristics</h2><p>Ethyl 4-(2-methylbenzamido)ethylamino)piperidine-1-carboxylate hydrochloride</p><div id="ml071.fu1" class="figure"><div class="graphic"><img src="/books/NBK47340/bin/ml071fu1.jpg" alt="Image ml071fu1" /></div></div><div id="ml071.tu1" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK47340/table/ml071.tu1/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml071.tu1_lrgtbl__"><table><thead><tr><th id="hd_h_ml071.tu1_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">CID</th><th id="hd_h_ml071.tu1_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Target Name</th><th id="hd_h_ml071.tu1_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">IC50/EC50 (nM) [SID,
|
||
AID]</th><th id="hd_h_ml071.tu1_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Anti-target Name(s)</th><th id="hd_h_ml071.tu1_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">IC50/EC50 (μM)
|
||
[SID, AID]</th><th id="hd_h_ml071.tu1_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Selectivity</th><th id="hd_h_ml071.tu1_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary<br /><br />Assay(s) Name:
|
||
IC50/EC50</th></tr></thead><tbody><tr><td headers="hd_h_ml071.tu1_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">
|
||
<b>25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>)</b>
|
||
</td><td headers="hd_h_ml071.tu1_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">
|
||
<b>M1</b>
|
||
</td><td headers="hd_h_ml071.tu1_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">
|
||
<b>198</b>
|
||
<br />
|
||
<br />
|
||
<b>[<a href="https://pubchem.ncbi.nlm.nih.gov/substance/56353039" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-56353039</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/626" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-626</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1488" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1488</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1741" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1741</a>,
|
||
<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1744" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1744</a>]</b>
|
||
</td><td headers="hd_h_ml071.tu1_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">
|
||
<b>M2–M5</b>
|
||
</td><td headers="hd_h_ml071.tu1_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><b>>30</b>
|
||
μ<b>M</b><br /><br /><b>[<a href="https://pubchem.ncbi.nlm.nih.gov/substance/56353039" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-56353039</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/626" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-626</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1488" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1488</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1741" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1741</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1744" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1744</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1470" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1470</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1767" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1767</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1764" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1764</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1757" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1757</a>,
|
||
<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1508" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1508</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1788" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1788</a>]</b></td><td headers="hd_h_ml071.tu1_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">
|
||
<b>>263-fold</b>
|
||
</td><td headers="hd_h_ml071.tu1_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">
|
||
<b>M1 Y381A 379 nM [<a href="https://pubchem.ncbi.nlm.nih.gov/substance/56353039" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-56353039</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1743" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1743</a>]</b>
|
||
</td></tr></tbody></table></div></div></div><div id="ml071.s3"><h2 id="_ml071_s3_">Recommendations for the scientific use of this probe</h2><p>This probe can be used for both <i>in vitro</i> and <i>in
|
||
vivo</i> to study the role of selective M1 receptor activation. The
|
||
compound possesses unprecedented selectivity versus M2–M5 and against a
|
||
large panel of GPCRs, ion channels and transporters. Moreover, the probe is
|
||
centrally penetrant and soluble in saline (>25 mg/mL).</p><div id="ml071.s4"><h3>Specific Aim</h3><p>To identify small molecule agonists of M1 muscarinic receptor that are cell
|
||
permeable, exhibit submicromolar potency, and show greater than 10 fold
|
||
selectivity over other muscarinic family members M2, M3, M4 and M5. This probe
|
||
report describes a potent, highly selective M1 allosteric agonist (CID 25010775
|
||
(<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>), <a href="https://pubchem.ncbi.nlm.nih.gov/substance/56353039" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-56353039</a>) that activates M1 by virtue of binding at an allosteric
|
||
site in the third extracellular loop of the M1 receptor. The probe is
|
||
‘best in class’ providing unprecedented mAChR
|
||
selectivity, clean ancillary pharmacology, soluble in saline (~25 mg/mL) and
|
||
centrally penetrant.</p></div><div id="ml071.s5"><h3>Significance</h3><p>Previous attempts to develop compounds that are highly selective for M1 or other
|
||
specific mAChR subtypes have failed because of the high conservation of the Ach
|
||
binding site and difficulty in developing truly specific compounds (<a class="bk_pop" href="#ml071.r2">Bonner et al. 1997</a>, <a class="bk_pop" href="#ml071.r3">1998</a>; <a class="bk_pop" href="#ml071.r7">Felder et al. 2000</a>; <a class="bk_pop" href="#ml071.r4">Bymaster et al. 2003</a>). The lack of highly selective
|
||
compounds has made it impossible to definitively determine the behavioral and
|
||
clinical effects of these receptors. In numerous Phase II and III clinical
|
||
trails, pan-mAChR agonists were shown to improve cognitive performance in AD
|
||
patients, but the GI-and/or cardiovascular side effects, resulting from
|
||
activation of peripheral mAChRs, were deemed intolerable and the trials were
|
||
discontinued (Eglen et al. 2001; Tarsy et al. 2006). Importantly, several
|
||
pan-mAChR agonists demonstrated decline of Aβ42 in the cerebral
|
||
spinal fluid of AD patients, suggesting that mAChR activation has the potential
|
||
to be disease modifying as well as providing palliative cognitive therapy (<a class="bk_pop" href="#ml071.r1">Bodick et al. 1997</a>). More recent
|
||
studies in 3xTg-AD mice further support a disease modifying role for mAChR
|
||
activation, and several Ph III trials demonstrated that mAChR activation lowered
|
||
Aβ42 in patients (<a class="bk_pop" href="#ml071.r5">Caccamo et
|
||
al. 2006</a>). Interestingly, the M1/M4 preferring xanomeline, in
|
||
addition to improving cognitive performance, had robust therapeutic effects on
|
||
the psychotic symptoms and behavioral disturbances associated with AD and
|
||
recently published clinical trial data indicates efficacy in schizophrenic
|
||
patients (<a class="bk_pop" href="#ml071.r1">Bodick et al. 1997</a>; <a class="bk_pop" href="#ml071.r10">Shekhar et al. 2008</a>). Probes
|
||
developed from these efforts will greatly advance the current state of the art
|
||
by aiding in the understanding of M1’s role in cell-based physiology
|
||
and may extend the clinical understanding of psychotic and cognitive symptoms
|
||
associated with neurodegenerative disorders like Alzheimer’s Disease
|
||
and schizophrenia.</p></div><div id="ml071.s6"><h3>Rationale</h3><p>In recent years, major advances have been made in the discovery of highly
|
||
selective agonists of other G protein-coupled receptors (GPCRs) that act at an
|
||
allosteric site rather than the orthosteric binding site (<a class="bk_pop" href="#ml071.r9">May et al. 2003</a>). By screening for compounds that act
|
||
at an allosteric site on the receptor, it is anticipated that compounds that
|
||
selectively activate M1 versus the other muscarinic subtypes may be identified.
|
||
While allosteric M1 agonists have been identified, AC-42 and TBPB, they both
|
||
suffer from undesirable ancillary pharmacology, poor physicochemical properties,
|
||
poor pharmacokinetics and/or limited CNS penetration (<a class="bk_pop" href="#ml071.r11">Spalding et al. 2002</a>; <a class="bk_pop" href="#ml071.r8">Jones et al. 2008</a>). Thus, to truly enable the
|
||
biomedical community to dissect the relative contributions of selective M1
|
||
activation in preclinical models of AD and schizophrenia and to understand the
|
||
role of M1 in the pronounced efficacy of the M1/M4 preferring xanomeline,
|
||
improved M1 probes are required.</p><p><b><i>Screening center information</i></b></p><p>Assay Implementation and Screening</p><p><b>PubChem Bioassay Name:</b> Discovery of novel allosteric modulators of the M1 muscarinic
|
||
receptor: Agonist</p><p><b>List of PubChem bioassay identifiers generated for this screening
|
||
project (AIDS):</b>
|
||
<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/626" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-626</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1488" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1488</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1741" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1741</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1508" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1508</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1470" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1470</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1744" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1744</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1767" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1767</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1764" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1764</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1757" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1757</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1743" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1743</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1788" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID-1788</a>.</p></div></div><div id="ml071.s11"><h2 id="_ml071_s11_">PubChem Primary Assay Description</h2><ol><li class="half_rhythm"><div>Hamster Ovary (CHO) cells containing M1 receptor (ATCC#CRL-1985)
|
||
were plated at 10,000 cells/well in assay media (F12 (Ham), 10%
|
||
FBS, 2 millimolar GlutaMAX (Invitrogen), 20mM HEPES) in 384 well plates.</div></li><li class="half_rhythm"><div>The plates were incubated overnight at 37 degrees C in 5%
|
||
CO2.</div></li><li class="half_rhythm"><div>Media was removed and assay buffer (Hanks Balanced Salt Solution, 20
|
||
millimolar HEPES, 2.5 millimolar Probenecid, pH 7.4) containing 4.0
|
||
micromolar Fluo4-AM dye (Invitrogen) was added.</div></li><li class="half_rhythm"><div>Cells were incubated for 45 minutes (37 degrees C, 5% CO2) for
|
||
dye loading.</div></li><li class="half_rhythm"><div>Cell plates were loaded into the Hamamatsu FDSS equipped with 480 nanometer
|
||
excitation and 540 nanometer emission filters.</div></li><li class="half_rhythm"><div>10 micromolar test compound in assay buffer + 0.1%
|
||
DMSO was added at 5 seconds; simultaneously, the plate was kinetically
|
||
imaged.</div></li><li class="half_rhythm"><div>Subsequently, 8 nanomolar acetylcholine (EC80) in assay buffer was added at
|
||
197 seconds and imaging continued for a total of 4 minutes acquisition
|
||
time.</div></li><li class="half_rhythm"><div>0.1% DMSO, compound vehicle, and 80nM acetylcholine (ECMAX) were
|
||
added to each plate as controls.</div></li><li class="half_rhythm"><div>Compounds that stimulated an increase in intracellular calcium upon their
|
||
addition deviating by more than three standard deviations from the test
|
||
populations were identified as “hits”.</div></li></ol><div id="ml071.s12"><h3>Summary of Screen</h3><p>For the discovery of novel allosteric agonists of the M1 muscarinic receptor, we
|
||
completed the primary screen using a real-time cell-based assay against the full
|
||
65K library. The assay performed very well (Z′ averaged 0.7) and we
|
||
identified approximately 2000 putative agonist primary hits. The compounds
|
||
clustered nicely into different structural classes with multiple representative
|
||
analogs in each cluster. The agonist compounds were chosen as the focus of this
|
||
probe effort, but we previously filed a probe report for the first truly
|
||
selective small molecule M1 antagonist, <a href="https://pubchem.ncbi.nlm.nih.gov/substance/56373925" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-56373925</a>.</p><p>For the confirmation screen, 1666 agonist hits were reordered from Biofocus-DPI.
|
||
The compounds were tested in duplicate against M1/CHO cells, and in parallel,
|
||
counter screened against M4/Gqi5 CHO cells. Compounds showing initial
|
||
selectivity for M1 over M4 were tested in triplicate in a 10 point dose-response
|
||
series against the muscarinic panel (M1–M5). Compounds showing
|
||
selectivity for M1 were then tested as a concentration series in the presence of
|
||
atropine. Pubchem CID 644390 and CID 647412 were selective (>50-fold) for
|
||
M1 vs. M4 with M1 EC<sub>50</sub>s from the DPI DMSO stock of ~ 1 μM
|
||
(<a class="figpopup" href="/books/NBK47340/figure/ml071.f1/?report=objectonly" target="object" rid-figpopup="figml071f1" rid-ob="figobml071f1">Figure 1</a>) and sensitive to block
|
||
by atropine.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml071f1" co-legend-rid="figlgndml071f1"><a href="/books/NBK47340/figure/ml071.f1/?report=objectonly" target="object" title="Figure 1" class="img_link icnblk_img figpopup" rid-figpopup="figml071f1" rid-ob="figobml071f1"><img class="small-thumb" src="/books/NBK47340/bin/ml071f1.gif" src-large="/books/NBK47340/bin/ml071f1.jpg" alt="Figure 1. M1 vs. M4 selective Agonist Leads from the HTS." /></a><div class="icnblk_cntnt" id="figlgndml071f1"><h4 id="ml071.f1"><a href="/books/NBK47340/figure/ml071.f1/?report=objectonly" target="object" rid-ob="figobml071f1">Figure 1</a></h4><p class="float-caption no_bottom_margin">M1 vs. M4 selective Agonist Leads from the HTS. </p></div></div><p>We then resynthesized the two HTS leads to confirm structure and activity (<a class="figpopup" href="/books/NBK47340/figure/ml071.f11/?report=objectonly" target="object" rid-figpopup="figml071f11" rid-ob="figobml071f11">Scheme 1</a>). In the event, commercial
|
||
mono- <i>N</i>-Boc diaminoethane was acylated with either
|
||
2-chlorobenzoyl chloride (<b>2</b>) or 2-thienyl acyl chloride
|
||
(<b>3</b>) to deliver <b>4</b> and <b>5</b>, respectively.
|
||
The Boc group was removed by exposure to 4N HCl/dioxane to provide the
|
||
corresponding HCl salts <b>6</b> and <b>7</b>. Finally, a reductive
|
||
amination sequence employing ethyl-4-oxopiperidine-1-carboxylate (8) with a
|
||
polymer-supported hydride delivered the original HTS leads, CID 644390 and CID
|
||
647412 in overall yields for the three step sequence in excess of
|
||
70%.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml071f11" co-legend-rid="figlgndml071f11"><a href="/books/NBK47340/figure/ml071.f11/?report=objectonly" target="object" title="Scheme 1" class="img_link icnblk_img figpopup" rid-figpopup="figml071f11" rid-ob="figobml071f11"><img class="small-thumb" src="/books/NBK47340/bin/ml071f11.gif" src-large="/books/NBK47340/bin/ml071f11.jpg" alt="Scheme 1. Resynthesis of M1 vs. M4 selective Agonist Leads from the HTS; 70% overall yield." /></a><div class="icnblk_cntnt" id="figlgndml071f11"><h4 id="ml071.f11"><a href="/books/NBK47340/figure/ml071.f11/?report=objectonly" target="object" rid-ob="figobml071f11">Scheme 1</a></h4><p class="float-caption no_bottom_margin">Resynthesis of M1 vs. M4 selective Agonist Leads from the HTS;
|
||
70% overall yield. </p></div></div><p>The resynthesized CID 644390 and CID 647412 confirmed, with M1 EC<sub>50</sub>s
|
||
of 804 nM and 1.74 μM, respectively. Importantly, both compounds
|
||
proved to be highly selective versus M2–M5, affording no activation
|
||
at concentrations exceeding 50 μM (<a class="figpopup" href="/books/NBK47340/figure/ml071.f2/?report=objectonly" target="object" rid-figpopup="figml071f2" rid-ob="figobml071f2">Figure 2</a>). Based on this unprecedented mAChR
|
||
selectivity, we assumed these ligands must be binding at an allosteric
|
||
sitetoelicit receptor activation. Competition binding studies with the
|
||
orthosteric antagonist [<sup>3</sup>H]-NMS demonstrated
|
||
that CID 644390 (<a href="https://pubchem.ncbi.nlm.nih.gov/substance/842134" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-842134</a>) and CID 647412 (<a href="https://pubchem.ncbi.nlm.nih.gov/substance/845074" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-845074</a>) displace the
|
||
radioligand only at very high concentrations, suggesting they do in fact bind at
|
||
an allosteric site (<a class="figpopup" href="/books/NBK47340/figure/ml071.f3/?report=objectonly" target="object" rid-figpopup="figml071f3" rid-ob="figobml071f3">Figure 3</a>).
|
||
However, these initial hits do not meet MLPCN probe criteria, so we initiated a
|
||
chemical lead optimization campaign to develop a selective M1 allosteric agonist
|
||
that meets MLPCN criteria.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml071f2" co-legend-rid="figlgndml071f2"><a href="/books/NBK47340/figure/ml071.f2/?report=objectonly" target="object" title="Figure 2" class="img_link icnblk_img figpopup" rid-figpopup="figml071f2" rid-ob="figobml071f2"><img class="small-thumb" src="/books/NBK47340/bin/ml071f2.gif" src-large="/books/NBK47340/bin/ml071f2.jpg" alt="Figure 2. In vitro pharmacological profile of resynthesized CID 644390 and CID 647412 on M1–M5." /></a><div class="icnblk_cntnt" id="figlgndml071f2"><h4 id="ml071.f2"><a href="/books/NBK47340/figure/ml071.f2/?report=objectonly" target="object" rid-ob="figobml071f2">Figure 2</a></h4><p class="float-caption no_bottom_margin">In vitro pharmacological profile of resynthesized CID 644390 and CID
|
||
647412 on M1–M5. </p></div></div><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml071f3" co-legend-rid="figlgndml071f3"><a href="/books/NBK47340/figure/ml071.f3/?report=objectonly" target="object" title="Figure 3" class="img_link icnblk_img figpopup" rid-figpopup="figml071f3" rid-ob="figobml071f3"><img class="small-thumb" src="/books/NBK47340/bin/ml071f3.gif" src-large="/books/NBK47340/bin/ml071f3.jpg" alt="Figure 3. Competition Binding with [3H]-NMS." /></a><div class="icnblk_cntnt" id="figlgndml071f3"><h4 id="ml071.f3"><a href="/books/NBK47340/figure/ml071.f3/?report=objectonly" target="object" rid-ob="figobml071f3">Figure 3</a></h4><p class="float-caption no_bottom_margin">Competition Binding with
|
||
[<sup>3</sup>H]-NMS. </p></div></div></div></div><div id="ml071.s13"><h2 id="_ml071_s13_">Probe Chemical Lead Optimization Strategy</h2><p>We employed an iterative parallel synthesis approach for the optimization of CID
|
||
644390 and CID 647412, as the scaffolds were modular and readily amenable to this
|
||
approach. We simultaneously investigated three areas of the scaffold (<a class="figpopup" href="/books/NBK47340/figure/ml071.f4/?report=objectonly" target="object" rid-figpopup="figml071f4" rid-ob="figobml071f4">Figure 4</a>) employing the synthetic route
|
||
depicted in <a class="figpopup" href="/books/NBK47340/figure/ml071.f11/?report=objectonly" target="object" rid-figpopup="figml071f11" rid-ob="figobml071f11">Scheme 1</a>, and quickly
|
||
noticed (<a class="figpopup" href="/books/NBK47340/table/ml071.t1/?report=objectonly" target="object" rid-figpopup="figml071t1" rid-ob="figobml071t1">Table 1</a>), that responsive SAR
|
||
was only observed for the amide moiety (blue). No modifications to the linker region
|
||
(yellow) were tolerated, and this included substitutions along the ethyl chain,
|
||
capping either terminal nitrogen with a methyl, or homologation of the linker.
|
||
Similar flat SAR was observed for the ethyl carbamate moiety (green). No
|
||
alternatives (amides, alkyl/phenyl or differen carbamates) were tolerated. Of 40
|
||
analogs prepared, only six (15%) displayed any measurable M1 activity.
|
||
However, the six active compounds all possessed sub-micromolar EC<sub>50</sub>s at
|
||
M1 (152 nM to 459 nM), and all were completely selective versus M2–M5
|
||
(EC<sub>50</sub>s >50 μM). Thus, all six met the minimum
|
||
criteria for an MLPCN probe. We then counter-screened the six compounds versus D2 as
|
||
a mean to distinguish the probe candidates, as activity at D2 would greatly diminish
|
||
the value of an M1 probe to explore the role of selective M1 activation in
|
||
modulating the pathophysiology of schizophrenia. This counter-screen eliminated four
|
||
potential probe compounds (low micromolar D2 IC<sub>50</sub>s) and left CID 650889
|
||
and CID 25010776 as contenders (D2 IC<sub>50</sub>s >10 μM). The
|
||
two potential probes were then resynthesized and screened as the corresponding HCl
|
||
salts 650889 (25010774) and 25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>). After completing n=3 CRCs
|
||
for the potential probes, the M1 EC<sub>50</sub>s for CID 25010774 and CID 25010775
|
||
(<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>) were 152±8.4 nM (85.1±3.3% ACh Max) and
|
||
198±13.2 nM (80.52±7.6% ACh Max) with complete
|
||
selectivity versus M2–M5 (<a class="figpopup" href="/books/NBK47340/figure/ml071.f5/?report=objectonly" target="object" rid-figpopup="figml071f5" rid-ob="figobml071f5">Figure
|
||
5a</a>). To confirm that these compounds are indeed allosteric agonists, we
|
||
evaluated them on a Y381A mutant M1 cell line. Allosteric agonists of M1 can be
|
||
differentiated from orthosteric agonists by their ability to activate the receptor
|
||
in which there is a single point mutation (Y381A) in the orthosteric binding site
|
||
that renders the receptor insensitive to acetylcholine or orthosteric agonists. As
|
||
shown in <a class="figpopup" href="/books/NBK47340/figure/ml071.f5/?report=objectonly" target="object" rid-figpopup="figml071f5" rid-ob="figobml071f5">Figure 5b</a>, the Y381A mutation
|
||
causes three order of magnitude right-shift in the ACh CRC and TBPB, the
|
||
prototypical M1 allosteric agonist, retains some efficacy on this line. In contrast,
|
||
the initial HTS leads (CID 644390 and CID 647412) and the two probe candidates (CID
|
||
25010774 and CID 25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>)) remain fully efficacious on this mutant line. In
|
||
fact, the EC<sub>50</sub>s for CID 650889 and CID 25010776 shift less than 2-fold
|
||
(Y381A M1 EC<sub>50</sub>s = 304±56 nM and
|
||
379±91 nM, respectively). These data, coupled with competition binding
|
||
studies with [<sup>3</sup>H]-NMS (<a class="figpopup" href="/books/NBK47340/figure/ml071.f6/?report=objectonly" target="object" rid-figpopup="figml071f6" rid-ob="figobml071f6">Figure 6</a>), clearly indicate that these new M1 agonists
|
||
activate the receptor through binding at an allosteric site.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml071f4" co-legend-rid="figlgndml071f4"><a href="/books/NBK47340/figure/ml071.f4/?report=objectonly" target="object" title="Figure 4" class="img_link icnblk_img figpopup" rid-figpopup="figml071f4" rid-ob="figobml071f4"><img class="small-thumb" src="/books/NBK47340/bin/ml071f4.gif" src-large="/books/NBK47340/bin/ml071f4.jpg" alt="Figure 4. Chemical Optimization Plan for CID 644390 and CID 647412." /></a><div class="icnblk_cntnt" id="figlgndml071f4"><h4 id="ml071.f4"><a href="/books/NBK47340/figure/ml071.f4/?report=objectonly" target="object" rid-ob="figobml071f4">Figure 4</a></h4><p class="float-caption no_bottom_margin">Chemical Optimization Plan for CID 644390 and CID 647412. </p></div></div><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml071t1"><a href="/books/NBK47340/table/ml071.t1/?report=objectonly" target="object" title="Table 1" class="img_link icnblk_img figpopup" rid-figpopup="figml071t1" rid-ob="figobml071t1"><img class="small-thumb" src="/books/NBK47340/table/ml071.t1/?report=thumb" src-large="/books/NBK47340/table/ml071.t1/?report=previmg" alt="Table 1. Structure-Activity Relationships for Analogs of CID 644390 and CID 647412." /></a><div class="icnblk_cntnt"><h4 id="ml071.t1"><a href="/books/NBK47340/table/ml071.t1/?report=objectonly" target="object" rid-ob="figobml071t1">Table 1</a></h4><p class="float-caption no_bottom_margin">Structure-Activity Relationships for Analogs of CID 644390 and CID
|
||
647412. </p></div></div><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml071f5" co-legend-rid="figlgndml071f5"><a href="/books/NBK47340/figure/ml071.f5/?report=objectonly" target="object" title="Figure 5" class="img_link icnblk_img figpopup" rid-figpopup="figml071f5" rid-ob="figobml071f5"><img class="small-thumb" src="/books/NBK47340/bin/ml071f5.gif" src-large="/books/NBK47340/bin/ml071f5.jpg" alt="Figure 5. A) Full CRCs for CID 25010774 and CID 25010775 for wt M1–M5; B) CRCs for TBPB, ACh, the initial HTS leads CID 644390 and CID 647412 and the probe candidates CID 25010774 and CID 25010775." /></a><div class="icnblk_cntnt" id="figlgndml071f5"><h4 id="ml071.f5"><a href="/books/NBK47340/figure/ml071.f5/?report=objectonly" target="object" rid-ob="figobml071f5">Figure 5</a></h4><p class="float-caption no_bottom_margin">A) Full CRCs for CID 25010774 and CID 25010775 for wt M1–M5;
|
||
B) CRCs for TBPB, ACh, the initial HTS leads CID 644390 and CID 647412 and
|
||
the probe candidates CID 25010774 and CID 25010775. </p></div></div><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml071f6" co-legend-rid="figlgndml071f6"><a href="/books/NBK47340/figure/ml071.f6/?report=objectonly" target="object" title="Figure 6" class="img_link icnblk_img figpopup" rid-figpopup="figml071f6" rid-ob="figobml071f6"><img class="small-thumb" src="/books/NBK47340/bin/ml071f6.gif" src-large="/books/NBK47340/bin/ml071f6.jpg" alt="Figure 6. Competition [3H]-NMS binding for the initial HTS leads CID 644390 and CID 647412 and the probe candidates CID 25010774 and CID 25010775 relative to the orthosteric antagonist atropine." /></a><div class="icnblk_cntnt" id="figlgndml071f6"><h4 id="ml071.f6"><a href="/books/NBK47340/figure/ml071.f6/?report=objectonly" target="object" rid-ob="figobml071f6">Figure 6</a></h4><p class="float-caption no_bottom_margin">Competition [<sup>3</sup>H]-NMS binding for the
|
||
initial HTS leads CID 644390 and CID 647412 and the probe candidates CID
|
||
25010774 and CID 25010775 relative to the orthosteric antagonist
|
||
atropine. </p></div></div><p>A major liability with TBPB, the leading allosteric M1 agonist in the literature to
|
||
date, is the fact that TBPB is a pan-mAChR antagonist at higher concentrations at
|
||
M2–M5. This undesired ancillary pharmacology greatly limits the utility
|
||
of TBPB to cleanly dissect the role of selective M1 activation <i>in
|
||
vitro</i> and <i>in vivo</i>. Gratifyingly (<a class="figpopup" href="/books/NBK47340/figure/ml071.f7/?report=objectonly" target="object" rid-figpopup="figml071f7" rid-ob="figobml071f7">Figure 7</a>), the initial HTS leads (CID 644390 and CID
|
||
647412) and the two probe candidates (CID 25010774 and CID 25010776) show no
|
||
significant antagonism of M2–M5 and further distinguish themselves as
|
||
superior M1 allosteric agonist probes.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml071f7" co-legend-rid="figlgndml071f7"><a href="/books/NBK47340/figure/ml071.f7/?report=objectonly" target="object" title="Figure 7" class="img_link icnblk_img figpopup" rid-figpopup="figml071f7" rid-ob="figobml071f7"><img class="small-thumb" src="/books/NBK47340/bin/ml071f7.gif" src-large="/books/NBK47340/bin/ml071f7.jpg" alt="Figure 7. Functional antagonism comparison of TBPB and for the initial HTS leads CID 644390 and CID 647412 and the probe candidates CID 25010774 and CID 25010775." /></a><div class="icnblk_cntnt" id="figlgndml071f7"><h4 id="ml071.f7"><a href="/books/NBK47340/figure/ml071.f7/?report=objectonly" target="object" rid-ob="figobml071f7">Figure 7</a></h4><p class="float-caption no_bottom_margin">Functional antagonism comparison of TBPB and for the initial HTS leads
|
||
CID 644390 and CID 647412 and the probe candidates CID 25010774 and CID
|
||
25010775. </p></div></div><p>In parallel, to further confirm that the two probe candidates (CID 25010774 and CID
|
||
25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>)) are indeed activating M1 at an allosteric site, we evaluated both
|
||
probe candidates against a mutant line we had in house targeting the third
|
||
extracellular loop (e3) that is far removed from the orthosteric binding site. A
|
||
(K392D/E397V/E410H) triple mutant abolishes the ability of the two probe candidates
|
||
(CID 25010774 and CID 25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>)) to activate M1 (<a class="figpopup" href="/books/NBK47340/figure/ml071.f8/?report=objectonly" target="object" rid-figpopup="figml071f8" rid-ob="figobml071f8">Figure 8</a>). Evaluation of the three individual single point
|
||
mutants demonstrated that the E401H is critical for M1 activation with the two probe
|
||
candidates (CID 25010774 and CID 25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>)), suggesting they bind to the e3
|
||
loop of M1, far removed from the orthosteric binding site.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml071f8" co-legend-rid="figlgndml071f8"><a href="/books/NBK47340/figure/ml071.f8/?report=objectonly" target="object" title="Figure 8" class="img_link icnblk_img figpopup" rid-figpopup="figml071f8" rid-ob="figobml071f8"><img class="small-thumb" src="/books/NBK47340/bin/ml071f8.gif" src-large="/books/NBK47340/bin/ml071f8.jpg" alt="Figure 8. Further data with an M1 triple mutant (K392D/E397V/E401H) for the probe candidates CID 25010774 and CID 25010775, and identification of a lone single point mutant (E401H) that diminishes M1 activation." /></a><div class="icnblk_cntnt" id="figlgndml071f8"><h4 id="ml071.f8"><a href="/books/NBK47340/figure/ml071.f8/?report=objectonly" target="object" rid-ob="figobml071f8">Figure 8</a></h4><p class="float-caption no_bottom_margin">Further data with an M1 triple mutant (K392D/E397V/E401H) for the probe
|
||
candidates CID 25010774 and CID 25010775, and identification of a lone
|
||
single point mutant (E401H) that diminishes M1 activation. </p></div></div><p>At this point, the Lead Profiling Screen (68 GPCRs, ion channels and transporters)
|
||
from MDS Pharma was performed on the two probe candidates (CID 25010774 and CID
|
||
25010776) to attempt to distinguish which would be promoted to probe status. CID
|
||
25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>) (<a href="https://pubchem.ncbi.nlm.nih.gov/substance/56353039" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-56353039</a>) possessed superior ancillary profile to both TBPB
|
||
and CID 25010774 (<a class="figpopup" href="/books/NBK47340/table/ml071.f9/?report=objectonly" target="object" rid-figpopup="figml071f9" rid-ob="figobml071f9">Figure 9</a>), with only
|
||
three activities >50% at 10 μM, and was thus declared
|
||
an MLPCN probe. CID 25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>) has the following IUPAC nomenclature: ethyl
|
||
4-(2-methylbenzamido)ethylamino) piperidine-1-carboxylate.</p><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml071f9"><a href="/books/NBK47340/table/ml071.f9/?report=objectonly" target="object" title="Figure 9" class="img_link icnblk_img figpopup" rid-figpopup="figml071f9" rid-ob="figobml071f9"><img class="small-thumb" src="/books/NBK47340/table/ml071.f9/?report=thumb" src-large="/books/NBK47340/table/ml071.f9/?report=previmg" alt="Figure 9. MDS Pharma Lead Profiling Screen of 68 GPCRs, ion channels and transporters against CID 25010775 (SID-56353039) at a concentration of 10 μM." /></a><div class="icnblk_cntnt"><h4 id="ml071.f9"><a href="/books/NBK47340/table/ml071.f9/?report=objectonly" target="object" rid-ob="figobml071f9">Figure 9</a></h4><p class="float-caption no_bottom_margin">MDS Pharma Lead Profiling Screen of 68 GPCRs, ion channels and transporters against CID 25010775 (SID-56353039) at a concentration of 10 μM. </p></div></div><div id="ml071.fu2" class="figure bk_fig"><div class="graphic"><img src="/books/NBK47340/bin/ml071fu8.jpg" alt="CID 25010776 A Best-in-Class M1 allosteric agonist probe." /></div><h3><span class="title">CID 25010776 A Best-in-Class M1 allosteric agonist probe</span></h3></div><p>Finally, a brain/plasma study was conducted to determine if our M1 allosteric agonist
|
||
probe (CID 25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>)) was centrally penetrant after systemic dosing, as this
|
||
would add significant value to the probe for the biomedical research community. When
|
||
converted to the mono- HCl salt, CID 25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>) displayed excellent solubility
|
||
across pharmaceutically acceptable vehicles, as well as DMSO (> 100 mM) and
|
||
water (homogeneous solutions up to 25 mg/mL). For the brain/plasma study, CID
|
||
25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>) (<a href="https://pubchem.ncbi.nlm.nih.gov/substance/56353039" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID-56353039</a>) as the mono-HCl salt, was dosed in water at 10 mg/kg
|
||
i.p. to Sprauge-Dawley rats (<a class="figpopup" href="/books/NBK47340/figure/ml071.f10/?report=objectonly" target="object" rid-figpopup="figml071f10" rid-ob="figobml071f10">Figure 10</a>).
|
||
A brain/plasma ratio of 4.2 was observed, with the compound preferentially
|
||
portioning into the brain. This is an excellent profile for a CNS agent, with brain
|
||
levels in the μM range for up to 5 hours post dose - >8-fold
|
||
above the EC<sub>50</sub> for M1 activation. Importantly, the animals were closely
|
||
monitored, and were healthy, with no signs of classical pan-mAChR activation (SLUD
|
||
– salivation, lacrimation, urination and defecation) indicating that the
|
||
in vitro mAChR selectivity profile was mirrored in vivo. Thus, CID 25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>)
|
||
has utility as both an <i>in vitro</i> and <i>in vivo</i> probe
|
||
for selective M1 activation by an allosteric mechanism.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml071f10" co-legend-rid="figlgndml071f10"><a href="/books/NBK47340/figure/ml071.f10/?report=objectonly" target="object" title="Figure 10" class="img_link icnblk_img figpopup" rid-figpopup="figml071f10" rid-ob="figobml071f10"><img class="small-thumb" src="/books/NBK47340/bin/ml071f10.gif" src-large="/books/NBK47340/bin/ml071f10.jpg" alt="Figure 10. Brain/plasma study with SID-56353039." /></a><div class="icnblk_cntnt" id="figlgndml071f10"><h4 id="ml071.f10"><a href="/books/NBK47340/figure/ml071.f10/?report=objectonly" target="object" rid-ob="figobml071f10">Figure 10</a></h4><p class="float-caption no_bottom_margin">Brain/plasma study with SID-56353039. </p></div></div><p><a class="figpopup" href="/books/NBK47340/figure/ml071.f12/?report=objectonly" target="object" rid-figpopup="figml071f12" rid-ob="figobml071f12">Scheme 2</a> below highlights the optimized
|
||
route to prepare CID 25010776 as the mono-HCl salt CID 25010775 (<a href="/pcsubstance/?term=ML071[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML071</a>) in
|
||
82.5% overall yield. All of the reagents are commercially available from
|
||
Aldrich Chemical company. <b>MLS#s:</b> MLS002279948,
|
||
MLS002279949, MLS002279950, MLS002279951, MLS002279952, MLS002279953</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml071f12" co-legend-rid="figlgndml071f12"><a href="/books/NBK47340/figure/ml071.f12/?report=objectonly" target="object" title="Scheme 2" class="img_link icnblk_img figpopup" rid-figpopup="figml071f12" rid-ob="figobml071f12"><img class="small-thumb" src="/books/NBK47340/bin/ml071f12.gif" src-large="/books/NBK47340/bin/ml071f12.jpg" alt="Scheme 2. Optimized synthesis of CID 25010776 as free base and mono-HCl salt, 25010775: overall yield of 82.5%." /></a><div class="icnblk_cntnt" id="figlgndml071f12"><h4 id="ml071.f12"><a href="/books/NBK47340/figure/ml071.f12/?report=objectonly" target="object" rid-ob="figobml071f12">Scheme 2</a></h4><p class="float-caption no_bottom_margin">Optimized synthesis of CID 25010776 as free base and mono-HCl salt,
|
||
25010775: overall yield of 82.5%. </p></div></div></div><div id="ml071.rl1"><h2 id="_ml071_rl1_">Bibliography</h2><ol><li><div class="bk_ref" id="ml071.r1">Bodick NC, Ofen WW, Levey AI, Cutler NR, Gauthier SG, Satlin A, Shannon HE, Tollefson GD, Rasmussen K, Bymster FP, Hurley DJ, Potter WZ, Paul SM. Effects of xanomeline, a selective muscarinic receptor agonist,
|
||
on cognitive function and behavioral symptoms in Alzheimer’s
|
||
disease. <span><span class="ref-journal">Arch. Neurol. </span>1997;<span class="ref-vol">54</span>:465–473.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/9109749" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 9109749</span></a>]</div></li><li><div class="bk_ref" id="ml071.r2">Bonner TI, Buckley NJ, Young AC, Brann MR. Identification of a family of muscarinic acetylcholine receptor
|
||
gene. <span><span class="ref-journal">Science. </span><span class="ref-vol">1987</span><span class="ref-vol">237</span>:527–532.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/3037705" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 3037705</span></a>]</div></li><li><div class="bk_ref" id="ml071.r3">Bonner TI, Young AC, Buckley NJ, Brann MR. Cloning and expression of the human and rat m5 muscarinic
|
||
receptor gene. <span><span class="ref-journal">Neuron. </span>1988;<span class="ref-vol">1</span>:403–410.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/3272174" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 3272174</span></a>]</div></li><li><div class="bk_ref" id="ml071.r4">Bymaster FP, McKinzie DL, Felder CC, Wess J. Use of M1–M5 muscarinic receptor knockout mice as
|
||
novel tools to delineate the physiological roles of the muscarinic
|
||
cholinergic system. <span><span class="ref-journal">Neurochem. Res. </span>2003;<span class="ref-vol">28</span>:437–442.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/12675128" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 12675128</span></a>]</div></li><li><div class="bk_ref" id="ml071.r5">Caccamo A, Oddo S, Billings LM, Green KN, Martinez-Coria H, Fisher A, LaFerla FM. M1 receptors play a cnetral role in modulating AD-like pathology
|
||
in transgenic mice. <span><span class="ref-journal">Neuron. </span>2006;<span class="ref-vol">49</span>:671–682.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/16504943" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 16504943</span></a>]</div></li><li><div class="bk_ref" id="ml071.r6">Eglen RM, Choppin A, Dillon MP, Hedge S. Muscarinic receptor ligands and their therapeutic
|
||
potential. <span><span class="ref-journal">Curr. Opin. Chem. Biol. </span>1999;<span class="ref-vol">3</span>:426–432.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/10419852" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 10419852</span></a>]</div></li><li><div class="bk_ref" id="ml071.r7">Felder CC, Bymaster FP, Ward J, DeLapp N. Therapeutic opportunities for muscarinic receptors in the central
|
||
nervous system. <span><span class="ref-journal">J. Med. Chem. </span>2000;<span class="ref-vol">43</span>:4333–4353.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/11087557" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11087557</span></a>]</div></li><li><div class="bk_ref" id="ml071.r8">Jones CK, Brady AE, Davis AA, Xiang Z, Bubser M, Tantawy MN, Kane A, Bridges TM, Kennedy JP, Bradley SR, Peterson T, Baldwin RM, Kessler R, Deutch A, Lah JL, Levey AI, Lindsley CW, Conn PJ. Novel selective allosteric activator of the M1 muscarinic
|
||
acetylcholine receptor reduces amyloid processing and produces
|
||
antipsychotic-like activity in rats. <span><span class="ref-journal">J. Neurosci. </span>2008;<span class="ref-vol">28</span>(41):10422–10433.</span> [<a href="/pmc/articles/PMC2577155/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC2577155</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/18842902" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 18842902</span></a>]</div></li><li><div class="bk_ref" id="ml071.r9">May LT, Christopoulos A. Allosteric modulators of G-protein–coupled
|
||
receptors. <span><span class="ref-journal">Curr Opin Pharmacol. </span>2003;<span class="ref-vol">3</span>(5):551–6.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/14559102" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 14559102</span></a>]</div></li><li><div class="bk_ref" id="ml071.r10">Shekhar A, Potter WZ, Lightfoot J, Lienemann J, Dube S, Mallinckrodt C, Bymaster FP, McKinzie DL, Felder CC. Selective muscarinic receptor agonist xanomeline as a novel
|
||
treatment approach for schizophrenia. <span><span class="ref-journal">Am. J. Psychiatry. </span>2008;<span class="ref-vol">165</span>:1033–1039.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/18593778" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 18593778</span></a>]</div></li><li><div class="bk_ref" id="ml071.r11">Spalding TA, Trotter C, Skajaerbaek N, Messier TL, Currier EA, Burstein ES, Li D, Hacksell U, Brann MR. Discovery of an ectopic site activation site on the M1 muscarinic
|
||
receptor. <span><span class="ref-journal">Mol. Pharm. </span>2002;<span class="ref-vol">61</span>:1297–1302.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/12021390" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 12021390</span></a>]</div></li></ol></div><div id="bk_toc_contnr"></div></div></div>
|
||
<div class="post-content"><div><div class="half_rhythm"><a href="/books/about/copyright/">Copyright Notice</a></div><div class="small"><span class="label">Bookshelf ID: NBK47340</span><span class="label">PMID: <a href="https://pubmed.ncbi.nlm.nih.gov/21433356" title="PubMed record of this page" ref="pagearea=meta&targetsite=entrez&targetcat=link&targettype=pubmed">21433356</a></span></div><div style="margin-top:2em" class="bk_noprnt"><a class="bk_cntns" href="/books/n/mlprobe/">Contents</a><div class="pagination bk_noprnt"><a class="active page_link prev" href="/books/n/mlprobe/ml075/" title="Previous page in this title">< Prev</a><span class="inactive page_link next">Next ></span></div></div></div></div>
|
||
|
||
</div>
|
||
|
||
<!-- Custom content below content -->
|
||
<div class="col4">
|
||
|
||
</div>
|
||
|
||
|
||
<!-- Book content -->
|
||
|
||
<!-- Custom contetnt below bottom nav -->
|
||
<div class="col5">
|
||
|
||
</div>
|
||
</div>
|
||
|
||
<div id="rightcolumn" class="four_col col last">
|
||
<!-- Custom content above discovery portlets -->
|
||
<div class="col6">
|
||
<div id="ncbi_share_book"><a href="#" class="ncbi_share" data-ncbi_share_config="popup:false,shorten:true" ref="id=NBK47340&db=books">Share</a></div>
|
||
|
||
</div>
|
||
<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/NBK47340/?report=reader">PubReader</a></li><li><a href="/books/NBK47340/?report=printable">Print View</a></li><li><a data-jig="ncbidialog" href="#_ncbi_dlg_citbx_NBK47340" data-jigconfig="width:400,modal:true">Cite this Page</a><div id="_ncbi_dlg_citbx_NBK47340" style="display:none" title="Cite this Page"><div class="bk_tt">Lindsley C, Lewis M, Weaver D. Discovery of a Highly Selective in vitro and in vivo M1 Allosteric Agonist Probe. 2008 Dec 3 [Updated 2010 Oct 4]. 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/NBK47340/pdf/Bookshelf_NBK47340.pdf">PDF version of this page</a> (609K)</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="#ml071.s2" ref="log$=inpage&link_id=inpage">Probe Structure & Characteristics</a></li><li><a href="#ml071.s3" ref="log$=inpage&link_id=inpage">Recommendations for the scientific use of this probe</a></li><li><a href="#ml071.s11" ref="log$=inpage&link_id=inpage">PubChem Primary Assay Description</a></li><li><a href="#ml071.s13" ref="log$=inpage&link_id=inpage">Probe Chemical Lead Optimization Strategy</a></li><li><a href="#ml071.rl1" ref="log$=inpage&link_id=inpage">Bibliography</a></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Related information</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="discovery_db_links" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pmc&DbFrom=books&Cmd=Link&LinkName=books_pmc_refs&IdsFromResult=2358500" ref="log$=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=2358500" ref="log$=recordlinks">PubChem BioAssay for Chemical Probe</a><div class="brieflinkpop offscreen_noflow">PubChem BioAssay records reporting screening data for the development of the chemical probe(s) described in this book chapter</div></li><li class="brieflinkpopper"><a class="brieflinkpopperctrl" href="/books/?Db=pcsubstance&DbFrom=books&Cmd=Link&LinkName=books_pcsubstance&IdsFromResult=2358500" ref="log$=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=2358500" ref="log$=recordlinks">PubMed</a><div class="brieflinkpop offscreen_noflow">Links to PubMed</div></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Similar articles in PubMed</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="PBooksDiscovery_RA" id="Shutter"></a></div><div class="portlet_content"><ul><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/21433387" ref="ordinalpos=1&linkpos=1&log$=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> Discovery and development of the a highly selective M(1) Positive Allosteric Modulator (PAM).</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> Discovery and development of the a highly selective M(1) Positive Allosteric Modulator (PAM).<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Bridges TM, Lewis LM, Dawson ES, Weaver CD, Lindsley CW. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Probe Reports from the NIH Molecular Libraries Program. 2010</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/21433394" ref="ordinalpos=1&linkpos=2&log$=relatedreviews&logdbfrom=pubmed"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> Discovery and development of a second highly selective M(1) Positive Allosteric Modulator (PAM).</a><span class="source">[Probe Reports from the NIH Mol...]</span><div class="brieflinkpop offscreen_noflow"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> Discovery and development of a second highly selective M(1) Positive Allosteric Modulator (PAM).<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Bridges TM, Reid PR, Lewis LM, Dawson ES, Weaver CD, Wood MR, Lindsley CW. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Probe Reports from the NIH Molecular Libraries Program. 2010</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/21433380" ref="ordinalpos=1&linkpos=3&log$=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> Discovery of a Highly Selective in vitro and in vivo M4 Positive Allosteric Modulator (PAM).</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> Discovery of a Highly Selective in vitro and in vivo M4 Positive Allosteric Modulator (PAM).<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Lewis LM, Bridges TM, Niswender CM, Weaver CD, Lindsley CW. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Probe Reports from the NIH Molecular Libraries Program. 2010</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/21433383" ref="ordinalpos=1&linkpos=4&log$=relatedreviews&logdbfrom=pubmed"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> Discovery of the first mAChR 5 (M5) selective ligand, an M5 Positive Allosteric Modulator (PAM).</a><span class="source">[Probe Reports from the NIH Mol...]</span><div class="brieflinkpop offscreen_noflow"><span xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="invert">Review</span> Discovery of the first mAChR 5 (M5) selective ligand, an M5 Positive Allosteric Modulator (PAM).<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Bridges TM, Lewis LM, Weaver CD, Lindsley CW. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">Probe Reports from the NIH Molecular Libraries Program. 2010</em></div></div></li><li class="brieflinkpopper two_line"><a class="brieflinkpopperctrl" href="/pubmed/21558436" ref="ordinalpos=1&linkpos=5&log$=relatedarticles&logdbfrom=pubmed">Pharmacological characterization of LY593093, an M1 muscarinic acetylcholine receptor-selective partial orthosteric agonist.</a><span class="source">[J Pharmacol Exp Ther. 2011]</span><div class="brieflinkpop offscreen_noflow">Pharmacological characterization of LY593093, an M1 muscarinic acetylcholine receptor-selective partial orthosteric agonist.<div class="brieflinkpopdesc"><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="author">Watt ML, Schober DA, Hitchcock S, Liu B, Chesterfield AK, McKinzie D, Felder CC. </em><em xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="cit">J Pharmacol Exp Ther. 2011 Aug; 338(2):622-32. Epub 2011 May 10.</em></div></div></li></ul><a class="seemore" href="/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed_reviews&uid=21433356" ref="ordinalpos=1&log$=relatedreviews_seeall&logdbfrom=pubmed">See reviews...</a><a class="seemore" href="/sites/entrez?db=pubmed&cmd=link&linkname=pubmed_pubmed&uid=21433356" ref="ordinalpos=1&log$=relatedarticles_seeall&logdbfrom=pubmed">See all...</a></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Recent Activity</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="recent_activity" id="Shutter"></a></div><div class="portlet_content"><div xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" id="HTDisplay" class=""><div class="action"><a href="javascript:historyDisplayState('ClearHT')">Clear</a><a href="javascript:historyDisplayState('HTOff')" class="HTOn">Turn Off</a><a href="javascript:historyDisplayState('HTOn')" class="HTOff">Turn On</a></div><ul id="activity"><li class="ra_rcd ralinkpopper two_line"><a class="htb ralinkpopperctrl" ref="log$=activity&linkpos=1" href="/portal/utils/pageresolver.fcgi?recordid=67d66c3c84f3725e592e5210">Discovery of a Highly Selective in vitro and in vivo M1 Allosteric Agonist Probe...</a><div class="ralinkpop offscreen_noflow">Discovery of a Highly Selective in vitro and in vivo M1 Allosteric Agonist Probe - Probe Reports from the NIH Molecular Libraries Program<div class="brieflinkpopdesc"></div></div><div class="tertiary"></div></li><li class="ra_rcd ralinkpopper two_line"><a class="htb ralinkpopperctrl" ref="log$=activity&linkpos=2" href="/portal/utils/pageresolver.fcgi?recordid=67d66c3b67c23b31e0b42d82">Probe Report for NPY-Y2 Receptor Antagonists - Probe Reports from the NIH Molecu...</a><div class="ralinkpop offscreen_noflow">Probe Report for NPY-Y2 Receptor Antagonists - Probe Reports from the NIH Molecular Libraries Program<div class="brieflinkpopdesc"></div></div><div class="tertiary"></div></li><li class="ra_rcd ralinkpopper two_line"><a class="htb ralinkpopperctrl" ref="log$=activity&linkpos=3" href="/portal/utils/pageresolver.fcgi?recordid=67d66c3a84f3725e592e4d28">Inhibitors of Protein Folding: DnaK - Probe Reports from the NIH Molecular Libra...</a><div class="ralinkpop offscreen_noflow">Inhibitors of Protein Folding: DnaK - Probe Reports from the NIH Molecular Libraries Program<div class="brieflinkpopdesc"></div></div><div class="tertiary"></div></li><li class="ra_rcd ralinkpopper two_line"><a class="htb ralinkpopperctrl" ref="log$=activity&linkpos=4" href="/portal/utils/pageresolver.fcgi?recordid=67d66c392f30673f7bf96462">Discovery of a Highly Selective KCC2 Antagonist - Probe Reports from the NIH Mol...</a><div class="ralinkpop offscreen_noflow">Discovery of a Highly Selective KCC2 Antagonist - Probe Reports from the NIH Molecular Libraries Program<div class="brieflinkpopdesc"></div></div><div class="tertiary"></div></li><li class="ra_rcd ralinkpopper two_line"><a class="htb ralinkpopperctrl" ref="log$=activity&linkpos=5" href="/portal/utils/pageresolver.fcgi?recordid=67d66c3884f3725e592e45c3">Identification of Functionally Selective Small Molecule Antagonists of the Neuro...</a><div class="ralinkpop offscreen_noflow">Identification of Functionally Selective Small Molecule Antagonists of the Neuropeptide-S Receptor: Naphthopyranopyrimidines - Probe Reports from the NIH Molecular Libraries Program<div class="brieflinkpopdesc"></div></div><div class="tertiary"></div></li></ul><p class="HTOn">Your browsing activity is empty.</p><p class="HTOff">Activity recording is turned off.</p><p id="turnOn" class="HTOff"><a href="javascript:historyDisplayState('HTOn')">Turn recording back on</a></p><a class="seemore" href="/sites/myncbi/recentactivity">See more...</a></div></div></div>
|
||
|
||
<!-- Custom content below discovery portlets -->
|
||
<div class="col7">
|
||
|
||
</div>
|
||
</div>
|
||
</div>
|
||
|
||
<!-- Custom content after all -->
|
||
<div class="col8">
|
||
|
||
</div>
|
||
<div class="col9">
|
||
|
||
</div>
|
||
|
||
<script type="text/javascript" src="/corehtml/pmc/js/jquery.scrollTo-1.4.2.js"></script>
|
||
<script type="text/javascript">
|
||
(function($){
|
||
$('.skiplink').each(function(i, item){
|
||
var href = $($(item).attr('href'));
|
||
href.attr('tabindex', '-1').addClass('skiptarget'); // ensure the target can receive focus
|
||
$(item).on('click', function(event){
|
||
event.preventDefault();
|
||
$.scrollTo(href, 0, {
|
||
onAfter: function(){
|
||
href.focus();
|
||
}
|
||
});
|
||
});
|
||
});
|
||
})(jQuery);
|
||
</script>
|
||
</div>
|
||
<div class="bottom">
|
||
|
||
<div id="NCBIFooter_dynamic">
|
||
<!--<component id="Breadcrumbs" label="breadcrumbs"/>
|
||
<component id="Breadcrumbs" label="helpdesk"/>-->
|
||
|
||
</div>
|
||
|
||
<div class="footer" id="footer">
|
||
<section class="icon-section">
|
||
<div id="icon-section-header" class="icon-section_header">Follow NCBI</div>
|
||
<div class="grid-container container">
|
||
<div class="icon-section_container">
|
||
<a class="footer-icon" id="footer_twitter" href="https://twitter.com/ncbi" aria-label="Twitter"><svg xmlns="http://www.w3.org/2000/svg" data-name="Layer 1" viewBox="0 0 300 300">
|
||
<defs>
|
||
<style>
|
||
.cls-11 {
|
||
fill: #737373;
|
||
}
|
||
</style>
|
||
</defs>
|
||
<title>Twitter</title>
|
||
<path class="cls-11" d="M250.11,105.48c-7,3.14-13,3.25-19.27.14,8.12-4.86,8.49-8.27,11.43-17.46a78.8,78.8,0,0,1-25,9.55,39.35,39.35,0,0,0-67,35.85,111.6,111.6,0,0,1-81-41.08A39.37,39.37,0,0,0,81.47,145a39.08,39.08,0,0,1-17.8-4.92c0,.17,0,.33,0,.5a39.32,39.32,0,0,0,31.53,38.54,39.26,39.26,0,0,1-17.75.68,39.37,39.37,0,0,0,36.72,27.3A79.07,79.07,0,0,1,56,223.34,111.31,111.31,0,0,0,116.22,241c72.3,0,111.83-59.9,111.83-111.84,0-1.71,0-3.4-.1-5.09C235.62,118.54,244.84,113.37,250.11,105.48Z">
|
||
</path>
|
||
</svg></a>
|
||
<a class="footer-icon" id="footer_facebook" href="https://www.facebook.com/ncbi.nlm" aria-label="Facebook"><svg xmlns="http://www.w3.org/2000/svg" data-name="Layer 1" viewBox="0 0 300 300">
|
||
<title>Facebook</title>
|
||
<path class="cls-11" d="M210.5,115.12H171.74V97.82c0-8.14,5.39-10,9.19-10h27.14V52l-39.32-.12c-35.66,0-42.42,26.68-42.42,43.77v19.48H99.09v36.32h27.24v109h45.41v-109h35Z">
|
||
</path>
|
||
</svg></a>
|
||
<a class="footer-icon" id="footer_linkedin" href="https://www.linkedin.com/company/ncbinlm" aria-label="LinkedIn"><svg xmlns="http://www.w3.org/2000/svg" data-name="Layer 1" viewBox="0 0 300 300">
|
||
<title>LinkedIn</title>
|
||
<path class="cls-11" d="M101.64,243.37H57.79v-114h43.85Zm-22-131.54h-.26c-13.25,0-21.82-10.36-21.82-21.76,0-11.65,8.84-21.15,22.33-21.15S101.7,78.72,102,90.38C102,101.77,93.4,111.83,79.63,111.83Zm100.93,52.61A17.54,17.54,0,0,0,163,182v61.39H119.18s.51-105.23,0-114H163v13a54.33,54.33,0,0,1,34.54-12.66c26,0,44.39,18.8,44.39,55.29v58.35H198.1V182A17.54,17.54,0,0,0,180.56,164.44Z">
|
||
</path>
|
||
</svg></a>
|
||
<a class="footer-icon" id="footer_github" href="https://github.com/ncbi" aria-label="GitHub"><svg xmlns="http://www.w3.org/2000/svg" data-name="Layer 1" viewBox="0 0 300 300">
|
||
<defs>
|
||
<style>
|
||
.cls-11,
|
||
.cls-12 {
|
||
fill: #737373;
|
||
}
|
||
|
||
.cls-11 {
|
||
fill-rule: evenodd;
|
||
}
|
||
</style>
|
||
</defs>
|
||
<title>GitHub</title>
|
||
<path class="cls-11" d="M151.36,47.28a105.76,105.76,0,0,0-33.43,206.1c5.28,1,7.22-2.3,7.22-5.09,0-2.52-.09-10.85-.14-19.69-29.42,6.4-35.63-12.48-35.63-12.48-4.81-12.22-11.74-15.47-11.74-15.47-9.59-6.56.73-6.43.73-6.43,10.61.75,16.21,10.9,16.21,10.9,9.43,16.17,24.73,11.49,30.77,8.79,1-6.83,3.69-11.5,6.71-14.14C108.57,197.1,83.88,188,83.88,147.51a40.92,40.92,0,0,1,10.9-28.39c-1.1-2.66-4.72-13.42,1-28,0,0,8.88-2.84,29.09,10.84a100.26,100.26,0,0,1,53,0C198,88.3,206.9,91.14,206.9,91.14c5.76,14.56,2.14,25.32,1,28a40.87,40.87,0,0,1,10.89,28.39c0,40.62-24.74,49.56-48.29,52.18,3.79,3.28,7.17,9.71,7.17,19.58,0,14.15-.12,25.54-.12,29,0,2.82,1.9,6.11,7.26,5.07A105.76,105.76,0,0,0,151.36,47.28Z">
|
||
</path>
|
||
<path class="cls-12" d="M85.66,199.12c-.23.52-1.06.68-1.81.32s-1.2-1.06-.95-1.59,1.06-.69,1.82-.33,1.21,1.07.94,1.6Zm-1.3-1">
|
||
</path>
|
||
<path class="cls-12" d="M90,203.89c-.51.47-1.49.25-2.16-.49a1.61,1.61,0,0,1-.31-2.19c.52-.47,1.47-.25,2.17.49s.82,1.72.3,2.19Zm-1-1.08">
|
||
</path>
|
||
<path class="cls-12" d="M94.12,210c-.65.46-1.71,0-2.37-.91s-.64-2.07,0-2.52,1.7,0,2.36.89.65,2.08,0,2.54Zm0,0"></path>
|
||
<path class="cls-12" d="M99.83,215.87c-.58.64-1.82.47-2.72-.41s-1.18-2.06-.6-2.7,1.83-.46,2.74.41,1.2,2.07.58,2.7Zm0,0">
|
||
</path>
|
||
<path class="cls-12" d="M107.71,219.29c-.26.82-1.45,1.2-2.64.85s-2-1.34-1.74-2.17,1.44-1.23,2.65-.85,2,1.32,1.73,2.17Zm0,0">
|
||
</path>
|
||
<path class="cls-12" d="M116.36,219.92c0,.87-1,1.59-2.24,1.61s-2.29-.68-2.3-1.54,1-1.59,2.26-1.61,2.28.67,2.28,1.54Zm0,0">
|
||
</path>
|
||
<path class="cls-12" d="M124.42,218.55c.15.85-.73,1.72-2,1.95s-2.37-.3-2.52-1.14.73-1.75,2-2,2.37.29,2.53,1.16Zm0,0"></path>
|
||
</svg></a>
|
||
<a class="footer-icon" id="footer_blog" href="https://ncbiinsights.ncbi.nlm.nih.gov/" aria-label="Blog">
|
||
<svg xmlns="http://www.w3.org/2000/svg" id="Layer_1" data-name="Layer 1" viewBox="0 0 40 40">
|
||
<defs><style>.cls-1{fill:#737373;}</style></defs>
|
||
<title>NCBI Insights Blog</title>
|
||
<path class="cls-1" d="M14,30a4,4,0,1,1-4-4,4,4,0,0,1,4,4Zm11,3A19,19,0,0,0,7.05,15a1,1,0,0,0-1,1v3a1,1,0,0,0,.93,1A14,14,0,0,1,20,33.07,1,1,0,0,0,21,34h3a1,1,0,0,0,1-1Zm9,0A28,28,0,0,0,7,6,1,1,0,0,0,6,7v3a1,1,0,0,0,1,1A23,23,0,0,1,29,33a1,1,0,0,0,1,1h3A1,1,0,0,0,34,33Z"></path>
|
||
</svg>
|
||
</a>
|
||
</div>
|
||
</div>
|
||
</section>
|
||
|
||
<section class="container-fluid bg-primary">
|
||
<div class="container pt-5">
|
||
<div class="row mt-3">
|
||
<div class="col-lg-3 col-12">
|
||
<p><a class="text-white" href="https://www.nlm.nih.gov/socialmedia/index.html">Connect with NLM</a></p>
|
||
<ul class="list-inline social_media">
|
||
<li class="list-inline-item"><a href="https://twitter.com/NLM_NIH" aria-label="Twitter" target="_blank" rel="noopener noreferrer"><svg xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink" version="1.1" x="0px" y="0px" viewBox="0 0 249 249" style="enable-background:new 0 0 249 249;" xml:space="preserve">
|
||
<style type="text/css">
|
||
.st20 {
|
||
fill: #FFFFFF;
|
||
}
|
||
|
||
.st30 {
|
||
fill: none;
|
||
stroke: #FFFFFF;
|
||
stroke-width: 8;
|
||
stroke-miterlimit: 10;
|
||
}
|
||
</style>
|
||
<title>Twitter</title>
|
||
<g>
|
||
<g>
|
||
<g>
|
||
<path class="st20" d="M192.9,88.1c-5,2.2-9.2,2.3-13.6,0.1c5.7-3.4,6-5.8,8.1-12.3c-5.4,3.2-11.4,5.5-17.6,6.7 c-10.5-11.2-28.1-11.7-39.2-1.2c-7.2,6.8-10.2,16.9-8,26.5c-22.3-1.1-43.1-11.7-57.2-29C58,91.6,61.8,107.9,74,116 c-4.4-0.1-8.7-1.3-12.6-3.4c0,0.1,0,0.2,0,0.4c0,13.2,9.3,24.6,22.3,27.2c-4.1,1.1-8.4,1.3-12.5,0.5c3.6,11.3,14,19,25.9,19.3 c-11.6,9.1-26.4,13.2-41.1,11.5c12.7,8.1,27.4,12.5,42.5,12.5c51,0,78.9-42.2,78.9-78.9c0-1.2,0-2.4-0.1-3.6 C182.7,97.4,189.2,93.7,192.9,88.1z"></path>
|
||
</g>
|
||
</g>
|
||
<circle class="st30" cx="124.4" cy="128.8" r="108.2"></circle>
|
||
</g>
|
||
</svg></a></li>
|
||
<li class="list-inline-item"><a href="https://www.facebook.com/nationallibraryofmedicine" aria-label="Facebook" rel="noopener noreferrer" target="_blank">
|
||
<svg xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink" version="1.1" x="0px" y="0px" viewBox="0 0 249 249" style="enable-background:new 0 0 249 249;" xml:space="preserve">
|
||
<style type="text/css">
|
||
.st10 {
|
||
fill: #FFFFFF;
|
||
}
|
||
|
||
.st110 {
|
||
fill: none;
|
||
stroke: #FFFFFF;
|
||
stroke-width: 8;
|
||
stroke-miterlimit: 10;
|
||
}
|
||
</style>
|
||
<title>Facebook</title>
|
||
<g>
|
||
<g>
|
||
<path class="st10" d="M159,99.1h-24V88.4c0-5,3.3-6.2,5.7-6.2h16.8V60l-24.4-0.1c-22.1,0-26.2,16.5-26.2,27.1v12.1H90v22.5h16.9 v67.5H135v-67.5h21.7L159,99.1z"></path>
|
||
</g>
|
||
</g>
|
||
<circle class="st110" cx="123.6" cy="123.2" r="108.2"></circle>
|
||
</svg>
|
||
</a></li>
|
||
<li class="list-inline-item"><a href="https://www.youtube.com/user/NLMNIH" aria-label="Youtube" target="_blank" rel="noopener noreferrer"><svg xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink" version="1.1" x="0px" y="0px" viewBox="0 0 249 249" style="enable-background:new 0 0 249 249;" xml:space="preserve">
|
||
<title>Youtube</title>
|
||
<style type="text/css">
|
||
.st4 {
|
||
fill: none;
|
||
stroke: #FFFFFF;
|
||
stroke-width: 8;
|
||
stroke-miterlimit: 10;
|
||
}
|
||
|
||
.st5 {
|
||
fill: #FFFFFF;
|
||
}
|
||
</style>
|
||
<circle class="st4" cx="124.2" cy="123.4" r="108.2"></circle>
|
||
<g transform="translate(0,-952.36218)">
|
||
<path class="st5" d="M88.4,1037.4c-10.4,0-18.7,8.3-18.7,18.7v40.1c0,10.4,8.3,18.7,18.7,18.7h72.1c10.4,0,18.7-8.3,18.7-18.7 v-40.1c0-10.4-8.3-18.7-18.7-18.7H88.4z M115.2,1058.8l29.4,17.4l-29.4,17.4V1058.8z"></path>
|
||
</g>
|
||
</svg></a></li>
|
||
</ul>
|
||
</div>
|
||
<div class="col-lg-3 col-12">
|
||
<p class="address_footer text-white">National Library of Medicine<br />
|
||
<a href="https://www.google.com/maps/place/8600+Rockville+Pike,+Bethesda,+MD+20894/@38.9959508,-77.101021,17z/data=!3m1!4b1!4m5!3m4!1s0x89b7c95e25765ddb:0x19156f88b27635b8!8m2!3d38.9959508!4d-77.0988323" class="text-white" target="_blank" rel="noopener noreferrer">8600 Rockville Pike<br />
|
||
Bethesda, MD 20894</a></p>
|
||
</div>
|
||
<div class="col-lg-3 col-12 centered-lg">
|
||
<p><a href="https://www.nlm.nih.gov/web_policies.html" class="text-white">Web Policies</a><br />
|
||
<a href="https://www.nih.gov/institutes-nih/nih-office-director/office-communications-public-liaison/freedom-information-act-office" class="text-white">FOIA</a><br />
|
||
<a href="https://www.hhs.gov/vulnerability-disclosure-policy/index.html" class="text-white" id="vdp">HHS Vulnerability Disclosure</a></p>
|
||
</div>
|
||
<div class="col-lg-3 col-12 centered-lg">
|
||
<p><a class="supportLink text-white" href="https://support.nlm.nih.gov/">Help</a><br />
|
||
<a href="https://www.nlm.nih.gov/accessibility.html" class="text-white">Accessibility</a><br />
|
||
<a href="https://www.nlm.nih.gov/careers/careers.html" class="text-white">Careers</a></p>
|
||
</div>
|
||
</div>
|
||
<div class="row">
|
||
<div class="col-lg-12 centered-lg">
|
||
<nav class="bottom-links">
|
||
<ul class="mt-3">
|
||
<li>
|
||
<a class="text-white" href="//www.nlm.nih.gov/">NLM</a>
|
||
</li>
|
||
<li>
|
||
<a class="text-white" href="https://www.nih.gov/">NIH</a>
|
||
</li>
|
||
<li>
|
||
<a class="text-white" href="https://www.hhs.gov/">HHS</a>
|
||
</li>
|
||
<li>
|
||
<a class="text-white" href="https://www.usa.gov/">USA.gov</a>
|
||
</li>
|
||
</ul>
|
||
</nav>
|
||
</div>
|
||
</div>
|
||
</div>
|
||
</section>
|
||
<script type="text/javascript" src="/portal/portal3rc.fcgi/rlib/js/InstrumentOmnitureBaseJS/InstrumentNCBIConfigJS/InstrumentNCBIBaseJS/InstrumentPageStarterJS.js?v=1"> </script>
|
||
<script type="text/javascript" src="/portal/portal3rc.fcgi/static/js/hfjs2.js"> </script>
|
||
</div>
|
||
</div>
|
||
</div>
|
||
<!--/.page-->
|
||
</div>
|
||
<!--/.wrap-->
|
||
</div><!-- /.twelve_col -->
|
||
</div>
|
||
<!-- /.grid -->
|
||
|
||
<span class="PAFAppResources"></span>
|
||
|
||
<!-- BESelector tab -->
|
||
|
||
|
||
|
||
<noscript><img alt="statistics" src="/stat?jsdisabled=true&ncbi_db=books&ncbi_pdid=book-part&ncbi_acc=NBK47340&ncbi_domain=mlprobe&ncbi_report=record&ncbi_type=fulltext&ncbi_objectid=&ncbi_pcid=/NBK47340/&ncbi_pagename=Discovery of a Highly Selective in vitro and in vivo M1 Allosteric Agonist Probe - Probe Reports from the NIH Molecular Libraries Program - NCBI Bookshelf&ncbi_bookparttype=chapter&ncbi_app=bookshelf" /></noscript>
|
||
|
||
|
||
<!-- usually for JS scripts at page bottom -->
|
||
<!--<component id="PageFixtures" label="styles"></component>-->
|
||
|
||
|
||
<!-- CE8B5AF87C7FFCB1_0191SID /projects/books/PBooks@9.11 portal107 v4.1.r689238 Tue, Oct 22 2024 16:10:51 -->
|
||
<span id="portal-csrf-token" style="display:none" data-token="CE8B5AF87C7FFCB1_0191SID"></span>
|
||
|
||
<script type="text/javascript" src="//static.pubmed.gov/portal/portal3rc.fcgi/4216699/js/3879255/4121861/3501987/4008961/3893018/3821238/4062932/4209313/4212053/4076480/3921943/3400083/3426610.js" snapshot="books"></script></body>
|
||
</html> |