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<script type="text/javascript" src="/corehtml/pmc/jatsreader/ptpmc_3.22/js/jr.boots.min.js"> </script><title>Chemical Genetic Analysis of Platelet Granule Secretion-Probe 3 - Probe Reports from the NIH Molecular Libraries Program - NCBI Bookshelf</title>
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<meta name="citation_inbook_title" content="Probe Reports from the NIH Molecular Libraries Program [Internet]">
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<meta name="citation_title" content="Chemical Genetic Analysis of Platelet Granule Secretion-Probe 3">
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<meta name="citation_publisher" content="National Center for Biotechnology Information (US)">
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<meta name="citation_date" content="2011/02/10">
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<meta name="citation_author" content="Lynn VerPlank">
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<meta name="citation_author" content="Chris Dockendorff">
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<meta name="citation_author" content="Joseph Negri">
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<meta name="citation_author" content="Jose R Perez">
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<meta name="citation_author" content="James Dilks">
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<meta name="citation_author" content="Lawrence MacPherson">
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<meta name="citation_author" content="Michelle Palmer">
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<meta name="citation_author" content="Robert Flaumenhaft">
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<meta name="citation_author" content="Stuart L Schreiber">
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<meta name="citation_pmid" content="21634078">
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<meta name="citation_fulltext_html_url" content="https://www.ncbi.nlm.nih.gov/books/NBK55066/">
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<meta name="DC.Title" content="Chemical Genetic Analysis of Platelet Granule Secretion-Probe 3">
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<meta name="DC.Type" content="Text">
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<meta name="DC.Publisher" content="National Center for Biotechnology Information (US)">
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<meta name="DC.Contributor" content="Lynn VerPlank">
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<meta name="DC.Contributor" content="Chris Dockendorff">
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<meta name="DC.Contributor" content="Joseph Negri">
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<meta name="DC.Contributor" content="Jose R Perez">
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<meta name="DC.Contributor" content="James Dilks">
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<meta name="DC.Contributor" content="Lawrence MacPherson">
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<meta name="DC.Contributor" content="Michelle Palmer">
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<meta name="DC.Contributor" content="Robert Flaumenhaft">
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<meta name="DC.Contributor" content="Stuart L Schreiber">
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<meta name="DC.Date" content="2011/02/10">
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<meta name="DC.Identifier" content="https://www.ncbi.nlm.nih.gov/books/NBK55066/">
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<meta name="description" content="Platelet activation occurs in response to vascular injury, which triggers an array of cascading signaling events that potentiate the formation of thrombi to control bleeding. One downstream effect of activation is the secretion of granules, which plays a role in the formation of thrombi. Platelet activation inhibitors that target various surface receptors and cytosolic pathways within platelets are well validated for inhibiting thrombosis, clinically preventing adverse cardiovascular events such as intermittent claudication and stroke, and reducing mortality and morbidity in acute myocardial infarction. These inhibitors have provided a greater understanding of the mechanisms of platelet biology. Some inhibitors are currently in use in the clinical setting or in development for the treatment of thrombotic conditions. Unfortunately, many of the characterized platelet inhibitors can cause undesired side effects in patients. None of the characterized inhibitors are known to target the granule secretion machinery. We report the outcome of a high-throughput chemical library screen and a series of secondary conformation assays to identify novel, small-molecule inhibitors of platelet activation, specifically fitting into three possible attribute classes: 1) inhibitors of the pathways required for dense granule secretion, 2) inhibitors of pathways required for granule secretion, and 3) inhibitors of upstream G protein-coupled receptor (GPCR) signaling. A primary assay was developed to measure the release of granules from platelets activated through the thrombin GPCR PAR1. Of 302,457 screened compounds, 28 prioritized compounds were selected for additional characterization in secondary specificity assays, which yielded three probe candidates. These three probe candidates met the criteria as possible modulators of GPCR signaling in platelet activation. Of these, a diaminophenyl scaffold (CID 1048267/ML161) showed acceptable potency (av. 11.2 μM ± 1.3) and inhibited P-selectin expression in a concentration-dependent manner (EC50 0.3 μM), as well as inhibiting SFLLRN-induced thrombus formation. This new GPCR-specific probe will be useful in future cell-based investigations of the complex mechanisms of platelet activation and in vivo studies as an antithrombotic agent in murine models.">
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<meta name="og:title" content="Chemical Genetic Analysis of Platelet Granule Secretion-Probe 3">
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<meta name="og:description" content="Platelet activation occurs in response to vascular injury, which triggers an array of cascading signaling events that potentiate the formation of thrombi to control bleeding. One downstream effect of activation is the secretion of granules, which plays a role in the formation of thrombi. Platelet activation inhibitors that target various surface receptors and cytosolic pathways within platelets are well validated for inhibiting thrombosis, clinically preventing adverse cardiovascular events such as intermittent claudication and stroke, and reducing mortality and morbidity in acute myocardial infarction. These inhibitors have provided a greater understanding of the mechanisms of platelet biology. Some inhibitors are currently in use in the clinical setting or in development for the treatment of thrombotic conditions. Unfortunately, many of the characterized platelet inhibitors can cause undesired side effects in patients. None of the characterized inhibitors are known to target the granule secretion machinery. We report the outcome of a high-throughput chemical library screen and a series of secondary conformation assays to identify novel, small-molecule inhibitors of platelet activation, specifically fitting into three possible attribute classes: 1) inhibitors of the pathways required for dense granule secretion, 2) inhibitors of pathways required for granule secretion, and 3) inhibitors of upstream G protein-coupled receptor (GPCR) signaling. A primary assay was developed to measure the release of granules from platelets activated through the thrombin GPCR PAR1. Of 302,457 screened compounds, 28 prioritized compounds were selected for additional characterization in secondary specificity assays, which yielded three probe candidates. These three probe candidates met the criteria as possible modulators of GPCR signaling in platelet activation. Of these, a diaminophenyl scaffold (CID 1048267/ML161) showed acceptable potency (av. 11.2 μM ± 1.3) and inhibited P-selectin expression in a concentration-dependent manner (EC50 0.3 μM), as well as inhibiting SFLLRN-induced thrombus formation. This new GPCR-specific probe will be useful in future cell-based investigations of the complex mechanisms of platelet activation and in vivo studies as an antithrombotic agent in murine models.">
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id="jr-fip-info-p"><a id="jr-fip-prev" class="wsprkl btn" title="Jump to previuos match">◀</a><button id="jr-fip-matches">no matches yet</button><a id="jr-fip-next" class="wsprkl btn" title="Jump to next match">▶</a></nav></nav></div><div id="jr-epub-interstitial" class="hidden"></div><div id="jr-content"><article data-type="main"><div class="main-content lit-style" itemscope="itemscope" itemtype="http://schema.org/CreativeWork"><div class="meta-content fm-sec"><div class="fm-sec"><h1 id="_NBK55066_"><span class="title" itemprop="name">Chemical Genetic Analysis of Platelet Granule Secretion-Probe 3</span></h1><p class="contribs">VerPlank L, Dockendorff C, Negri J, et al.</p><p class="fm-aai"><a href="#_NBK55066_pubdet_">Publication Details</a></p></div></div><div class="jig-ncbiinpagenav body-content whole_rhythm" data-jigconfig="allHeadingLevels: ['h2'],smoothScroll: false" itemprop="text"><div id="_abs_rndgid_" itemprop="description"><p>Platelet activation occurs in response to vascular injury, which triggers an array of cascading signaling events that potentiate the formation of thrombi to control bleeding. One downstream effect of activation is the secretion of granules, which plays a role in the formation of thrombi. Platelet activation inhibitors that target various surface receptors and cytosolic pathways within platelets are well validated for inhibiting thrombosis, clinically preventing adverse cardiovascular events such as intermittent claudication and stroke, and reducing mortality and morbidity in acute myocardial infarction. These inhibitors have provided a greater understanding of the mechanisms of platelet biology. Some inhibitors are currently in use in the clinical setting or in development for the treatment of thrombotic conditions. Unfortunately, many of the characterized platelet inhibitors can cause undesired side effects in patients. None of the characterized inhibitors are known to target the granule secretion machinery. We report the outcome of a high-throughput chemical library screen and a series of secondary conformation assays to identify novel, small-molecule inhibitors of platelet activation, specifically fitting into three possible attribute classes: 1) inhibitors of the pathways required for dense granule secretion, 2) inhibitors of pathways required for granule secretion, and 3) inhibitors of upstream G protein-coupled receptor (GPCR) signaling. A primary assay was developed to measure the release of granules from platelets activated through the thrombin GPCR PAR1. Of 302,457 screened compounds, 28 prioritized compounds were selected for additional characterization in secondary specificity assays, which yielded three probe candidates. These three probe candidates met the criteria as possible modulators of GPCR signaling in platelet activation. Of these, a diaminophenyl scaffold (CID 1048267/<a href="/pcsubstance/?term=ML161[synonym]" ref="pagearea=abstract&targetsite=entrez&targetcat=term&targettype=pubchem">ML161</a>) showed acceptable potency (av. 11.2 μM ± 1.3) and inhibited P-selectin expression in a concentration-dependent manner (EC<sub>50</sub> 0.3 μM), as well as inhibiting SFLLRN-induced thrombus formation. This new GPCR-specific probe will be useful in future cell-based investigations of the complex mechanisms of platelet activation and <i>in vivo</i> studies as an antithrombotic agent in murine models.</p></div><div class="h2"></div><p><b>Assigned Assay Grant No:</b> 1 RO3 DA026209-01</p><p><b>Screening Center Name and PI:</b> Broad Institute Probe Development Center, Stuart Schreiber, PhD</p><p><b>Chemistry Center Name and PI:</b> Broad Institute Probe Development Center, Stuart Schreiber, PhD</p><p><b>Assay Submitter and Institution:</b> Robert Flaumenhaft, Beth Israel Deaconess Medical Center, Boston, MA</p><p><b>PubChem Summary Bioassay Identifier (AID):</b>
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<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1678" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 1678</a></p><div id="ml161.s1"><h2 id="_ml161_s1_">Probe Structure and Characteristics</h2><div id="ml161.fu1" class="figure"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu1.jpg" alt="Image ml161fu1" /></div></div><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml161tu1"><a href="/books/NBK55066/table/ml161.tu1/?report=objectonly" target="object" title="Table" class="img_link icnblk_img figpopup" rid-figpopup="figml161tu1" rid-ob="figobml161tu1"><img class="small-thumb" src="/books/NBK55066/table/ml161.tu1/?report=thumb" src-large="/books/NBK55066/table/ml161.tu1/?report=previmg" alt="Image " /></a><div class="icnblk_cntnt"><h4 id="ml161.tu1"><a href="/books/NBK55066/table/ml161.tu1/?report=objectonly" target="object" rid-ob="figobml161tu1">Table</a></h4></div></div><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml161tu2"><a href="/books/NBK55066/table/ml161.tu2/?report=objectonly" target="object" title="Table" class="img_link icnblk_img figpopup" rid-figpopup="figml161tu2" rid-ob="figobml161tu2"><img class="small-thumb" src="/books/NBK55066/table/ml161.tu2/?report=thumb" src-large="/books/NBK55066/table/ml161.tu2/?report=previmg" alt="Image " /></a><div class="icnblk_cntnt"><h4 id="ml161.tu2"><a href="/books/NBK55066/table/ml161.tu2/?report=objectonly" target="object" rid-ob="figobml161tu2">Table</a></h4></div></div></div><div id="ml161.s2"><h2 id="_ml161_s2_">Recommendations for Scientific Use of the Probe</h2><p>The goal of this project is to identify novel probes that inhibit platelet activation. This project focuses on the pathways involved in the activation of platelets initiated through the thrombin receptor pathway. The SFLLRN peptide activates platelets through the G protein-coupled receptor (GPCR) protease-activated receptor-1 (PAR1). The SFLLRN peptide is derived from the cleavage of the N-terminus of the PAR1 receptor by thrombin (<a class="bibr" href="#ml161.r1" rid="ml161.r1">1</a>). Upon activation, the platelets undergo many changes induced by multiple signaling cascades. One downstream effect of activation is the secretion of granules, which then potentiate platelets in controlling bleeding. Granule secretion also contributes to the growth of thrombi, which if not controlled properly can cause blockage of blood vessels and trigger myocardial infarction (MI). Probes from this project are defined as fitting into three possible attribute classes: 1) inhibitors of the pathways required for dense granule secretion, 2) inhibitors of the pathways required for granule secretion, which includes dense granules, alpha-granules, and lysosomes, and 3) inhibitors of upstream GPCR signaling. The first and second classes of inhibitors target distal mechanisms of secretion (i.e., the platelet second messenger systems) (<a class="bibr" href="#ml161.r2" rid="ml161.r2">2</a>). The third class of inhibitors targets the proximal cell-surface receptors.</p><p>The probes identified in this project will be used to further investigate the complex mechanisms of platelet activation. As these probes have been classified as GPCR-specific, additional assays developed in the Assay Provider’s laboratory (Robert Flaumenhaft, MD, PhD) will be performed to determine the specific GPCR target and site of action on the target GPCR signaling pathway. Depending on the results of these additional assays, these probes could be evaluated for their potential as antithrombotic agents in an <i>in vivo</i> murine model.</p><p>The GPCR-specific probe presented in this report is a valuable tool compound with unique attributes. Additional experimentation carried out after identification of the probe has demonstrated that this compound acts specifically on the GPCR PAR1. It exhibits several characteristics that distinguish it from published orthosteric inhibitors of PAR1. This area of investigation is of particular interest to the Assay Provider and others studying clinical aspects of platelet biology as platelets represent a good cellular target for pharmacological manipulation via allosteric modulation (<a class="bibr" href="#ml161.r3" rid="ml161.r3">3</a>). The central challenge in developing an antiplatelet agent is how to achieve a potent antiplatelet effect while avoiding hemorrhagic complications. Development of modulators of platelet function with novel modes of action, such as the probe described here, could limit bleeding complications associated with current antiplatelet agents (<a class="bibr" href="#ml161.r4" rid="ml161.r4">4</a>).</p></div><div id="ml161.s3"><h2 id="_ml161_s3_">1. Introduction</h2><div id="ml161.s4"><h3>Scientific Rationale</h3><p>Platelets are small, anucleate cells that circulate in the blood of mammals, and are required for maintaining hemostasis. The number of platelets circulating and their relative activity requires strict control. Too few platelets or too little activity can cause uncontrolled bleeding, while too many platelets and too much activity can cause acute thrombosis formation. Thrombi formation can block blood vessels causing myocardial and cerebral infarctions. Platelets undergo dramatic phenotypic changes in response to vascular injury that lead to the formation of thrombosis. These phenotypic changes are the result of an activation process induced by a diverse array of agonists (e.g., thrombin, adenosine diphosphate [ADP], epinephrine, and thromboxane A<sub>2</sub>) that act on platelet surface receptors. Phenotypic changes include granule secretion, shape change, and expression of specific cell-surface proteins. Platelet activation inhibitors are well validated for the inhibition of thrombosis and the clinical prevention of adverse cardiovascular events, including MI and stroke (<a class="bibr" href="#ml161.r5" rid="ml161.r5">5</a>). These inhibitors target various platelet G protein-coupled receptors (GPCRs), as well as certain cytosolic enzymes, which comprise key parts of the platelet activation pathway. Examples of important drugs within the category of GPCR inhibitors include: clopidogrel (Plavix<sup>®</sup>, Bristol-Myers Squibb/Sanofi Aventis), the metabolism of which leads to irreversible inhibition of the ADP receptor P2Y<sub>12</sub>; and SCH 530348 (Schering-Plough), a potent inhibitor of protease activated receptor (PAR1), which was in Phase 3 clinical trials prior to January 2011. Examples of platelet enzyme inhibitors include: aspirin, an irreversible inhibitor of COX1; and cilostazol (Pletal<sup>®</sup>, Otsuka), a cyclic nucleotide phosphodiesterase 3 (PDE3) inhibitor in clinical use for intermittent claudication. The structures of these compounds are depicted in <a class="figpopup" href="/books/NBK55066/figure/ml161.f1/?report=objectonly" target="object" rid-figpopup="figml161f1" rid-ob="figobml161f1">Figure 1</a>.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml161f1" co-legend-rid="figlgndml161f1"><a href="/books/NBK55066/figure/ml161.f1/?report=objectonly" target="object" title="Figure 1" class="img_link icnblk_img figpopup" rid-figpopup="figml161f1" rid-ob="figobml161f1"><img class="small-thumb" src="/books/NBK55066/bin/ml161f1.gif" src-large="/books/NBK55066/bin/ml161f1.jpg" alt="Figure 1. Examples of Platelet Activation Inhibitors." /></a><div class="icnblk_cntnt" id="figlgndml161f1"><h4 id="ml161.f1"><a href="/books/NBK55066/figure/ml161.f1/?report=objectonly" target="object" rid-ob="figobml161f1">Figure 1</a></h4><p class="float-caption no_bottom_margin">Examples of Platelet Activation Inhibitors. </p></div></div><p>Unfortunately, many of these drugs and others within the broader category of antithrombotics suffer from significant liabilities. For example, P2Y<sub>12</sub> inhibition leads to a significant risk of dangerous hemorrhagic side effects (<a class="bibr" href="#ml161.r6" rid="ml161.r6">6</a>). Key platelet enzymes such as COX1 and PDE3 are also present in many other cell types, and their inhibition can lead to undesired complications (<a class="bibr" href="#ml161.r7" rid="ml161.r7">7</a>). For this reason, novel inhibitors of platelet activation are of great interest, particularly those that modulate new targets, those that inhibit select features of platelet activation, and those that demonstrate novel pharmacological properties such as allosteric inhibition (<a class="bibr" href="#ml161.r3" rid="ml161.r3">3</a>).</p><p>Platelets are best studied via chemical genetic analysis due to their anucleate state rendering them unsusceptible to direct genetic manipulation. Platelets in plasma are a readily available reagent in the form of expired donor-collected units of platelet-rich plasma (PRP) from blood centers nationwide. The Assay Provider’s laboratory previously developed and executed a small molecule screen to study platelet activation in PRP using granule secretion as a measurement of platelet activation (<a class="bibr" href="#ml161.r2" rid="ml161.r2">2</a>). The assay measures the amount of ATP released into the plasma via secreted dense granules upon activation (<a class="bibr" href="#ml161.r8" rid="ml161.r8">8</a>).</p><p>The purpose of these studies is to use a cell-based platelet assay to identify novel inhibitors of platelet activation. Inhibitors identified from this project are defined as fitting into three possible attribute classes: 1) inhibitors of the pathways required for dense granule secretion, 2) inhibitors of the pathways required for granule secretion, which includes dense granules, alpha-granules, and lysosomes, and 3) inhibitors of upstream GPCR signaling. The first and second classes of inhibitors target distal mechanisms of secretion (i.e., the platelet second messenger systems) (<a class="bibr" href="#ml161.r2" rid="ml161.r2">2</a>). There are no known compounds that selectively inhibit granule secretion in either the first or second class. The third class of inhibitors targets the proximal cell-surface receptors. One important platelet GPCR mentioned above is PAR1, which undergoes a cleavage reaction by the serine protease thrombin that allows its tethered peptide ligand to agonize the receptor (<a class="bibr" href="#ml161.r9" rid="ml161.r9">9</a>).</p><p>The primary screen was adapted from the cell-based phenotypic screen previously executed by the Assay Provider’s laboratory with an aim to identify inhibitors of platelet activation (<a class="bibr" href="#ml161.r2" rid="ml161.r2">2</a>). Platelets were activated with the thrombin receptor activation peptide SFLLRN, which binds the PAR1 GPCR (<a class="bibr" href="#ml161.r1" rid="ml161.r1">1</a>). Upon activation, the platelets undergo many changes induced by multiple signaling cascades, which results in the secretion of granules. Platelet activation was quantified by measurement of ATP from the secreted granules, as measured by a luciferin-luciferase assay. Cilostazol, a known platelet aggregation inhibitor that targets PDE3, was used as a positive control in the primary assay (see <a class="figpopup" href="/books/NBK55066/figure/ml161.f1/?report=objectonly" target="object" rid-figpopup="figml161f1" rid-ob="figobml161f1">Figure 1</a>). Even though it acts on a target that is not relevant to the project, cilostazol was chosen as a positive control for the high level of inhibition seen in the primary assay.</p><p>The secondary assays in this project served to determine the specificity of the inhibitors identified in the primary assay. Specifically, the first secondary assay distinguishes between inhibitors of granule secretion and those acting nonspecifically on the ATP-detection system. From a list of compounds acting specifically to decrease the levels of secreted ATP-rich granules from platelets, those compounds known to inhibit intracellular platelet targets were filtered out. The final secondary assays were designed to differentiate between inhibitors of the distal granule secretion events and those inhibiting proximal GPCRs.</p></div></div><div id="ml161.s5"><h2 id="_ml161_s5_">2. Materials and Methods</h2><div id="ml161.s6"><h3>Materials and Reagents</h3><p>Expired units of platelets were acquired from BloodSource (Sacramento, CA), Rhode Island Blood Center (Providence, RI), Blood Center of New Jersey (East Orange, NJ), and Blood Center of Wisconsin (Milwaukee, WI). CellTiter-Glo was purchased from Promega (Madison, WI). SFLLRN peptide was purchased from Bachem (Torrance, CA). PBS was acquired from Broad Institute Supply and Quality Management Group (SQM), and Cilostazol, ATP, and Resveratrol were acquired from Sigma. Phycoerythrin-labeled anti-P-selectin antibody was purchased from BD Biosciences. White, solid-bottom, flat, opaque 384-well plates were purchased from Aurora Biotechnologies (Catalog no. 00030721).</p></div><div id="ml161.s7"><h3>2.1. Assays</h3><p>A summary listing of completed assays and corresponding PubChem AID numbers is provided in <a href="#ml161.app1">Appendix A</a> (see <a class="figpopup" href="/books/NBK55066/table/ml161.t4/?report=objectonly" target="object" rid-figpopup="figml161t4" rid-ob="figobml161t4">Table A1</a>). Refer to <a href="#ml161.app2">Appendix B</a> for the detailed assay protocols.</p><div id="ml161.s8"><h4>2.1.1. Primary Screen and Conformation at Dose: Granule Secretion</h4><p>Platelet volumes from a single donor ranged from 200 mL to 700 mL. Samples were taken for counting platelets/mL and tested for activation activity by the addition of SFLLRN and CellTiter-Glo. Only platelets demonstrating at least a 500-fold level of activation were used in the assay. The 384-well plates were filled with platelets (20 μL/well), using a Thermo Multidrop Combi liquid handler in a biosafety cabinet. The volume of platelets determined the number plates used in a run. The filled plates were loaded into an automated incubator set at 30°C, 95% humidity, 5% CO<sub>2</sub>, docked to an enclosed automated screening system (HighRes Biosolutions, Woburn, MA). Unsealed compound plates were loaded into another automated incubator on the same system, set at 22°C, 15% humidity. CellTiter-Glo substrate was resuspended in PBS (100 mL/bottle, 1:1) and the SFLLRN peptide was reconstituted in PBS to 10 mM. The CellTiter-Glo/SFLLRN screening solution was prepared by dilution of CellTiter-Glo solution to 1:4 with PBS and the addition of reconstituted SFLLRN to a final concentration of 15 μM. Each run was initiated by the software application Broad Chemical Biology Informatics Platform (CBIP) and scheduled with Cellario (HighRes Biosolutions, Woburn, MA). Robotic arms moved plates from the different instruments on the screening system. Next, 100nL of compounds were transferred from DMSO daughter plates into assay plates using a MicroPin (HighRes Biosolutions, Woburn, MA). After compound addition, the assay plates were returned to the incubator for 30-minute incubation. At the completion of incubation, the plates were moved to a MultiDrop Combi for addition of 10 μL CellTiter-Glo/SFLLRN solution per well, to a final well concentration of 0.083X CellTiter-Glo and 5 μM SFLLRN. The plates were incubated on the deck of the screening system for 15 minutes, at room temperature. At the completion of incubation, the plates were read for luminescence in an EnVision 2104 Multilabel Reader (Perkin Elmer, Waltham, MA). The ultra sensitive detection was used, with the 1536 aperture in place, to decrease bleed-through from adjacent wells. Read time was 0.1 s/well.</p><p>The same assay protocol was used for a conformation at dose response. The 1584 cherry-pick compounds were received from BioFocus and powder compounds tested in this assay were either synthesized (BIPDeC) or purchased from various vendors.</p></div><div id="ml161.s9"><h4>2.1.2. Luciferase Counterscreen</h4><p>ATP was diluted to 1.5 μM in PBS from 10-mM aliquots in PBS. Next, 1:1 aliquots of CellTiter-Glo reconstituted in PBS, were diluted to 1:4 with PBS. A MultiDrop Combi (Thermo) was used to add 20 μL of 1.5 μM ATP in PBS. Then, 50 nL of compounds at dose were added to the assay plates using a CyBi-Well (CyBio Inc.). Upon completion of compound addition, 10 μL of CellTiter-Glo (1:4) in PBS was added to every well, to a final concentration of 0.083X. The plates were transferred to an EnVison Plate Reader (PerkinElmer, Waltham, MA) to measure luminescence, as described in the primary screen (<a href="#ml161.s8">Section 2.1.1</a>).</p></div><div id="ml161.s10"><h4>2.1.3. Secondary Assay for <sup>14</sup>C -Serotonin Release</h4><p>This assay was performed in the Assay Provider’s laboratory with 28 cherry-pick compounds acquired from BioFocus, selected from a narrowed list of putative platelet activation inhibitory compounds. Platelet rich plasma (PRP; 4.5 mL, approximately 3×10<sup>8</sup> platelets/mL) was incubated in the presence of <sup>14</sup>C-serotonin (0.225 μCi) for 45 minutes at 37°C. Next, 15% acid-citrate-dextrose (ACD)+ 150 nM prostaglandin E1 (PGE1) was added to the PRP and spun at 1000 g for 10 minutes to maintain the resting state of platelets while removing unincorporated <sup>14</sup>C-serotonin. The platelets were resuspended in HEPES-Tyrodes buffer (10 mM HEPES/NaOH, pH 7.4, 5.56 mM glucose, 137 mM NaCl, 12 mM NaHCO3, 2.7 mM KCl, 0.36 mM NaH2PO4, 1 mM MgCl2) to a platelet concentration of 2×10<sup>8</sup> platelets/ml. Compounds were added to resuspended platelets at 10 μM and incubated for 15 minutes. Platelets were subsequently stimulated with 5 μM SFLLRN peptide and 5 μM imipramine (to prevent <sup>14</sup>C-serotonin reuptake) for 15 minutes. The platelets were separated from medium by centrifugation at 8000xg. Supernatants were collected and counted using a beta liquid scintillation counter. Disintegrations per minute (DPM) were collected for 50 μL of platelets.</p></div><div id="ml161.s11"><h4>2.1.4. Secondary Assay for SFLLRN-induced P-selectin Surface Expression</h4><p>This assay was also performed in the Assay Provider’s laboratory with 28 cherry-pick compounds acquired from BioFocus, selected from a narrowed list of putative platelet activation inhibitory compounds. In addition to the 28 cherry-pick compounds acquired from BioFocus, powder compounds synthesized (BIPDeC) or purchased from various sources were screened in this and the following assays in the Assay Provider’s laboratory. Washed platelets obtained from individual donors were treated with a selection of compounds resourced as powders found to be active in secondary assays. Platelet samples were tested at a range of concentrations from 0 μM to 100 μM of compounds. Following compound addition, the platelets were subsequently stimulated with 5 mM SFLLRN. After a 15-minute incubation, phycoerythrin-labeled anti-P-selectin antibody was added for a 20-minute incubation. The samples were analyzed by flow cytometry to determine P-selectin expression on the surface of the platelets as a response to activation. Geometric mean values were collected for each sample.</p></div><div id="ml161.s12"><h4>2.1.5. Counterscreen for the Identification of Inducers of cAMP in Platelets</h4><p>Washed platelets obtained from individual donors were treated with a selection of compounds that were chosen based on activity in an SFLLRN-induced P-selectin expression assay. Prostaglandin E1 (PGE1) at 1 μM was used as a positive control to elevate platelet cAMP levels. Compounds were tested at 10 μM. The platelets were lysed following compound or control incubation, and lysates were analyzed with a cAMP competitive ELISA kit (Thermo Scientific). Values were converted to cAMP concentration (pmol/mL) based on a cAMP standard curve measured in parallel.</p></div><div id="ml161.s13"><h4>2.1.6. Secondary Assay for the Identification of Inhibitors SFLLRN-mediated Actin Polymerization in Platelets</h4><p>Platelets were obtained from individual donors, gel-filtered, and treated with DMSO (neutral control) or compounds (10 μM) that did not induce elevation of cAMP in platelets as measured in a cAMP assay. Following incubation with compounds, the platelets were either treated or not treated with the thrombin receptor peptide agonist, SFLLRN. Both populations of platelets were fixed, and then incubated in the presence of FITC-phalloidin and Triton X-100. The samples were analyzed by flow cytometry to determine levels of actin polymerization upon activation. Actin filament formation results in higher fluorescent measurements. Geometric mean values were collected for each sample.</p></div><div id="ml161.s14"><h4>2.1.7. Counter Assay for Identification of the Inhibitors of Ca2+-ionophore (A23187)-induced P-selectin Surface Expression</h4><p>Washed platelets obtained from individual donors were treated with compounds found to show less than 50% inhibition in an SFLLRN-induced FITC phalloidin assay. The platelet samples were incubated with 30 μM of compound. Following compound addition, platelets were stimulated with 10 μM A23187. After a 15-minute incubation, phycoerythrin-labeled anti-P-selectin antibody was added for a 20-minute incubation. The samples were analyzed by flow cytometry to determine P-selectin expression on the surface of the platelets as a response to activation. Geometric mean values were collected for each sample.</p></div><div id="ml161.s15"><h4>2.1.8. Counter Assay for the Identification of Inhibitors of Phorbol Myristate Acetate (PMA)-induced P-selectin Surface Expression</h4><p>Washed platelets obtained from individual donors were treated with compounds found to show less than 50% inhibition in an SFLLRN-induced FITC phalloidin assay. The platelet samples were treated with 30 μM of compound. Following compound addition, the platelets were stimulated with 5 nM PMA. After a 15-minute incubation, phycoerythrin-labeled anti-P-selectin antibody was added for a 20-minute incubation. The samples were analyzed by flow cytometry to determine P-selectin expression on the surface of the platelets as a response to activation. Geometric mean values were collected for each sample.</p></div></div><div id="ml161.s16"><h3>2.2. Probe Chemical Characterization</h3><p>The probe (CID 1048267/<a href="/pcsubstance/?term=ML161[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML161</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457110" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID 87457110</a> and <a href="https://pubchem.ncbi.nlm.nih.gov/substance/87219021" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID 87219021</a>, MLS002699906) was purchased from ChemBridge Corp. It was analyzed by HPLC, <sup>1</sup>H and <sup>13</sup>C NMR, and high-resolution mass spectrometry. The data were consistent with the structure of the probe, and HPLC indicated a purity of 98%. The relevant data is provided in <a href="#ml161.s17">Section 2.3</a>. The observed solubility of the probe in PBS buffer (pH 7.4) was determined to be 83 μM. No decomposition of the probe was observed over a 48-hour period in PBS, indicating excellent aqueous stability (data not shown). Plasma protein binding (PPB) was determined to be 97% bound in human plasma. The probe is stable in human plasma with 80% remaining after a 5-hour incubation period, but the stability of the probe and most its analogs are unstable in murine plasma (less than 2% of the probe remains after 5 hours). The solubility, PPB, and plasma stability results are summarized in <a class="figpopup" href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object" rid-figpopup="figml161t2" rid-ob="figobml161t2">Table 2</a> and <a class="figpopup" href="/books/NBK55066/table/ml161.t3/?report=objectonly" target="object" rid-figpopup="figml161t3" rid-ob="figobml161t3">Table 3</a> (see <a href="#ml161.s24">Section 3.4</a>).</p><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml161t2"><a href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object" title="Table 2" class="img_link icnblk_img figpopup" rid-figpopup="figml161t2" rid-ob="figobml161t2"><img class="small-thumb" src="/books/NBK55066/table/ml161.t2/?report=thumb" src-large="/books/NBK55066/table/ml161.t2/?report=previmg" alt="Table 2. SAR Analysis and Properties of Butanoic Acid Derivatives." /></a><div class="icnblk_cntnt"><h4 id="ml161.t2"><a href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object" rid-ob="figobml161t2">Table 2</a></h4><p class="float-caption no_bottom_margin">SAR Analysis and Properties of Butanoic Acid Derivatives. </p></div></div><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml161t3"><a href="/books/NBK55066/table/ml161.t3/?report=objectonly" target="object" title="Table 3" class="img_link icnblk_img figpopup" rid-figpopup="figml161t3" rid-ob="figobml161t3"><img class="small-thumb" src="/books/NBK55066/table/ml161.t3/?report=thumb" src-large="/books/NBK55066/table/ml161.t3/?report=previmg" alt="Table 3. Search Strings and Databases Employed in the Prior Art Search." /></a><div class="icnblk_cntnt"><h4 id="ml161.t3"><a href="/books/NBK55066/table/ml161.t3/?report=objectonly" target="object" rid-ob="figobml161t3">Table 3</a></h4><p class="float-caption no_bottom_margin">Search Strings and Databases Employed in the Prior Art Search. </p></div></div><p>The probe and eight additional analogs were submitted to the SMR collection [MLS002699906 (probe), MLS002699907, MLS002699908, MLS002699909, MLS002699910, MLS002699911, MLS002699912, MLS002699922, and MLS002699924 (analogs)].</p><p>The <sup>1</sup>H and <sup>13</sup>C NMR spectra and LC-MS chromatograms of the probe (CID 1048267/<a href="/pcsubstance/?term=ML161[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML161</a>) and analogs are provided in <a href="#ml161.app3">Appendix C</a>.</p></div><div id="ml161.s17"><h3>2.3. Analog Preparation</h3><p>Most analogs of the probe were prepared using the simple sequence outlined in <a class="figpopup" href="/books/NBK55066/figure/ml161.f2/?report=objectonly" target="object" rid-figpopup="figml161f2" rid-ob="figobml161f2">Scheme 1</a> for the 2-chlorophenyl analog. The final step can be performed using a variety of standard peptide coupling conditions; the conditions and yields in <a class="figpopup" href="/books/NBK55066/figure/ml161.f2/?report=objectonly" target="object" rid-figpopup="figml161f2" rid-ob="figobml161f2">Scheme 1</a> are not optimized.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml161f2" co-legend-rid="figlgndml161f2"><a href="/books/NBK55066/figure/ml161.f2/?report=objectonly" target="object" title="Scheme 1" class="img_link icnblk_img figpopup" rid-figpopup="figml161f2" rid-ob="figobml161f2"><img class="small-thumb" src="/books/NBK55066/bin/ml161f2.gif" src-large="/books/NBK55066/bin/ml161f2.jpg" alt="Scheme 1. Synthesis of 1,3-Diaminophenyl Analog." /></a><div class="icnblk_cntnt" id="figlgndml161f2"><h4 id="ml161.f2"><a href="/books/NBK55066/figure/ml161.f2/?report=objectonly" target="object" rid-ob="figobml161f2">Scheme 1</a></h4><p class="float-caption no_bottom_margin">Synthesis of 1,3-Diaminophenyl Analog. </p></div></div><div id="ml161.s18"><h4>Chemistry Experimental Methods</h4><p><b>General Details.</b> All reagents and solvents were purchased from commercial vendors and used as received. NMR spectra were recorded on a Bruker 300 MHz or Varian UNITY INOVA 500 MHz spectrometer as indicated. Proton and carbon chemical shifts are reported in parts per million (ppm;δ) relative to tetramethylsilane, CDCl<sub>3</sub> solvent, or d<sub>6</sub>-DMSO (<sup>1</sup>H δ 0, <sup>13</sup>C δ 77.0, or <sup>13</sup>C δ 39.5, respectively). NMR data are reported as follows: chemical shifts, multiplicity (obs. = obscured, br = broad, s = singlet, d = doublet, t = triplet, m = multiplet, comp = complex overlapping signals); coupling constant(s) in Hz; integration. Unless otherwise indicated, NMR data were collected at 25°C. Flash chromatography was performed using 40–60 μm Silica Gel (60 Å mesh) on a Teledyne Isco Combiflash R<i><sub>f</sub></i> system. Tandem liquid chromatography/mass spectrometry (LCMS) was performed on a Waters 2795 separations module and Waters 3100 mass detector. Analytical thin layer chromatography (TLC) was performed on EM Reagent 0.25 mm silica gel 60-F plates. Visualization was accomplished with UV light and aqueous potassium permanganate (KMnO<sub>4</sub>) stain followed by heating. High-resolution mass spectra were obtained at the MIT Mass Spectrometry Facility with a Bruker Daltonics APEXIV 4.7 Tesla Fourier Transform Ion Cyclotron Resonance mass spectrometer. Compound purity and identity were determined by UPLC-MS (Waters, Milford, MA). Purity was measured by UV absorbance at 210 nm. Identity was determined on a SQ mass spectrometer by positive electrospray ionization. Mobile Phase A consisted of either 0.1% ammonium hydroxide or 0.1% trifluoroacetic acid in water, while mobile Phase B consisted of the same additives in acetonitrile. The gradient ran from 5% to 95% mobile Phase B over 0.8 minutes at 0.45 mL/min. An Acquity BEH C18, 1.7 μm, 1.0 × 50 mm column was used with column temperature maintained at 65°C. Compounds were dissolved in DMSO at a nominal concentration of 1 mg/mL, and 0.25 μL of this solution was injected.</p><p><b>Synthesis of</b>
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<b><i>N</i></b><b>-(3-nitrophenyl)butyramide:</b> 3-Nitroaniline (6.00 g, 43.4 mmol) was sealed in a flask under nitrogen, then dichloromethane (145 mL) was added and the solution was cooled on ice. (<i>i</i>-Pr)<sub>2</sub>NEt (8.35 mL, 47.8 mmol) and butyryl chloride (5.00 mL, 47.8 mmol) were then added, and the reaction was removed from the ice bath and stirred for 16 hours. The reaction mixture was then washed twice with 1 M aqueous HCl, then with brine. The organic phase was dried over MgSO<sub>4</sub>, filtered, and concentrated to give the title compound as a yellow oil (8.60 g, 95%). <b><sup>1</sup>H NMR (300 MHz, CDCl<sub>3</sub>):</b> δ 8.45 (br s, 1 H), 8.19 (br s, 1 H), 7.94 (comp, 2 H), 7.46 (dd, <i>J</i> = 8.2, 8.2 Hz, 1 H), 2.43 (t, <i>J</i> = 7.4 Hz, 2 H), 1.77 (m, 2 H), 1.00 (t, <i>J</i> = 7.4 Hz, 3 H). <b>MS (ESI<sup>−</sup>):</b> 207.06.</p><p><b>Synthesis of</b>
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<b><i>N</i></b><b>-(3-aminophenyl)butyramide:</b> A solution of <i>N</i>-(3-nitrophenyl)butyramide (8.60 g, 41.3 mmol) in methanol (207 mL) was divided between two 500-mL flasks with stir bars. Both flasks were flushed well with nitrogen, before 5% palladium on carbon (4.40 g, 2.07 mmol) was added, approximately half of the measured amount to each flask. A glass 3-way adapter with attached large hydrogen-filled balloon was secured to each flask. The flask was evacuated and back-filled with hydrogen five times while stirring vigorously. The reaction was stirred for 6 hours, after which time LC-MS analysis showed complete consumption of starting material. The reaction mixture was filtered through a funnel packed with Celite and washed with methanol, then concentrated. The resulting crude semi-solid was dissolved with dichloromethane and purified by column chromatography (SiO<sub>2</sub>, 0 to 50% ethyl acetate/hexanes). A white solid was obtained after concentrating the resulting oil with hexanes (5.87 g, 80%). <b><sup>1</sup>H NMR (300 MHz, CDCl<sub>3</sub>):</b> δ 7.19 (br s, 2 H), 7.06 (t, <i>J</i> = 8.0, 1 H), 6.66 (d, <i>J</i> = 7.8, 1 H), 6.42 (d, <i>J</i> = 6.9, 1 H), 3.69 (s, 2 H), 2.30 (t, <i>J</i> = 7.4, 2 H), 1.84 – 1.61 (m, 3 H), 0.99 (t, <i>J</i> = 7.4, 3 H). <b>MS (ESI<sup>+</sup>):</b> 179.18.</p><p><b>Synthesis of</b>
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<b><i>N</i></b><b>-(3-butyramidophenyl)-2-chlorobenzamide:</b>
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<i>N</i>-(3-aminophenyl)butyramide (30 mg, 0.19 mmol), 2-chlorobenzoic acid (31.6 mg, 0.20 mmol), HOBt (28.4 mg, 0.19 mmol), and silica-supported-dicyclohexylcarbodiimide (DCC) (362 mg, 0.34 mmol, supplied by Silicycle) were added to a 13 × 100-mm test tube with cross-shaped stir bar. The tube was sealed with a septum and dry dichloromethane (3.4 mL) was added. The reaction was stirred for 15 hours, after which time some precipitate was present. Next, 1:1 acetonitrile:dichloromethane (4 mL) was added to ensure the precipitate dissolved, then silica supported-carbonate (534 mg, 0.34 mmol, supplied by Silicycle) was added and stirred for 2 hours. The reaction mixture was filtered through Celite, washed down with acetonitrile:dichloromethane (4 mL), then concentrated. The compound was adsorbed on SiO<sub>2</sub> and purified by column chromatography (0 to 50% ethyl acetate/hexanes), yielding a colorless oil (24 mg, 44%). <b><sup>1</sup>H NMR (300 MHz, CDCl<sub>3</sub>):</b> δ 10.50 (s, 1 H), 9.93 (s, 1 H), 8.06 (s, 1 H), 7.64 – 7.36 (m, 5 H), 7.36 – 7.17 (m, 2 H), 2.28 (t, <i>J</i> = 7.3, 2 H), 1.70 – 1.51 (m, 2 H), 0.91 (t, <i>J</i> = 7.4, 3 H). <b>MS (ESI<sup>+</sup>):</b> 317.22.</p><p><b>2-Bromo-</b><b><i>N</i></b><b>-(3-butyramidophenyl)benzamide (PROBE):</b> Purchased from ChemBridge Corp. <b><sup>1</sup>H NMR (300 MHz, CDCl<sub>3</sub>):</b> δ 7.95 (s, 1 H), 7.75 (s, 1 H), 7.62 (t, <i>J</i> = 7.6, 2 H), 7.37 (comp, 6 H), 2.31 (t, <i>J</i> = 7.4, 2 H), 1.74 (m, 2 H), 1.00 (t, <i>J</i> = 7.4, 3 H). <b><sup>13</sup>C NMR (75 MHz, CDCl<sub>3</sub>):</b> δ 171.3, 165.6, 138.8, 138.1, 137.7, 133.6, 131.6, 129.7, 127.8, 119.3, 116.0, 115.5, 111.1, 39.7, 19.0, 13.7. <b>HRMS (ESI<sup>+</sup>):</b> calculated mass for C<sub>17</sub>H<sub>17</sub>BrN<sub>2</sub>O<sub>2</sub> [M+H] 361.0546, found 361.0532.</p></div></div><div id="ml161.s19"><h3>2.4. Additional Analytical Analysis</h3><p><b>Plasma Protein Binding.</b> Plasma protein binding was determined by equilibrium dialysis using the Rapid Equilibrium Dialysis (RED) device (Pierce Biotechnology, Rockford, IL) for both human and mouse plasma. Each compound was prepared in duplicate at 5 μM in plasma (0.95% acetonitrile, 0.05% DMSO) and added to one side of the membrane (200 μL) with PBS pH 7.4 added to the other side (350 μL). Compounds were incubated at 37°C for 5 hours in a 250-rpm orbital shake. Following the incubation, the samples were analyzed by UPLC-MS (Waters, Milford, MA) with compounds detected by SIR detection on a single quadrupole mass spectrometer.</p><p><b>Plasma Stability.</b> Plasma stability was determined at 37°C for 5 hours in both human and mouse plasma. Each compound was prepared in duplicate at 5 μM in plasma diluted 50/50 (v/v) with PBS pH 7.4 (0.95% acetonitrile, 0.05% DMSO). The compounds were incubated at 37°C for 5 hours with a 250-rpm orbital shake with time points taken at 0 and 5 hours. The samples were analyzed by UPLC-MS (Waters, Milford, MA) with compounds detected by SIR detection on a single quadrupole mass spectrometer.</p></div></div><div id="ml161.s20"><h2 id="_ml161_s20_">3. Results</h2><div id="ml161.s21"><h3>3.1. Summary of Screening Results</h3><p>A high throughput screen (HTS) of 302,457 compounds (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1663" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 1663</a>) was performed in duplicate to measure platelet activation by monitoring granule secretion in PRP. Using expired PRP activated with the thrombin receptor activation peptide SFLLRN, platelet activation was quantified by measurement of ATP from secreted granules, as measured by a luciferin-luciferase assay. A new assay format was developed for this project to aid in scaling up to screen the full MLPCN library with the use of automation. A stable luciferin-luciferase reagent (CellTiter-Glo, Promega) was adapted to measure secreted granules without lysing the platelets. The compounds were screened at a final concentration of 7.5 μM. From the HTS data, 661 compounds were classified as hits, based on inhibiting granule secretion ≥ 50%. A set of 1584 compounds was chosen as cherry picks, including available positive compounds, compounds classified as inconclusive, and additional inactive compounds containing structures similar to active compounds to provide additional SAR information. Cherry-picked compounds were retested in a concentration-dependent manner in the primary assay using PRP activated with SFLLRN (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1889" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 1889</a>). Out of 1584 compounds, 651 compounds were confirmed as active in this assay with IC<sub>50</sub> values < 10 μM. The same set of dose-response compound plates were also tested in a biochemical counterscreen to rule out activity seen in the primary assay due to nonspecific inhibition from the luciferin-luciferase detection system (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1891" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 1891</a>). The 446 compounds showing an IC<sub>50</sub> > 112.2 μM in the counterscreen were considered inactive in this assay, thus considered specific for inhibiting platelet activation. The results from these secondary assays were taken together to narrow the list to specific active compounds by advancing only compounds that exhibited a ratio of IC<sub>50</sub> values of greater than 10-fold (i.e., counter/primary) from both assays. From this list, compounds that had been previously shown to inhibit platelet activation were also excluded. Of the remaining compounds, 30 compounds were prioritized for additional characterization in secondary assays. Of these 30 prioritized compounds, 28 were acquired thorough an additional cherry pick and tested in a suite of secondary specificity assays in the Assay Provider’s laboratory (see <a class="figpopup" href="/books/NBK55066/table/ml161.t4/?report=objectonly" target="object" rid-figpopup="figml161t4" rid-ob="figobml161t4">Table A1</a>, <a href="#ml161.app1">Appendix A</a>; <a class="figpopup" href="/books/NBK55066/figure/ml161.f3/?report=objectonly" target="object" rid-figpopup="figml161f3" rid-ob="figobml161f3">Figure 3</a>). These assays narrowed the list of compounds to four. The four probe candidates and analogs of each were acquired as dry powders and retested in the confirmatory and secondary assays. Of these, one of the four probe candidates did not retest and was removed from further consideration as a probe. Completion of the series of secondary assays confirmed that the three remaining compounds met the criteria for a Class III probe, possible modulators of GPCR signaling in platelet activation (see <a class="figpopup" href="/books/NBK55066/table/ml161.t4/?report=objectonly" target="object" rid-figpopup="figml161t4" rid-ob="figobml161t4">Table A1</a>, <a href="#ml161.app1">Appendix A</a>; <a class="figpopup" href="/books/NBK55066/figure/ml161.f3/?report=objectonly" target="object" rid-figpopup="figml161f3" rid-ob="figobml161f3">Figure 3</a>).</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml161f3" co-legend-rid="figlgndml161f3"><a href="/books/NBK55066/figure/ml161.f3/?report=objectonly" target="object" title="Figure 3" class="img_link icnblk_img figpopup" rid-figpopup="figml161f3" rid-ob="figobml161f3"><img class="small-thumb" src="/books/NBK55066/bin/ml161f3.gif" src-large="/books/NBK55066/bin/ml161f3.jpg" alt="Figure 3. Critical Path for Probe Development." /></a><div class="icnblk_cntnt" id="figlgndml161f3"><h4 id="ml161.f3"><a href="/books/NBK55066/figure/ml161.f3/?report=objectonly" target="object" rid-ob="figobml161f3">Figure 3</a></h4><p class="float-caption no_bottom_margin">Critical Path for Probe Development. </p></div></div><p>The third of the three probes to meet the defined probe criteria is a diaminophenyl (CID 1048267/<a href="/pcsubstance/?term=ML161[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML161</a>,). This probe reproducibly retests in the primary assay yielding IC<sub>50</sub> values of 10.3 μM and 12.1 μM (see <a class="figpopup" href="/books/NBK55066/figure/ml161.f4/?report=objectonly" target="object" rid-figpopup="figml161f4" rid-ob="figobml161f4">Figure 4</a>). However, the probe has been retested multiple times beyond the two tests shown in <a class="figpopup" href="/books/NBK55066/figure/ml161.f4/?report=objectonly" target="object" rid-figpopup="figml161f4" rid-ob="figobml161f4">Figure 4</a>, showing a range of IC<sub>50</sub> values from 4.9 μM to 12.1 μM (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2398" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 2398</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/493100" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 493100</a>; data not shown). The variability in IC<sub>50</sub> values observed across experiments was attributed to differences in platelet response from donor to donor; however, the rank ordering of potency within a probe series was maintained across multiple experiments. The diaminophenyl probe also inhibits P-selectin expression in a concentration-dependent manner, with an EC<sub>50</sub> less than 1 μM. This result demonstrated that the probe candidate met Class II or Class III probe definitions (see <a class="figpopup" href="/books/NBK55066/figure/ml161.f3/?report=objectonly" target="object" rid-figpopup="figml161f3" rid-ob="figobml161f3">Figure 3</a>).</p><p>A cyclic AMP (cAMP) assay performed on the diaminophenyl probe demonstrated that the compound did not have activity in modulating cAMP levels, as compared to the potent activity of the positive control compound cilostazol (see <a class="figpopup" href="/books/NBK55066/figure/ml161.f1/?report=objectonly" target="object" rid-figpopup="figml161f1" rid-ob="figobml161f1">Figure 1</a>). Additional testing of the probe candidate in a fluorescent actin polymerization assay determined that it has greater than 50% activity in inhibiting SFLLRN-induced actin filament formation. The results of this assay suggest the probe is acting as a potential Class III probe, at an upstream GPCR target. Counterscreening in two additional assays in which platelets were activated with phorbol myristate acetate (PMA) or Ca2+ ionophore, agents that stimulate activation outside the GPCR signaling pathways, supported that the diaminophenyl meets the criteria as a Class III probe (see <a class="figpopup" href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object" rid-figpopup="figml161t2" rid-ob="figobml161t2">Table 2</a>, <a href="#ml161.s24">Section 3.4</a>; <a class="figpopup" href="/books/NBK55066/figure/ml161.f3/?report=objectonly" target="object" rid-figpopup="figml161f3" rid-ob="figobml161f3">Figure 3</a>).</p></div><div id="ml161.s22"><h3>3.2. Dose Response Curves for Probe</h3><div id="ml161.f4" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161f4.jpg" alt="Figure 4. Diaminophenyl Probe (CID 1048267/ML161) Concentration-dependent Activity in the Primary Screen Retest (AID 493100)." /></div><h3><span class="label">Figure 4</span><span class="title">Diaminophenyl Probe (CID 1048267/<a href="/pcsubstance/?term=ML161[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML161</a>) Concentration-dependent Activity in the Primary Screen Retest (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/493100" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 493100</a>)</span></h3><div class="caption"><p>Primary assay retest measuring extracellular ATP as a measure of granule release: Retest 1, EC<sub>50</sub> 10.3 μM (<i>a);</i> Retest 2, EC<sub>50</sub> 12.1 μM (<i>b</i>)</p></div></div></div><div id="ml161.s23"><h3>3.3. Scaffold/Moiety Chemical Liabilities</h3><p>The scaffold does not have obvious chemical liabilities. It is stable over 48 hours in PBS buffer and shows excellent stability in both human and mouse plasma. The PBS stability data is shown in <a class="figpopup" href="/books/NBK55066/figure/ml161.f5/?report=objectonly" target="object" rid-figpopup="figml161f5" rid-ob="figobml161f5">Figure 5</a> below.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml161f5" co-legend-rid="figlgndml161f5"><a href="/books/NBK55066/figure/ml161.f5/?report=objectonly" target="object" title="Figure 5" class="img_link icnblk_img figpopup" rid-figpopup="figml161f5" rid-ob="figobml161f5"><img class="small-thumb" src="/books/NBK55066/bin/ml161f5.gif" src-large="/books/NBK55066/bin/ml161f5.jpg" alt="Figure 5. PBS Stability Data for the Probe (CID 1048267/ML161." /></a><div class="icnblk_cntnt" id="figlgndml161f5"><h4 id="ml161.f5"><a href="/books/NBK55066/figure/ml161.f5/?report=objectonly" target="object" rid-ob="figobml161f5">Figure 5</a></h4><p class="float-caption no_bottom_margin">PBS Stability Data for the Probe (CID 1048267/ML161. </p></div></div><p>As of January 2011, the probe (CID 1048267/<a href="/pcsubstance/?term=ML161[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML161</a>) was active in 8 assays out of 399 reported in PubChem, of which 6 assays are related to this platelet activation project. Therefore, the probe is considered not to be a promiscuous binder.</p></div><div id="ml161.s24"><h3>3.4. SAR Tables</h3><p>The structures and the activities of the probe (CID 1048267/<a href="/pcsubstance/?term=ML161[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML161</a>) and analogs in the primary assay and the suite of secondary assays (tested as dry powders of validated purity) are summarized in <a class="figpopup" href="/books/NBK55066/table/ml161.t1/?report=objectonly" target="object" rid-figpopup="figml161t1" rid-ob="figobml161t1">Table 1</a> and <a class="figpopup" href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object" rid-figpopup="figml161t2" rid-ob="figobml161t2">Table 2</a>. Good activity was obtained with the analog possessing a 2-substituted aryl bromide on the “eastern” side, and a 3-carbon chain on the “western” side of the molecule (entry 1, <a class="figpopup" href="/books/NBK55066/table/ml161.t1/?report=objectonly" target="object" rid-figpopup="figml161t1" rid-ob="figobml161t1">Table 1</a>). This compound was eventually designated as the probe, and its activity was later confirmed to be proximal to the GPCR PAR1. Analogs with shorter carbon chains were found to be less active] (entries 2 and 4, <a class="figpopup" href="/books/NBK55066/table/ml161.t1/?report=objectonly" target="object" rid-figpopup="figml161t1" rid-ob="figobml161t1">Table 1</a>), and the analog with a 4-carbon chain was also inactive (entry 3, <a class="figpopup" href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object" rid-figpopup="figml161t2" rid-ob="figobml161t2">Table 2</a>). An analog with a 4-chlorobenzyl substituent on the eastern side also proved to be inactive (entry 6, <a class="figpopup" href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object" rid-figpopup="figml161t2" rid-ob="figobml161t2">Table 2</a>).</p><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml161t1"><a href="/books/NBK55066/table/ml161.t1/?report=objectonly" target="object" title="Table 1" class="img_link icnblk_img figpopup" rid-figpopup="figml161t1" rid-ob="figobml161t1"><img class="small-thumb" src="/books/NBK55066/table/ml161.t1/?report=thumb" src-large="/books/NBK55066/table/ml161.t1/?report=previmg" alt="Table 1. SAR Analysis and Properties of Diaminophenyl Probe and Analogs." /></a><div class="icnblk_cntnt"><h4 id="ml161.t1"><a href="/books/NBK55066/table/ml161.t1/?report=objectonly" target="object" rid-ob="figobml161t1">Table 1</a></h4><p class="float-caption no_bottom_margin">SAR Analysis and Properties of Diaminophenyl Probe and Analogs. </p></div></div><p>To probe the SAR in more detail, a series of analogs were synthesized where the 3-carbon chain was held constant and the eastern aryl group was modified using the chemistry outlined in <a class="figpopup" href="/books/NBK55066/figure/ml161.f2/?report=objectonly" target="object" rid-figpopup="figml161f2" rid-ob="figobml161f2">Scheme 1</a> (<a class="figpopup" href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object" rid-figpopup="figml161t2" rid-ob="figobml161t2">Table 2</a>). The only compounds showing significant activity were those with 2-substituents on the arene (entries 1 to 4, <a class="figpopup" href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object" rid-figpopup="figml161t2" rid-ob="figobml161t2">Table 2</a>). The 2-chloro and 2-methyl analogs showed activity slightly better than the probe in the primary assay, but interestingly the 2-methoxy analog was inactive (entry 3 <a class="figpopup" href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object" rid-figpopup="figml161t2" rid-ob="figobml161t2">Table 2</a>). The 3- and 4-substituted analogs (entries 5 to 17, <a class="figpopup" href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object" rid-figpopup="figml161t2" rid-ob="figobml161t2">Table 2</a>)</p><p>A select number of heterocyclic analogs were also inactive (entries 18–20, <a class="figpopup" href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object" rid-figpopup="figml161t2" rid-ob="figobml161t2">Table 2</a>). All analogs generally had good human plasma stability but poor stability in mouse plasma. Measured human plasma protein binding was generally 85–99%. Additional characterization of the active analogs in a battery of secondary assays is underway in the lab of the Assay Provider. A summary of the current SAR data for this scaffold is provided in <a class="figpopup" href="/books/NBK55066/figure/ml161.f6/?report=objectonly" target="object" rid-figpopup="figml161f6" rid-ob="figobml161f6">Figure 6</a>.</p><div class="iconblock whole_rhythm clearfix ten_col fig" id="figml161f6" co-legend-rid="figlgndml161f6"><a href="/books/NBK55066/figure/ml161.f6/?report=objectonly" target="object" title="Figure 6" class="img_link icnblk_img figpopup" rid-figpopup="figml161f6" rid-ob="figobml161f6"><img class="small-thumb" src="/books/NBK55066/bin/ml161f6.gif" src-large="/books/NBK55066/bin/ml161f6.jpg" alt="Figure 6. Summary of Structure-Activity Relationships for 1,3-diaminophenyl Scaffold." /></a><div class="icnblk_cntnt" id="figlgndml161f6"><h4 id="ml161.f6"><a href="/books/NBK55066/figure/ml161.f6/?report=objectonly" target="object" rid-ob="figobml161f6">Figure 6</a></h4><p class="float-caption no_bottom_margin">Summary of Structure-Activity Relationships for 1,3-diaminophenyl Scaffold. </p></div></div></div><div id="ml161.s25"><h3>3.5. Cellular Activity</h3><p>The primary assay and many of the secondary assays were performed on platelets in plasma or washed platelets (see <a href="#ml161.s7">Section 2.1</a> for experimental details). Those compounds found to be toxic to platelets would have demonstrated no effect or an increased luminescence response in the primary assay and were not selected for additional analysis.</p></div><div id="ml161.s26"><h3>3.6. Profiling Assays</h3><p>As described in <a href="#ml161.s23">Section 3.3</a>, the diaminophenyl probe was active in eight assays out of 399 reported in PubChem, of which 6 assays are related to this platelet activation project. Other profiling screens (e.g., CEREP) have not been performed with this compound to date.</p></div></div><div id="ml161.s27"><h2 id="_ml161_s27_">4. Discussion</h2><div id="ml161.s28"><h3>4.1. Comparison to Existing Art and How the New Probe is an Improvement</h3><p>Additional studies beyond the scope of the outlined project have demonstrated that this probe exhibits several characteristics that distinguish it from existing inhibitors of the platelet GPCR PAR1. It inhibits platelet secretion and aggregation without inhibiting platelet shape change (data not shown), suggesting that it inhibits Gαq, but not Gα12/13 signaling (<a class="bibr" href="#ml161.r9" rid="ml161.r9">9</a>). Further studies, which are ongoing in the Assay Provider’s laboratory are required to determine the specific mode of action of this probe. It is likely that the probe is acting as an allosteric inhibitor of PAR1. Currently, there are no known allosteric inhibitors of PAR1 with the ability to selectively inhibit certain phenotypic changes (e.g., granule secretion, but not platelet shape change). This probe could have unique advantages over orthosteric GPCR inhibitors currently used as antithrombotic drugs (<a class="bibr" href="#ml161.r3" rid="ml161.r3">3</a>).</p><p>To support the novelty of our probes and their phenotypic effects, as well as their proposed mode of action, an investigation into the relevant prior art was performed by searching the following databases: SciFinder, Web of Science, Patent Lens, and PubMed. The search terms applied and hit statistics are provided in <a class="figpopup" href="/books/NBK55066/table/ml161.t3/?report=objectonly" target="object" rid-figpopup="figml161t3" rid-ob="figobml161t3">Table 3</a>. Abstracts were obtained for all references returned and were analyzed for relevance to the current project. The searches were performed on February 2, 2011.</p></div><div id="ml161.s29"><h3>4.2. Mechanism of Action Studies</h3><p>The Assay Provider’s laboratory is currently investigating the mechanism of action of this probe. The proposed allosteric mechanism of PAR1 inhibition will be studied by measuring the agonist dose/response with different probe concentrations. Decrease of maximum response to a PAR1 orthosteric agonist with increasing concentrations of inhibitor will support an allosteric, noncompetitive mechanism of inhibition. Mutational studies to determine the GPCR binding site(s) are also underway.</p></div><div id="ml161.s30"><h3>4.3. Planned Future Studies</h3><p>Concurrent with the mechanistic studies described above, the Broad Institute completed the synthesis and optimization of all analogs for this probe series. Further studies will include testing in assays designed and executed in the Assay Provider’s laboratory. Promising compounds will also be tested in mouse models of thrombus formation.</p></div></div><div id="ml161.reflist"><h2 id="_ml161_reflist_">5. References</h2><dl class="temp-labeled-list"><dl class="bkr_refwrap"><dt>1.</dt><dd><div class="bk_ref" id="ml161.r1">Vu TK, Hung DT, Wheaton VI, Coughlin SR. Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. <span><span class="ref-journal">Cell. </span>1991 Mar 22;<span class="ref-vol">64</span>(6):1057–1068.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/1672265" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 1672265</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>2.</dt><dd><div class="bk_ref" id="ml161.r2">Flaumenhaft R, Sim DS. The platelet as a model for chemical genetics. <span><span class="ref-journal">Chem Biol. </span>2003 Jun;<span class="ref-vol">10</span>(6):481–486.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/12837380" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 12837380</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>3.</dt><dd><div class="bk_ref" id="ml161.r3">Dowal L, Flaumenhaft R. Targeting platelet G-protein coupled receptors (GPCRs): looking beyond conventional GPCR antagonism. <span><span class="ref-journal">Curr Vasc Pharmacol. </span>2010 Mar;<span class="ref-vol">8</span>(2):140–154.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/19485898" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 19485898</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>4.</dt><dd><div class="bk_ref" id="ml161.r4">Shehab N, Sperling LS, Kegler SR, Budnitz DS. National estimates of emergency department visits for hemorrhage-related adverse events from clopidogrel plus aspirin and from warfarin. <span><span class="ref-journal">Arch Intern Med. </span>2010 Nov 22;<span class="ref-vol">170</span>(21):1926–1933.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/21098354" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 21098354</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>5.</dt><dd><div class="bk_ref" id="ml161.r5">Michelson AD. Antiplatelet therapies for the treatment of cardiovascular disease. <span><span class="ref-journal">Nat Rev Drug Discov. </span>2010 Feb;<span class="ref-vol">9</span>(2):154–169.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/20118963" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 20118963</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>6.</dt><dd><div class="bk_ref" id="ml161.r6">Wiviott SD, Braunwald E, McCabe CH, Montalescot G, Ruzyllo W, Gottlieb S, Neumann FJ, Ardissino D, De Servi S, Murphy SA, et al. for the TRITON-TIMI 38 Investigators. Prasugrel versus clopidogrel in patients with acute coronary syndromes. <span><span class="ref-journal">N Engl J Med. </span>2007 Nov 15;<span class="ref-vol">357</span>(20):2001–2015.</span> Epub 2007 Nov 4. [<a href="https://pubmed.ncbi.nlm.nih.gov/17982182" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 17982182</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>7.</dt><dd><div class="bk_ref" id="ml161.r7">Ding B, Abe J, Wei H, Huang Q, Walsh RA, Molina CA, Zhao A, Sadoshima J, Blaxall BC, Berk BC, et al. Functional role of phosphodiesterase 3 in cardiomyocyte apoptosis: implication in heart failure. <span><span class="ref-journal">Circulation. </span>2005 May 17;<span class="ref-vol">111</span>(19):2469–2476.</span> [<a href="/pmc/articles/PMC4108189/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC4108189</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/15867171" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 15867171</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>8.</dt><dd><div class="bk_ref" id="ml161.r8">McNicol A, Israels SJ. Platelet dense granules: structure, function, and implications for haemostasis. <span><span class="ref-journal">Thromb Res. </span>1999 Jul 1;<span class="ref-vol">95</span>(1):1–18.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/10403682" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 10403682</span></a>]</div></dd></dl><dl class="bkr_refwrap"><dt>9.</dt><dd><div class="bk_ref" id="ml161.r9">Coughlin SR. Thrombin signalling and protease-activated receptors. <span><span class="ref-journal">Nature. </span>2000 Sep 14;<span class="ref-vol">407</span>(6801):258–264.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/11001069" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 11001069</span></a>]</div></dd></dl></dl></div><div id="ml161.appgroup1"><h2 id="_ml161_appgroup1_">6. Appendices</h2><div id="ml161.app1"><h3>Appendix A. Assay Summary Table</h3><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml161t4"><a href="/books/NBK55066/table/ml161.t4/?report=objectonly" target="object" title="Table A1" class="img_link icnblk_img figpopup" rid-figpopup="figml161t4" rid-ob="figobml161t4"><img class="small-thumb" src="/books/NBK55066/table/ml161.t4/?report=thumb" src-large="/books/NBK55066/table/ml161.t4/?report=previmg" alt="Table A1. Summary of Completed Assays and AIDs." /></a><div class="icnblk_cntnt"><h4 id="ml161.t4"><a href="/books/NBK55066/table/ml161.t4/?report=objectonly" target="object" rid-ob="figobml161t4">Table A1</a></h4><p class="float-caption no_bottom_margin">Summary of Completed Assays and AIDs. </p></div></div></div><div id="ml161.app2"><h3>Appendix B. Detailed Assay Protocols</h3><div id="ml161.s35"><h4>Primary Screen and Conformation at Dose: Granule Secretion</h4><div id="ml161.s36"><h5>Materials</h5><p>White, opaque, flat bottom, 384-well plates (Aurora 00030721, Lot 010909-011009); Expired units of donor-collected platelets (various blood centers, see <a href="#ml161.s7">Section 2.1</a>); CellTiter-Glo (Promega, G7573, Lot H-2936.0025); SFLLRN (TRAP-6 amide peptide) (Bachem, H-8365, Lot 2500375); PBS (Supply and Quality Management (SQM), Broad Institute); Cilostazol (Sigma, C0737, Lot 042K4704, BRD-K67017579-001-04-2)</p><div id="ml161.s37"><h5>SFLLRN stock solution-10mM</h5><ul><li class="half_rhythm"><div>Dissolve 100 mg of SFLLRN in 13.32 ml of PBS.</div></li><li class="half_rhythm"><div>Filter, sterilize, aliquot into 1-ml tubes.</div></li><li class="half_rhythm"><div>Store at −20°C. Thaw prior to use.</div></li></ul></div><div id="ml161.s38"><h5>CellTiter-Glo stock solution-1:1</h5><ul><li class="half_rhythm"><div>Warm CellTiter-Glo substrate to room temperature.</div></li><li class="half_rhythm"><div>Add 100 ml of PBS to resuspend substrate.</div></li><li class="half_rhythm"><div>Aliquot solution into 25-ml tubes.</div></li><li class="half_rhythm"><div>Store at −20°C. Thaw prior to use.</div></li></ul></div><div id="ml161.s39"><h5>CellTiter-Glo (1:4)/SFLLRN (15 μM) solution</h5><ul><li class="half_rhythm"><div>Prepare a sufficient to complete a day’s worth of runs.</div></li><li class="half_rhythm"><div>For 400 ml, mix together 100 ml CellTiter-Glo (1:1), 600 μL 10 mM SFLLRN, and 300 ml PBS in a 500-ml centrifuge bottle, wrapped in foil to shield from light.</div></li></ul></div></div><div id="ml161.s40"><h5>Protocol</h5><ol><li class="half_rhythm"><div>Empty one plasma bag into a sterile bottle (250 or 500 ml, depending on volume of plasma) in the hood. If multiple bags exist from one donor (matching barcodes), combine into one bottle.</div></li><li class="half_rhythm"><div>Prepare Combi in biosafety cabinet. Use to fill plates with 20 μl per well. Fill as many plates allowable with volume of plasma. While filling, keep platelets in homogeneous suspension by gentle agitation.</div></li><li class="half_rhythm"><div>Load plates into racks for placement into Liconic incubator set at 25°C, 95% humidity, 5% CO2. Load incubator on BL2+ screening system.</div></li><li class="half_rhythm"><div>Prepare CellTiter-Glo/SFLLRN reagent to volume to accommodate the number of assay plates used in run. Prepare and prime Combi on BL2+ system for run.</div></li><li class="half_rhythm"><div>Initiate run on BL2+ system with Chemical Biology Informatics Platform (CBIP) and Cellario (HighRes Biosolutions). Next, 50 nL of compounds (stored in Greiner or Abgene polystyrene compound storage plates) are pinned into the assay plates, using 25-nL head programmed to deliver 50 nL.</div></li><li class="half_rhythm"><div>Return plates to incubator for 30-minute incubation.</div></li><li class="half_rhythm"><div>At completion of incubation, move plates to Combi for addition of 10 μl CellTiter-Glo (1:4 dilution)/SFLLRN (15 μM) solution. Use a standard cassette.</div></li><li class="half_rhythm"><div>Move plates to plate hotel on deck for 15-minute incubation.</div></li><li class="half_rhythm"><div>At completion of incubation, move plates to EnVision for luminescence detection. Use the ultra sensitive detection, with the 1536 aperture in place, to decrease bleed-through from adjacent wells. Read time is 0.1 s/well.</div></li></ol></div></div><div id="ml161.s41"><h4>Luciferase Counterscreen</h4><div id="ml161.s42"><h5>Materials</h5><p>White, opaque, flat bottom, 384-well plates (Aurora 00030721, Lot 010909-011009); CellTiter-Glo (Promega, G7573, Lot H-2936.0025); PBS (Supply and Quality Management (SQM), Broad Institute); ATP (Sigma, A1852, Lot 077K70121); Resveratrol (Sigma, R5010, Lot 038K5202, BRD-K80738081-001-10-4)</p><div id="ml161.s43"><h5>ATP stock solution (10 mM)</h5><ul><li class="half_rhythm"><div>Add 5.44 ml of PBS to 30 mg of ATP.</div></li><li class="half_rhythm"><div>Filter, sterilize, aliquot into 1-ml tubes.</div></li><li class="half_rhythm"><div>Store at −20°C. Thaw prior to use.</div></li></ul></div><div id="ml161.s44"><h5>CellTiter-Glo stock solution (1:1)</h5><ul><li class="half_rhythm"><div>Warm CellTiter-Glo substrate to room temperature.</div></li><li class="half_rhythm"><div>Add 100 mL of PBS to resuspend substrate.</div></li><li class="half_rhythm"><div>Aliquot solution into 25-mL tubes.</div></li><li class="half_rhythm"><div>Store at −20°C. Thaw prior to use.</div></li></ul></div><div id="ml161.s45"><h5>CellTiter-Glo solution (1:4)</h5><ul><li class="half_rhythm"><div>Prepare a volume prepared sufficient to complete a day’s worth of runs.</div></li><li class="half_rhythm"><div>For 100 ml, mix together 25 ml CellTiter-Glo (1:1) and 75 ml PBS in a centrifuge bottle, wrapped in foil to shield from light.</div></li></ul></div><div id="ml161.s46"><h5>1.5. μM ATP solution</h5><ul><li class="half_rhythm"><div>Add 1.5 μl of 10 mM ATP to 10 ml of PBS, in a 25-ml conical tube.</div></li></ul></div></div><div id="ml161.s47"><h5>Protocol</h5><ol><li class="half_rhythm"><div>Prepare reagents as follows:</div><ul><li class="half_rhythm"><div>Dilute 10 mM ATP in PBS to 1.5 μM in PBS. Prepare enough reagent for the run (approx. 8 ml/plate + extra dead volume).</div></li><li class="half_rhythm"><div>Thaw frozen aliquots of CTG in PBS and dilute in PBS 1:4. Prepare enough reagent for the run (approx. 4 ml/plate + extra dead volume).</div></li></ul></li><li class="half_rhythm"><div>Prepare a Thermo MultiDrop Combi with a standard cassette, washed, and primed. Fill plates with 20 μl of 1.5 μM ATP in PBS.</div></li><li class="half_rhythm"><div>Add compounds using the CyBiWell (CyBio) Pin tool. Next, 50 nL are pinned using the 25 nL pin array, programmed with 2 dips. Separate each compound addition by a pin wash cycle.</div></li><li class="half_rhythm"><div>During compound addition, prepare a Combi for second reagent addition by washing a standard cassette and priming with new reagent. Add 10 μl of CTG (1:4) in PBS to every well.</div></li><li class="half_rhythm"><div>Transfer plates to an EnVison Plate Reader (Perkin Elmer) to read luminescence. Use the ultra sensitive detection, with the 1536 aperture in place, to decrease bleed-through from adjacent wells. Read time is 0.1s/well.</div></li></ol></div></div></div><div id="ml161.app3"><h3>Appendix C. NMR and LC Data of Probe and Analogs</h3><div id="ml161.fu2" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20CDCl3)%20of%20probe%20CID%201048267.&p=BOOKS&id=55066_ml161fu28.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu28.jpg" alt="1H NMR spectra (300 MHz, CDCl3) of probe CID 1048267." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, CDCl<sub>3</sub>) of probe CID 1048267</span></h3></div><div id="ml161.fu3" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu29.jpg" alt="LC-MS chromatogram of probe CID 1048267." /></div><h3><span class="title">LC-MS chromatogram of probe CID 1048267</span></h3></div><div id="ml161.s51"><h4>Spectroscopic Data for SAR Analogs</h4><div id="ml161.fu4" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%20877874.&p=BOOKS&id=55066_ml161fu30.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu30.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 877874." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 877874</span></h3></div><div id="ml161.fu5" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=LC-MS%20chromatogram%20of%20analog%20CID%20877874.&p=BOOKS&id=55066_ml161fu31.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu31.jpg" alt="LC-MS chromatogram of analog CID 877874." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title">LC-MS chromatogram of analog CID 877874</span></h3></div><div id="ml161.fu6" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%202186104.&p=BOOKS&id=55066_ml161fu32.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu32.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 2186104." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 2186104</span></h3></div><div id="ml161.fu7" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu33.jpg" alt="LC-MS chromatogram of analog CID 2186104." /></div><h3><span class="title">LC-MS chromatogram of analog CID 2186104</span></h3></div><div id="ml161.fu8" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%20792012.&p=BOOKS&id=55066_ml161fu34.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu34.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 792012." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 792012</span></h3></div><div id="ml161.fu9" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu35.jpg" alt="LC-MS chromatogram of analog CID 792012." /></div><h3><span class="title">LC-MS chromatogram of analog CID 792012</span></h3></div><div id="ml161.fu10" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%20669190.&p=BOOKS&id=55066_ml161fu36.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu36.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 669190." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 669190</span></h3></div><div id="ml161.fu11" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu37.jpg" alt="LC-MS chromatogram of analog CID 669190." /></div><h3><span class="title">LC-MS chromatogram of analog CID 669190</span></h3></div><div id="ml161.fu12" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%202274454.&p=BOOKS&id=55066_ml161fu38.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu38.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 2274454." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 2274454</span></h3></div><div id="ml161.fu13" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu39.jpg" alt="LC-MS chromatogram of analog CID 2274454." /></div><h3><span class="title">LC-MS chromatogram of analog CID 2274454</span></h3></div><div id="ml161.fu14" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20CD3OD)%20of%20analog%20CID%2017187609.&p=BOOKS&id=55066_ml161fu40.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu40.jpg" alt="1H NMR spectra (300 MHz, CD3OD) of analog CID 17187609." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, CD<sub>3</sub>OD) of analog CID 17187609</span></h3></div><div id="ml161.fu15" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu41.jpg" alt="LC-MS chromatogram of analog CID 17187609." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187609</span></h3></div><div id="ml161.fu16" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2017187534.&p=BOOKS&id=55066_ml161fu42.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu42.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 17187534." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 17187534</span></h3></div><div id="ml161.fu17" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu43.jpg" alt="LC-MS chromatogram of analog CID 17187534." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187534</span></h3></div><div id="ml161.fu18" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%203438308.&p=BOOKS&id=55066_ml161fu44.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu44.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 3438308." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 3438308</span></h3></div><div id="ml161.fu19" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu45.jpg" alt="LC-MS chromatogram of analog CID 3438308." /></div><h3><span class="title">LC-MS chromatogram of analog CID 3438308</span></h3></div><div id="ml161.fu20" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2017187522.&p=BOOKS&id=55066_ml161fu46.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu46.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 17187522." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 17187522</span></h3></div><div id="ml161.fu21" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu47.jpg" alt="LC-MS chromatogram of analog CID 17187522." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187522</span></h3></div><div id="ml161.fu22" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2017187580.&p=BOOKS&id=55066_ml161fu48.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu48.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 17187580." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 17187580</span></h3></div><div id="ml161.fu23" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu49.jpg" alt="LC-MS chromatogram of analog CID 17187580." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187580</span></h3></div><div id="ml161.fu24" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2049766548.&p=BOOKS&id=55066_ml161fu50.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu50.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 49766548." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 49766548</span></h3></div><div id="ml161.fu25" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu51.jpg" alt="LC-MS chromatogram of analog CID 49766548." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766548</span></h3></div><div id="ml161.fu26" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2049766516.&p=BOOKS&id=55066_ml161fu52.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu52.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 49766516." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 49766516</span></h3></div><div id="ml161.fu27" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu53.jpg" alt="LC-MS chromatogram of analog CID 49766516." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766516</span></h3></div><div id="ml161.fu28" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2017187536.&p=BOOKS&id=55066_ml161fu54.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu54.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 17187536." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 17187536</span></h3></div><div id="ml161.fu29" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu55.jpg" alt="LC-MS chromatogram of analog CID 17187536." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187536</span></h3></div><div id="ml161.fu30" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2049766528.&p=BOOKS&id=55066_ml161fu56.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu56.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 49766528." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 49766528</span></h3></div><div id="ml161.fu31" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu57.jpg" alt="LC-MS chromatogram of analog CID 49766528." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766528</span></h3></div><div id="ml161.fu32" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20CD3OD)%20of%20analog%20CID%2049766546.&p=BOOKS&id=55066_ml161fu58.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu58.jpg" alt="1H NMR spectra (300 MHz, CD3OD) of analog CID 49766546." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, CD<sub>3</sub>OD) of analog CID 49766546</span></h3></div><div id="ml161.fu33" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu59.jpg" alt="LC-MS chromatogram of analog CID 49766546." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766546</span></h3></div><div id="ml161.fu34" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2049766543.&p=BOOKS&id=55066_ml161fu60.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu60.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 49766543." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 49766543</span></h3></div><div id="ml161.fu35" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu61.jpg" alt="LC-MS chromatogram of analog CID 49766543." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766543</span></h3></div><div id="ml161.fu36" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2017187529.&p=BOOKS&id=55066_ml161fu62.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu62.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 17187529." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 17187529</span></h3></div><div id="ml161.fu37" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu63.jpg" alt="LC-MS chromatogram of analog CID 17187529." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187529</span></h3></div><div id="ml161.fu38" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2017187535.&p=BOOKS&id=55066_ml161fu64.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu64.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 17187535." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 17187535</span></h3></div><div id="ml161.fu39" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu65.jpg" alt="LC-MS chromatogram of analog CID 17187535." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187535</span></h3></div><div id="ml161.fu40" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20CD3OD)%20of%20analog%20CID%2049766525.&p=BOOKS&id=55066_ml161fu66.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu66.jpg" alt="1H NMR spectra (300 MHz, CD3OD) of analog CID 49766525." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, CD<sub>3</sub>OD) of analog CID 49766525</span></h3></div><div id="ml161.fu41" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu67.jpg" alt="LC-MS chromatogram of analog CID 49766525." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766525</span></h3></div><div id="ml161.fu42" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2049766536.&p=BOOKS&id=55066_ml161fu68.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu68.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 49766536." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 49766536</span></h3></div><div id="ml161.fu43" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu69.jpg" alt="LC-MS chromatogram of analog CID 49766536." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766536</span></h3></div><div id="ml161.fu44" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%20960686.&p=BOOKS&id=55066_ml161fu70.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu70.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 960686." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 960686</span></h3></div><div id="ml161.fu45" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu71.jpg" alt="LC-MS chromatogram of analog CID 960686." /></div><h3><span class="title">LC-MS chromatogram of analog CID 960686</span></h3></div><div id="ml161.fu46" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20CD3OD)%20of%20analog%20CID%2017187546.&p=BOOKS&id=55066_ml161fu72.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu72.jpg" alt="1H NMR spectra (300 MHz, CD3OD) of analog CID 17187546." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, CD<sub>3</sub>OD) of analog CID 17187546</span></h3></div><div id="ml161.fu47" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu73.jpg" alt="LC-MS chromatogram of analog CID 17187546." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187546</span></h3></div><div id="ml161.fu48" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20CD3OD)%20of%20analog%20CID%20876023.&p=BOOKS&id=55066_ml161fu74.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu74.jpg" alt="1H NMR spectra (300 MHz, CD3OD) of analog CID 876023." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, CD<sub>3</sub>OD) of analog CID 876023</span></h3></div><div id="ml161.fu49" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu75.jpg" alt="LC-MS chromatogram of analog CID 876023." /></div><h3><span class="title">LC-MS chromatogram of analog CID 876023</span></h3></div><div id="ml161.fu50" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%20798120.&p=BOOKS&id=55066_ml161fu76.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu76.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 798120." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 798120</span></h3></div><div id="ml161.fu51" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu77.jpg" alt="LC-MS chromatogram of analog CID 798120." /></div><h3><span class="title">LC-MS chromatogram of analog CID 798120</span></h3></div><div id="ml161.fu52" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2049766507.&p=BOOKS&id=55066_ml161fu78.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img src="/books/NBK55066/bin/ml161fu78.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 49766507." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 49766507</span></h3></div><div id="ml161.fu53" class="figure bk_fig"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu79.jpg" alt="LC-MS chromatogram of analog CID 49766507." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766507</span></h3></div></div></div><div id="ml161.app4"><h3>Appendix D. Compounds Provided to BioFocus</h3><div class="iconblock whole_rhythm clearfix ten_col table-wrap" id="figml161t5"><a href="/books/NBK55066/table/ml161.t5/?report=objectonly" target="object" title="Table A2" class="img_link icnblk_img figpopup" rid-figpopup="figml161t5" rid-ob="figobml161t5"><img class="small-thumb" src="/books/NBK55066/table/ml161.t5/?report=thumb" src-large="/books/NBK55066/table/ml161.t5/?report=previmg" alt="Table A2. Probe and Analog Information." /></a><div class="icnblk_cntnt"><h4 id="ml161.t5"><a href="/books/NBK55066/table/ml161.t5/?report=objectonly" target="object" rid-ob="figobml161t5">Table A2</a></h4><p class="float-caption no_bottom_margin">Probe and Analog Information. </p></div></div></div></div><div id="ml161.ack"><h2 id="_ml161_ack_">Acknowledgements</h2><p>The authors thank Patti Aha, Tyler Aldredge, Frank An, PJ Aspesi, Doug Barker, Jason Burbank, Josh Bittker, Dave DeCaprio, Melanie de Silva, Karen Emmith, Robert Gould, Mary Pat Happ, Kate Hartland, Tom Hasaka, Stephen Johnston, Corrie Lade, Hanh Le, John McGrath, Chris Moore, Evan Mulligan, Myra O’Leary, Gil Walzer, Greg Wendel, and other members of BIPDeC for their assistance with this project. The authors also acknowledge and thank BloodSource (CA), Rhode Island Blood Center, Blood Center of New Jersey, and Blood Center of Wisconsin for supplying platelets for the assays performed at BIPDeC.</p></div><div style="display:none"><div style="display:none" id="figml161f4"><img alt="Image ml161f4" src-large="/books/NBK55066/bin/ml161f4.jpg" /></div></div><div id="bk_toc_contnr"></div></div></div><div class="fm-sec"><h2 id="_NBK55066_pubdet_">Publication Details</h2><h3>Author Information and Affiliations</h3><p class="contrib-group"><h4>Authors</h4><span itemprop="author">Lynn VerPlank</span>,<sup>1</sup><sup>,*</sup> <span itemprop="author">Chris Dockendorff</span>,<sup>1</sup> <span itemprop="author">Joseph Negri</span>,<sup>1</sup> <span itemprop="author">Jose R Perez</span>,<sup>1</sup> <span itemprop="author">James Dilks</span>,<sup>2</sup> <span itemprop="author">Lawrence MacPherson</span>,<sup>1</sup> <span itemprop="author">Michelle Palmer</span>,<sup>1</sup> <span itemprop="author">Robert Flaumenhaft</span>,<sup>2</sup> and <span itemprop="author">Stuart L Schreiber</span><sup>1,3</sup>.</p><h4>Affiliations</h4><div class="affiliation"><sup>1</sup>
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The Broad Institute Probe Development Center, Cambridge, MA</div><div class="affiliation"><sup>2</sup>
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Beth Israel Deaconess Medical Center, Boston, MA</div><div class="affiliation"><sup>3</sup>
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Howard Hughes Medical Institute, Chemistry and Chemical Biology, Harvard University, Cambridge, MA</div><div class="affiliation">
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<sup>*</sup> Corresponding author email:
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<a href="mailto:dev@null" data-email="gro.etutitsnidaorb@knalprev" class="oemail">gro.etutitsnidaorb@knalprev</a></div><h3>Publication History</h3><p class="small">Received: <span itemprop="datePublished">March 25, 2010</span>; Last Update: <span itemprop="dateModified">February 10, 2011</span>.</p><h3>Copyright</h3><div><div class="half_rhythm"><a href="/books/about/copyright/">Copyright Notice</a></div></div><h3>Publisher</h3><p>National Center for Biotechnology Information (US), Bethesda (MD)</p><h3>NLM Citation</h3><p>VerPlank L, Dockendorff C, Negri J, et al. Chemical Genetic Analysis of Platelet Granule Secretion-Probe 3. 2010 Mar 25 [Updated 2011 Feb 10]. 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></p></div><div class="small-screen-prev"><a href="/books/n/mlprobe/ml162/?report=reader"><svg xmlns="http://www.w3.org/2000/svg" viewBox="0 0 100 100" preserveAspectRatio="none"><path d="M75,30 c-80,60 -80,0 0,60 c-30,-60 -30,0 0,-60"></path><text x="20" y="28" textLength="60" style="font-size:25px">Prev</text></svg></a></div><div class="small-screen-next"><a href="/books/n/mlprobe/ml160/?report=reader"><svg xmlns="http://www.w3.org/2000/svg" viewBox="0 0 100 100" preserveAspectRatio="none"><path d="M25,30c80,60 80,0 0,60 c30,-60 30,0 0,-60"></path><text x="20" y="28" textLength="60" style="font-size:25px">Next</text></svg></a></div></article><article data-type="fig" id="figobml161fu1"><div id="ml161.fu1" class="figure"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu1.jpg" alt="Image ml161fu1" /></div></div></article><article data-type="table-wrap" id="figobml161tu1"><div id="ml161.tu1" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK55066/table/ml161.tu1/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml161.tu1_lrgtbl__"><table><tbody><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><b>IUPAC Chemical Name</b></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">2-bromo-N-[3-(butanoylamino)phenyl]benzamide</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><b>PubChem CID</b></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">1048267</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><b>Molecular Weight</b></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">361.23308</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><b>Molecular Formula</b></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">C<sub>17</sub>H<sub>17</sub>BrN<sub>2</sub>O<sub>2</sub></td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><b>AlogP</b></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">3.5</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><b>H-Bond Donor</b></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">2</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><b>H-Bond Acceptor</b></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">2</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><b>Rotatable Bond Count</b></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">5</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><b>Exact Mass</b></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">360.04734</td></tr><tr><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><b>Topological Polar Surface Area</b></td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">58.2</td></tr></tbody></table></div></div></article><article data-type="table-wrap" id="figobml161tu2"><div id="ml161.tu2" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK55066/table/ml161.tu2/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml161.tu2_lrgtbl__"><table><thead><tr><th id="hd_h_ml161.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:bottom;">CID/ML No.</th><th id="hd_h_ml161.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Target Name</th><th id="hd_h_ml161.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">IC<sub>50</sub>/EC<sub>50</sub> (nM) [SID, AID]</th><th id="hd_h_ml161.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Anti-target Name(s)</th><th id="hd_h_ml161.tu2_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">IC<sub>50</sub>/EC<sub>50</sub> (μM) [SID, AID]</th><th id="hd_h_ml161.tu2_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Fold Selective</th><th id="hd_h_ml161.tu2_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary Assay(s) Name: IC<sub>50</sub>/EC<sub>50</sub> (nM) [SID, AID]</th></tr></thead><tbody><tr><td headers="hd_h_ml161.tu2_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">CID 1048267/<a href="/pcsubstance/?term=ML161[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML161</a></td><td headers="hd_h_ml161.tu2_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Granule secretion</td><td headers="hd_h_ml161.tu2_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">11,200 [<a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457110" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID 87457110</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/493100" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 493100</a>]</td><td headers="hd_h_ml161.tu2_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Luciferase</td><td headers="hd_h_ml161.tu2_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">112 [<a href="https://pubchem.ncbi.nlm.nih.gov/substance/1738663" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID 1738663</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1891" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 1891</a>]</td><td headers="hd_h_ml161.tu2_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">10</td><td headers="hd_h_ml161.tu2_1_1_1_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P-selectin 300 [<a href="https://pubchem.ncbi.nlm.nih.gov/substance/87219021" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID 87219021</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2511" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 2511</a>];<br />cAMP [<a href="https://pubchem.ncbi.nlm.nih.gov/substance/87219021" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID 87219021</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2646" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 2646</a>];<br />FITC Phalloidin [<a href="https://pubchem.ncbi.nlm.nih.gov/substance/87219021" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID 87219021</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2655" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 2655</a>];<br />PMA [<a href="https://pubchem.ncbi.nlm.nih.gov/substance/87219021" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID 87219021</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2529" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 2529</a>];<br />Ca Ionophore [<a href="https://pubchem.ncbi.nlm.nih.gov/substance/87219021" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">SID 87219021</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2527" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 2527</a>]</td></tr></tbody></table></div></div></article><article data-type="fig" id="figobml161f1"><div id="ml161.f1" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Figure%201.%20Examples%20of%20Platelet%20Activation%20Inhibitors.&p=BOOKS&id=55066_ml161f1.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161f1.jpg" alt="Figure 1. Examples of Platelet Activation Inhibitors." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="label">Figure 1</span><span class="title">Examples of Platelet Activation Inhibitors</span></h3></div></article><article data-type="fig" id="figobml161f2"><div id="ml161.f2" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161f2.jpg" alt="Scheme 1. Synthesis of 1,3-Diaminophenyl Analog." /></div><h3><span class="label">Scheme 1</span><span class="title">Synthesis of 1,3-Diaminophenyl Analog</span></h3></div></article><article data-type="fig" id="figobml161f3"><div id="ml161.f3" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=Figure%203.%20Critical%20Path%20for%20Probe%20Development.&p=BOOKS&id=55066_ml161f3.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161f3.jpg" alt="Figure 3. Critical Path for Probe Development." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="label">Figure 3</span><span class="title">Critical Path for Probe Development</span></h3></div></article><article data-type="fig" id="figobml161f4"><div id="ml161.f4" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161f4.jpg" alt="Figure 4. Diaminophenyl Probe (CID 1048267/ML161) Concentration-dependent Activity in the Primary Screen Retest (AID 493100)." /></div><h3><span class="label">Figure 4</span><span class="title">Diaminophenyl Probe (CID 1048267/<a href="/pcsubstance/?term=ML161[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML161</a>) Concentration-dependent Activity in the Primary Screen Retest (<a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/493100" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 493100</a>)</span></h3><div class="caption"><p>Primary assay retest measuring extracellular ATP as a measure of granule release: Retest 1, EC<sub>50</sub> 10.3 μM (<i>a);</i> Retest 2, EC<sub>50</sub> 12.1 μM (<i>b</i>)</p></div></div></article><article data-type="fig" id="figobml161f5"><div id="ml161.f5" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161f5.jpg" alt="Figure 5. PBS Stability Data for the Probe (CID 1048267/ML161." /></div><h3><span class="label">Figure 5</span><span class="title">PBS Stability Data for the Probe (CID 1048267/<a href="/pcsubstance/?term=ML161[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML161</a></span></h3></div></article><article data-type="fig" id="figobml161f6"><div id="ml161.f6" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161f6.jpg" alt="Figure 6. Summary of Structure-Activity Relationships for 1,3-diaminophenyl Scaffold." /></div><h3><span class="label">Figure 6</span><span class="title">Summary of Structure-Activity Relationships for 1,3-diaminophenyl Scaffold</span></h3></div></article><article data-type="table-wrap" id="figobml161t1"><div id="ml161.t1" class="table"><h3><span class="label">Table 1</span><span class="title">SAR Analysis and Properties of Diaminophenyl Probe and Analogs</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK55066/table/ml161.t1/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml161.t1_lrgtbl__"><table class="no_margin"><thead><tr><th id="hd_h_ml161.t1_1_1_1_1" colspan="3" rowspan="1" style="text-align:center;vertical-align:top;">SAR Analysis of Diaminophenyl Probe and Analogs</th><th id="hd_h_ml161.t1_1_1_1_2" colspan="2" rowspan="1" style="text-align:center;vertical-align:top;">Structure</th><th id="hd_h_ml161.t1_1_1_1_3" colspan="3" rowspan="1" style="text-align:center;vertical-align:top;">Target Activity</th><th id="hd_h_ml161.t1_1_1_1_4" colspan="2" rowspan="1" style="text-align:center;vertical-align:top;">Anti-target Activity (@ 30 μM)</th></tr><tr><th headers="hd_h_ml161.t1_1_1_1_1" id="hd_h_ml161.t1_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">No.</th><th headers="hd_h_ml161.t1_1_1_1_1" id="hd_h_ml161.t1_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">CID/SID</th><th headers="hd_h_ml161.t1_1_1_1_1" id="hd_h_ml161.t1_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Broad ID</th><th headers="hd_h_ml161.t1_1_1_1_2" id="hd_h_ml161.t1_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><sup>*</sup></th><th headers="hd_h_ml161.t1_1_1_1_2" id="hd_h_ml161.t1_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"></th><th headers="hd_h_ml161.t1_1_1_1_3" id="hd_h_ml161.t1_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">EC<sub>50</sub> (μM) (Dense Granule Release)<sup>a</sup></th><th headers="hd_h_ml161.t1_1_1_1_3" id="hd_h_ml161.t1_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">EC<sub>50</sub> (μM) (P-Selectin expression) actin polym.</th><th headers="hd_h_ml161.t1_1_1_1_3" id="hd_h_ml161.t1_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Inhib. of SFLLRN-induced @10 μM</th><th headers="hd_h_ml161.t1_1_1_1_4" id="hd_h_ml161.t1_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Inhib. of PMA-induced P-selectin expression</th><th headers="hd_h_ml161.t1_1_1_1_4" id="hd_h_ml161.t1_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Inhib. of Ca-ionophore-induced P-selectin expression</th></tr></thead><tbody><tr><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">1</td><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">1048267<br />87457110</td><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K59036917-001-05-0</td><td headers="hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P</td><td headers="hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu2.jpg" alt="Image ml161fu2.jpg" /></div></td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">10.8</td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><1</td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">80%</td><td headers="hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">13%</td><td headers="hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">2</td></tr><tr><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_2 hd_h_ml161.t1_1_1_2_3 hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_4 hd_h_ml161.t1_1_1_2_5 hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_6 hd_h_ml161.t1_1_1_2_7 hd_h_ml161.t1_1_1_2_8 hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_9 hd_h_ml161.t1_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 83 μM; Plasma Protein Binding (Human, Mouse): 97%, ND; Plasma Stability (Human, Mouse): 80%, 1.7%; Purity: 98%</td></tr><tr><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">2</td><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">877874<br />87457111</td><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K38012662-001-05-4</td><td headers="hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P</td><td headers="hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu3.jpg" alt="Image ml161fu3.jpg" /></div></td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">21.2</td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_2 hd_h_ml161.t1_1_1_2_3 hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_4 hd_h_ml161.t1_1_1_2_5 hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_6 hd_h_ml161.t1_1_1_2_7 hd_h_ml161.t1_1_1_2_8 hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_9 hd_h_ml161.t1_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 145 μM; Plasma Protein Binding (Human, Mouse): 93%, 38%; Plasma Stability (Human, Mouse): 36%, 1%; Purity: >95%</td></tr><tr><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">3</td><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">2186104<br />87457112</td><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K75903201-001-05-6</td><td headers="hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P</td><td headers="hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu4.jpg" alt="Image ml161fu4.jpg" /></div></td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_2 hd_h_ml161.t1_1_1_2_3 hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_4 hd_h_ml161.t1_1_1_2_5 hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_6 hd_h_ml161.t1_1_1_2_7 hd_h_ml161.t1_1_1_2_8 hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_9 hd_h_ml161.t1_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 14 μM; Plasma Protein Binding (Human, Mouse): 95%, ND; Plasma Stability (Human, Mouse): 97%, 1.3%; Purity: >95%</td></tr><tr><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">4</td><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">792012<br />87457115</td><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K98991361-001-01-9</td><td headers="hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P</td><td headers="hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu5.jpg" alt="Image ml161fu5.jpg" /></div></td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_2 hd_h_ml161.t1_1_1_2_3 hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_4 hd_h_ml161.t1_1_1_2_5 hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_6 hd_h_ml161.t1_1_1_2_7 hd_h_ml161.t1_1_1_2_8 hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_9 hd_h_ml161.t1_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 361 μM; Plasma Protein Binding (Human, Mouse): 94%, 14%; Plasma Stability (Human, Mouse): >99%, 6%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">5</td><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">669190<br />87457116</td><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K61217870-001-01-6</td><td headers="hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P</td><td headers="hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu6.jpg" alt="Image ml161fu6.jpg" /></div></td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_2 hd_h_ml161.t1_1_1_2_3 hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_4 hd_h_ml161.t1_1_1_2_5 hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_6 hd_h_ml161.t1_1_1_2_7 hd_h_ml161.t1_1_1_2_8 hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_9 hd_h_ml161.t1_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 63 μM; Plasma Protein Binding (Human, Mouse): 90%, 80%; Plasma Stability (Human, Mouse): >99%, >99%; Purity: >95%</td></tr><tr><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">6</td><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">2274454<br />87457118</td><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K88013557-001-01-8</td><td headers="hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P</td><td headers="hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu7.jpg" alt="Image ml161fu7.jpg" /></div></td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t1_1_1_1_1 hd_h_ml161.t1_1_1_2_2 hd_h_ml161.t1_1_1_2_3 hd_h_ml161.t1_1_1_1_2 hd_h_ml161.t1_1_1_2_4 hd_h_ml161.t1_1_1_2_5 hd_h_ml161.t1_1_1_1_3 hd_h_ml161.t1_1_1_2_6 hd_h_ml161.t1_1_1_2_7 hd_h_ml161.t1_1_1_2_8 hd_h_ml161.t1_1_1_1_4 hd_h_ml161.t1_1_1_2_9 hd_h_ml161.t1_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 10 μM; Plasma Protein Binding (Human, Mouse): NT, NT; Plasma Stability (Human, Mouse): NT, NT; Purity: >95%</td></tr></tbody></table></div><div class="tblwrap-foot"><div><dl class="temp-labeled-list small"><dl class="bkr_refwrap"><dt></dt><dd><div id="ml161.tfn2"><p class="no_margin">ND=Not determined; NT = not tested;</p></div></dd></dl><dl class="bkr_refwrap"><dt>*</dt><dd><div id="ml161.tfn3"><p class="no_margin">S = synthesized; P= purchased;</p></div></dd></dl><dl class="bkr_refwrap"><dt>a</dt><dd><div id="ml161.tfn4"><p class="no_margin">Primary HTS assay (average of two different batches of platelets, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/493100" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 493100</a>).</p></div></dd></dl></dl></div></div></div></article><article data-type="table-wrap" id="figobml161t2"><div id="ml161.t2" class="table"><h3><span class="label">Table 2</span><span class="title">SAR Analysis and Properties of Butanoic Acid Derivatives</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK55066/table/ml161.t2/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml161.t2_lrgtbl__"><table class="no_margin"><thead><tr><th id="hd_h_ml161.t2_1_1_1_1" colspan="3" rowspan="1" style="text-align:center;vertical-align:top;">SAR Analysis of Butanoic Acid Derivatives</th><th id="hd_h_ml161.t2_1_1_1_2" colspan="2" rowspan="1" style="text-align:center;vertical-align:top;">Structure</th><th id="hd_h_ml161.t2_1_1_1_3" colspan="3" rowspan="1" style="text-align:center;vertical-align:top;">Target Activity</th><th id="hd_h_ml161.t2_1_1_1_4" colspan="2" rowspan="1" style="text-align:center;vertical-align:top;">Anti-target Activity (@ 30 μM)</th></tr><tr><th headers="hd_h_ml161.t2_1_1_1_1" id="hd_h_ml161.t2_1_1_2_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">No.</th><th headers="hd_h_ml161.t2_1_1_1_1" id="hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">CID/SID</th><th headers="hd_h_ml161.t2_1_1_1_1" id="hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Broad ID</th><th headers="hd_h_ml161.t2_1_1_1_2" id="hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><sup>*</sup></th><th headers="hd_h_ml161.t2_1_1_1_2" id="hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"></th><th headers="hd_h_ml161.t2_1_1_1_3" id="hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">EC<sub>50</sub> (μM) (Dense Granule Release)<sup>a</sup></th><th headers="hd_h_ml161.t2_1_1_1_3" id="hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">EC<sub>50</sub> (μM) (P-Selectin expression)</th><th headers="hd_h_ml161.t2_1_1_1_3" id="hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Inhib. of SFLLRN-induced actin polym. @10 μM</th><th headers="hd_h_ml161.t2_1_1_1_4" id="hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Inhib. of PMA-induced P-selectin expression</th><th headers="hd_h_ml161.t2_1_1_1_4" id="hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Inhib. of Ca-ionophore-induced P-selectin expression</th></tr></thead><tbody><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">1</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">17187609<br />103023297</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K90331903-001-01-1</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu8.jpg" alt="Image ml161fu8.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">31.6</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 133 μM; Plasma Protein Binding (Human, Mouse): 92%, ND; Plasma Stability (Human, Mouse): >99%, 0%; Purity: 99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">2</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">17187534<br />103023303</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K27923933-001-01-7</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu9.jpg" alt="Image ml161fu9.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">6.1</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 123 μM; Plasma Protein Binding (Human, Mouse): 84%, ND; Plasma Stability (Human, Mouse): 77%, 0.8%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">3</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">3438308<br />103023307</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K34858469-001-01-8</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu10.jpg" alt="Image ml161fu10.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 124 μM; Plasma Protein Binding (Human, Mouse): 95%, ND; Plasma Stability (Human, Mouse): >99%, 3%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">4</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">17187522<br />103023309</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K67103402-001-01-9</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu11.jpg" alt="Image ml161fu11.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">5.5</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 225 μM; Plasma Protein Binding (Human, Mouse): 90%, ND; Plasma Stability (Human, Mouse): 89, 0%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">5</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">17187580<br />103023312</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K11361517-001-01-2</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu12.jpg" alt="Image ml161fu12.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 7 μM; Plasma Protein Binding (Human, Mouse): 98%, ND; Plasma Stability (Human, Mouse): >99%, 0%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">6</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">49766548<br />103023317</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K22734993-001-01-0</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu13.jpg" alt="Image ml161fu13.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): <1 μM; Plasma Protein Binding (Human, Mouse): 71%, 58%; Plasma Stability (Human, Mouse): >99%, 46%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">7</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">49766516<br />103023310</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K15547084-001-01-5</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu14.jpg" alt="Image ml161fu14.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 12 μM; Plasma Protein Binding (Human, Mouse): 91%, 44%; Plasma Stability (Human, Mouse): >99%, 7%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">8</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">17187536<br />103023306</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K65412348-001-01-3</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu15.jpg" alt="Image ml161fu15.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 13 μM; Plasma Protein Binding (Human, Mouse): 94%, 50%; Plasma Stability (Human, Mouse): >99%, 10%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">9</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">49766528<br />103023324</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K13763013-001-01-6</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu16.jpg" alt="Image ml161fu16.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): NT; Plasma Protein Binding (Human, Mouse): 79%, 60%; Plasma Stability (Human, Mouse): >99%, 21%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">10</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">49766546<br />103023319</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K91104611-001-01-4</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu17.jpg" alt="Image ml161fu17.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): <1 μM; Plasma Protein Binding (Human, Mouse): 99.0%, 98.7%; Plasma Stability (Human, Mouse): >99%, 31%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">11</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">49766543<br />103023313</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K88072051-001-01-5</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu18.jpg" alt="Image ml161fu18.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): <1 μM; Plasma Protein Binding (Human, Mouse): 98.9%, 78; Plasma Stability (Human, Mouse): >99%, 1.2%; Purity: 99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">12</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">17187529<br />103023315</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K15684313-001-01-9</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu19.jpg" alt="Image ml161fu19.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 34 μM; Plasma Protein Binding (Human, Mouse): 96%, ND; Plasma Stability (Human, Mouse): >99%, 1.3%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">13</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">17187535<br />103023318</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K80676054-001-01-8</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu20.jpg" alt="Image ml161fu20.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 2.4 μM; Plasma Protein Binding (Human, Mouse): 97%, 22%; Plasma Stability (Human, Mouse): >99%, 4%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">14</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">49766525<br />103023323</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K16260147-001-01-9</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu21.jpg" alt="Image ml161fu21.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): NT; Plasma Protein Binding (Human, Mouse): 96%, 92%; Plasma Stability (Human, Mouse): >99%, 39%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">15</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">49766536<br />103023321</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K71348315-001-01-0</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu22.jpg" alt="Image ml161fu22.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 14 μM; Plasma Protein Binding (Human, Mouse): 63%, 66%; Plasma Stability (Human, Mouse): >99%, 42%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">16</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">960686<br />103023305</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K90518988-001-01-1</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu23.jpg" alt="Image ml161fu23.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 9 μM; Plasma Protein Binding (Human, Mouse): 94%, 38%; Plasma Stability (Human, Mouse): >99%, 8%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">17</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">17187546<br />103023304</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K21329444-001-01-5</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu24.jpg" alt="Image ml161fu24.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 11 μM; Plasma Protein Binding (Human, Mouse): 98.5%, ND; Plasma Stability (Human, Mouse): >99%, 1.7%; Purity: 99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">18</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">876023<br />103023302</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K96615411-001-01-6</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu25.jpg" alt="Image ml161fu25.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 497 μM; Plasma Protein Binding (Human, Mouse): 76%, ND; Plasma Stability (Human, Mouse): >99%, <1%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">19</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">798120<br />103023298</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K28436739-001-02-3</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu26.jpg" alt="Image ml161fu26.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): 460 μM; Plasma Protein Binding (Human, Mouse): 64%, 4%; Plasma Stability (Human, Mouse): >99%, 23%; Purity: >99%</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_1" rowspan="2" colspan="1" style="text-align:center;vertical-align:top;">20</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">49766507<br />103023300</td><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">BRD-K95667704-001-01-4</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">S</td><td headers="hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><div class="graphic"><img src="/books/NBK55066/bin/ml161fu27.jpg" alt="Image ml161fu27.jpg" /></div></td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">inactive</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_7" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_8" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td><td headers="hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_10" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NT</td></tr><tr><td headers="hd_h_ml161.t2_1_1_1_1 hd_h_ml161.t2_1_1_2_2 hd_h_ml161.t2_1_1_2_3 hd_h_ml161.t2_1_1_1_2 hd_h_ml161.t2_1_1_2_4 hd_h_ml161.t2_1_1_2_5 hd_h_ml161.t2_1_1_1_3 hd_h_ml161.t2_1_1_2_6 hd_h_ml161.t2_1_1_2_7 hd_h_ml161.t2_1_1_2_8 hd_h_ml161.t2_1_1_1_4 hd_h_ml161.t2_1_1_2_9 hd_h_ml161.t2_1_1_2_10" colspan="9" rowspan="1" style="text-align:center;vertical-align:top;">Solubility (PBS): <1 μM; Plasma Protein Binding (Human, Mouse): 97%, 64%; Plasma Stability (Human, Mouse): >99%, 4%; Purity: >99%</td></tr></tbody></table></div><div class="tblwrap-foot"><div><dl class="temp-labeled-list small"><dl class="bkr_refwrap"><dt></dt><dd><div id="ml161.tfn5"><p class="no_margin">ND=Not determined; NT = not tested;</p></div></dd></dl><dl class="bkr_refwrap"><dt>*</dt><dd><div id="ml161.tfn6"><p class="no_margin">S = synthesized; P= purchased;</p></div></dd></dl><dl class="bkr_refwrap"><dt>a</dt><dd><div id="ml161.tfn7"><p class="no_margin">Primary HTS assay (average of two different batches of platelets, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/493100" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">AID 493100</a>).</p></div></dd></dl></dl></div></div></div></article><article data-type="table-wrap" id="figobml161t3"><div id="ml161.t3" class="table"><h3><span class="label">Table 3</span><span class="title">Search Strings and Databases Employed in the Prior Art Search</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK55066/table/ml161.t3/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml161.t3_lrgtbl__"><table class="no_top_margin"><thead><tr><th id="hd_h_ml161.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Search String</th><th id="hd_h_ml161.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Database</th><th id="hd_h_ml161.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">Hits Found</th></tr></thead><tbody><tr><td headers="hd_h_ml161.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">“platelet granule secretion inhibitor”</td><td headers="hd_h_ml161.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Sci Finder</td><td headers="hd_h_ml161.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">151</td></tr><tr><td headers="hd_h_ml161.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">“allosteric PAR1”</td><td headers="hd_h_ml161.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Sci Finder</td><td headers="hd_h_ml161.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">6</td></tr><tr><td headers="hd_h_ml161.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">“platelet granule secretion inhibitor”</td><td headers="hd_h_ml161.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Web of Science</td><td headers="hd_h_ml161.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">130</td></tr><tr><td headers="hd_h_ml161.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">“allosteric PAR1”</td><td headers="hd_h_ml161.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Web of Science</td><td headers="hd_h_ml161.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">7</td></tr><tr><td headers="hd_h_ml161.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">“granule secretion inhibitor”</td><td headers="hd_h_ml161.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Patent Lens</td><td headers="hd_h_ml161.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">7 (>75% score)</td></tr><tr><td headers="hd_h_ml161.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">“allosteric PAR1”</td><td headers="hd_h_ml161.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Patent Lens</td><td headers="hd_h_ml161.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">1 (>75% score)</td></tr><tr><td headers="hd_h_ml161.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">“platelet granule secretion inhibitor”</td><td headers="hd_h_ml161.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">PubMed</td><td headers="hd_h_ml161.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">127</td></tr><tr><td headers="hd_h_ml161.t3_1_1_1_1" rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">“allosteric PAR1”</td><td headers="hd_h_ml161.t3_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">PubMed</td><td headers="hd_h_ml161.t3_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">7</td></tr></tbody></table></div></div></article><article data-type="table-wrap" id="figobml161t4"><div id="ml161.t4" class="table"><h3><span class="label">Table A1</span><span class="title">Summary of Completed Assays and AIDs</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK55066/table/ml161.t4/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml161.t4_lrgtbl__"><table class="no_top_margin"><thead><tr><th id="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">PubChem AID No.</th><th id="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Type</th><th id="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Target</th><th id="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Concentration Range (μM)</th><th id="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Samples Tested</th></tr></thead><tbody><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1663" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1663</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Primary</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Granule Release</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">7.5 (average)</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">302,457</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1889" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1889</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">DR in Primary (DMSO)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Granule Release</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">11.22-0.09</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">1584</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1891" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1891</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Counterscreen</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Luciferase</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">11.22-0.09</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">1584</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2398" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2398</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2519" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2519</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/493100" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">493100</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">DR in Primary (Powder)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Granule Release</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">35-0.005</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">52</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2518" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2518</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary (DMSO)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P-selectin expression</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">100-0.3</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">28</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2511" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2511</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/493031" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">493031</a>, <a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/493068" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">493068</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary (Powder)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P-selectin expression</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">12.5-0.3</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">34</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2656" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2656</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary (DMSO)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><sup>14</sup>C-serotonin release</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">10</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">7</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2645" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2645</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary (DMSO)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">cAMP</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">10</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">16</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2646" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2646</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary (Powder)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">cAMP</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">10</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">1</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2657" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2657</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary (DMSO)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Actin (SFLLRN-induced)</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">10</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">2</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2655" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2655</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary (Powder)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Actin (SFLLRN-induced)</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">10</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">2</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2509" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2509</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary (DMSO)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P-selectin (PMA-induced)</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">30</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">2</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2529" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2529</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary (Powder)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P-selectin (PMA-induced)</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">30</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">2</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2522" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2522</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary (DMSO)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P-selectin (Ionophore-induced)</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">30</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">2</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/2527" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">2527</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Secondary (Powder)</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">P-selectin (Ionophore-induced)</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">30</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">2</td></tr><tr><td headers="hd_h_ml161.t4_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;"><a href="https://pubchem.ncbi.nlm.nih.gov/bioassay/1678" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">1678</a></td><td headers="hd_h_ml161.t4_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">Summary</td><td headers="hd_h_ml161.t4_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NA</td><td headers="hd_h_ml161.t4_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NA</td><td headers="hd_h_ml161.t4_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:top;">NA</td></tr></tbody></table></div></div></article><article data-type="fig" id="figobml161fu2"><div id="ml161.fu2" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20CDCl3)%20of%20probe%20CID%201048267.&p=BOOKS&id=55066_ml161fu28.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu28.jpg" alt="1H NMR spectra (300 MHz, CDCl3) of probe CID 1048267." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, CDCl<sub>3</sub>) of probe CID 1048267</span></h3></div></article><article data-type="fig" id="figobml161fu3"><div id="ml161.fu3" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu29.jpg" alt="LC-MS chromatogram of probe CID 1048267." /></div><h3><span class="title">LC-MS chromatogram of probe CID 1048267</span></h3></div></article><article data-type="fig" id="figobml161fu4"><div id="ml161.fu4" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%20877874.&p=BOOKS&id=55066_ml161fu30.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu30.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 877874." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 877874</span></h3></div></article><article data-type="fig" id="figobml161fu5"><div id="ml161.fu5" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=LC-MS%20chromatogram%20of%20analog%20CID%20877874.&p=BOOKS&id=55066_ml161fu31.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu31.jpg" alt="LC-MS chromatogram of analog CID 877874." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title">LC-MS chromatogram of analog CID 877874</span></h3></div></article><article data-type="fig" id="figobml161fu6"><div id="ml161.fu6" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%202186104.&p=BOOKS&id=55066_ml161fu32.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu32.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 2186104." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 2186104</span></h3></div></article><article data-type="fig" id="figobml161fu7"><div id="ml161.fu7" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu33.jpg" alt="LC-MS chromatogram of analog CID 2186104." /></div><h3><span class="title">LC-MS chromatogram of analog CID 2186104</span></h3></div></article><article data-type="fig" id="figobml161fu8"><div id="ml161.fu8" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%20792012.&p=BOOKS&id=55066_ml161fu34.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu34.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 792012." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 792012</span></h3></div></article><article data-type="fig" id="figobml161fu9"><div id="ml161.fu9" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu35.jpg" alt="LC-MS chromatogram of analog CID 792012." /></div><h3><span class="title">LC-MS chromatogram of analog CID 792012</span></h3></div></article><article data-type="fig" id="figobml161fu10"><div id="ml161.fu10" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%20669190.&p=BOOKS&id=55066_ml161fu36.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu36.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 669190." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 669190</span></h3></div></article><article data-type="fig" id="figobml161fu11"><div id="ml161.fu11" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu37.jpg" alt="LC-MS chromatogram of analog CID 669190." /></div><h3><span class="title">LC-MS chromatogram of analog CID 669190</span></h3></div></article><article data-type="fig" id="figobml161fu12"><div id="ml161.fu12" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%202274454.&p=BOOKS&id=55066_ml161fu38.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu38.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 2274454." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 2274454</span></h3></div></article><article data-type="fig" id="figobml161fu13"><div id="ml161.fu13" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu39.jpg" alt="LC-MS chromatogram of analog CID 2274454." /></div><h3><span class="title">LC-MS chromatogram of analog CID 2274454</span></h3></div></article><article data-type="fig" id="figobml161fu14"><div id="ml161.fu14" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20CD3OD)%20of%20analog%20CID%2017187609.&p=BOOKS&id=55066_ml161fu40.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu40.jpg" alt="1H NMR spectra (300 MHz, CD3OD) of analog CID 17187609." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, CD<sub>3</sub>OD) of analog CID 17187609</span></h3></div></article><article data-type="fig" id="figobml161fu15"><div id="ml161.fu15" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu41.jpg" alt="LC-MS chromatogram of analog CID 17187609." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187609</span></h3></div></article><article data-type="fig" id="figobml161fu16"><div id="ml161.fu16" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2017187534.&p=BOOKS&id=55066_ml161fu42.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu42.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 17187534." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 17187534</span></h3></div></article><article data-type="fig" id="figobml161fu17"><div id="ml161.fu17" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu43.jpg" alt="LC-MS chromatogram of analog CID 17187534." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187534</span></h3></div></article><article data-type="fig" id="figobml161fu18"><div id="ml161.fu18" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%203438308.&p=BOOKS&id=55066_ml161fu44.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu44.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 3438308." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 3438308</span></h3></div></article><article data-type="fig" id="figobml161fu19"><div id="ml161.fu19" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu45.jpg" alt="LC-MS chromatogram of analog CID 3438308." /></div><h3><span class="title">LC-MS chromatogram of analog CID 3438308</span></h3></div></article><article data-type="fig" id="figobml161fu20"><div id="ml161.fu20" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2017187522.&p=BOOKS&id=55066_ml161fu46.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu46.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 17187522." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 17187522</span></h3></div></article><article data-type="fig" id="figobml161fu21"><div id="ml161.fu21" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu47.jpg" alt="LC-MS chromatogram of analog CID 17187522." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187522</span></h3></div></article><article data-type="fig" id="figobml161fu22"><div id="ml161.fu22" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2017187580.&p=BOOKS&id=55066_ml161fu48.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu48.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 17187580." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 17187580</span></h3></div></article><article data-type="fig" id="figobml161fu23"><div id="ml161.fu23" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu49.jpg" alt="LC-MS chromatogram of analog CID 17187580." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187580</span></h3></div></article><article data-type="fig" id="figobml161fu24"><div id="ml161.fu24" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2049766548.&p=BOOKS&id=55066_ml161fu50.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu50.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 49766548." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 49766548</span></h3></div></article><article data-type="fig" id="figobml161fu25"><div id="ml161.fu25" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu51.jpg" alt="LC-MS chromatogram of analog CID 49766548." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766548</span></h3></div></article><article data-type="fig" id="figobml161fu26"><div id="ml161.fu26" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2049766516.&p=BOOKS&id=55066_ml161fu52.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu52.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 49766516." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 49766516</span></h3></div></article><article data-type="fig" id="figobml161fu27"><div id="ml161.fu27" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu53.jpg" alt="LC-MS chromatogram of analog CID 49766516." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766516</span></h3></div></article><article data-type="fig" id="figobml161fu28"><div id="ml161.fu28" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2017187536.&p=BOOKS&id=55066_ml161fu54.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu54.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 17187536." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 17187536</span></h3></div></article><article data-type="fig" id="figobml161fu29"><div id="ml161.fu29" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu55.jpg" alt="LC-MS chromatogram of analog CID 17187536." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187536</span></h3></div></article><article data-type="fig" id="figobml161fu30"><div id="ml161.fu30" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2049766528.&p=BOOKS&id=55066_ml161fu56.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu56.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 49766528." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 49766528</span></h3></div></article><article data-type="fig" id="figobml161fu31"><div id="ml161.fu31" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu57.jpg" alt="LC-MS chromatogram of analog CID 49766528." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766528</span></h3></div></article><article data-type="fig" id="figobml161fu32"><div id="ml161.fu32" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20CD3OD)%20of%20analog%20CID%2049766546.&p=BOOKS&id=55066_ml161fu58.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu58.jpg" alt="1H NMR spectra (300 MHz, CD3OD) of analog CID 49766546." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, CD<sub>3</sub>OD) of analog CID 49766546</span></h3></div></article><article data-type="fig" id="figobml161fu33"><div id="ml161.fu33" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu59.jpg" alt="LC-MS chromatogram of analog CID 49766546." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766546</span></h3></div></article><article data-type="fig" id="figobml161fu34"><div id="ml161.fu34" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2049766543.&p=BOOKS&id=55066_ml161fu60.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu60.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 49766543." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 49766543</span></h3></div></article><article data-type="fig" id="figobml161fu35"><div id="ml161.fu35" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu61.jpg" alt="LC-MS chromatogram of analog CID 49766543." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766543</span></h3></div></article><article data-type="fig" id="figobml161fu36"><div id="ml161.fu36" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2017187529.&p=BOOKS&id=55066_ml161fu62.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu62.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 17187529." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 17187529</span></h3></div></article><article data-type="fig" id="figobml161fu37"><div id="ml161.fu37" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu63.jpg" alt="LC-MS chromatogram of analog CID 17187529." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187529</span></h3></div></article><article data-type="fig" id="figobml161fu38"><div id="ml161.fu38" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2017187535.&p=BOOKS&id=55066_ml161fu64.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu64.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 17187535." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 17187535</span></h3></div></article><article data-type="fig" id="figobml161fu39"><div id="ml161.fu39" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu65.jpg" alt="LC-MS chromatogram of analog CID 17187535." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187535</span></h3></div></article><article data-type="fig" id="figobml161fu40"><div id="ml161.fu40" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20CD3OD)%20of%20analog%20CID%2049766525.&p=BOOKS&id=55066_ml161fu66.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu66.jpg" alt="1H NMR spectra (300 MHz, CD3OD) of analog CID 49766525." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, CD<sub>3</sub>OD) of analog CID 49766525</span></h3></div></article><article data-type="fig" id="figobml161fu41"><div id="ml161.fu41" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu67.jpg" alt="LC-MS chromatogram of analog CID 49766525." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766525</span></h3></div></article><article data-type="fig" id="figobml161fu42"><div id="ml161.fu42" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2049766536.&p=BOOKS&id=55066_ml161fu68.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu68.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 49766536." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 49766536</span></h3></div></article><article data-type="fig" id="figobml161fu43"><div id="ml161.fu43" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu69.jpg" alt="LC-MS chromatogram of analog CID 49766536." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766536</span></h3></div></article><article data-type="fig" id="figobml161fu44"><div id="ml161.fu44" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%20960686.&p=BOOKS&id=55066_ml161fu70.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu70.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 960686." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 960686</span></h3></div></article><article data-type="fig" id="figobml161fu45"><div id="ml161.fu45" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu71.jpg" alt="LC-MS chromatogram of analog CID 960686." /></div><h3><span class="title">LC-MS chromatogram of analog CID 960686</span></h3></div></article><article data-type="fig" id="figobml161fu46"><div id="ml161.fu46" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20CD3OD)%20of%20analog%20CID%2017187546.&p=BOOKS&id=55066_ml161fu72.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu72.jpg" alt="1H NMR spectra (300 MHz, CD3OD) of analog CID 17187546." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, CD<sub>3</sub>OD) of analog CID 17187546</span></h3></div></article><article data-type="fig" id="figobml161fu47"><div id="ml161.fu47" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu73.jpg" alt="LC-MS chromatogram of analog CID 17187546." /></div><h3><span class="title">LC-MS chromatogram of analog CID 17187546</span></h3></div></article><article data-type="fig" id="figobml161fu48"><div id="ml161.fu48" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20CD3OD)%20of%20analog%20CID%20876023.&p=BOOKS&id=55066_ml161fu74.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu74.jpg" alt="1H NMR spectra (300 MHz, CD3OD) of analog CID 876023." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, CD<sub>3</sub>OD) of analog CID 876023</span></h3></div></article><article data-type="fig" id="figobml161fu49"><div id="ml161.fu49" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu75.jpg" alt="LC-MS chromatogram of analog CID 876023." /></div><h3><span class="title">LC-MS chromatogram of analog CID 876023</span></h3></div></article><article data-type="fig" id="figobml161fu50"><div id="ml161.fu50" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%20798120.&p=BOOKS&id=55066_ml161fu76.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu76.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 798120." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 798120</span></h3></div></article><article data-type="fig" id="figobml161fu51"><div id="ml161.fu51" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu77.jpg" alt="LC-MS chromatogram of analog CID 798120." /></div><h3><span class="title">LC-MS chromatogram of analog CID 798120</span></h3></div></article><article data-type="fig" id="figobml161fu52"><div id="ml161.fu52" class="figure bk_fig"><div class="graphic"><a href="/core/lw/2.0/html/tileshop_pmc/tileshop_pmc_inline.html?title=1H%20NMR%20spectra%20(300%20MHz%2C%20d6-DMSO)%20of%20analog%20CID%2049766507.&p=BOOKS&id=55066_ml161fu78.jpg" target="tileshopwindow" class="inline_block pmc_inline_block ts_canvas img_link" title="Click on image to zoom"><div class="ts_bar small" title="Click on image to zoom"></div><img data-src="/books/NBK55066/bin/ml161fu78.jpg" alt="1H NMR spectra (300 MHz, d6-DMSO) of analog CID 49766507." class="tileshop" title="Click on image to zoom" /></a></div><h3><span class="title"><sup>1</sup>H NMR spectra (300 MHz, d<sub>6</sub>-DMSO) of analog CID 49766507</span></h3></div></article><article data-type="fig" id="figobml161fu53"><div id="ml161.fu53" class="figure bk_fig"><div class="graphic"><img data-src="/books/NBK55066/bin/ml161fu79.jpg" alt="LC-MS chromatogram of analog CID 49766507." /></div><h3><span class="title">LC-MS chromatogram of analog CID 49766507</span></h3></div></article><article data-type="table-wrap" id="figobml161t5"><div id="ml161.t5" class="table"><h3><span class="label">Table A2</span><span class="title">Probe and Analog Information</span></h3><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK55066/table/ml161.t5/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__ml161.t5_lrgtbl__"><table class="no_margin"><thead><tr><th id="hd_h_ml161.t5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">BRD</th><th id="hd_h_ml161.t5_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">SID</th><th id="hd_h_ml161.t5_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">CID</th><th id="hd_h_ml161.t5_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">P/A</th><th id="hd_h_ml161.t5_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS</th><th id="hd_h_ml161.t5_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">ML</th></tr></thead><tbody><tr><td headers="hd_h_ml161.t5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">BRD-K59036917-001-05-0</td><td headers="hd_h_ml161.t5_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457110" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457110</a></td><td headers="hd_h_ml161.t5_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">1048267</td><td headers="hd_h_ml161.t5_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">P</td><td headers="hd_h_ml161.t5_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699906</td><td headers="hd_h_ml161.t5_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="/pcsubstance/?term=ML161[synonym]" ref="pagearea=body&targetsite=entrez&targetcat=term&targettype=pubchem">ML161</a></td></tr><tr><td headers="hd_h_ml161.t5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">BRD-K38012662-001-05-4</td><td headers="hd_h_ml161.t5_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457111" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457111</a></td><td headers="hd_h_ml161.t5_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">877874</td><td headers="hd_h_ml161.t5_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">A</td><td headers="hd_h_ml161.t5_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699907</td><td headers="hd_h_ml161.t5_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NA</td></tr><tr><td headers="hd_h_ml161.t5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">BRD-K75903201-001-05-6</td><td headers="hd_h_ml161.t5_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457112" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457112</a></td><td headers="hd_h_ml161.t5_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">2186104</td><td headers="hd_h_ml161.t5_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">A</td><td headers="hd_h_ml161.t5_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699908</td><td headers="hd_h_ml161.t5_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NA</td></tr><tr><td headers="hd_h_ml161.t5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">BRD-K04156788-001-01-7</td><td headers="hd_h_ml161.t5_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457113" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457113</a></td><td headers="hd_h_ml161.t5_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">25126</td><td headers="hd_h_ml161.t5_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">A</td><td headers="hd_h_ml161.t5_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699909</td><td headers="hd_h_ml161.t5_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NA</td></tr><tr><td headers="hd_h_ml161.t5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">BRD-K63175663-001-01-5</td><td headers="hd_h_ml161.t5_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457114" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457114</a></td><td headers="hd_h_ml161.t5_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">669117</td><td headers="hd_h_ml161.t5_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">A</td><td headers="hd_h_ml161.t5_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699910</td><td headers="hd_h_ml161.t5_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NA</td></tr><tr><td headers="hd_h_ml161.t5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">BRD-K98991361-001-01-9</td><td headers="hd_h_ml161.t5_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457115" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457115</a></td><td headers="hd_h_ml161.t5_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">792012</td><td headers="hd_h_ml161.t5_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">A</td><td headers="hd_h_ml161.t5_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699911</td><td headers="hd_h_ml161.t5_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NA</td></tr><tr><td headers="hd_h_ml161.t5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">BRD-K61217870-001-01-6</td><td headers="hd_h_ml161.t5_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457116" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457116</a></td><td headers="hd_h_ml161.t5_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">669190</td><td headers="hd_h_ml161.t5_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">A</td><td headers="hd_h_ml161.t5_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699912</td><td headers="hd_h_ml161.t5_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NA</td></tr><tr><td headers="hd_h_ml161.t5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">BRD-K88013557-001-01-8</td><td headers="hd_h_ml161.t5_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457118" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457118</a></td><td headers="hd_h_ml161.t5_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">2274454</td><td headers="hd_h_ml161.t5_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">A</td><td headers="hd_h_ml161.t5_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699922</td><td headers="hd_h_ml161.t5_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NA</td></tr><tr><td headers="hd_h_ml161.t5_1_1_1_1" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">BRD-K96213252-001-01-2</td><td headers="hd_h_ml161.t5_1_1_1_2" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;"><a href="https://pubchem.ncbi.nlm.nih.gov/substance/87457117" ref="pagearea=body&targetsite=entrez&targetcat=link&targettype=pubchem">87457117</a></td><td headers="hd_h_ml161.t5_1_1_1_3" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">727425</td><td headers="hd_h_ml161.t5_1_1_1_4" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">A</td><td headers="hd_h_ml161.t5_1_1_1_5" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">MLS002699924</td><td headers="hd_h_ml161.t5_1_1_1_6" rowspan="1" colspan="1" style="text-align:center;vertical-align:middle;">NA</td></tr></tbody></table></div><div class="tblwrap-foot"><div><dl class="temp-labeled-list small"><dl class="bkr_refwrap"><dt></dt><dd><div id="ml161.tfn8"><p class="no_margin">A=analog; NA= not applicable; P=probe</p></div></dd></dl></dl></div></div></div></article></div><div id="jr-scripts"><script src="/corehtml/pmc/jatsreader/ptpmc_3.22/js/libs.min.js"> </script><script src="/corehtml/pmc/jatsreader/ptpmc_3.22/js/jr.min.js"> </script></div></div>
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