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<meta name="robots" content="INDEX,FOLLOW,NOARCHIVE" /><meta name="citation_inbook_title" content="Molecular Imaging and Contrast Agent Database (MICAD) [Internet]" /><meta name="citation_title" content="Pyro-Gly-Pro-Leu-Gly-Leu-Ala-Arg-Lys(BHQ3)" /><meta name="citation_publisher" content="National Center for Biotechnology Information (US)" /><meta name="citation_date" content="2008/07/21" /><meta name="citation_author" content="Huiming Zhang" /><meta name="citation_pmid" content="20641316" /><meta name="citation_fulltext_html_url" content="https://www.ncbi.nlm.nih.gov/books/NBK23111/" /><link rel="schema.DC" href="http://purl.org/DC/elements/1.0/" /><meta name="DC.Title" content="Pyro-Gly-Pro-Leu-Gly-Leu-Ala-Arg-Lys(BHQ3)" /><meta name="DC.Type" content="Text" /><meta name="DC.Publisher" content="National Center for Biotechnology Information (US)" /><meta name="DC.Contributor" content="Huiming Zhang" /><meta name="DC.Date" content="2008/07/21" /><meta name="DC.Identifier" content="https://www.ncbi.nlm.nih.gov/books/NBK23111/" /><meta name="description" content="Photodynamic therapy (PDT), also known as photochemotherapy, uses light-activated photosensitizers (PS) in the presence of oxygen to kill cells (1). PDT has become a promising modality to treat skin, esophagus, and lung cancers, as well as other diseases such as atherosclerosis, macular degeneration, and rheumatoid arthritis (2). In PDT, light excites the singlet state of the PS, followed by intersystem transition from the singlet state to the triplet state; then, the energy is transferred from the triplet state of the PS to the triplet ground state of oxygen, 3O2(X3Σg-) (3O2 triplet state quenching) to generate singlet oxygen, 1O2(a1Δg) (3). The produced 1O2 is a major cytotoxic agent that has a short life time (<200 ns) and an average diffusion range (~20 nm, which is smaller than the diameter of a cell) (2). Such a short diffusion range requires the delivery of target-specific PS agents into subcellular compartments such as cytoskeletal tubulin, lysosomes, mitochondria, plasma membrane, and the nucleus, where they can generate 1O2 efficiently (2). A novel type of PS agent, called a photodynamic molecular beacon (PMB) or killer beacon, has been developed to meet this requirement (2, 4). A typical PMB consists of four modular components: a fluorescent PS, a quencher, a linker, and a delivery vehicle. The target-specific linkers keep the fluorescent PS and the quencher within effective distance of the Föster radius (3–6 nm) (2), which allows efficient fluorescence resonance energy transfer between the fluorescent PS and the quencher. As a result, the fluorescent PS is silent until the PMB meets the target, where the enzyme cleaves the linker and activates the fluorescence of the PS (4). Thus, the PS performs two functions by producing 1O2 to kill cells and by illuminating detectable fluorescence to image its own therapeutic outcome (5)." /><meta name="og:title" content="Pyro-Gly-Pro-Leu-Gly-Leu-Ala-Arg-Lys(BHQ3)" /><meta name="og:type" content="book" /><meta name="og:description" content="Photodynamic therapy (PDT), also known as photochemotherapy, uses light-activated photosensitizers (PS) in the presence of oxygen to kill cells (1). PDT has become a promising modality to treat skin, esophagus, and lung cancers, as well as other diseases such as atherosclerosis, macular degeneration, and rheumatoid arthritis (2). In PDT, light excites the singlet state of the PS, followed by intersystem transition from the singlet state to the triplet state; then, the energy is transferred from the triplet state of the PS to the triplet ground state of oxygen, 3O2(X3Σg-) (3O2 triplet state quenching) to generate singlet oxygen, 1O2(a1Δg) (3). The produced 1O2 is a major cytotoxic agent that has a short life time (<200 ns) and an average diffusion range (~20 nm, which is smaller than the diameter of a cell) (2). Such a short diffusion range requires the delivery of target-specific PS agents into subcellular compartments such as cytoskeletal tubulin, lysosomes, mitochondria, plasma membrane, and the nucleus, where they can generate 1O2 efficiently (2). A novel type of PS agent, called a photodynamic molecular beacon (PMB) or killer beacon, has been developed to meet this requirement (2, 4). A typical PMB consists of four modular components: a fluorescent PS, a quencher, a linker, and a delivery vehicle. The target-specific linkers keep the fluorescent PS and the quencher within effective distance of the Föster radius (3–6 nm) (2), which allows efficient fluorescence resonance energy transfer between the fluorescent PS and the quencher. As a result, the fluorescent PS is silent until the PMB meets the target, where the enzyme cleaves the linker and activates the fluorescence of the PS (4). Thus, the PS performs two functions by producing 1O2 to kill cells and by illuminating detectable fluorescence to image its own therapeutic outcome (5)." /><meta name="og:url" content="https://www.ncbi.nlm.nih.gov/books/NBK23111/" /><meta name="og:site_name" content="NCBI Bookshelf" /><meta name="og:image" content="https://www.ncbi.nlm.nih.gov/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-micad-lrg.png" /><meta name="twitter:card" content="summary" /><meta name="twitter:site" content="@ncbibooks" /><meta name="bk-non-canon-loc" content="/books/n/micad/PMB/" /><link rel="canonical" href="https://www.ncbi.nlm.nih.gov/books/NBK23111/" /><link rel="stylesheet" href="/corehtml/pmc/css/figpopup.css" type="text/css" media="screen" /><link rel="stylesheet" href="/corehtml/pmc/css/bookshelf/2.26/css/books.min.css" type="text/css" /><link rel="stylesheet" href="/corehtml/pmc/css/bookshelf/2.26/css/books_print.min.css" type="text/css" media="print" /><style type="text/css">p a.figpopup{display:inline !important} .bk_tt {font-family: monospace} .first-line-outdent .bk_ref {display: inline} .body-content h2, .body-content .h2 {border-bottom: 1px solid #97B0C8} .body-content h2.inline {border-bottom: none} a.page-toc-label , .jig-ncbismoothscroll a {text-decoration:none;border:0 !important} .temp-labeled-list .graphic {display:inline-block !important} .temp-labeled-list img{width:100%}</style><script type="text/javascript" src="/corehtml/pmc/js/jquery.hoverIntent.min.js"> </script><script type="text/javascript" src="/corehtml/pmc/js/common.min.js?_=3.18"> </script><script type="text/javascript" src="/corehtml/pmc/js/large-obj-scrollbars.min.js"> </script><script type="text/javascript">window.name="mainwindow";</script><script type="text/javascript" src="/corehtml/pmc/js/bookshelf/2.26/book-toc.min.js"> </script><script type="text/javascript" src="/corehtml/pmc/js/bookshelf/2.26/books.min.js"> </script><meta name="book-collection" content="NONE" />
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<div class="pre-content"><div><div class="bk_prnt"><p class="small">NCBI Bookshelf. A service of the National Library of Medicine, National Institutes of Health.</p><p>Molecular Imaging and Contrast Agent Database (MICAD) [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2004-2013. </p></div><div class="iconblock clearfix whole_rhythm no_top_margin bk_noprnt"><a class="img_link icnblk_img" title="Table of Contents Page" href="/books/n/micad/"><img class="source-thumb" src="/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-micad-lrg.png" alt="Cover of Molecular Imaging and Contrast Agent Database (MICAD)" height="100px" width="80px" /></a><div class="icnblk_cntnt eight_col"><h2>Molecular Imaging and Contrast Agent Database (MICAD) [Internet].</h2><a data-jig="ncbitoggler" href="#__NBK23111_dtls__">Show details</a><div style="display:none" class="ui-widget" id="__NBK23111_dtls__"><div>Bethesda (MD): <a href="https://www.ncbi.nlm.nih.gov/" ref="pagearea=page-banner&targetsite=external&targetcat=link&targettype=publisher">National Center for Biotechnology Information (US)</a>; 2004-2013.</div></div><div class="half_rhythm"><ul class="inline_list"><li style="margin-right:1em"><a class="bk_cntns" href="/books/n/micad/">Contents</a></li></ul></div><div class="bk_noprnt"><form method="get" action="/books/n/micad/" id="bk_srch"><div class="bk_search"><label for="bk_term" class="offscreen_noflow">Search term</label><input type="text" title="Search this book" id="bk_term" name="term" value="" data-jig="ncbiclearbutton" /> <input type="submit" class="jig-ncbibutton" value="Search this book" submit="false" style="padding: 0.1em 0.4em;" /></div></form></div></div><div class="icnblk_cntnt two_col"><div class="pagination bk_noprnt"><a class="active page_link prev" href="/books/n/micad/PPB/" title="Previous page in this title">< Prev</a><a class="active page_link next" href="/books/n/micad/sosquantumdotqd/" title="Next page in this title">Next ></a></div></div></div></div></div>
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<div class="main-content lit-style" itemscope="itemscope" itemtype="http://schema.org/CreativeWork"><div class="meta-content fm-sec"><h1 id="_NBK23111_"><span class="title" itemprop="name">Pyro-Gly-Pro-Leu-Gly-Leu-Ala-Arg-Lys(BHQ3) </span></h1><div itemprop="alternativeHeadline" class="subtitle whole_rhythm">PP<sub>MMP7</sub>B</div><p class="contrib-group"><span itemprop="author">Huiming Zhang</span>, PhD.</p><a data-jig="ncbitoggler" href="#__NBK23111_ai__" style="border:0;text-decoration:none">Author Information and Affiliations</a><div style="display:none" class="ui-widget" id="__NBK23111_ai__"><div class="contrib half_rhythm"><span itemprop="author">Huiming Zhang</span>, PhD<div class="affiliation small">
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National Center for Biotechnology Information, NLM, NIH, Bethesda, MD,
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<span class="before-email-separator"></span><span class="email-label">Email: </span><a href="mailto:dev@null" data-email="vog.hin.mln.ibcn@dacim" class="oemail">vog.hin.mln.ibcn@dacim</a>
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</div></div></div><p class="small">Created: <span itemprop="datePublished">May 22, 2008</span>; Last Update: <span itemprop="dateModified">July 21, 2008</span>.</p></div><div class="jig-ncbiinpagenav body-content whole_rhythm" data-jigconfig="allHeadingLevels: ['h2'],smoothScroll: false" itemprop="text"><div id="PMB.T1" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK23111/table/PMB.T1/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__PMB.T1_lrgtbl__"><table><tbody><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Chemical name:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Pyro-Gly-Pro-Leu-Gly-Leu-Ala-Arg-Lys(BHQ3)</td><td rowspan="9" colspan="1" style="text-align:left;vertical-align:middle;"></td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Abbreviated name:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">PP<sub>MMP7</sub>B</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Synonym:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"></td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Agent category:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Peptide</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Target:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Matrix metalloproteinase subtype-7 (MMP-7)</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Target category:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Enzyme</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Method of detection:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Optical imaging, near-infrared (NIR) fluorescence</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Source of signal/contrast:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Pyropheophorbide α (Pyro)</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Activation:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Yes</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
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<b>Studies:</b>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
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<ul class="simple-list"><li class="half_rhythm"><div>
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<img alt="Checkbox" src="/corehtml/pmc/css/bookshelf/2.26/img/studies.checkbox.png" />
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<i>In vitro</i>
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</div></li></ul>
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<ul class="simple-list"><li class="half_rhythm"><div>
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<img alt="Checkbox" src="/corehtml/pmc/css/bookshelf/2.26/img/studies.checkbox.png" /> Rodents
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</div></li></ul>
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</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">No structure is currently available in <a href="http://pubchem.ncbi.nlm.nih.gov" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubChem</a>.</td></tr></tbody></table></div></div><div id="PMB.Background"><h2 id="_PMB_Background_">Background</h2><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=PPMMP7B" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>Photodynamic therapy (PDT), also known as photochemotherapy, uses light-activated photosensitizers (PS) in the presence of oxygen to kill cells (<a class="bk_pop" href="#PMB.REF.1">1</a>). PDT has become a promising modality to treat skin, esophagus, and lung cancers, as well as other diseases such as atherosclerosis, macular degeneration, and rheumatoid arthritis (<a class="bk_pop" href="#PMB.REF.2">2</a>). In PDT, light excites the singlet state of the PS, followed by intersystem transition from the singlet state to the triplet state; then, the energy is transferred from the triplet state of the PS to the triplet ground state of oxygen, <sup>3</sup>O<sub>2</sub>(X<sup>3</sup>Σ<sub>g</sub><sup>-</sup>) (<sup>3</sup>O<sub>2</sub> triplet state quenching) to generate singlet oxygen, <sup>1</sup>O<sub>2</sub>(a<sup>1</sup>Δ<sub>g</sub>) (<a class="bk_pop" href="#PMB.REF.3">3</a>). The produced <sup>1</sup>O<sub>2</sub> is a major cytotoxic agent that has a short life time (<200 ns) and an average diffusion range (~20 nm, which is smaller than the diameter of a cell) (<a class="bk_pop" href="#PMB.REF.2">2</a>). Such a short diffusion range requires the delivery of target-specific PS agents into subcellular compartments such as cytoskeletal tubulin, lysosomes, mitochondria, plasma membrane, and the nucleus, where they can generate <sup>1</sup>O<sub>2</sub> efficiently (<a class="bk_pop" href="#PMB.REF.2">2</a>). A novel type of PS agent, called a photodynamic molecular beacon (PMB) or killer beacon, has been developed to meet this requirement (<a class="bk_pop" href="#PMB.REF.2">2</a>, <a class="bk_pop" href="#PMB.REF.4">4</a>). A typical PMB consists of four modular components: a fluorescent PS, a quencher, a linker, and a delivery vehicle. The target-specific linkers keep the fluorescent PS and the quencher within effective distance of the Föster radius (3–6 nm) (<a class="bk_pop" href="#PMB.REF.2">2</a>), which allows efficient fluorescence resonance energy transfer between the fluorescent PS and the quencher. As a result, the fluorescent PS is silent until the PMB meets the target, where the enzyme cleaves the linker and activates the fluorescence of the PS (<a class="bk_pop" href="#PMB.REF.4">4</a>). Thus, the PS performs two functions by producing <sup>1</sup>O<sub>2</sub> to kill cells and by illuminating detectable fluorescence to image its own therapeutic outcome (<a class="bk_pop" href="#PMB.REF.5">5</a>).</p><p>Matrix metalloproteinases (MMPs) have been pharmaceutical targets for many years because they play important roles in many diseases such as atherosclerosis, lung pulmonary fibrosis, and cancer (<a class="bk_pop" href="#PMB.REF.4">4</a>). MMPs in tumors aid the degradation of extracellular matrix, facilitate neoplastic cell motility, and direct cell invasion (<a class="bk_pop" href="#PMB.REF.4">4</a>). Also known as matrilysin, MMP subtype-7 (MMP7) is one of only a few MMPs that are actually secreted by tumor cells (<a class="bk_pop" href="#PMB.REF.6">6</a>). Pyro-Gly-Pro-Leu-Gly-Leu-Ala-Arg-Lys(BHQ3) (PP<sub>MMP7</sub>B) is a PMB specific for MMP7, and it is detectable with near-infrared (NIR) fluorescence imaging (<a class="bk_pop" href="#PMB.REF.4">4</a>). PP<sub>MMP7</sub>B consists of the infrared fluorescence PS pyropheophorbide α (Pyro), a black hole quencher 3 (BHQ3), and a peptide linker (Gly-Pro-Leu-Gly-Leu-Ala-Arg-Lys (GPLGLARK)) (<a class="bk_pop" href="#PMB.REF.4">4</a>). The peptide contains the tripeptide motif Pro-Leu-Gly for MMP7 recognition, and the cleavage site is located between the Gly and Leu residues. Pyro acts as the intracellular delivery vehicle and as the PS (absorption, 665 nm; emission, 675 nm and 720 nm) with good <sup>1</sup>O<sub>2</sub> production (50%). Pyro lacks dark toxicity (toxicity in absence of light) because of its low absorption between 450–600 nm. BHQ3 (absorption, 672 nm) can efficiently quench Pyro fluorescence <i>via</i> fluorescence resonance energy transfer (FRET). The cleavage of Pyro-Gly-Asp-Glu-Val-Asp-Gly-Ser-Gly-Lys(BHQ3) (PPB) by MMP7 separates the PS (Pyro) from the quencher (BHQ3) and restores the Pyro fluorescence for detection.</p></div><div id="PMB.Synthesis"><h2 id="_PMB_Synthesis_">Synthesis</h2><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20PPMMP7B)+AND+synthesis%0D%0A" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>Zheng et al. reported the detailed synthesis of PP<sub>MMP7</sub>B (<a class="bk_pop" href="#PMB.REF.4">4</a>). A peptide of Fmoc-GPLGLAR(Pbf)K(Mtt)-Sieber resin (Pbf is 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl; Mtt is methyltrityl) was synthesized by the manual, Fmoc, solid-phase, peptide synthesis protocol with the use of commercially available N-α-protected amino acids as building blocks, Sieber amide resin as a solid support, and N-hydroxybenzotriazole (HOBt)/2-(benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU) as amino acid activators. After the last Fmoc group cleavage, Pyro-acid was coupled to the N-terminal glycine of the peptide-resin at a molar ratio of 3:1. The produced peptide resin was treated with 3% trifluoroacetic acid (TFA) and 5% triisopropylsilane (Tis) in dichloromethane to give Pyro-GPLGLAR(Pbf)K, followed by reaction with BHQ3-N-hydroxysuccinimide (NHS) to produce Pyro-GPLGLAR(Pbf)K(BHQ3). The resulting peptide was treated with 95% trifluoroacetic acid and 5% triisopropylsilane to produce PP<sub>MMP7</sub>B. The labeling was confirmed with ultraviolet-visible spectroscopy by identifying Pyro-specific absorbance at 414 nm and BHQ3-specific absorbance at 676 nm.</p></div><div id="PMB.In_Vitro_Studies_Tes"><h2 id="_PMB_In_Vitro_Studies_Tes_"><i>In Vitro</i> Studies: Testing in Cells and Tissues</h2><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20PPMMP7B)+AND+%22in+vitro%22" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>The specificity of PP<sub>MMP7</sub>B to MMP-7 was tested in aqueous solution by measuring the fluorescence emitted from the Pyro in PP<sub>MMP7</sub>B and the production of singlet oxygen <sup>1</sup>O<sub>2</sub> induced by the activated Pyro (<a class="bk_pop" href="#PMB.REF.4">4</a>). PP<sub>MMP7</sub>B emitted 15-fold less fluorescence than PP<sub>MMP7</sub> (the positive control without attached BHQ3), which demonstrated an efficient quenching of Pyro fluorescence by BHQ3. In the presence of the enzyme MMP7, an immediate increase in the Pyro fluorescence of PP<sub>MMP7</sub>B was observed, which reached a plateau at 3 h with a 12-fold increase in fluorescence. However, in the presence of MMP2 or the co-presence of MMP7 and its inhibitor, no noticeable increase in Pyro fluorescence was observed. No cleavage by MMP7 occurred when PP<sub>MMP7</sub>B was replaced with its scrambled sequence (Pyro-GDEVDGSGK-BHQ3). The results were further confirmed with matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF). The production of <sup>1</sup>O<sub>2</sub> was evaluated through the NIR luminescence of <sup>1</sup>O<sub>2</sub> at 1,270 nm. <sup>1</sup>O<sub>2</sub> production in PP<sub>MMP7</sub>B was 18-fold lower than in PP<sub>MMP7</sub> or in a mixture of PP<sub>MMP7</sub> and BHQ3. This demonstrated that the close proximity of Pyro and BHQ3 to the self-folding of the MMP7-cleavable peptide effectively inhibited the <sup>1</sup>O<sub>2</sub> production of Pyro. Adding MMP7 to PP<sub>MMP7</sub>B restored the quenched <sup>1</sup>O<sub>2</sub> production by 19-fold, whereas the presence of MMP7 and its inhibitor prevented this restoration. This confirmed that MMP7 is suitable specifically for cleavage of PP<sub>MMP7</sub>B, which leads to the separation of Pyro and BHQ3 and the photo-activation of Pyro.</p><p>Zheng et al. used PP<sub>MMP7</sub>B to study MMP-7–triggered PDT in cancer cells <i>in vitro</i> (<a class="bk_pop" href="#PMB.REF.4">4</a>). KB cells (human nasopharyngeal epidermoid carcinoma cells) with high MMP7 expression were used as MMP7<sup>+</sup> cells, and BT20 cells (human breast cancer cell line) with deficient MMP7 expression were used as MMP7<sup>-</sup> cells. After incubation with PP<sub>MMP7</sub>B or its scrambled sequence (Pyro-GDEVDGSGK-BHQ3 (C-PPB)), confocal microscopy was performed with excitation at 633 nm and detection at >650 nm. A strong fluorescence signal was observed in KB cells incubated with PP<sub>MMP7</sub>B, whereas minimal fluorescence was found in BT20 cells incubated with PP<sub>MMP7</sub>B and in KB cells or BT20 cells incubated with C-PPB. High-performance liquid chromatography was used to analyze the cell medium collected at the end of drug incubation, which demonstrated that the cleavage of MMP7 occurred in the KB cell medium and produced two fragments as found in the solution studies. No cleavages were found in the BT20 cell medium. These results suggest that the Pyro on PP<sub>MMP7</sub>B also serves as a delivery vehicle to cross the cell membrane in addition to being a fluorescence dye and a PS. The subcellular location of PP<sub>MMP7</sub>B and its cleaved fragments was examined with confocal microscopy <i>via</i> the measurement of Pyro fluorescence and MitoTracker (stain for mitochondria) fluorescence (<a class="bk_pop" href="#PMB.REF.4">4</a>). The Pyro fragment of PP<sub>MMP7</sub>B was located in nearby mitochondria but was absent from the nucleus. The <sup>1</sup>O<sub>2</sub> production was evaluated through the measurement cell viability with the use of 3-(4,5-<a href="http://en.wikipedia.org/wiki/Di-" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri"><u>di</u></a><a href="http://en.wikipedia.org/wiki/Methyl" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri"><u>methyl</u></a><a href="http://en.wikipedia.org/wiki/Thiazole" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri"><u>thiazol</u></a>-2-yl)-2,5-di<a href="http://en.wikipedia.org/wiki/Phenyl" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri"><u>phenyl</u></a>tetrazolium bromide (MTT assay) in KB cells and BT20 cells before and after PDT treatment. Compared with cells not treated with drugs or light, no noticeable dark toxicity was found for treatment with up to 4 μM PP<sub>MMP7</sub>B or C-PPB. Upon PDT treatment, only PP<sub>MMP7</sub>B reduced the viability of the KB cells. To further confirm whether this PDT-induced cytotoxicity occurred <i>via</i> the apoptotic mechanism, Apoptag Plus <i>in situ</i> fluorescein was used to stain the KB cells. KB cells with PP<sub>MMP7</sub>B and PDT treatment demonstrated a strong signal in the Apoptag fluorescein channel (excitation, 488 nm; emission, 497–580 nm), whereas KB cells with PP<sub>MMP7</sub>B but without PDT treatment exhibited no significant apoptosis, which suggests that apoptosis may be the primary cytotoxic mechanism.</p></div><div id="PMB.Animal_Studies"><h2 id="_PMB_Animal_Studies_">Animal Studies</h2><div id="PMB.Rodents"><h3>Rodents</h3><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20PPMMP7B)%20+AND++rodentia" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>Zheng et al. studied the cleavage of PP<sub>MMP7</sub>B <i>in vivo</i> with fluorescence imaging (<a class="bk_pop" href="#PMB.REF.4">4</a>). A mouse bearing two KB tumors (one on each flank) was injected intravenously with 80 nmol PP<sub>MMP7</sub>B. Compared with the prescan images or with the drug-free mouse, no increase in Pyro fluorescence was observed immediately after injection. However, the fluorescence in the tumor increased 20 min later and peaked at 3 h, indicating MMP7-triggered PP<sub>MMP7</sub>B activation. At 3 h, PDT treatment was administered to the tumor on the left flank, whereas the tumor on the right flank served as a dark control. One hour after PDT treatment (4 h after injection), the treated tumor became edematous, whereas the untreated tumor showed no changes in size or fluorescence. For comparison, a drug-free mouse bearing two tumors was treated in the same way; no changes in size or fluorescence were observed in either tumor. Three days after PDT, the treated tumor in the drugged mouse decreased in size, whereas the untreated tumor and both tumors in the drug-free mouse continued to grow. These data demonstrate that PP<sub>MMP7</sub>B that accumulated in MMP7<sup>+</sup> tumors is photodynamically activatable.</p></div><div id="PMB.Other_NonPrimate_Mam"><h3>Other Non-Primate Mammals</h3><p>[<a href="/entrez/query.fcgi?cmd=PureSearch&db=pubmed&details_term=%22%20SUBSTANCENAME%22%5BSubstance%20Name%5D%20AND%20%28dog%20OR%20rabbit%20OR%20pig%20OR%20sheep%29" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>No publication is currently available.</p></div><div id="PMB.NonHuman_Primates"><h3>Non-Human Primates</h3><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20PPMMP7B)%20+AND+(primate+non+human)" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>No publication is currently available.</p></div></div><div id="PMB.Human_Studies"><h2 id="_PMB_Human_Studies_">Human Studies</h2><p>[<a href="/sites/entrez?Db=pubmed&Cmd=DetailsSearch&Term=(%20PPMMP7B)%20+AND+human" ref="pagearea=body&targetsite=external&targetcat=link&targettype=uri">PubMed</a>]</p><p>No publication is currently available.</p><div id="PMB.NIH_Support"><h3>NIH Support</h3><p>CA 95330</p></div></div><div id="PMB.references"><h2 id="_PMB_references_">References</h2><dl class="temp-labeled-list"><dt>1.</dt><dd><div class="bk_ref" id="PMB.REF.1">Dougherty T.J. , Gomer C.J. , Henderson B.W. , Jori G. , Kessel D. , Korbelik M. , Moan J. , Peng Q. Photodynamic therapy. <span><span class="ref-journal">J Natl Cancer Inst. </span>1998;<span class="ref-vol">
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<strong>90</strong>
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</span>(12):889–905.</span> [<a href="/pmc/articles/PMC4592754/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC4592754</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/9637138" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 9637138</span></a>]</div></dd><dt>2.</dt><dd><div class="bk_ref" id="PMB.REF.2">Stefflova K. , Chen J. , Zheng G. Killer beacons for combined cancer imaging and therapy. <span><span class="ref-journal">Curr Med Chem. </span>2007;<span class="ref-vol">
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<strong>14</strong>
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</span>(20):2110–25.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/17691951" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 17691951</span></a>]</div></dd><dt>3.</dt><dd><div class="bk_ref" id="PMB.REF.3">Clo E. , Snyder J.W. , Voigt N.V. , Ogilby P.R. , Gothelf K.V. DNA-programmed control of photosensitized singlet oxygen production. <span><span class="ref-journal">J Am Chem Soc. </span>2006;<span class="ref-vol">
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<strong>128</strong>
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</span>(13):4200–1.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/16568974" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 16568974</span></a>]</div></dd><dt>4.</dt><dd><div class="bk_ref" id="PMB.REF.4">Zheng G. , Chen J. , Stefflova K. , Jarvi M. , Li H. , Wilson B.C. Photodynamic molecular beacon as an activatable photosensitizer based on protease-controlled singlet oxygen quenching and activation. <span><span class="ref-journal">Proc Natl Acad Sci U S A. </span>2007;<span class="ref-vol">
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<strong>104</strong>
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</span>(21):8989–94.</span> [<a href="/pmc/articles/PMC1868591/" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pmc">PMC free article<span class="bk_prnt">: PMC1868591</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/17502620" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 17502620</span></a>]</div></dd><dt>5.</dt><dd><div class="bk_ref" id="PMB.REF.5">Stefflova K. , Chen J. , Marotta D. , Li H. , Zheng G. Photodynamic therapy agent with a built-in apoptosis sensor for evaluating its own therapeutic outcome in situ. <span><span class="ref-journal">J Med Chem. </span>2006;<span class="ref-vol">
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<strong>49</strong>
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</span>(13):3850–6.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/16789741" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 16789741</span></a>]</div></dd><dt>6.</dt><dd><div class="bk_ref" id="PMB.REF.6">Overall C.M. , Kleifeld O. Tumour microenvironment - opinion: validating matrix metalloproteinases as drug targets and anti-targets for cancer therapy. <span><span class="ref-journal">Nat Rev Cancer. </span>2006;<span class="ref-vol">
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<strong>6</strong>
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</span>(3):227–39.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/16498445" ref="pagearea=cite-ref&targetsite=entrez&targetcat=link&targettype=pubmed">PubMed<span class="bk_prnt">: 16498445</span></a>]</div></dd></dl></div><div id="bk_toc_contnr"></div></div></div>
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<div class="post-content"><div><div class="half_rhythm"><a href="/books/about/copyright/">Copyright Notice</a></div><div class="small"><span class="label">Bookshelf ID: NBK23111</span><span class="label">PMID: <a href="https://pubmed.ncbi.nlm.nih.gov/20641316" title="PubMed record of this page" ref="pagearea=meta&targetsite=entrez&targetcat=link&targettype=pubmed">20641316</a></span></div><div style="margin-top:2em" class="bk_noprnt"><a class="bk_cntns" href="/books/n/micad/">Contents</a><div class="pagination bk_noprnt"><a class="active page_link prev" href="/books/n/micad/PPB/" title="Previous page in this title">< Prev</a><a class="active page_link next" href="/books/n/micad/sosquantumdotqd/" title="Next page in this title">Next ></a></div></div></div></div>
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