Name: split-and-pool synthesis and characterization of peptide tertiary amide library
Uploaded: 2014-06-20T13:00:00
Description: Watch this Scientific Journal Video about Split-and-pool Synthesis and Characterization of Peptide Tertiary Amide Library at JoVE.com

De auteurs willen graag Dr Jumpei Morimoto en Dr Todd Doran bedanken voor waardevolle hulp. Dit werk werd ondersteund door een contract van de NHLBI (NO1-HV-00242).

1. Basis van Split-en-zwembad Synthese Om efficiënt genereren van een groot aantal verbindingen op vaste fase wordt split-and-pool synthese vaak gebruikt als een algemene strategie. Zoals getoond in figuur 4, TentaGel kralen worden eerst gesplitst in drie porties. Elk deel wordt omgezet met een ander reagens, genereren van de eerste rest van kralen. Na de eerste reactie worden drie porties samengevoegd, gemengd en opnieuw verdeeld in drie porties. Elk deel zal weer reageren m...

<ol>
	<li>Xiao, X., Yu, P., Lim, H. -. S., Sikder, D., Kodadek, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+Synthesis+of+a+Cell-Permeable+Synthetic+Transcription+Factor+Mimic.">Design and Synthesis of a Cell-Permeable Synthetic Transcription Factor Mimic.</a> Journal of Combinatorial Chemistry. 9, 592-600 (2007).</li><li>Miller, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proteolytic+Studies+of+Homologous+Peptide+and+N-Substituted+Glycine+Peptoid+Oligomers.">Proteolytic Studies of Homologous Peptide and N-Substituted Glycine Peptoid Oligomers.</a> Bioorganic &amp; Medicinal Chemistry Letters. 4, 2657-2662 (1994).</li><li>Grauer, A., Konig, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peptidomimetics+-+A+Versatile+Route+to+Biologically+Active+Compounds.">Peptidomimetics - A Versatile Route to Biologically Active Compounds.</a> European Journal of Organic Chemistry. 30, 5099-5111 (2009).</li><li>Zuckermann, R. N., Kerr, J. M., Kent, S. B. H., Moos, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+method+for+the+preparation+of+peptoids+[oligo(N-substituted+glycines)]+by+submonomer+solid-phase+synthesis.">Efficient method for the preparation of peptoids [oligo(N-substituted glycines)] by submonomer solid-phase synthesis.</a> Journal of the American Chemical Society. 114, 10646-10647 (1992).</li><li>Figliozzi, G. M., Goldsmith, R., Ng, S. C., Banville, S. C., Zuckermann, R. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+N-substituted+glycine+peptoid+libraries.">Synthesis of N-substituted glycine peptoid libraries.</a> Methods in Enzymology. 267, 437-447 (1996).</li><li>Seebach, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=beta-peptides:+Synthesis+by+Arndt-Eistert+homologation+with+concomitant+peptide+coupling.+Structure+determination+by+NMR+and+CD+spectroscopy+and+by+X-ray+crystallography.+Helical+secondary+structure+of+a+beta-hexapeptide+in+solution+and+its+stability+towards+pepsin.">beta-peptides: Synthesis by Arndt-Eistert homologation with concomitant peptide coupling. Structure determination by NMR and CD spectroscopy and by X-ray crystallography. Helical secondary structure of a beta-hexapeptide in solution and its stability towards pepsin.</a> Helv Chim Acta. 79, 913-941 (1996).</li><li>Lam, K. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+New+Type+of+Synthetic+Peptide+Library+for+Identifying+Ligand-Binding+Activity.">A New Type of Synthetic Peptide Library for Identifying Ligand-Binding Activity.</a> Nature. 354, 82-84 (1991).</li><li>Simon, R. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peptoids+-+a+Modular+Approach+to+Drug+Discovery.">Peptoids - a Modular Approach to Drug Discovery.</a> Proceedings of the National Academy of Sciences of the United States of America. 89, 9367-9371 (1992).</li><li>Burkoth, T. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toward+the+synthesis+of+artificial+proteins:+the+discovery+of+an+amphiphilic+helical+peptoid+assembly.">Toward the synthesis of artificial proteins: the discovery of an amphiphilic helical peptoid assembly.</a> Chem Biol. 9, 647-654 (2002).</li><li>Alluri, P. G., Reddy, M. M., Bachhawat-Sikder, K., Olivos, H. J., Kodadek, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+protein+ligands+from+large+peptoid+libraries.">Isolation of protein ligands from large peptoid libraries.</a> Journal of the American Chemical Society. 125, 13995-14004 (2003).</li><li>Lim, H. S., Archer, C. T., Kodadek, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+a+peptoid+inhibitor+of+the+proteasome+19S+regulatory+particle.">Identification of a peptoid inhibitor of the proteasome 19S regulatory particle.</a> Journal of the American Chemical Society. 129, 7750-7751 (2007).</li><li>Wrenn, S. J., Weisinger, R. M., Halpin, D. R., Harbury, P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthetic+ligands+discovered+by+in+vitro+selection.">Synthetic ligands discovered by in vitro selection.</a> Journal of the American Chemical Society. 129, 13137-13143 (2007).</li><li>Aina, O. H., Marik, J., Liu, R. W., Lau, D. H., Lam, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+novel+targeting+peptides+for+human+ovarian+cancer+cells+using+&amp;#34;one-bead+one-compound&amp;#34;+combinatorial+libraries.">Identification of novel targeting peptides for human ovarian cancer cells using &amp;#34;one-bead one-compound&amp;#34; combinatorial libraries.</a> Mol Cancer Ther. 4, 806-813 (2005).</li><li>Udugamasooriya, D. G., Dineen, S. P., Brekken, R. A., Kodadek, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Peptoid+&#8220;Antibody+Surrogate&#8221;+That+Antagonizes+VEGF+Receptor+2+Activity.">A Peptoid &#8220;Antibody Surrogate&#8221; That Antagonizes VEGF Receptor 2 Activity.</a> Journal of the American Chemical Society. 130, 5744-5752 (2008).</li><li>Shah, N. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oligo(+N-aryl+glycines):+A+New+Twist+on+Structured+Peptoids.">Oligo( N-aryl glycines): A New Twist on Structured Peptoids.</a> Journal of the American Chemical Society. 130, 16622-16632 (2008).</li><li>Chongsiriwatana, N. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peptoids+that+mimic+the+structure,+function,+and+mechanism+of+helical+antimicrobial+peptides.">Peptoids that mimic the structure, function, and mechanism of helical antimicrobial peptides.</a> Proceedings of the National Academy of Sciences of the United States of America. 105, 2794-2799 (2008).</li><li>Paul, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=N-Naphthyl+Peptoid+Foldamers+Exhibiting+Atropisomerism.">N-Naphthyl Peptoid Foldamers Exhibiting Atropisomerism.</a> Organic Letters. 14, 926-929 (2012).</li><li>Crapster, J. A., Guzei, I. A., Blackwell, H. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+peptoid+ribbon+secondary+structure.">A peptoid ribbon secondary structure.</a> Angewandte Chemie. 52, 5079-5084 (2013).</li><li>Gorske, B. C., Stringer, J. R., Bastian, B. L., Fowler, S. A., Blackwell, H. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+strategies+for+the+design+of+folded+peptoids+revealed+by+a+survey+of+noncovalent+interactions+in+model+systems.">New strategies for the design of folded peptoids revealed by a survey of noncovalent interactions in model systems.</a> J Am Chem Soc. 131, 16555-16567 (2009).</li><li>Stringer, J. R., Crapster, J. A., Guzei, I. A., Blackwell, H. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extraordinarily+robust+polyproline+type+I+peptoid+helices+generated+via+the+incorporation+of+alpha-chiral+aromatic+N-1-naphthylethyl+side+chains.">Extraordinarily robust polyproline type I peptoid helices generated via the incorporation of alpha-chiral aromatic N-1-naphthylethyl side chains.</a> J Am Chem Soc. 133, 15559-15567 (2011).</li><li>Huang, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+threaded+loop+conformation+adopted+by+a+family+of+peptoid+nonamers.">A threaded loop conformation adopted by a family of peptoid nonamers.</a> Journal of the American Chemical Society. 128, 1733-1738 (2006).</li><li>Lee, J. H., Kim, H. S., Lim, H. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Design+and+Facile+Solid-Phase+Synthesis+of+Conformationally+Constrained+Bicyclic+Peptoids.">Design and Facile Solid-Phase Synthesis of Conformationally Constrained Bicyclic Peptoids.</a> Organic Letters. 13, 5012-5015 (2011).</li><li>Gao, Y., Kodadek, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+and+Screening+of+Stereochemically+Diverse+Combinatorial+Libraries+of+Peptide+Tertiary+Amides.">Synthesis and Screening of Stereochemically Diverse Combinatorial Libraries of Peptide Tertiary Amides.</a> Chem Biol. 20, 360-369 (2013).</li><li>Urban, J., Vaisar, T., Shen, R., Lee, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lability+of+N-alkylated+peptides+towards+TFA+cleavage.">Lability of N-alkylated peptides towards TFA cleavage.</a> Int J Pept Protein Res. 47, 182-189 (1996).</li><li>Rzuczek, S. G., Gao, Y., Tang, Z., Thornton, C. A., Kodadek, T., Disney, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Features+of+Modularly+Assembled+Compounds+That+Impart+Bioactivity+Against+an+RNA+Target.">Features of Modularly Assembled Compounds That Impart Bioactivity Against an RNA Target.</a> ACS Chemical Biology. 8 (10), 2312-2321 (2013).</li><li>Thern, B., Rudolph, J., Jung, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Triphosgene+as+highly+efficient+reagent+for+the+solid-phase+coupling+of+N-alkylated+amino+acids&#8212;total+synthesis+of+cyclosporin+O.">Triphosgene as highly efficient reagent for the solid-phase coupling of N-alkylated amino acids&#8212;total synthesis of cyclosporin O.</a> Tetrahedron Letters. 43, 5013-5016 (2002).</li><li>Sleebs, M. M., Scanlon, D., Karas, J., Maharani, R., Hughes, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Total+Synthesis+of+the+Antifungal+Depsipeptide+Petriellin+A.">Total Synthesis of the Antifungal Depsipeptide Petriellin A.</a> J Org Chem. 76, 6686-6693 (2011).</li><li>Vaisar, T., Urban, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gas-phase+fragmentation+of+protonated+mono-N-methylated+peptides.+Analogy+with+solution-phase+acid-catalyzed+hydrolysis.">Gas-phase fragmentation of protonated mono-N-methylated peptides. Analogy with solution-phase acid-catalyzed hydrolysis.</a> Journal of Mass Spectrometry. 33, 505-524 (1998).</li><li>Creighton, C. J., Romoff, T. T., Bu, J. H., Goodman, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanistic+studies+of+an+unusual+amide+bond+scission.">Mechanistic studies of an unusual amide bond scission.</a> Journal of the American Chemical Society. 121, 6786-6791 (1999).</li><li>Sewald, N., Sewald, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient,+racemization-free+peptide+coupling+of+N-alkyl+amino+acids+by+using+amino+acid+chlorides+generated+in+situ--total+syntheses+of+the+cyclopeptides+cyclosporin+O+and+omphalotin+A.">Efficient, racemization-free peptide coupling of N-alkyl amino acids by using amino acid chlorides generated in situ--total syntheses of the cyclopeptides cyclosporin O and omphalotin A.</a> Angewandte Chemie (International ed. in English). 41, 4661-4663 (2002).</li><li>Astle, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Seamless+Bead+to+Microarray+Screening:+Rapid+Identification+of+the+Highest+Affinity+Protein+Ligands+from+Large+Combinatorial+Libraries.">Seamless Bead to Microarray Screening: Rapid Identification of the Highest Affinity Protein Ligands from Large Combinatorial Libraries.</a> Chem Biol. 17, 38-45 (2010).</li><li>Strohalm, M., Kavan, D., Novak, P., Volny, M., Havlicek, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=mMass+3:+a+cross-platform+software+environment+for+precise+analysis+of+mass+spectrometric+data.">mMass 3: a cross-platform software environment for precise analysis of mass spectrometric data.</a> Anal Chem. 82, 4648-4651 (2010).</li></ol>

Peptide tertiaire amiden (PTA&#39;s) zijn een superfamilie van peptidomimetische oligomeren. Naast de goed bestudeerde peptiden, peptoïden en N-gemethyleerd peptiden, een groot deel van verbindingen binnen deze familie blijft understudied, majorly gebrek aan synthesemethode om algemene N-gealkyleerde peptiden. Hier een efficiënte methode beschrijven we PTA synthetiseren met chirale bouwstenen afgeleid van aminozuren. Eerder hebben we gemeld dat zij een nieuwe sub-monomeer route naar synthese bibliotheken van PTA molec...

Peptidomimetica zijn verbindingen die de structuur van natuurlijke peptiden nabootsen. Ze zijn ontworpen om de biologische activiteit te behouden terwijl het overwinnen van enkele van de problemen in verband met natuurlijke peptiden, waaronder celdoorlaatbaarheid en stabiliteit tegen proteolyse 1-3. Vanwege de oligomere aard van deze verbindingen kunnen grote synthetische bibliotheken gemakkelijk toegankelijk via monomere of sub-monomere syntheseroutes 4-7. Een van de meest bestudeerde klassen van peptidomimetica is peptoïden. Peptoïden zijn oligomeren van N-gealkyleerde glycines die gemakkelijk kunnen worden gesynthetiseerd met behulp van een sub-monomeer strategie 8, 9. Veel nuttige eiwitliganden zijn succes geïdentificeerd door screenen van grote bibliotheken van synthetische peptoïde tegen eiwitdoelstellingen 1, 10-14. Niettemin, &quot;hits&quot; geïdentificeerd uit peptoïde bibliotheken zelden archiveren zeer hoge affiniteit voor eiwit targets 1,10-14,22. Een major verschil tussen peptoïden en natuurlijke peptiden is dat de meeste peptoids algemeen niet de mogelijkheid om de secundaire structuur te vormen door het ontbreken van chirale centra en conformationele beperkingen. Om dit probleem op te lossen, zijn verschillende strategieën ontwikkeld in de afgelopen tien jaar, grotendeels gericht op de modificatie van zijketens die op de hoofdketen stikstofatomen 15-22. Onlangs hebben we een nieuwe synthetische route naar natuurlijke aminozuren zijketens introduceren op een peptoïde ruggengraat peptide tertiaire amides 23 te creëren ontwikkeld. Peptide tertiaire amides (PTA) zijn een superfamilie van peptidomimetica die omvatten maar zijn niet beperkt tot peptiden (R2 = H), peptoïden (R1 = H) en N-gemethyleerd peptiden (R1 ≠ H, R2 = Me) . (Zie figuur 1) De syntheseweg wendt natuurlijk voorkomende aminozuren als de bron van chiraliteit en zijketens op de45, koolstof, en commercieel verkrijgbare primaire aminen N-substituties verschaffen. Daarom kan een grotere chemische ruimte dan eenvoudige peptiden, peptoiden of N-gemethyleerd peptiden onderzocht. Circulaire dichroïsme spectra blijkt dat PTA moleculen zeer gestructureerd in oplossing. Karakterisering van een van de PTA-eiwitcomplexen duidelijk dat de conformationele beperkingen PTA vereist voor binding. Recent hebben wij ook ontdekt dat sommige van de PTA moleculen bezitten verbeterde celdoorlaatbaarheid dan hun peptoïde en peptide tegenhangers. Wij geloven dat deze PTA bibliotheken een goede bron van liganden met hoge affiniteit voor eiwit targets kunnen zijn. In dit artikel gaan we de synthese van een steekproef een-tje een-verbinding (OBOC) PTA bibliotheek in detail samen met een aantal verbeterde voorwaarden voor de koppeling en de splitsing van deze verbindingen te bespreken.

<table border="0" cellpadding="0" cellspacing="0" width="889">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">2,4,6 trimethylpyridine</td>
			<td style="width:169px;">ACROS</td>
			<td style="width:160px;">161950010</td>
			<td style="width:311px;">CAS:108-75-8</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">2-morpholinoethanamine</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">06680&#160;</td>
			<td>CAS:2038-03-1&#160;&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">48% HBr Water solution</td>
			<td style="width:169px;">ALFA AESAR</td>
			<td style="width:160px;">AA14036AT</td>
			<td>CAS:10035-10-6</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Acetaldehyde</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">402788</td>
			<td>CAS:75-07-0&#160;&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Acetonitrile</td>
			<td style="width:169px;">Fisher</td>
			<td style="width:160px;">SR015AA-19PS</td>
			<td>CAS:75-05-8</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Anhydrous Tetrahydrofuran (THF)</td>
			<td style="width:169px;">EMD</td>
			<td style="width:160px;">EM-TX0277-6&#160;</td>
			<td>CAS:109-99-9</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Benzylamine</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">185701</td>
			<td>CAS:100-46-9</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">bis(trichloromethyl) carbonate (BTC)</td>
			<td style="width:169px;">ACROS</td>
			<td style="width:160px;">258950050</td>
			<td>CAS:32315-10-9</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Bromoacetic acid</td>
			<td style="width:169px;">ACROS</td>
			<td style="width:160px;">106570010</td>
			<td>CAS:79-08-3</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Chloranil</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">23290</td>
			<td>CAS:118-75-2</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Cyclohexanemethylamine</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">101842</td>
			<td>CAS:3218-02-8</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;">D2O</td>
			<td style="width:169px;">Cambridge Isotope</td>
			<td style="width:160px;">DLM-4-99.8-1000</td>
			<td>CAS:7789-20-0</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">D-alanine</td>
			<td style="width:169px;">Anaspec</td>
			<td style="width:160px;">61387-100</td>
			<td>CAS:338-69-2&#160;&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">Dichloromethane (DCM)</td>
			<td style="width:169px;">Fisher</td>
			<td style="width:160px;">BJ-NS300-20</td>
			<td>CAS:75-09-2</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">Dimethylformamide (DMF)</td>
			<td style="width:169px;">Fisher</td>
			<td style="width:160px;">BJ-076-4</td>
			<td>CAS:68-12-2</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">Ethylene glycol</td>
			<td style="width:169px;">Oakwood</td>
			<td style="width:160px;">44710</td>
			<td>CAS:107-21-1</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">Isopentylamine</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">W321907</td>
			<td>CAS:107-85-7</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">KBr</td>
			<td style="width:169px;">ACROS</td>
			<td style="width:160px;">424070025</td>
			<td>CAS:7758-02-3</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">L-alanine</td>
			<td style="width:169px;">Anaspec</td>
			<td style="width:160px;">61385-100</td>
			<td>CAS:56-41-7&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">3-Methoxypropylamine</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">M25007</td>
			<td>CAS:5332-73-0</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">2-Methoxyethylamine</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">143693</td>
			<td>CAS:109-85-3</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">N-(3-Aminopropyl)-2-pyrrolidinone</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">136565&#160;</td>
			<td>CAS:7663-77-6&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">N,N&#39;-Diisopropylcarbodiimide (DIC)</td>
			<td style="width:169px;">ACROS</td>
			<td style="width:160px;">115211000</td>
			<td>CAS:693-13-0</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">N,N-Diisopropylethylamine (DIPEA)</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">D125806</td>
			<td>CAS:7087-68-5</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;">NaNO2</td>
			<td style="width:169px;">ACROS</td>
			<td style="width:160px;">424340010</td>
			<td>CAS:7631-99-4</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">NAOD 40% solution in water</td>
			<td style="width:169px;">ACROS</td>
			<td style="width:160px;">200058-506</td>
			<td>CAS:7732-18-5</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Piperidine</td>
			<td style="width:169px;">ALFA AESAR</td>
			<td style="width:160px;">A12442-AE</td>
			<td>CAS:110-89-4</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Piperonylamine</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">P49503&#160;</td>
			<td>CAS:2620-50-0</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Propylamine</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">240958</td>
			<td>CAS:107-10-8</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Trifluoroacetic acid</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">299537</td>
			<td>CAS:76-05-1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">&#945;-Cyano-4-hydroxycinnamic acid&#160;</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">39468</td>
			<td>CAS:28166-41-8&#160;&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">&#945;-ketoglutarate</td>
			<td style="width:169px;">ALFA AESAR</td>
			<td style="width:160px;">AAA10256-22</td>
			<td style="width:311px;">CAS:328-50-7</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Tentagel Resin with RINK linker</td>
			<td style="width:169px;">Rapp-Polymere</td>
			<td style="width:160px;">S30023</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Alanine transaminase</td>
			<td style="width:169px;">Roche</td>
			<td style="width:160px;">10105589001</td>
			<td>AKA: Glutamate-Pyruvate Transaminase (GPT)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:249px;">Incubator</td>
			<td style="width:169px;">New Brunswick Scientific</td>
			<td style="width:160px;">Innova44</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">NMR</td>
			<td style="width:169px;">Bruker</td>
			<td style="width:160px;">400MHz</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">MALDI mass spectrometer</td>
			<td style="width:169px;">Applied Biosystems&#160;</td>
			<td style="width:160px;">4800 MALDI-TOF/TOF</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Lyophilizer</td>
			<td style="width:169px;">SP Scientific</td>
			<td style="width:160px;">VirTis benchtop K</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Syringe reactor</td>
			<td style="width:169px;">INTAVIS&#160;</td>
			<td style="width:160px;">Reaction Column</td>
			<td>3ml, 5ml, 10ml, 20ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Vacuum manifold&#160;</td>
			<td style="width:169px;">Promega</td>
			<td style="width:160px;">A7231</td>
			<td>Vac-Man</td>
		</tr>
	</tbody>
</table>

split-and-pool synthesis and characterization of peptide tertiary amide library

Peptidomimetica zijn grote bronnen van eiwit liganden. De oligomere aard van deze verbindingen stelt ons in staat om grote synthetische bibliotheken op vaste fase met behulp van combinatorische chemie. Een van de meest bestudeerde klassen van peptidomimetics is peptoïden. Peptoïden zijn gemakkelijk te synthetiseren en is aangetoond dat proteolyse-resistente en celpermeable zijn. In de afgelopen tien jaar hebben veel nuttige eiwitliganden geïdentificeerd door middel van screening van peptoïde bibliotheken. Echter, de meeste van de liganden geïdentificeerd uit peptoïde bibliotheken hebben hoge affiniteit niet weergegeven, met zeldzame uitzonderingen. Dit kan ten dele, het gebrek aan chirale centra en conformationele beperkingen in peptoïde moleculen. Onlangs beschreven we een nieuwe synthetische route voor toegang peptide tertiaire amiden (PTA). PTA zijn een superfamilie van peptidomimetica die omvatten maar zijn niet beperkt tot, peptiden, peptoïden en N-gemethyleerd peptiden. Met zijketens aan beide α-koolstof en hoofdketen stikstofatomen,de conformatie van deze moleculen sterk beperkt door sterische hindering en allylische 1,3 stam. (Figuur 1) Onze studie suggereert dat deze PTA moleculen zeer gestructureerd in oplossing en kunnen worden gebruikt om eiwit liganden te identificeren. Wij geloven dat deze moleculen een toekomstige bron van hoge-affiniteit eiwitliganden zijn. Hier beschrijven we de synthetische methode combineert de kracht van beide split-en-zwembad en sub-monomeer strategieën om een ​​monster een-tje een-verbinding (OBOC) bibliotheek van PTA synthetiseren.

Hier laten we drie representatieve MALDI spectra van een PTA trimeer met linker. Zoals getoond in figuur 6A, wanneer gesplitst bij kamertemperatuur met 50% TFA / DCM oplossing, wordt significante afbraak waargenomen. In figuur 6A, pieken 593 en 484 corresponderen met de linker en PTA trimeer respectievelijk blijkt dat het hele molecuul met succes gesynthetiseerd op bead maar afgebroken tijdens splitsing. Wanneer gesplitst bij lage temperatuuromstandigheden zoals hierboven beschreven, wo...

Watch this Scientific Journal Video about Split-and-pool Synthesis and Characterization of Peptide Tertiary Amide Library at JoVE.com

Split-and-pool Synthesis and Characterization of Peptide Tertiary Amide Library

Peptide tertiaire amides (PTA) zijn een superfamilie van peptidomimetica die omvatten maar zijn niet beperkt tot, peptiden, peptoïden en N-gemethyleerd peptiden. Hier beschrijven we een synthetische methode die zowel split-en-zwembad en sub-monomeer strategieën combineert tot een kraal een-verbinding bibliotheek van PTA synthetiseren.

Split-en-zwembad Synthese en karakterisatie van peptide Tertiaire amide Bibliotheek

Peptidomimetics are great sources of protein ligands. The oligomeric nature of these compounds enables us to access large synthetic libraries on solid ...

Research

Education

JoVE Core

JoVE Journal

Organic Chemistry

Chemistry

Unit 1

Carboxylic Acid Derivatives

SCIENTISTS IN ACTION

Nucleoside Triphosphates - From Synthesis to Biochemical Characterization

Nucleosidetrifosfaten - Van Synthese tot biochemische karakterisering

The  traditional strategy for the introduction of chemical functionalities is the  use of solid-phase synthesis by appending suitably modified ...

Amide Coupling Reaction for the Synthesis of Bispyridine-based Ligands and Their Complexation to Platinum as Dinuclear Anticancer Agents

Amidekoppeling reactie voor de synthese van Bispyridine-gebaseerde liganden en hun Complexatie naar Platinum als dinucleair antikankermiddelen

Amide coupling reactions can be used to synthesize bispyridine-based ligands for use as bridging linkers in multinuclear platinum anticancer drugs. ...

Synthesis and Characterization of Functionalized Metal-organic Frameworks

Synthese en karakterisering van gefunctionaliseerde metaal-organische roosters

Metal-organic frameworks have attracted extraordinary amounts of research attention, as they are attractive candidates for numerous industrial and ...

The Synthesis, Characterization and Reactivity of a Series of Ruthenium N-triphosPh Complexes

The Synthesis, Characterization and Reactivity of a Series of Ruthenium N-triphosPh Complexes

De Synthese, karakterisering en reactiviteit van een serie van ruthenium<em&gt; N</em&gt; -triphos<sup&gt; Ph</sup&gt; Complexen

Herein we report the synthesis of a tridentate phosphine ligand N(CH2PPh2)3 (N-triphosPh) (1) via a phosphorus based Mannich reaction of the ...

Synthesis, Characterization, and Functionalization of Hybrid Au/CdS and Au/ZnS Core/Shell Nanoparticles

Synthese, karakterisering en functionalisering van de hybride AU / CdS en Au / ZnS Core / Shell Nanodeeltjes

Plasmonic nanoparticles are an attractive material for light harvesting applications due to their easily modified surface, high surface area and large ...

Development of a Backbone Cyclic Peptide Library as Potential Antiparasitic Therapeutics Using Microwave Irradiation

Ontwikkeling van een Backbone cyclisch peptide bibliotheek als potentieel antiparasitaire Therapeutics behulp van microgolfstraling

Protein-protein interactions (PPIs) are intimately involved in almost all biological processes and are linked to many human diseases. Therefore, there is ...

Synthesis and Characterization of Supramolecular Colloids

Synthese en karakterisering van Supramoleculaire colloïden

Control over colloidal assembly is of utmost importance for the development of functional colloidal materials with tailored structural and mechanical ...

Synthesis of Cyclic Polymers and Characterization of Their Diffusive Motion in the Melt State at the Single Molecule Level

Synthese van cyclische polymeren en karakterisering van hun Diffusive Motion in de Smelt staat bij de Single Molecule niveau

We demonstrate a method for the synthesis of cyclic polymers and a protocol for characterizing their diffusive motion in a melt state at the single ...

Synthesis and Characterization of Fe-doped Aluminosilicate Nanotubes with Enhanced Electron Conductive Properties

Synthese en karakterisering van Fe gedoteerde aluminosilicaten nanobuisjes met Enhanced Electron geleidende eigenschappen

The goal of the protocol is to synthesize Fe-doped aluminosilicate nanotubes of the imogolite type with the formula (OH)3Al2-xFexO3SiOH. Doping with Fe ...

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Protocol voor de vaste fase synthese van oligomeren van RNA die een 2&#39;-<em&gt; O</em&gt; -thiophenylmethylmodificatie en karakterisering via cirkelvormige dichroïsme

Solid-phase synthesis has been used to obtain canonical and modified polymers of nucleic acids, specifically of DNA or RNA, which has made it a popular ...