using-fluorescent-proteins-to-visualize-quantitate-chlamydia-vacuole

Using Fluorescent Proteins to Visualize and Quantitate <em>Chlamydia </em>Vacuole Growth Dynamics in Living Cells

The obligate intracellular bacterium Chlamydia elicits a great burden on global public health. C.&#160;trachomatis is the leading bacterial cause of ...

University of Washington

University of California at Berkeley

2-Dichloroamino-2-methyl-propane-1-sulfonic acid sodium salt (2a), a stable derivative of endogenous N,N-dichlorotaurine (1), has been identified and is under development as a topical antimicrobial agent. Structure-activity relationships of analogs were explored to achieve optimal antimicrobial activity with minimal mammalian toxicity while maintaining the desired stability. All the analogs synthesized showed antimicrobial activity against Staphylococcus aureus, Escherichia coli, and Candida albicans in the range of 1-128 microg/mL and cytotoxicity against mammalian L929 cells in the range 80-1900 microg/mL.

N,N-Dichloroaminosulfonic acids as novel topical antimicrobial agents.

&#x8FEB;&#x5207;&#x9700;&#x8981;&#x5177;&#x6709;&#x5E7F;&#x8C31;&#x6D3B;&#x6027;&#x53CA;&#x6297;&#x751F;&#x7D20;&#x8010;&#x836F;&#x7684;&#x6700;&#x5C11;&#x6F5C;&#x529B;&#x6297;&#x83CC;&#x5316;&#x5408;&#x7269;&#x3002;&#x4E3A;&#x8FBE;&#x5230;&#x6B64;&#x76EE;&#x7684;&#xFF0C;&#x6211;&#x4EEC;&#x7F16;&#x5199;&#x548C;&#x8C03;&#x67E5;&#x7684; N chloroheterocycles &#x7CFB;&#x5217;&#x5C0F;&#x8BF4;&#x3002;&#x5BA1;&#x67E5;&#x7684;&#x5316;&#x5408;&#x7269;&#xFF0C;N chloroamine &#x7CFB;&#x5217;&#x88AB;&#x53D1;&#x73B0;&#x4E0A;&#x7EA7;&#x5BF9; N chloroamide &#x7CFB;&#x5217;&#x5728;&#x5C55;&#x793A;&#x6297;&#x83CC;&#x6D3B;&#x6027;&#x9AD8;&#x3001; &#x4F4E;&#x6BD2;&#x6027;&#x548C;&#x6C34;&#x7684;&#x957F;&#x671F;&#x7A33;&#x5B9A;&#x7684;&#x95EE;&#x5019;&#x3002;

&#x65B0;&#x578B; N chloroheterocyclic &#x6297;&#x83CC;&#x836F;&#x7269;&#x3002;

Antimicrobial compounds with broad-spectrum activity and minimal potential for antibiotic resistance are urgently needed. Toward this end, we prepared and investigated a novel series of N-chloroheterocycles. Of the compounds examined, the N-chloroamine series were found superior over N-chloroamide series in regards to exhibiting high antimicrobial activity, low cytotoxicity, and long-term aqueous stability.

Novel N-chloroheterocyclic antimicrobials.

Compos&eacute;s antimicrobiens avec activit&eacute; &agrave; large spectre et un potentiel minimal pour la r&eacute;sistance aux antibiotique sont n&eacute;cessaires d&#39;urgence. &Agrave; cette fin, nous avons pr&eacute;par&eacute; et &eacute;tudi&eacute; une nouvelle s&eacute;rie de N-chloroheterocycles. Des compos&eacute;s examin&eacute;s, la s&eacute;rie N-chloroamines trouv&eacute;e sup&eacute;rieure par rapport aux s&eacute;ries N-chloroamide en ce qui concerne pr&eacute;sentant une forte activit&eacute; antimicrobienne, faible cytotoxicit&eacute; et stabilit&eacute; aqueuse &agrave; long terme.

Nouveaux antimicrobiens de N-chloroheterocyclic.

Antimikrobielle Compounds mit Breitspektrum-Aktivit&auml;t und minimal Potenzial f&uuml;r Antibiotikaresistenz sind dringend erforderlich. Dazu haben wir vorbereitet und untersucht eine neuartige Reihe von N-Chloroheterocycles. Der untersuchten Verbindungen die N-Chloroamine-Serie &uuml;berlegen &uuml;ber N-Chloroamide-Serie in Bezug auf die ausstellenden hohe antimikrobielle Aktivit&auml;t, geringe Zytotoxizit&auml;t und w&auml;ssrige Langzeitstabilit&auml;t gefunden.

Neuartige N-Chloroheterocyclic Antibiotika.

Composti antimicrobici con attivit&agrave; ad ampio spettro e minimo potenziale di resistenza agli antibiotici sono urgentemente necessari. Verso questo fine, abbiamo preparato e studiato una serie di romanzi di N-chloroheterocycles. Dei composti esaminati, la serie N-cloroammine trovata superiore sopra serie N-chloroamide per quanto riguarda il esibendo un&#39;elevata attivit&agrave; antimicrobica, bassa citotossicit&agrave; e stabilit&agrave; a lungo termine acquosa.

Romanzo N-chloroheterocyclic antimicrobici.

&#x6297;&#x83CC;&#x6027;&#x5316;&#x5408;&#x7269;&#x306E;&#x5E83;&#x57DF;&#x30B9;&#x30DA;&#x30AF;&#x30C8;&#x30EB;&#x6D3B;&#x6027;&#x3068;&#x6297;&#x751F;&#x7269;&#x8CEA;&#x8010;&#x6027;&#x306E;&#x53EF;&#x80FD;&#x6027;&#x3092;&#x6700;&#x5C0F;&#x9650;&#x306B;&#x306F;&#x7DCA;&#x6025;&#x306B;&#x5FC5;&#x8981;&#x3068;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x5E74;&#x672B;&#x306B;&#x5411;&#x3051;&#x3066;&#x6E96;&#x5099;&#x3057;&#x3001;N chloroheterocycles &#x306E;&#x5C0F;&#x8AAC;&#x30B7;&#x30EA;&#x30FC;&#x30BA;&#x3092;&#x691C;&#x8A0E;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x691C;&#x8A0E;&#x3057;&#x305F;&#x5316;&#x5408;&#x7269;&#x306E; N chloroamine &#x30B7;&#x30EA;&#x30FC;&#x30BA; N chloroamide &#x30B7;&#x30EA;&#x30FC;&#x30BA;&#x306E;&#x9AD8;&#x6297;&#x83CC;&#x4F5C;&#x7528;&#x3001;&#x4F4E;&#x6BD2;&#x6027;&#x3001;&#x6C34;&#x6EB6;&#x6DB2;&#x4E2D;&#x306E;&#x9577;&#x671F;&#x5B89;&#x5B9A;&#x3092;&#x51FA;&#x5C55;&#x306B;&#x95A2;&#x3057;&#x3066;&#x4EE5;&#x4E0A;&#x512A;&#x308C;&#x305F;&#x898B;&#x3064;&#x304B;&#x308A;&#x307E;&#x3057;&#x305F;&#x3002;

&#x65B0;&#x898F; N chloroheterocyclic &#x306E;&#x6297;&#x83CC;&#x85AC;&#x3002;

&#xC2A4;&#xD399;&#xD2B8;&#xB7FC; &#xD65C;&#xB3D9; &#xBC0F; &#xD56D;&#xC0DD;&#xC81C; &#xB0B4;&#xC131;&#xC5D0; &#xB300; &#xD55C; &#xCD5C;&#xC18C;&#xD55C;&#xC758; &#xC7A0;&#xC7AC;&#xB825;&#xACFC; &#xD56D;&#xADE0; &#xC131; &#xD654;&#xD569;&#xBB3C; &#xAE34;&#xAE09; &#xD558; &#xAC8C; &#xD544;&#xC694; &#xD569;&#xB2C8;&#xB2E4;. &#xC774; &#xB05D;&#xC744; &#xD5A5;&#xD574; &#xC900;&#xBE44; &#xD558; &#xACE0; N-chloroheterocycles&#xC758; &#xC18C;&#xC124; &#xC2DC;&#xB9AC;&#xC988;&#xB97C; &#xC870;&#xC0AC;. &#xAC80;&#xC0AC; &#xD558;&#xB294; &#xD654;&#xD569;&#xBB3C;&#xC758; N-chloroamine &#xC2DC;&#xB9AC;&#xC988; &#xC804;&#xC2DC; &#xB192;&#xC740; &#xD56D;&#xADE0; &#xD65C;&#xC131;, &#xB0AE;&#xC740; &#xC138;&#xD3EC; &#xB3C5;&#xC131;, &#xADF8;&#xB9AC;&#xACE0; &#xC7A5;&#xAE30; &#xC218;&#xC131; &#xC548;&#xC815;&#xC131;&#xC5D0; &#xAD00;&#xD574;&#xC11C;&#xC5D0;&#xC11C; N chloroamide &#xC2DC;&#xB9AC;&#xC988;&#xB97C; &#xD1B5;&#xD574; &#xC6B0;&#xC218;&#xD55C; &#xBC1C;&#xACAC; &#xB410;&#xB2E4;.

&#xC18C;&#xC124; N chloroheterocyclic antimicrobials&#xC785;&#xB2C8;&#xB2E4;.

Se necesitan urgentemente compuestos antimicrobianos con actividad de amplio espectro y potencial m&iacute;nimo de resistencia a los antibi&oacute;ticos. Con este fin, preparamos e investiga una serie de novelas de N-chloroheterocycles. De los compuestos examinados, la serie N-cloraminas se encontraron superior sobre la serie de N-chloroamide en lo que respecta a la alta actividad antimicrobiana, baja citotoxicidad, y estabilidad a largo plazo acuosa.

Nuevos antimicrobianos de N-chloroheterocyclic.

Antimicrobial resistance against many known therapeutics is on the rise. We examined derivatives of 3-chlorooxazolidin-2-one 1a (X=H) as antibacterial and antifungal agents. The key findings were that the activity and apparent in vitro cytotoxicity could be controlled by the substitution of charged solubilizers at the 4- and 5- positions. These changes both significantly increase the antifungal potency and decrease cytotoxicity. Particularly effective were trialkylammonium groups which led to 400- to 600-fold increases in the antifungal therapeutic index when compared to their unsubstituted counterparts.

Novel 3-chlorooxazolidin-2-ones as antimicrobial agents.

Stabilit&agrave;/activity relationships (SXR) della struttura di una nuova classe di N, N-dichloroamine composti sono stati esplorati per migliorare l&#39;attivit&agrave; antimicrobica contro Escherichia coli, Staphylococcus aureus e Candida albicans mantenendo la stabilit&agrave; in soluzione acquosa. Questo studio identificato una nuova classe di agenti antimicrobici soluzione stabile e di attualit&agrave;. Questi agenti sono stabilizzato solfone e possiedono un ammonio quaternario o appendici solfonato come acqua solubilizzanti gruppo. Diverse sfide uniche sono state confrontate nella sintesi di questi nuovi composti che sono evidenziate nella discussione.

Struttura relazioni attivit&agrave; stabilit&agrave; del solfone stabilizzato N, N-dichloroamines.

&#x7ED3;&#x6784;&#x7A33;&#x5B9A;&#x6027;/&#x6D3B;&#x52A8;&#x5173;&#x7CFB; (SXR) &#x7684;&#x4E00;&#x7C7B;&#x65B0;&#x7684; N&#xFF0C;&#x63A2;&#x8BA8;&#x4E86; N dichloroamine &#x5316;&#x5408;&#x7269;&#x6C34;&#x6EB6;&#x6DB2;&#x7A33;&#x5B9A;&#x6027;&#x7684;&#x540C;&#x65F6;&#x63D0;&#x9AD8;&#x5BF9;&#x5927;&#x80A0;&#x6746;&#x83CC;&#x3001; &#x91D1;&#x9EC4;&#x8272;&#x8461;&#x8404;&#x7403;&#x83CC;&#x548C;&#x767D;&#x8272;&#x5FF5;&#x73E0;&#x83CC;&#x7684;&#x6297;&#x83CC;&#x6D3B;&#x6027;&#x3002;&#x8FD9;&#x9879;&#x7814;&#x7A76;&#x53D1;&#x73B0;&#x4E00;&#x7C7B;&#x65B0;&#x7684;&#x89E3;&#x51B3;&#x65B9;&#x6848;&#x7A33;&#x5B9A;&#x548C;&#x5C40;&#x90E8;&#x6297;&#x83CC;&#x836F;&#x7269;&#x3002;&#x8FD9;&#x4E9B;&#x4EE3;&#x7406;&#x781C;&#x7A33;&#x5B9A;&#xFF0C;&#x4F5C;&#x4E3A;&#x6C34;&#x589E;&#x6EB6;&#x96C6;&#x56E2;&#x62E5;&#x6709;&#x5B63;&#x94F5;&#x76D0;&#x6216;&#x78FA;&#x9178;&#x9644;&#x80A2;&#x3002;&#x5728;&#x8BA8;&#x8BBA;&#x4E2D;&#x5F3A;&#x8C03;&#x4E86;&#x8FD9;&#x4E9B;&#x65B0;&#x578B;&#x5316;&#x5408;&#x7269;&#x7684;&#x5408;&#x6210;&#x9762;&#x4E34;&#x51E0;&#x9879;&#x72EC;&#x7279;&#x7684;&#x6311;&#x6218;&#x3002;

&#x7ED3;&#x6784;&#x7A33;&#x5B9A;&#x6027;/&#x6D3B;&#x52A8;&#x5173;&#x7CFB;&#x7684;&#x781C;&#x7A33;&#x5B9A;&#x7684; N&#x3001; N dichloroamines&#x3002;

Structure stability/activity relationships (SXR) of a new class of N,N-dichloroamine compounds were explored to improve antimicrobial activity against Escherichia coli, Staphylococcus aureus, and Candida albicans while maintaining aqueous solution stability. This study identified a new class of solution-stable and topical antimicrobial agents. These agents are sulfone-stabilized and possess either a quaternary ammonium or sulfonate appendages as a water solubilizing group. Several unique challenges were confronted in the synthesis of these novel compounds which are highlighted in the discussion.

Structure stability/activity relationships of sulfone stabilized N,N-dichloroamines.

Structurer les relations stabilit&eacute;/activit&eacute; (SXR) d&#39;une nouvelle classe de N, N-dichloroamine compos&eacute;s ont &eacute;t&eacute; &eacute;tudi&eacute;s afin d&#39;am&eacute;liorer l&#39;activit&eacute; antimicrobienne contre Escherichia coli, Staphylococcus aureus et Candida albicans tout en maintenant la stabilit&eacute; de la solution aqueuse. Cette &eacute;tude a identifi&eacute; une nouvelle classe d&#39;agents antimicrobiens solution stable et d&#39;actualit&eacute;. Ces agents sont stabilis&eacute;s sulfone et poss&egrave;dent un ammonium quaternaire ou sulfonate appendices comme groupe de solubilisation de l&#39;eau. Plusieurs d&eacute;fis uniques ont &eacute;t&eacute; confront&eacute;s dans la synth&egrave;se de ces nouveaux compos&eacute;s qui sont mises en &eacute;vidence au cours du d&eacute;bat.

Structurer les relations stabilit&eacute;/activit&eacute; de sulfone stabilis&eacute;e de N, N-dichloroamines.

Stabilit&auml;t/Aktivit&auml;ts-Beziehungen (SXR) Struktur einer neuen Klasse von N, N-Dichloroamine-Verbindungen wurden untersucht, um antimikrobielle Aktivit&auml;t gegen Escherichia coli, Staphylococcus Aureus und Candida Albicans zu verbessern und gleichzeitig die Stabilit&auml;t der w&auml;ssrigen L&ouml;sung. Dieser Studie wurden eine neue Klasse von L&ouml;sung-Stable und topischen antimikrobiellen. Diese Agenten sind Sulfon-stabilisiert und besitzen entweder eine Quatern&auml;re Ammonium oder sulfonat Anh&auml;ngseln wie ein Wasser L&ouml;sungsvermittlung Gruppe. Mehrere einzigartige Herausforderungen wurden bei der Synthese dieser neuartigen Verbindungen konfrontiert, die in der Diskussion hervorgehoben werden.

Stabilit&auml;t/Aktivit&auml;ts-Beziehungen von Sulfon-Struktur stabilisiert N, N-Dichloroamines.

&#x69CB;&#x9020;&#x5B89;&#x5B9A;&#x6027;/&#x30A2;&#x30AF;&#x30C6;&#x30A3;&#x30D3;&#x30C6;&#x30A3; &#x30EA;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x30B7;&#x30C3;&#x30D7; (&#x4ECB;&#x3059;&#x308B;) N &#x306E;&#x65B0;&#x3057;&#x3044;&#x30AF;&#x30E9;&#x30B9;&#x306F;&#x3001;N-dichloroamine &#x5316;&#x5408;&#x7269;&#x6C34;&#x6EB6;&#x6DB2;&#x306E;&#x5B89;&#x5B9A;&#x6027;&#x3092;&#x7DAD;&#x6301;&#x3057;&#x306A;&#x304C;&#x3089;&#x3001;&#x5927;&#x8178;&#x83CC;&#x3001;&#x9EC4;&#x8272;&#x30D6;&#x30C9;&#x30A6;&#x7403;&#x83CC;&#x3001;&#x30AB;&#x30F3;&#x30B8;&#x30C0; &#x30FB; &#x30A2;&#x30EB;&#x30D3;&#x30AB;&#x30F3;&#x30B9;&#x306B;&#x5BFE;&#x3059;&#x308B;&#x6297;&#x83CC;&#x6D3B;&#x6027;&#x3092;&#x6539;&#x5584;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x691C;&#x8A0E;&#x3055;&#x308C;&#x307E;&#x3057;&#x305F;&#x3002;&#x672C;&#x7814;&#x7A76;&#x306F;&#x3001;&#x65B0;&#x3057;&#x3044;&#x30AF;&#x30E9;&#x30B9; &#x30BD;&#x30EA;&#x30E5;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x5B89;&#x5B9A;&#x304A;&#x3088;&#x3073;&#x5C40;&#x6240;&#x6297;&#x83CC;&#x5264;&#x8B58;&#x5225;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x30A8;&#x30FC;&#x30B8;&#x30A7;&#x30F3;&#x30C8;&#x306F;&#x3001;&#x30B9;&#x30EB;&#x30DB;&#x30F3;&#x304C;&#x5B89;&#x5B9A;&#x5316;&#x3057;&#x3001;&#x7B2C; 4 &#x7D1A;&#x30A2;&#x30F3;&#x30E2;&#x30CB;&#x30A6;&#x30E0;&#x307E;&#x305F;&#x306F;&#x30B9;&#x30EB;&#x30DB;&#x30F3;&#x9178;&#x57FA;&#x306E;&#x4ED8;&#x5C5E;&#x80A2;&#x30B0;&#x30EB;&#x30FC;&#x30D7;&#x53EF;&#x6EB6;&#x5316;&#x3068;&#x3057;&#x3066;&#x306E;&#x6C34;&#x3092;&#x6301;&#x3063;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x3044;&#x304F;&#x3064;&#x304B;&#x306E;&#x30E6;&#x30CB;&#x30FC;&#x30AF;&#x306A;&#x8AB2;&#x984C;&#x306E;&#x8B70;&#x8AD6;&#x3067;&#x5F37;&#x8ABF;&#x8868;&#x793A;&#x3055;&#x308C;&#x307E;&#x3059;&#x3053;&#x308C;&#x3089;&#x306E;&#x5C0F;&#x8AAC;&#x306E;&#x5316;&#x5408;&#x7269;&#x306E;&#x5408;&#x6210;&#x306B;&#x76F4;&#x9762;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;

&#x69CB;&#x9020;&#x5B89;&#x5B9A;&#x6027;/&#x30A2;&#x30AF;&#x30C6;&#x30A3;&#x30D3;&#x30C6;&#x30A3; &#x30EA;&#x30EC;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x30B7;&#x30C3;&#x30D7; &#x30B9;&#x30EB;&#x30D5;&#x30A9;&#x30F3;&#x306E;&#x5B89;&#x5B9A;&#x5316;&#x306E; N, N-dichloroamines&#x3002;

&#xAD6C;&#xC870; &#xC548;&#xC815;&#xC131;/&#xD65C;&#xB3D9; &#xAD00;&#xACC4; (SXR) N&#xC758; &#xC0C8;&#xB85C;&#xC6B4; &#xD074;&#xB798;&#xC2A4;&#xC758; N-dichloroamine &#xD654;&#xD569;&#xBB3C; &#xC218; &#xC6A9;&#xC561; &#xC548;&#xC815;&#xC131;&#xC744; &#xC720;&#xC9C0; &#xD558;&#xBA74;&#xC11C; &#xB300;&#xC7A5;&#xADE0;, &#xD3EC;&#xB3C4; &#xC0C1; &#xAD6C; &#xADE0; &#xBC0F; Candida albicans&#xC5D0; &#xB300; &#xD55C; &#xD56D;&#xADE0; &#xD65C;&#xC131;&#xC744; &#xD5A5;&#xC0C1; &#xC2DC;&#xD0A4;&#xAE30; &#xC704;&#xD574; &#xD0D0;&#xD5D8; &#xD588;&#xB2E4;. &#xC774; &#xC5F0;&#xAD6C; &#xC194;&#xB8E8;&#xC158;-&#xC548;&#xC815;&#xC801;&#xC774; &#xACE0; &#xAD6D; &#xC18C; &#xD56D;&#xADE0; &#xC131; &#xB300;&#xB9AC;&#xC778;&#xC758; &#xC0C8;&#xB85C;&#xC6B4; &#xD074;&#xB798;&#xC2A4;&#xB97C; &#xC2DD;&#xBCC4;&#xD569;&#xB2C8;&#xB2E4;. &#xC774;&#xB7EC;&#xD55C; &#xC5D0;&#xC774;&#xC804;&#xD2B8; &#xD3F0; &#xC548;&#xC815;&#xD654; &#xB418; &#xACE0; 4 &#xAE09; &#xC554;&#xBAA8;&#xB284; &#xB610;&#xB294; &#xB41C; &#xBD80;&#xC18D; &#xADF8;&#xB8F9; solubilizing &#xBB3C;&#xC73C;&#xB85C; &#xD569;&#xB2C8;&#xB2E4;. &#xD1A0;&#xB860;&#xC5D0;&#xC11C; &#xAC15;&#xC870; &#xD558;&#xB294;&#xC774; &#xC0C8;&#xB85C;&#xC6B4; &#xD654;&#xD569;&#xBB3C;&#xC758; &#xD569;&#xC131;&#xC5D0;&#xB294; &#xBA87; &#xAC00;&#xC9C0; &#xB3C5;&#xD2B9;&#xD55C; &#xB3C4;&#xC804; &#xC9C1;&#xBA74; &#xD588;&#xB2E4;.

&#xD3F0;&#xC758; &#xC548;&#xC815;&#xC131;/&#xD65C;&#xB3D9; &#xAD00;&#xACC4; &#xAD6C;&#xC870; &#xC548;&#xC815;&#xB41C; N, N-dichloroamines.

Relaciones de estabilidad/actividad (SXR) de la estructura de una nueva clase de N, N-dichloroamine compuestos fueron explorados para mejorar actividad antimicrobiana contra Escherichia coli, Staphylococcus aureus y Candida albicans y manteniendo la estabilidad de la soluci&oacute;n acuosa. Este estudio identific&oacute; una nueva clase de agentes antimicrobianos t&oacute;picos y soluci&oacute;n estable. Estos agentes son sulfona-estabilizado y poseen un amonio cuaternario o sulfonato ap&eacute;ndices como un agua solubilizaci&oacute;n del grupo. Varios retos fueron confrontados en la s&iacute;ntesis de estos compuestos novedosos que destacan en la discusi&oacute;n.

Estructura de las relaciones de estabilidad y actividad de la sulfona estabilizado N, N-dichloroamines.

Human adenoviral conjunctivitis is a highly contagious eye infection affecting millions of people world-wide. If untreated, it can further develop into keratitis, corneal ulceration, scarring and possible blindness. Despite the significant patient morbidity and socio-economic costs, it is an unmet medical need with no FDA approved treatment. Here, we demonstrate the virucidal activity of NVC-422 (N,N-dichloro-2,2-dimethyltaurine) against adenovirus type 5 (Ad5) and investigated its mechanism of action of Ad5 inactivation. NVC-422 inhibits Ad5-induced loss of cell viability in vitro with 50% inhibitory concentration (IC(50)) ranging from 9 to 23 μM. NVC-422 does not cause any cytotoxicity at concentrations as high as 250 μM. Invitro, NVC-422 inactivates Ad5 but does not interfere with viral replication, indicating that NVC-422 acts on the extracellular adenovirus as a virucidal agent. NVC-422 inactivates Ad5 by oxidative inactivation of key viral proteins such as fiber and hexon as evidenced by SDS-PAGE, Western blotting and reversed-phase HPLC. These data, combined with measurements of the kinetics of the NVC-422 reactivity with selected amino acids, indicate that the changes in the viral proteins are caused by the selective oxidation of sulfur-containing amino acids. The conformational changes of the viral proteins result in the destruction of the viral morphology as shown by transmission electron microscopy. In summary, NVC-422 exhibits virucidal activity against Ad5 by the oxidative inactivation of key viral proteins, leading to the loss of viral integrity and infectivity.

Virucidal mechanism of action of NVC-422, a novel antimicrobial drug for the treatment of adenoviral conjunctivitis.

N-chlorotaurine (NCT) and its analogues N-monochloro-2,2-dimethyltaurine (NVC-612) and N-dichloro-2,2-dimethyltaurine (NVC-422) are new anti-infectives for topical treatment for conjunctivitis. The aim of this study was to show that these compounds are safe in an EpiOcular model and effective in corneas infected ex vivo.

N-chlorotaurine and its analogues N,N-dichloro-2,2-dimethyltaurine and N-monochloro-2,2-dimethyltaurine are safe and effective bactericidal agents in ex vivo corneal infection models.

The ability to exit host cells at the end of their developmental growth is a critical step for the intracellular bacterium Chlamydia. One exit strategy, extrusion, is mediated by host signaling pathways involved with actin polymerization. Here, we show that actin is recruited to the chlamydial inclusion as a late event, occurring after 20 hours post-infection (hpi) and only within a subpopulation of cells. This event increases significantly in prevalence and extent from 20 to 68 hpi, and actin coats strongly correlated with extrusions. In contrast to what has been reported for other intracellular pathogens, actin nucleation on Chlamydia inclusions did not 'flash', but rather exhibited moderate depolymerization dynamics. By using small molecule agents to selectively disrupt host signaling pathways involved with actin nucleation, modulate actin polymerization dynamics and also to disable the synthesis and secretion of chlamydial proteins, we further show that host and bacterial proteins are required for actin coat formation. Transient disruption of either host or bacterial signaling pathways resulted in rapid loss of coats in all infected cells and a reduction in extrusion formation. Inhibition of Chlamydia type III secretion also resulted in rapid loss of actin association on inclusions, thus implicating chlamydial effector proteins(s) as being central factors for engaging with host actin nucleating factors, such as formins. In conclusion, our data illuminate the host and bacterial driven process by which a dense actin matrix is dynamically nucleated and maintained on the Chlamydia inclusion. This late stage event is not ubiquitous for all infected cells in a population, and escalates in prevalence and extent throughout the developmental cycle of Chlamydia, culminating with their exit from the host cell by extrusion. The initiation of actin recruitment by Chlamydia appears to be novel, and may serve as an upstream determinant of the extrusion mechanism.

Actin recruitment to the Chlamydia inclusion is spatiotemporally regulated by a mechanism that requires host and bacterial factors.

Bacterial pathogens have specific virulence factors (e.g., toxins) that contribute significantly to the virulence and infectivity of microorganisms within the human hosts. Virulence factors are molecules expressed by pathogens that enable colonization, immunoevasion, and immunosuppression, obtaining nutrients from the host or gaining entry into host cells. They can cause pathogenesis by inhibiting or stimulating certain host functions. For example, in systemic Staphylococcus aureus infections, virulence factors such as toxic shock syndrome toxin 1 (TSST-1), staphylococcal enterotoxin A (SEA), and staphylococcal enterotoxin B (SEB) cause sepsis or toxic shock by uncontrolled stimulation of T lymphocytes and by triggering a cytokine storm. In vitro, these superantigens stimulate the proliferation of human peripheral blood mononuclear cells (PBMC) and the release of many cytokines. NVC-422 (N,N-dichloro-2,2-dimethyltaurine) is a broad-spectrum, fast-acting topical anti-infective agent against microbial pathogens, including antibiotic-resistant microbes. Using mass spectrometry, we demonstrate here that NVC-422 oxidizes methionine residues of TSST-1, SEA, SEB, and exfoliative toxin A (ETA). Exposure of virulence factors to 0.1% NVC-422 for 1 h prevented TSST-1-, SEA-, SEB-, and ETA-induced cell proliferation and cytokine release. Moreover, NVC-422 also delayed and reduced the protein A- and clumping factor-associated agglutination of S. aureus cultures. These results show that, in addition to its well-described direct microbicidal activity, NVC-422 can inactivate S. aureus virulence factors through rapid oxidation of methionines.

NVC-422 inactivates Staphylococcus aureus toxins.

Viral conjunctivitis is a highly contagious infection often causing major epidemics. A safe broad-spectrum antiviral agent is needed to treat this unmet medical need. The purpose of this study is to demonstrate that in vitro NVC-422 is a safe, broad-spectrum topical virucidal agent with activity against ophthalmic viral pathogens.

Broad-spectrum virucidal activity of (NVC-422) N,N-dichloro-2,2-dimethyltaurine against viral ocular pathogens in vitro.

La conjonctivite virale est une infection tr&egrave;s contagieuse, souvent responsable d&#39;&eacute;pid&eacute;mies majeures. Un agent antiviral &agrave; large spectre s&eacute;curitaire est n&eacute;cessaire pour traiter ce besoin m&eacute;dical non combl&eacute;. Le but de cette &eacute;tude est de d&eacute;montrer qu&#39;in vitro NVC-422 est un agent topique virucide s&eacute;curitaire, &agrave; large spectre ayant une activit&eacute; contre des pathog&egrave;nes viraux ophtalmiques.

Activit&eacute; virucide &agrave; large spectre (NVC-422) n, N-dichloro-2, 2-dimethyltaurine contre les agents pathog&egrave;nes oculaires virales in vitro.

Congiuntivite virale &egrave; un&#39;infezione altamente contagiosa che spesso causano grandi epidemie. Un sicuro agente antivirale ad ampio spettro &egrave; necessaria per trattare questa necessit&agrave; mediche insoddisfatte. Lo scopo di questo studio &egrave; quello di dimostrare che in vitro NVC-422 &egrave; un agente virucida topico ad ampio spettro, sicuro, con attivit&agrave; contro gli agenti patogeni virali oftalmici.

Attivit&agrave; virucida ad ampio spettro (NVC-422) n, N-dicloro-2,2-dimethyltaurine contro patogeni oculari virali in vitro.

The discovery and development of antimicrobial agents that do not give rise to resistance remains an ongoing challenge. Our efforts in this regard continue to reveal new potential therapeutic agents with differing physicochemical properties while retaining the effective N,N-dichloroamine pharmacophore as the key antimicrobial warhead. In this Letter, we disclose agents containing polyol units as a water solubilizing group. These sulfonyl-polyol agents show broad spectrum bactericidal and virucidal activity. These compounds show 1 h MBC's of 16-512 μg/mL against Escherichia coli and 4-256 μg/mL against Staphylococcus aureus at neutral pH, and 1-h IC50's of 4.5-32 μM against Adenovirus 5 and 0.7-3.0 μM against Herpes simplex virus 1. The lead compounds were tested in a tissue culture irritancy assay and showed only minimal irritation at the highest concentrations tested.

Article	Year
N,N-Dichloroaminosulfonic acids as novel topical antimicrobial agents. Bioorganic & medicinal chemistry letters\| PubMed ID: 19010674	2009
Novel N-chloroheterocyclic antimicrobials. Bioorganic & medicinal chemistry letters\| PubMed ID: 21459574	2011
Novel 3-chlorooxazolidin-2-ones as antimicrobial agents. Bioorganic & medicinal chemistry letters\| PubMed ID: 21470861	2011
Structure stability/activity relationships of sulfone stabilized N,N-dichloroamines. Bioorganic & medicinal chemistry letters\| PubMed ID: 21570284	2011
Virucidal mechanism of action of NVC-422, a novel antimicrobial drug for the treatment of adenoviral conjunctivitis. Antiviral research\| PubMed ID: 22024427	2011
N-chlorotaurine and its analogues N,N-dichloro-2,2-dimethyltaurine and N-monochloro-2,2-dimethyltaurine are safe and effective bactericidal agents in ex vivo corneal infection models. Acta ophthalmologica\| PubMed ID: 22971221	2012
Actin recruitment to the Chlamydia inclusion is spatiotemporally regulated by a mechanism that requires host and bacterial factors. PloS one\| PubMed ID: 23071671	2012
NVC-422 inactivates Staphylococcus aureus toxins. Antimicrobial agents and chemotherapy\| PubMed ID: 23208720	2013
Broad-spectrum virucidal activity of (NVC-422) N,N-dichloro-2,2-dimethyltaurine against viral ocular pathogens in vitro. Investigative ophthalmology & visual science\| PubMed ID: 23341010	2013
Sulfonyl-polyol N,N-dichloroamines with rapid, broad-spectrum antimicrobial activity. Bioorganic & medicinal chemistry letters\| PubMed ID: 23999044	2013

Meghan Zuck

Biography

JoVE Journal Publications

Other Publications