Safety and tolerability of repeated administration of pyridoxal 2-chlorobenzoyl hydrazone in rabbits
Identifieur interne : 002769 ( Istex/Corpus ); précédent : 002768; suivant : 002770Safety and tolerability of repeated administration of pyridoxal 2-chlorobenzoyl hydrazone in rabbits
Auteurs : M. Sterba ; T. Simunek ; Y. Mazurová ; M. Adamcová ; O. Popelová ; J. Kaplanová ; P. Ponka ; V. GerslSource :
- Human and Experimental Toxicology [ 0960-3271 ] ; 2005-11.
English descriptors
- Teeft :
- Animal behaviour, Anova, Antioxidant effects, Biochem cell biol, Biochem pharmacol, Biochemical parameters, Blood pressure, Blue trichrome, Body weight gain, Cardiac, Cardiac damage, Cardiac troponin, Castle hill, Charles university, Chelation, Chelator, Chelators, Control group, Control vehicle, Cremophor, Czech republic, Detection limit, Eosinophilic cytoplasm, Essential interface, Fifth administration, First administration, Gross changes, Haematological, Haematological analyses, Haematological parameters, Heart rate, Highest dose, Histopathological, Histopathological examination, Hradec, Hradec kralove, Hradec krdlove, Hydrazone, Hypertransfused rats, Inconsistent changes, Initial values, Invasive haemodynamic measurements, Iron chelation, Iron chelator, Iron chelators, Iron overload, Isonicotinoyl, Isovolumic phase, Novel iron chelator, Physiological limits, Plasma biochemistry, Plasma concentrations, Ponka, Premature deaths, Present study, Promising drug, Pyridoxal, Pyridoxal hydrazone, Pyridoxal isonicotinoyl hydrazone, Pyridoxal isonicotinoyl hydrazone analogs, Radioiron excretion, Reversible changes, Safety pharmacology, Significant changes, Software chart, Statistical significance, Sterba, Tolerability, Total plasma protein, Troponin, Vehicle group, Vehicle groups, Ventricular ejection fraction, Whole study.
Abstract
Recently, pyridoxal 2-chlorobenzoyl hydrazone (o-108) has been identified as an effective iron chelator [Link et al., Blood 2003; 101: 4172–79]. Since chronic treatment would be necessary in its potential indications, in the present study, the safety and tolerability of this agent after repeated administration was determined. Three doses of o-108 (25, 50, 100 mg/kg, in 10% Cremophor EL) were administered intraperitoneally, once weekly, for 10 weeks to three groups (n–5 each) of Chinchilla male rabbits. The effects on biochemical, haematological and cardiovascular parameters were examined during the experiment; histopathological examination was performed at the end of the experiment. Results were compared with control (saline 2 mL/kg, n–11) and vehicle groups (10% Cremophor EL, 2 mL/kg, n–12). No premature deaths occurred; the well-being of animals was evidenced by their body weight gain, although lower gain was observed with the highest dose (100 mg/kg). Significant elevations of cardiac troponin T plasma concentrations were observed with the highest dose of o-108, but no abnormalities were found in the cardiovascular function and only minor and inconsistent changes in haematological and biochemical parameters were observed. Histopathological examinations of selected organs revealed only weak and reversible changes through all studied groups. Thus, the data from this study suggest that o-108 remains a promising drug from the standpoint of the possibility of its repeated administration and warrants further investigation.
Url:
DOI: 10.1191/0960327105ht571oa
Links to Exploration step
ISTEX:3AD3773662DBDF3BE7D698E92759B971E77D6C64Le document en format XML
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Sterba</name><affiliation><mods:affiliation>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králové, Czech Republic; Department of Pharmacology, Faculty of Medicine in Hradec Králové, Charles University in Prague, Simkova 870, 500 38 Hradec Králové 1, Czech Republic</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: sterbam@lfhk.cuni.cz</mods:affiliation></affiliation></author><author wicri:is="90%"><name sortKey="Simunek, T" sort="Simunek, T" uniqKey="Simunek T" first="T" last="Simunek">T. Simunek</name><affiliation><mods:affiliation>Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Hradec Králová, Czech Republic</mods:affiliation></affiliation></author><author wicri:is="90%"><name sortKey="Mazurova, Y" sort="Mazurova, Y" uniqKey="Mazurova Y" first="Y" last="Mazurová">Y. Mazurová</name></author><author wicri:is="90%"><name sortKey="Adamcova, M" sort="Adamcova, M" uniqKey="Adamcova M" first="M" last="Adamcová">M. 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Gersl</name><affiliation><mods:affiliation>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králové, Czech Republic</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Human and Experimental Toxicology</title><idno type="ISSN">0960-3271</idno><idno type="eISSN">1477-0903</idno><imprint><publisher>Sage Publications</publisher><pubPlace>Sage CA: Thousand Oaks, CA</pubPlace><date type="published" when="2005-11">2005-11</date><biblScope unit="volume">24</biblScope><biblScope unit="issue">11</biblScope><biblScope unit="page" from="581">581</biblScope><biblScope unit="page" to="589">589</biblScope></imprint><idno type="ISSN">0960-3271</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0960-3271</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Animal behaviour</term><term>Anova</term><term>Antioxidant effects</term><term>Biochem cell biol</term><term>Biochem pharmacol</term><term>Biochemical parameters</term><term>Blood pressure</term><term>Blue trichrome</term><term>Body weight gain</term><term>Cardiac</term><term>Cardiac damage</term><term>Cardiac troponin</term><term>Castle hill</term><term>Charles university</term><term>Chelation</term><term>Chelator</term><term>Chelators</term><term>Control group</term><term>Control vehicle</term><term>Cremophor</term><term>Czech republic</term><term>Detection limit</term><term>Eosinophilic cytoplasm</term><term>Essential interface</term><term>Fifth administration</term><term>First administration</term><term>Gross changes</term><term>Haematological</term><term>Haematological analyses</term><term>Haematological parameters</term><term>Heart rate</term><term>Highest dose</term><term>Histopathological</term><term>Histopathological examination</term><term>Hradec</term><term>Hradec kralove</term><term>Hradec krdlove</term><term>Hydrazone</term><term>Hypertransfused rats</term><term>Inconsistent changes</term><term>Initial values</term><term>Invasive haemodynamic measurements</term><term>Iron chelation</term><term>Iron chelator</term><term>Iron chelators</term><term>Iron overload</term><term>Isonicotinoyl</term><term>Isovolumic phase</term><term>Novel iron chelator</term><term>Physiological limits</term><term>Plasma biochemistry</term><term>Plasma concentrations</term><term>Ponka</term><term>Premature deaths</term><term>Present study</term><term>Promising drug</term><term>Pyridoxal</term><term>Pyridoxal hydrazone</term><term>Pyridoxal isonicotinoyl hydrazone</term><term>Pyridoxal isonicotinoyl hydrazone analogs</term><term>Radioiron excretion</term><term>Reversible changes</term><term>Safety pharmacology</term><term>Significant changes</term><term>Software chart</term><term>Statistical significance</term><term>Sterba</term><term>Tolerability</term><term>Total plasma protein</term><term>Troponin</term><term>Vehicle group</term><term>Vehicle groups</term><term>Ventricular ejection fraction</term><term>Whole study</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Recently, pyridoxal 2-chlorobenzoyl hydrazone (o-108) has been identified as an effective iron chelator [Link et al., Blood 2003; 101: 4172–79]. Since chronic treatment would be necessary in its potential indications, in the present study, the safety and tolerability of this agent after repeated administration was determined. Three doses of o-108 (25, 50, 100 mg/kg, in 10% Cremophor EL) were administered intraperitoneally, once weekly, for 10 weeks to three groups (n–5 each) of Chinchilla male rabbits. The effects on biochemical, haematological and cardiovascular parameters were examined during the experiment; histopathological examination was performed at the end of the experiment. Results were compared with control (saline 2 mL/kg, n–11) and vehicle groups (10% Cremophor EL, 2 mL/kg, n–12). No premature deaths occurred; the well-being of animals was evidenced by their body weight gain, although lower gain was observed with the highest dose (100 mg/kg). Significant elevations of cardiac troponin T plasma concentrations were observed with the highest dose of o-108, but no abnormalities were found in the cardiovascular function and only minor and inconsistent changes in haematological and biochemical parameters were observed. Histopathological examinations of selected organs revealed only weak and reversible changes through all studied groups. Thus, the data from this study suggest that o-108 remains a promising drug from the standpoint of the possibility of its repeated administration and warrants further investigation.</div></front></TEI><istex><corpusName>sage</corpusName><keywords><teeft><json:string>chelator</json:string><json:string>hydrazone</json:string><json:string>pyridoxal</json:string><json:string>vehicle group</json:string><json:string>tolerability</json:string><json:string>chelators</json:string><json:string>vehicle groups</json:string><json:string>ponka</json:string><json:string>haematological</json:string><json:string>novel iron chelator</json:string><json:string>initial values</json:string><json:string>isonicotinoyl</json:string><json:string>chelation</json:string><json:string>hradec</json:string><json:string>highest dose</json:string><json:string>cremophor</json:string><json:string>troponin</json:string><json:string>sterba</json:string><json:string>anova</json:string><json:string>histopathological</json:string><json:string>control group</json:string><json:string>statistical significance</json:string><json:string>iron chelators</json:string><json:string>pyridoxal isonicotinoyl hydrazone</json:string><json:string>iron chelation</json:string><json:string>czech republic</json:string><json:string>first administration</json:string><json:string>control vehicle</json:string><json:string>pyridoxal hydrazone</json:string><json:string>plasma concentrations</json:string><json:string>cardiac</json:string><json:string>cardiac troponin</json:string><json:string>hradec krdlove</json:string><json:string>blue trichrome</json:string><json:string>blood pressure</json:string><json:string>premature deaths</json:string><json:string>physiological limits</json:string><json:string>charles university</json:string><json:string>heart rate</json:string><json:string>haematological parameters</json:string><json:string>fifth administration</json:string><json:string>biochem pharmacol</json:string><json:string>invasive haemodynamic measurements</json:string><json:string>present study</json:string><json:string>pyridoxal isonicotinoyl hydrazone analogs</json:string><json:string>body weight gain</json:string><json:string>cardiac damage</json:string><json:string>detection limit</json:string><json:string>software chart</json:string><json:string>castle hill</json:string><json:string>ventricular ejection fraction</json:string><json:string>histopathological examination</json:string><json:string>inconsistent changes</json:string><json:string>isovolumic phase</json:string><json:string>antioxidant effects</json:string><json:string>biochemical parameters</json:string><json:string>reversible changes</json:string><json:string>promising drug</json:string><json:string>whole study</json:string><json:string>plasma biochemistry</json:string><json:string>hypertransfused rats</json:string><json:string>total plasma protein</json:string><json:string>iron chelator</json:string><json:string>iron overload</json:string><json:string>significant changes</json:string><json:string>gross changes</json:string><json:string>hradec kralove</json:string><json:string>safety pharmacology</json:string><json:string>essential interface</json:string><json:string>eosinophilic cytoplasm</json:string><json:string>biochem cell biol</json:string><json:string>haematological analyses</json:string><json:string>animal behaviour</json:string><json:string>radioiron excretion</json:string></teeft></keywords><author><json:item><name>M Sterba</name><affiliations><json:string>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králové, Czech Republic; Department of Pharmacology, Faculty of Medicine in Hradec Králové, Charles University in Prague, Simkova 870, 500 38 Hradec Králové 1, Czech Republic</json:string><json:string>E-mail: sterbam@lfhk.cuni.cz</json:string></affiliations></json:item><json:item><name>T Simunek</name><affiliations><json:string>Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Hradec Králová, Czech Republic</json:string></affiliations></json:item><json:item><name>Y Mazurová</name></json:item><json:item><name>M Adamcová</name></json:item><json:item><name>O Popelová</name></json:item><json:item><name>J Kaplanová</name><affiliations><json:string>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králová, Czech Republic</json:string></affiliations></json:item><json:item><name>P Ponka</name><affiliations><json:string>Lady Davis Institute for Medical Research, McGill University, Montreal, Canada</json:string></affiliations></json:item><json:item><name>V Gersl</name><affiliations><json:string>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králové, Czech Republic</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>iron chelator</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>o-108</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>pyridoxal 2-chlorobenzoyl hydrazone</value></json:item></subject><articleId><json:string>10.1191_0960327105ht571oa</json:string></articleId><arkIstex>ark:/67375/M70-Z3MT1N5P-9</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>research-article</json:string></originalGenre><abstract>Recently, pyridoxal 2-chlorobenzoyl hydrazone (o-108) has been identified as an effective iron chelator [Link et al., Blood 2003; 101: 4172–79]. Since chronic treatment would be necessary in its potential indications, in the present study, the safety and tolerability of this agent after repeated administration was determined. Three doses of o-108 (25, 50, 100 mg/kg, in 10% Cremophor EL) were administered intraperitoneally, once weekly, for 10 weeks to three groups (n–5 each) of Chinchilla male rabbits. The effects on biochemical, haematological and cardiovascular parameters were examined during the experiment; histopathological examination was performed at the end of the experiment. Results were compared with control (saline 2 mL/kg, n–11) and vehicle groups (10% Cremophor EL, 2 mL/kg, n–12). No premature deaths occurred; the well-being of animals was evidenced by their body weight gain, although lower gain was observed with the highest dose (100 mg/kg). Significant elevations of cardiac troponin T plasma concentrations were observed with the highest dose of o-108, but no abnormalities were found in the cardiovascular function and only minor and inconsistent changes in haematological and biochemical parameters were observed. Histopathological examinations of selected organs revealed only weak and reversible changes through all studied groups. Thus, the data from this study suggest that o-108 remains a promising drug from the standpoint of the possibility of its repeated administration and warrants further investigation.</abstract><qualityIndicators><score>9.393</score><pdfWordCount>4729</pdfWordCount><pdfCharCount>31626</pdfCharCount><pdfVersion>1.3</pdfVersion><pdfPageCount>9</pdfPageCount><pdfPageSize>612 x 792 pts (letter)</pdfPageSize><refBibsNative>true</refBibsNative><abstractWordCount>222</abstractWordCount><abstractCharCount>1545</abstractCharCount><keywordCount>3</keywordCount></qualityIndicators><title>Safety and tolerability of repeated administration of pyridoxal 2-chlorobenzoyl hydrazone in rabbits</title><pmid><json:string>16323575</json:string></pmid><genre><json:string>research-article</json:string></genre><host><title>Human and Experimental 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scheme="https://scientific-publisher.data.istex.fr">Sage Publications</publisher><pubPlace>Sage CA: Thousand Oaks, CA</pubPlace><availability><licence><p>sage</p></licence></availability><p scheme="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-0J1N7DQT-B"></p><date>2005</date></publicationStmt><notesStmt><note type="research-article" scheme="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</note><note type="journal" scheme="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</note></notesStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a" type="main" xml:lang="en">Safety and tolerability of repeated administration of pyridoxal 2-chlorobenzoyl hydrazone in rabbits</title><author xml:id="author-0000"><persName><forename type="first">M</forename><surname>Sterba</surname></persName><email>sterbam@lfhk.cuni.cz</email><affiliation>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králové, Czech Republic; Department of Pharmacology, Faculty of Medicine in Hradec Králové, Charles University in Prague, Simkova 870, 500 38 Hradec Králové 1, Czech Republic</affiliation></author><author xml:id="author-0001"><persName><forename type="first">T</forename><surname>Simunek</surname></persName><affiliation>Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Hradec Králová, Czech Republic</affiliation></author><author xml:id="author-0002"><persName><forename type="first">Y</forename><surname>Mazurová</surname></persName></author><author xml:id="author-0003"><persName><forename type="first">M</forename><surname>Adamcová</surname></persName></author><author xml:id="author-0004"><persName><forename type="first">O</forename><surname>Popelová</surname></persName></author><author xml:id="author-0005"><persName><forename type="first">J</forename><surname>Kaplanová</surname></persName><affiliation>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králová, Czech Republic</affiliation></author><author xml:id="author-0006"><persName><forename type="first">P</forename><surname>Ponka</surname></persName><affiliation>Lady Davis Institute for Medical Research, McGill University, Montreal, Canada</affiliation></author><author xml:id="author-0007"><persName><forename type="first">V</forename><surname>Gersl</surname></persName><affiliation>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králové, Czech Republic</affiliation></author><idno type="istex">3AD3773662DBDF3BE7D698E92759B971E77D6C64</idno><idno type="ark">ark:/67375/M70-Z3MT1N5P-9</idno><idno type="DOI">10.1191/0960327105ht571oa</idno><idno type="article-id">10.1191_0960327105ht571oa</idno></analytic><monogr><title level="j">Human and Experimental Toxicology</title><idno type="pISSN">0960-3271</idno><idno type="eISSN">1477-0903</idno><idno type="publisher-id">HET</idno><idno type="PublisherID-hwp">sphet</idno><imprint><publisher>Sage Publications</publisher><pubPlace>Sage CA: Thousand Oaks, CA</pubPlace><date type="published" when="2005-11"></date><biblScope unit="volume">24</biblScope><biblScope unit="issue">11</biblScope><biblScope unit="page" from="581">581</biblScope><biblScope unit="page" to="589">589</biblScope></imprint></monogr></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>2005</date></creation><langUsage><language ident="en">en</language></langUsage><abstract xml:lang="en"><p>Recently, pyridoxal 2-chlorobenzoyl hydrazone (o-108) has been identified as an effective iron chelator [Link et al., Blood 2003; 101: 4172–79]. Since chronic treatment would be necessary in its potential indications, in the present study, the safety and tolerability of this agent after repeated administration was determined. Three doses of o-108 (25, 50, 100 mg/kg, in 10% Cremophor EL) were administered intraperitoneally, once weekly, for 10 weeks to three groups (n–5 each) of Chinchilla male rabbits. The effects on biochemical, haematological and cardiovascular parameters were examined during the experiment; histopathological examination was performed at the end of the experiment. Results were compared with control (saline 2 mL/kg, n–11) and vehicle groups (10% Cremophor EL, 2 mL/kg, n–12). No premature deaths occurred; the well-being of animals was evidenced by their body weight gain, although lower gain was observed with the highest dose (100 mg/kg). Significant elevations of cardiac troponin T plasma concentrations were observed with the highest dose of o-108, but no abnormalities were found in the cardiovascular function and only minor and inconsistent changes in haematological and biochemical parameters were observed. Histopathological examinations of selected organs revealed only weak and reversible changes through all studied groups. Thus, the data from this study suggest that o-108 remains a promising drug from the standpoint of the possibility of its repeated administration and warrants further investigation.</p></abstract><textClass><keywords scheme="keyword"><list><head>keywords</head><item><term>iron chelator</term></item><item><term>o-108</term></item><item><term>pyridoxal 2-chlorobenzoyl hydrazone</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="2005-11">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/M70-Z3MT1N5P-9/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="corpus sage not found" wicri:toSee="no header"><istex:xmlDeclaration>version="1.0" encoding="UTF-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//NLM//DTD Journal Publishing DTD v2.3 20070202//EN" URI="journalpublishing.dtd" name="istex:docType"></istex:docType><istex:document><article article-type="research-article" dtd-version="2.3" xml:lang="EN"><front><journal-meta><journal-id journal-id-type="hwp">sphet</journal-id><journal-id journal-id-type="publisher-id">HET</journal-id><journal-title>Human and Experimental Toxicology</journal-title><issn pub-type="ppub">0960-3271</issn><publisher>
<publisher-name>Sage Publications</publisher-name>
<publisher-loc>Sage CA: Thousand Oaks, CA</publisher-loc>
</publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.1191/0960327105ht571oa</article-id><article-id pub-id-type="publisher-id">10.1191_0960327105ht571oa</article-id><article-categories><subj-group subj-group-type="heading"><subject>Articles</subject></subj-group></article-categories><title-group><article-title>Safety and tolerability of repeated administration of pyridoxal 2-chlorobenzoyl hydrazone in rabbits</article-title></title-group>
<contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Sterba</surname><given-names>M</given-names></name><aff>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králové, Czech Republic; Department of Pharmacology, Faculty of Medicine in Hradec Králové, Charles University in Prague, Simkova 870, 500 38 Hradec Králové 1, Czech Republic<email xlink:type="simple">sterbam@lfhk.cuni.cz</email>
</aff></contrib></contrib-group>
<contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Simunek</surname><given-names>T</given-names></name><aff>Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Hradec Králová, Czech Republic</aff></contrib></contrib-group>
<contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Mazurová</surname><given-names>Y</given-names></name></contrib></contrib-group>
<contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Adamcová</surname><given-names>M</given-names></name></contrib></contrib-group>
<contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Popelová</surname><given-names>O</given-names></name></contrib></contrib-group>
<contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Kaplanová</surname><given-names>J</given-names></name><aff>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králová, Czech Republic</aff></contrib></contrib-group>
<contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Ponka</surname><given-names>P</given-names></name><aff>Lady Davis Institute for Medical Research, McGill University, Montreal, Canada</aff></contrib></contrib-group>
<contrib-group><contrib contrib-type="author" xlink:type="simple"><name name-style="western"><surname>Gersl</surname><given-names>V</given-names></name><aff>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králové, Czech Republic</aff></contrib></contrib-group>
<pub-date pub-type="ppub"><month>11</month><year>2005</year></pub-date><volume>24</volume><issue>11</issue><fpage>581</fpage><lpage>589</lpage><abstract>
<p>Recently, pyridoxal 2-chlorobenzoyl hydrazone (o-108) has been identified as an effective iron chelator [Link et al., Blood 2003; 101: 4172–79]. Since chronic treatment would be necessary in its potential indications, in the present study, the safety and tolerability of this agent after repeated administration was determined. Three doses of o-108 (25, 50, 100 mg/kg, in 10% Cremophor EL) were administered intraperitoneally, once weekly, for 10 weeks to three groups (n–5 each) of Chinchilla male rabbits. The effects on biochemical, haematological and cardiovascular parameters were examined during the experiment; histopathological examination was performed at the end of the experiment. Results were compared with control (saline 2 mL/kg, n–11) and vehicle groups (10% Cremophor EL, 2 mL/kg, n–12). No premature deaths occurred; the well-being of animals was evidenced by their body weight gain, although lower gain was observed with the highest dose (100 mg/kg). Significant elevations of cardiac troponin T plasma concentrations were observed with the highest dose of o-108, but no abnormalities were found in the cardiovascular function and only minor and inconsistent changes in haematological and biochemical parameters were observed. Histopathological examinations of selected organs revealed only weak and reversible changes through all studied groups. Thus, the data from this study suggest that o-108 remains a promising drug from the standpoint of the possibility of its repeated administration and warrants further investigation.</p>
</abstract><kwd-group>
<kwd>iron chelator</kwd>
<kwd>o-108</kwd>
<kwd>pyridoxal 2-chlorobenzoyl hydrazone</kwd>
</kwd-group><custom-meta-wrap><custom-meta xlink:type="simple"><meta-name>sagemeta-type</meta-name><meta-value>Journal Article</meta-value></custom-meta><custom-meta xlink:type="simple"><meta-name>search-text</meta-name><meta-value> Human & Experimental Toxicology (2005) 24: 581 - 589 www.hetjournal.com Safety and tolerability of repeated a dminiis tra tion of pyr idoxa 2-ch oro benzoy hydrazone in rabbits M Sterba*I, T Simunek2, Y Mazurova1, M Adamcova1, 0 Popelova1, J Kaplanov 1, P Po'ka3 and V Gersl1 'Faculty of Medicine in Hradec Krdlove, Charles University in Prague, Hradec Krdlove, Czech Republic; 2Faculty of Pharmacy in Hradec Krdlove, Charles University in Prague, Hradec Krdlove, Czech Republic; 3Lady Davis Institute for Medical Research, McGill University, Montreal, Canada Recently, pyridoxal 2-chlorobenzoyl hydrazone (o-108) has been identified as an effective iron chelator [Link et al., Blood 2003; 101: 4172-79]. Since chronic treat- ment would be necessary in its potential indications, in the present study, the safety and tolerability of this agent after repeated administration was determined. Three doses of o-108 (25, 50, 100 mg/kg, in 10% Cremophor EL) were administered intraperitoneally, once weekly, for 10 weeks to three groups (n =5 each) of Chinchilla male rabbits. The effects on biochemical, haematological and cardiovascular parameters were examined during the experiment; histopathological examination was per- formed at the end of the experiment. Results were compared with control (saline 2 mL/kg, n =11) and vehicle groups (10% Cremophor EL, 2 mL/kg, n =12). No premature deaths occurred; the well-being of animals was evidenced by their body weight gain, although lower gain was observed with the highest dose (100 mg/kg). Significant elevations of cardiac troponin T plasma concentrations were observed with the highest dose of o-108, but no abnormalities were found in the cardiovas- cular function and only minor and inconsistent changes in haematological and biochemical parameters were observed. Histopathological examinations of selected organs revealed only weak and reversible changes through all studied groups. Thus, the data from this study suggest that o-108 remains a promising drug from the standpoint of the possibility of its repeated adminis- tration and warrants further investigation. Human & Experimental Toxicology (2005) 24, 581- 589 Key words: iron chelator; o-108; pyridoxal 2-chlorobenzoyl hydrazone Introduction While iron (Fe) is a vital element for virtually all living organisms, it also presents a risk of serious injury. The biological importance, as well as the toxicity of iron, is based on its ability to readily precede one electron transmission between its Fe3" and Fe2+ forms.1 Free iron is able to efficiently catalyze the formation of extremely toxic hydroxyl radicals via a process known as the Haber-Weiss reaction.2 Thus, either an excess of iron in the body or disruption of its tightly regulated homeostasis at the cellular level may have a severe impact on human health. Iron chelation has been shown to be an effective therapeutic approach in the treatment of patients *Correspondence: Martin Sterba, Department of Pharmacology, Faculty of Medicine in Hradec Krilov6, Charles University in Prague, Simkova 870, 500 38 Hradec Krdlov6 1, Czech Republic E-mail: sterbam~lfhk.cuni.cz Received 10 January 2005; revised 1 August 2005; accepted 22 August 2005 © 2005 Edward Arnold (Publishers) Ltd suffering from both acute iron intoxication and chronic iron overload.34 Iron overload devel- ops as a result of treatment with repeated transfu- sions, which are necessary for patients suffering from hereditary diseases of haematopoiesis, such as 3-thallassemia. This chronic iron burden is accompanied with severe damage and subsequent failure of the heart and parenchymal organs. To date, iron chelation is the only strategy that considerably prolongs the lifespan of these patients.5 In addition, iron chelation appears to be a successful approach in many non-overload condi- tions. Some iron chelators were shown to have various attractive pharmacological properties, e.g., cardioprotective, neuroprotective, anticancer, or antimicrobial.6-9 The antioxidant effects of iron chelators are based on their ability to prevent iron- catalyzed formation of hydroxyl radicals, which are known to be the most toxic form of reactive 10.1191/0960327105ht571oa Safety and tolerability of a novel iron chelator M Sttrba et al. 582 oxygen species (ROS) with a short half-life and a powerful destructive effect. Progress in this area is unfortunately hindered by a lack of selective and non-toxic ligands. Deferoxamin (DFO) remains the only widely used chelator, despite its drawbacks. This drug must be administered by long subcutaneous infusions and, because of its hydrophilic nature, it cannot easily permeate into the cells. The first orally active iron chelator, deferiprone, which was de- signed to overcome some DFO disadvantages, remains a controversial subject concerning its safety and efficacy.10'11 Thus, there is an urgent need to develop new effective and non-toxic iron chelators. Pyridoxal 2-chlorobenzoyl hydrazone (agent o-108) is a novel and selective iron chelator from the group of pyridoxal isonicotinoyl hydrazone (PIH) analogues.'2"3 Analogous to PIH, o-108 is a tridental iron chelator selectively forming 2:1 com- plexes with Fe3 . Due to the presence of a halogen moiety and the replacement of the pyridine ring by benzene, this agent is even more lipophilic than its parent compound. Therefore, o-108 can reach the intracellular targets more efficiently. Its ability to mobilize iron was demonstrated both in vitro and in vivo. Agent o-108 was even more effective than PIH in the mobilisation of 59Fe from reticulocytes and K506 cells.14"15 Ortho-halogenated derivatives of pyridoxal benzoyl hydrazone were shown to be less toxic in vitro then its meta or para substituted derivatives.15 In a recent study, oral administration of o-108 to hypertransfused 59Fe-ferritin-tagged rats led to linear dose-dependent cumulative (7 days) radioiron excretion, ranging from 2.3 + 1.8% at 25 mg/kg to 42.6+ 7.7% at 200 mg/kg, expressed as the percentage of injected radioactivity. The residual hepatic radioactivity observed in this experiment was negatively correlated to radioiron excretion induced by o-108.16 Although o-108 has been shown to possess interesting pharmacodynamic properties, the safety of this agent remains unknown and is the subject of the present study. The dosage of o-108 and the design of our experiments were based on the first in vivo study demonstrating its efficacy.16 Since the therapy of iron overload (as well as its use in other potential indications) would require repeated administration of this agent, a knowledge of the potential hazards in this schedule may be of value. This study was therefore designed to assess the preliminary tolerability and safety of repeated weekly administration of o-108 in three escalating doses in rabbits. Material and methods Chemicals Pyridoxal 2-chlorobenzoyl hydrazone (o-108) was synthesized by Schiff-base condensation of pyri- doxal and 2-chlorobenzoylhydrazide as previously described.'7 The structure and purity of the com- pound was confirmed using NMR and HPLC with UV detection. Cremophor EL (Sigma-Aldrich, Czech Republic), ketamine (Calypsol inj., Gedeon Richter, Hungary), Aqua pro injectione (Biotika, Slovakia), and pentobarbital (Nembutal Sodium, Abbott, USA) were used in the experiment. Animals Adult medium-size Chinchilla male rabbits with an average initial weight of 3.5 + 0.1 kg were housed under a 12-h light cycle, constant temperature and humidity. The animals had free access to water and a standard laboratory pellet diet. All experimental procedures were performed under ketamine anaes- thesia (50 mg/kg, i.m.). Final invasive haemody- namic measurements were carried out under pentobarbital anaesthesia (30 mg/kg, i.v.). The experiments were performed in accordance with the 'Guide for the Care and Use of Laboratory Animals' (1996) and under the supervision of the Ethical Committee of the Medical Faculty in Hradec Kralove. Experimental design Fifteen rabbits were randomly divided into three groups. Pyridoxal o-chlorobenzoyl hydrazone (o-108) was partially dissolved in 10% aqueous Cremophor EL and administered intraperitoneally to the three groups of animals (n = 5, each), once weekly for 10 weeks, in three escalating doses (25, 50 and 100 mg/kg). Standard biochemical and hae- matological analyses of blood samples, as well as non-invasive evaluations of cardiac function, were performed repeatedly during the experiment as described below. The body weight gain and animal behaviour were recorded weekly. Finally, 4 to 5 days after the administration of the last dose, the invasive haemodynamic measurements were performed. The animals were then overdosed with pentobarbital and autopsy followed by histopathological evalua- tion was carried out. The results were compared with groups receiving vehicle (10% aqueous solu- tion of Cremophor EL: 2 mL/kg, n = 12) or saline (control-saline: 2 mL/kg, n = 11). Biochemical and haematological analysis Blood was sampled for standard biochemical and haematological analyses instantly before and Safety and tolerability of a novel iron chelator M Sterba et al. 583 24 hours after the first administration and also before the fifth administration of the drug and at the end of the experiment. For the determination of standard plasma/serum biochemical parameters, a fully automatic clinical chemistry analyser (HITA- CHI 737, Hitachi, Tokyo, Japan) was used in accor- dance with the manufacturer's recommendations. Haematological parameters were measured by means of a Coulter T890 counter (Beckman Coulter, Fullerton, USA). Additionally, plasma concentra- tions of cardiac troponin T (cTnT) as a specific marker of cardiac damage were determined using the Elecsys Troponin T STAT immunoassay on an Elecsys 2010 immunoassay analyser (Roche Diag- nostics, Rotkreuz, Switzerland) with the detection limit of 0.010 ng/mL. The determination of troponin T was performed in the plasma samples taken before and 24 hours after the first and before the fifth, eighth and tenth administrations and at the end of the experiment. Non-invasive cardiac measurements Non-invasive cardiac function measurements (echo- cardiography; EGG) were carried out before the first and fifth administrations and at the end of the experiment. ECG was recorded with an ADI Power- Lab/8SP, software Chart for Windows v. 3.4.11 (Adinstruments, Castle Hill, Australia). Echocardio- graphic examination was performed using GE Vingmed CFM 800A echocardiograph (GE Vingmed Ultrasound, Horten, Norway) equipped with a stan- dard paediatric 7.5 MHz probe. The left ventricle was visualized in its long axis by means of the left parasternal approach and guided M-mode measure- ment at the tips of the mitral valve was performed. The end-systolic and end-diastolic diameters were obtained and used for the determination of the left ventricular ejection fraction (LVEF). Invasive haemodynamic measurements At the end of the experiment, heart rate (HR), mean blood pressure in the femoral artery (BP) and the index of contractility (dP/dtmax) were measured invasively using an ADI PowerLab/8SP (Adinstru- ments, Castle Hill, Australia) and software Chart for Windows (v. 3.4.11). The left carotid artery was prepared and a PE catheter filled-in with hepari- nized (10 IU/mL) saline was introduced into the left heart ventricle to measure the maximal pressure rise in the isovolumic phase of systole (maximum of the first derivative of LV pressure: dP/dtmax). For the arterial blood pressure measurement, a PE cannula was inserted into the right femoral artery. All parameters were recorded after a 15-min equilibra- tion period. Histopathological examination Tissue blocks of the transversely sectioned left and right cardiac ventricles, left kidney, left liver lobule, left caudal lung lobule, duodenum (approximately 3 cm below the pyloric sphincter), and striated muscle tissue (from the left quadriceps femoral muscle) were fixed by immersion in 4% formalde- hyde for 5 to 7 days. Paraffin sections (6 gm thick) were stained with haematoxylin-eosin (H&E) and Masson's blue trichrome. Photomicrographs were made with a Lucia G software version 4.51 (Labora- tory Imaging, Prague, Czech Republic) in the Depart- ment of Medical Biology, Faculty of Medicine in Hradec Kralove. Statistical analysis Data are expressed as means+ SEM. The statistical software SigmaStat for Windows 3.0 (Jandel, Germany) was used in this study. Significances of differences between groups were determined using the one way ANOVA unpaired test or Kruskal- Wallis ANOVA on ranks (data without normal distribution). Significances within the group (com- parison with the initial values) were evaluated using the paired t-test or the Wilcoxon Signed Rank test. P <0.05 was used as the level of statistical signifi- cance. Results No premature deaths relating to the treatment occurred throughout the experiments. No seizures, manifest changes in animal behaviour or overall appearance were observed during the study. How- ever, somewhat reduced locomotive activity and tendency to reduced feed intake were noted in the group treated with the highest dose (100 mg/kg) of o- 108. The relative body weight gain was significant in control and vehicle groups during the whole study. The animals treated with the highest dose of the drug had the lowest body weight gain (insignificant in five of 11 measurements). The body weights in the course of the study are shown in Figure 1. Plasma biochemistry (Table 1) revealed predomi- nantly minor and inconsistent changes in most of studied parameters through all the groups. The levels of plasma ions in animals treated with o-108 were always in physiological limits and mostly followed the fluctuation seen in control and vehicle groups. The total plasma protein did not change in the first half of the experiment. However, at the end of the study, a slight drop was observed in the vehicle group and also (somewhat more marked) in the groups treated with 50 and 100 mg/kg of o-108. This change was also significant when compared to Safety and tolerability of a novel iron chelator M Sterba et al. 584 ----- control I ---vehicle |I A--- o-1 08/25mg I . -*- ---0- o-1 08/50mg - -r )K--- o-108/100mg T2 ' . -i ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ---- ~ ~ ~ - ----- T 1 2 3 4 5 6 7 8 9 10 11 week Figure 1 Body weight changes during the study. Statistical significance: 'i' insignificant body weight gain (paired t-test, P < 0.05) in comparison with the initial values within one group - all other body weights were significantly increased, (ANOVA, P <0.05) comparison between the groups - no significant differences were observed. The absolute initial values at the beginning of the study were: 3.45 + 0.11 (control group), 3.42 + 0.09 (vehicle group), 3.55 + 0.03 (o-108/25 mg group), 3.54 + 0.09 (o-108/50 mg group), 3.38+0.07 (o-108/100 mg group) - P=NS. the control and vehicle groups. Similar results were obtained for plasma albumin. Nevertheless, the values of total plasma protein and albumin did not fall below the physiological limits in either group. No elevations in alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phospha- tase (ALP) or total serum bilirubin were observed either 24 hours after the administration of the first dose or later. Minor changes in lactate dehydrogen- ase (LDH) followed those seen in the control and vehicle groups. Plasma cholesterol levels were somewhat higher 24 hours after the first o-108 administration compared with the initial values. Similar, though less pronounced changes occurred in the vehicle group. Weak and transient fluctuation of plasma lipids had fully normalized during the experiment. While only little oscillation in creatini- naemia occurred in the control and vehicle groups, scattered elevation was seen in animals treated with o-108. This observation became significant only at the end of experiment with the lowest dose of the chelator when compared to initial values, whereas no significance was found in comparison with other groups. Moderate elevations in cardiac troponin T plasma concentrations above the detection limit (Figure 2) were found in the control and vehicle groups. Similar findings were also present in the case of lowest and middle dose of studied compound. However, in the group treated with 100 mg/kg of o- 108, the rise was more prominent and significantly different from both the initial values and the control and vehicle groups. No changes in blood cell count were observed in animals treated with any dose of the chelator. A mild, though significant rise in the mean cell volume was present in the group receiving middle and higher doses of o-108. Other haematological parameters oscillated similarly in all groups. The overview of selected haematological parameters is given in Table 2. Non-invasive echocardiographic measurements (Figure 3) showed no significant changes with the exception of a slightly higher ejection fraction at the end of the experiment in 25 and 100 mg/kg o-108 receiving animals, compared to both control and vehicle groups. Nevertheless, all these changes were well within physiological limits. No significant changes in cardiac contractility, blood pressure and heart rate were detected (Table 3). Furthermore, no gross changes in ECG were observed in any group. The histopathological study of selected organs revealed only weak and reversible changes through all studied groups. Groups of cardiomyocytes with eosinophilic (intensely stained) cytoplasm were also found in controls (Figure 4a), and represented in majority reversible changes. Scattered degener- ated myocytes were observed in both the o-108- treated/animals (Figure 4c) and the vehicle group (Figure 4b). The kidney parenchyma revealed slight changes, particularly in the epithelium of the proximal and distal tubules: increased eosino- philia and a small number of pyknotic (shrunk) nuclei were found in the group with the vehicle (Figure 5b), less in the group with o-108 (Figure 5c). Other organs under study were free of any patholo- gical changes. Discussion Iron chelation has recently become a promising concept of pharmacological intervention in a num- ber of pathologies. To date, it is the only way of prolonging the lifespan of patients suffering from iron overload. Iron selective ligands of higher lipophilicity may easily penetrate the cell and prevent the iron-catalysed intracellular formation of free radical species. Moreover, dexrazoxane, which is intracellularly hydrolysed to form the iron chelator ADR-925, is the only well-documented cardioprotectant against anthracycline-induced car- diac damage, in which ROS are believed to be the key players.18 Interesting antioxidant properties were also reported for the lead compound of the aroylhydrazone chelators - PIH.19-22 A significant degree of cardioprotection against hydrogen perox- ide induced injury was reported for another analo- gue of o-108.23'24 Furthermore, several other pharmacodynamic properties (e.g., antiproliferative 130 - ._ 125 - Xa 120- s~ 115- C .4 110 - 0 m 105 - 0 0 m 100 95 Safety and tolerability of a novel iron chelator M St~rba et al 585 Table 1 Selected biochemical parameters Parameter/group Before the first administration After the first administration Before the fifth administration End of study Sodium (mmol/L) Control 138.7 +1.2 140.5 + 0.9 * 139.1 +1.0 141.4 ± 0.5* Vehicle 140.4 + 1.3 139.9 +0.7 140.7 +0.9 142.9 ±0.8 o-108/25 mg 142.5 +0.7 142 +0.9 140.8 + 0.5 141.8 ±0.8 o-108/50 mg 140.6+ 0.5 142.4 ±0.5 139.4 + 1.2 140.6 +0.9 0-108/100 mg 140.2 + 0.7 141.8 + 0.7 * 143.4 + 1.0* 141.8 ±0.6 Potassium (mmol/L) Control 3.6+ 0.2 3.9 +0.2 * 3.5 +0.2 3.4 +0.1 Vehicle 3.7+0.1 3.9+0.1 3.6 +0.1 3.4 +0.3 o-108/25 mg 3.7+0.1 4.0+0.1 3.9+0.1 4.0±0.1 o-108/50 mg 3.8+0.1 4.3±+0.1 * 3.7 +0.1 3.2 +0.1* o-108/100 mg 3.9 +0.2 4.2 ±0.2 3.7 +0.2 3.4 +0.1 Calcium (mmol/L) Control 3.11 +0.05 3.61 ± 0.09 * 3.09 +0.05 3.15 +0.04 Vehicle 2.94 + 0.05 3.3 7 + 0.06 * 3.06 + 0.04 2.89 +0.04 u-108/25 mg 3.21+0.05 3.30±+0.10 3.26 +0.19 3.11 +0.29 o-108/50 mg 3.07 +0.06 3.47 + 0.09* 3.06 +0.09 2.98 + 0.08 0-108/100 mg 3.11±0.05 3.25 +0.13 3.12 +0.05 2.86±+0.06* Iron-serum (jimol/L) Control 40.9+ 3.4 31.6 ± 2.4* 34.9 + 2.6 46.1 + 1.8 Vehicle 34.2 + 1.7 26.5 + 2.4* 32.1 +2.1 43.7 + 2.7* o-108/25 mg 36.5 +4.1 22.2±+5.6 35.5 +4.6 42.8 +3.6 o-108/50 mg 29.5 +4.4 27.3 +3.5 35.8+3.2 411 1+2.9* 0-108/100 mg 30.1 ± 5.4 20.8 + 2.9 31.0 +2.8 46.8 + 2.7* Cholesterol (jimol/L) Control 1.81 ±0.48 1.41 + 0.27 1.53 + 0.31 0.97 + 0.17 Vehicle 1.50+0.15 1.80+0.17* 1.16+0.16* 1.15 +0.17 o-108/25 mg/kg 1.73 + 0.42 2.28 +0.32* 1.71 +0.37 1.19 +0.25 0-108/50 mg/kg 1.42 ± 0.26 2.18 ±0.37* 1.36 + 0.23 1.10±0.14 0-108/100 mg/kg 1.87± 0.22 2.86 ±0.20*cv 1.64 +0.15 1.08 +0.13* Creatinine (pmol/L) Control 112.7 +4.5 109.5 + 5.4 118.2 + 6.6 106.8 ±8.1 Vehicle 109.1 +4.7 100. 1 ± 5.5* 106.4+4.8 111.3 ±3.8 o-108/25 mg/kg 105.0 +1.5 97.3 + 3.7 115.0±5.6 150.1 +8.7* o-108/50 mg/kg 110.2 + 2.6 129.8 + 22.4 111.0 +3.8 117.6 + 7.8 o-108/100 mg/kg 103.0+ 6.4 144.2 ± 19.5 112.8 ±5.5* 127.8 + 14.3 ALT (jikat/L) Cuntrul 1.19±0.15 1.37±+0.13 1.22 LJ.18 0.99+0.18 Vehicle 1.51±0.24 1.77 +0.31 1.19+0.12 0. 92±+0.05 * o-108/25 mg/kg 1.91±0.31 1.94 +0.28 1.40+0.13 1. 13±+0.13 * o-108/50 mg/kg 1.69+0.35 1.50 + 0.25 1.06 +0.16 0.97 ± 0.18 o-108/100 mg/kg 1.28±0.14 1.41 +0.26 0.68±0.15* 0.70+0.15* AST (jikat/L) Control 0.60 +0.11 0.86 +0.07 0.75 ± 0.17 0.68 + 0.06 Vehicle 0.67+0.09 1.24 +0.31 0 52±+0.05 0.68 ±0.04 o-108/25 mg/kg 0.70 + 0.13 0. 55 + 0.06* 0.56 +0.09 0.70±0.05 o-108/50 mg/kg 0.55 + 0.15 0.53 + 0.04 0.49 +0.04 0.51 +0.06 o-108/100 mg/kg 0.71 +0.10 0.62 + 0.11 0.42 ± 0.06 0.66 ± 0.20 ALP (pkat/L) Control 2.16+ 0.32 1.32 +0.16* 2.10 +0.26 1.93 +0.21 Vehicle 2.73 +0.46 1.93 +0.36 2.10 +0.32 1. 73 +0.28 * o-108/25 mg/kg 2.27 + 0.23 1.51 + 0.20* 2.10 +0.31 1.76 + 0.26 o-108/50 mg/kg 2.76 + 0.48 1.61 + 0.25 2.46 + 0.4 8* 2.05 ± 0.50 * o-108/100 mg/kg 3.92 + 0.86 2.2 7 + 0.64* 2.81 + 0.65 * 2.3 7 + 0.64 * Bilirubin (j~tmol/L) Control 2.8+0.5 2.8±+0.3 2.5 +0.2 2.9±+0.3 Vehicle 2.8+ 0.5 2.1 +0.3 2.5±+0.5 2.5±+03 0-108/25 mg/kg 3.0+0.0 3.0 +0.0 3.0±+0.0 2.5±+0.3 o-108/50 mg/kg 2.6 + 0.3 2.4 +0.4 1.6 + 0.3 * 2.6 +0.3 o-108/100 mg/kg 3.0+ 0.0 3.0 + 0.0 2.0 ± 0.3 * 3.0 ±0.0 Albumin (g/L) Control 55.0 + 0.9 52.6 + 1.1 53.9 + 1.3 55.3 +0.9 Vehicle 56.5 + 0.8 52.1 +0.5* 55.5 ±0.8 5 3.4 ± 0.9 * 0-108/25 mg/kg 51.0+±0.9vh 49.5 +2.7* 52.6 +2.8 49.2 +3.3 *c o-108/50 mg/kg 56.3 + 0.6 5 3.4 ± 0.8 * 54.8+ 1.1 47.7 + 1.3*cv o-1 08/1 00 mg/kg 57.4 + 0.8c 54.1 + 0.7* 55.8± 1.0 50.5 + 1.3* LDH (jikat/L) Control 8.53 + 1.25 12.88 + 1.2 4* 7.00 + 1.33 7.80 + 0.62 Vehicle 5.85 +0.75 13.89 +3.11 * 13.89 +3.11* 8.98 +0.82 * o-108/25 mg/kg 6.71 +0.71 9.97 +1.91 9.97 +1.91 10. 7 +1.48 * o-108/50 mg/kg 6.56 + 1.26 9.99 + 1.11 9.99 +1.11 7.84 + 1.71 Statistical significance: *comparison with the initial values within one group (paired t-test, P <0.05); c: comparison with the control group; v: comparison with vehicle group; h: comparison with highest dose (100 mg/kg) group, (ANOVA, P <0.05). Safety and tolerability of a novel iron chelator M Sterba et al. 586 0.14- 0.12- 0.1 - m 0.08- 'E 0.06- t' 0.04- 0.02 - x Av 2 cv O control E vehicle E o-108/25mg 0 o-108/50mg * o-108/100mg before after before before before last administration 1 st 1 st 5th 8th 1 0th week Figure 2 Cardiac troponin T plasma concentrations. Statistical significance: *(paired t-test, P < 0.05) comparison with the initial values, (ANOVA, P <0.05) comparison between treatments: c, comparison with control group; v, comparison with vehicle group. and antimalarial) were already reported for some pyridoxal o-chlorobenzoyl hydrazone ana- logues.9'25 27 Thus, besides the interesting iron excretion properties of this novel chelator o-108,16 a number of other potential uses is opening. Besides the research focused on potentially ben- eficial pharmacodynamic properties of any new chemical entity, it is no less important to assess also its safety and tolerability. This approach is considered to be of value even in the early part of drug development, since it may save the budget and accelerate the whole process of identifying pharma- cologically active compounds with acceptable toxi- city. Much interest in this field is dedicated to a recently constituted discipline, safety pharmacol- ogy, which has become an essential interface of pharmacology and toxicology in non-clinical set- tings.28 This study was designed as a preliminarily safety pharmacological study of the novel iron chelating agent o-108 after its repeated administration. The dosage schedule used in this experiment was de- rived from the study focused on weekly cumulative iron excretion following single administration of the chelator.16 The study presented herein shows that this novel iron chelator was well tolerated, though at Table 2 Selected haematological parameters Parameter/group Leukocytes ( x 109/L) Control Vehicle o-108/25 mg/kg o-108/50 mg/kg o-108/100 mg/kg Erythrocytes (x 1012/L) Control Vehicle o-108/25 mg/kg o-108/50 mg/kg o-108/100 mg/kg MCV (fl) Control Vehicle o-108/25 mg/kg o-108/50 mg/kg o-108/100 mg/kg Haemoglobin (g/L) Control Vehicle o-108/25 mg/kg o-108/50 mg/kg o-108/100 mg/kg Haematocrit Control Vehicle o-108/25 mg/kg o-108/50 mg/kg o-108/100 mg/kg Thrombocytes ( x 109/L) Control Vehicle o-108/25 mg/kg o-108/50 mg/kg o-108/100 mg/kg Before the first administration 7.1+ 0.5 6.6+ 0.3 6.2 + 0.5 6.9+ 0.2 6.3+ 0.9 5.72 + 0.18 5.84 + 0.18 5.68 + 0.19 5.92 + 0.13 5.88 + 0.07 67.3 + 0.9 68.8 + 1.1 66.8 +0.4 66.3+ 1.0 66.4 + 0.4 125.3+ 3.4 128.1 + 3.1 120.3 + 5.5 127.2 + 2.6 125.4 + 1.9 0.385 + 0.012 0.399 + 0.009 0.379 + 0.013 0.392 + 0.008 0.391 + 0.004 469+ 58 330 + 21 327 + 23 305 + 34 306 + 34 Before the fifth administration 6.5 + 0.7 6.0+0.3* 7.5 + 0.7* 6.3 + 0.6 7.7+ 0.7 5.99 + 0.10 5.56 + 0.14* 5.60 + 0.15 5.86 + 0.16 5.56 + 0.09 69.6+ 1.1 70.9 + 0.8* 66.8+1.1 69.6 + 0.8* 70.1 + 1.1* 132.9+ 1.8* 126.7 + 2.9 118.8 + 2.7*c 132.6 + 2.7 127.8 + 1.3 0.417 + 0.008* 0.393 + 0.008 0.374 + 0.009c 0.407 + 0.010* 0.404 + 0.007* 399 + 28 352 + 19 448 + 72 341 + 25 357 + 43 Statistical significance: *(paired t-test, P <0.05) comparison with the initial values, (ANOVA, P <0.05); c: comparison with control group; v: comparison with vehicle group. End of study 5.1 +0.5* 5.5 + 0.7 6.0+ 0.5 5.1+ 0.7 5.2 + 0.3 5.83 + 0.12 5.68 + 0.12 5.34 + 0.18 5.38 + 0.15 5.68 + 0.12 69.0+ 0.7 69.0+ 1.0 69.9+ 1.3 70.4 + 1.4* 69.9+0.7* 130.4 + 1.8 129.5 + 2.2 118.3 + 5.2 120.4 + 4.0 123.4 + 3.7 0.402 + 0.007 0.391 ± 0.008 0.373 + 0.017 0.379 + 0.012 0.386 + 0.012 445 + 31 370 + 31 355 + 31 327 + 26 349 + 31 {, .-1 .I . 70- 60- Safety and tolerability of a novel iron chelator M St&ba et al. 587 Table 3 Invasive cardiovascular measurements 50 - 0 control U_ 0~~~~~~~~~~~ ol108/25mg * oi 08/50mg 20 - 0~~~~~~~~~ o1 08/1 00 mg 1 st week 5th week last week Figure 3 Left ventricular ejection fraction. Statistical signifi- cance: *(paired t-test, P <0.05) comparison with the initial values, (ANOVA, P <0.05) comparison between treatments: c, with control group; v, with vehicle group. the highest dose (100 mg/kg) it caused minor dis- turbances in animal activity, weight gain and plasma biochemistry. The selectivity of chelation was evi- denced by invariability of plasma levels of biologi- cally important ions (e.g., calcium). In intact (i.e., non-iron overloaded) animals, the majority of iron is present in haemoproteins (hemoglobin, myoglobin, etc.), transport protein (transferin) and storage pro- teins (ferritin), and hence is mostly inaccessible to the chelator. Thus, the absence of sideropenia is a positive observation and may be explained by the limited amount of iron, which is present in the chelatable pool in the intact animals. The determination of plasma enzymes (e.g., ALT, AST) as well as the histopathological examination of the liver did not reveal any abnormalities induced by the administration of the chelator. Changes in lipid metabolism were transient and should be interpreted as negligible. On the other hand, the scattered elevation of creatinaemia warrants further vigilance, despite the negative result of careful dP/dtmax (kPa.s-1) BP (mmHg) HR (min ') Control 1344.8 + 61.2 103.9+ 2.9 286.9 + 18.1 Vehicle 1152.6 + 88.7 99.2 +4.1 309.2 + 6.1 o-108/25 mg/kg 1457.1 + 37.6 95.2 + 7.3 320.5 + 6.5 o-108/50 mg/kg 1389.5 + 48.7 93.7 + 2.5 303.8 + 5.7 o-108/100 mg/kg 1357.8 + 123.5 87.4+4.6 312.8 + 2.9 dP/dtmax: maximal rate of the pressure rise in the isovolumic phase of the systole; BP: mean blood pressure in the right femoral artery; HR: heart rate. histological evaluation. At the level of light micro- scopic observation and with the use of basic histo- logical staining methods, it was not possible to distinguish clearly the changes either within the different dosage schedules of the chelator or in comparison with the control and vehicle groups. Mild changes occasionally found in the vehicle group might be associated with the previously described effects of Cremophor EL.2930 The pre- sence of very slight ultrastructural changes cannot be excluded at this time and may be the subject of further investigation. Somewhat higher cardiac tro- ponin T plasma concentrations in the highest dose of the chelator had no correlate in either myocardial histopathology or in functional cardiovascular para- meters. Although there were no marked gross changes in ECG, a specific telemetric study should be performed before the final statement. Nonethe- less, especially in high dose schedules of o-108, it might be relevant to monitor cardiac functions. In conclusion, the repeated 10-week administra- tion of the novel iron chelator pyridoxal o-chlor- obenzoyl hydrazone in three escalating doses (25, 50 and 100 mg/kg weekly) did not induce any - Figure 4 Myocardium of the left ventricular wall (a, control group; b, vehicle group; c, o-108/100 mg/kg group). Some cardiomyocytes revealed intensely eosinophilic cytoplasm (E) not only in the group treated with the vehicle (b) and o-108 (c) but also in control animals (a). As the number of those cells is low, the changes cannot be described as pathological. Scattered degenerated cells (D) were present in both tested groups (b, c). Since the number of those myocytes was low, no typical mononuclear infiltrate followed. C, capillary. Masson's blue trichrome. Bar 10 lim. Safety and tolerability of a novel iron chelator M Sterba et a. I Figure 5 Kidney (a, control group; b, vehicle group; c, o-108/100 mg group). Cortex with renal corpuscles (R); glomerular capillaries are marked by erythrocytes. Only proximal (P) and distal (D) tubules were slightly damaged in the group with o-108 (c), a little more in the vehicle group (b) compared with controls (a). Intensely eosinophilic cytoplasm appeared in some cells, mostly those of the proximal tubules in the group with 100 mg/kg o-108 (c); their nuclei became pyknotic (dark-stained) - arrows. Most of the proximal (P) and distal (D) tubules in the kidneys of rabbits in the vehicle group (b) showed higher eosinophilia and granulated cytoplasm, and also the number of pyknotic nuclei was rather higher (arrows). Masson's blue trichrome. Bar 10 gm. premature deaths and was generally well-tolerated in rabbits. No distinct abnormalities could be observed in most biochemical, haematological, his- topathological and functional parameters. 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</ref-list></back></article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Safety and tolerability of repeated administration of pyridoxal 2-chlorobenzoyl hydrazone in rabbits</title></titleInfo><titleInfo type="alternative" lang="en" contentType="CDATA"><title>Safety and tolerability of repeated administration of pyridoxal 2-chlorobenzoyl hydrazone in rabbits</title></titleInfo><name type="personal"><namePart type="given">M</namePart><namePart type="family">Sterba</namePart><affiliation>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králové, Czech Republic; Department of Pharmacology, Faculty of Medicine in Hradec Králové, Charles University in Prague, Simkova 870, 500 38 Hradec Králové 1, Czech Republic</affiliation><affiliation>E-mail: sterbam@lfhk.cuni.cz</affiliation></name><name type="personal"><namePart type="given">T</namePart><namePart type="family">Simunek</namePart><affiliation>Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Hradec Králová, Czech Republic</affiliation></name><name type="personal"><namePart type="given">Y</namePart><namePart type="family">Mazurová</namePart></name><name type="personal"><namePart type="given">M</namePart><namePart type="family">Adamcová</namePart></name><name type="personal"><namePart type="given">O</namePart><namePart type="family">Popelová</namePart></name><name type="personal"><namePart type="given">J</namePart><namePart type="family">Kaplanová</namePart><affiliation>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králová, Czech Republic</affiliation></name><name type="personal"><namePart type="given">P</namePart><namePart type="family">Ponka</namePart><affiliation>Lady Davis Institute for Medical Research, McGill University, Montreal, Canada</affiliation></name><name type="personal"><namePart type="given">V</namePart><namePart type="family">Gersl</namePart><affiliation>Faculty of Medicine in Hradec Králové, Charles University in Prague, Hradec Králové, Czech Republic</affiliation></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="research-article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</genre><originInfo><publisher>Sage Publications</publisher><place><placeTerm type="text">Sage CA: Thousand Oaks, CA</placeTerm></place><dateIssued encoding="w3cdtf">2005-11</dateIssued><copyrightDate encoding="w3cdtf">2005</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Recently, pyridoxal 2-chlorobenzoyl hydrazone (o-108) has been identified as an effective iron chelator [Link et al., Blood 2003; 101: 4172–79]. Since chronic treatment would be necessary in its potential indications, in the present study, the safety and tolerability of this agent after repeated administration was determined. Three doses of o-108 (25, 50, 100 mg/kg, in 10% Cremophor EL) were administered intraperitoneally, once weekly, for 10 weeks to three groups (n–5 each) of Chinchilla male rabbits. The effects on biochemical, haematological and cardiovascular parameters were examined during the experiment; histopathological examination was performed at the end of the experiment. Results were compared with control (saline 2 mL/kg, n–11) and vehicle groups (10% Cremophor EL, 2 mL/kg, n–12). No premature deaths occurred; the well-being of animals was evidenced by their body weight gain, although lower gain was observed with the highest dose (100 mg/kg). Significant elevations of cardiac troponin T plasma concentrations were observed with the highest dose of o-108, but no abnormalities were found in the cardiovascular function and only minor and inconsistent changes in haematological and biochemical parameters were observed. Histopathological examinations of selected organs revealed only weak and reversible changes through all studied groups. Thus, the data from this study suggest that o-108 remains a promising drug from the standpoint of the possibility of its repeated administration and warrants further investigation.</abstract><subject><genre>keywords</genre><topic>iron chelator</topic><topic>o-108</topic><topic>pyridoxal 2-chlorobenzoyl hydrazone</topic></subject><relatedItem type="host"><titleInfo><title>Human and Experimental Toxicology</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><identifier type="ISSN">0960-3271</identifier><identifier type="eISSN">1477-0903</identifier><identifier type="PublisherID">HET</identifier><identifier type="PublisherID-hwp">sphet</identifier><part><date>2005</date><detail type="volume"><caption>vol.</caption><number>24</number></detail><detail type="issue"><caption>no.</caption><number>11</number></detail><extent unit="pages"><start>581</start><end>589</end></extent></part></relatedItem><identifier type="istex">3AD3773662DBDF3BE7D698E92759B971E77D6C64</identifier><identifier type="ark">ark:/67375/M70-Z3MT1N5P-9</identifier><identifier type="DOI">10.1191/0960327105ht571oa</identifier><identifier type="ArticleID">10.1191_0960327105ht571oa</identifier><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-0J1N7DQT-B">sage</recordContentSource></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/M70-Z3MT1N5P-9/record.json</uri></json:item></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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