treatments-xml/data/68/75/87/68758797FFFD2A1DFCBEFAB44B42C2FE.xml

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<mods:title id="F15841AD6079A25340C265DE01C632AD">Metabolomic profiling of Zanthoxylum species: Identification of anticholinesterase alkaloids candidates</mods:title>
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<mods:namePart id="3D8A56C4C70063828F31ADFCE39C3B32">Plazas, Erika</mods:namePart>
<mods:affiliation id="F1327E2B11D17A9679BF0D27A2BEF881">∗ &amp; National University of Colombia, Chemistry Department, Cr 30 N ° 45 - 03, 111321, Bogotá, Colombia</mods:affiliation>
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<mods:namePart id="AFAF37D8931474FE5EC94EE3D7085ACB">Casoti, Rosana</mods:namePart>
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<mods:namePart id="0313E9C3553884D858C17F97F752F3D9">Murillo, Monica Avila</mods:namePart>
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<mods:namePart id="834F2D9ABC465A28E653EC3F11A1DE7A">Costa, Fernando Batista Da</mods:namePart>
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<mods:namePart id="4A72C5002C5502889E9589C0D3D2B7FC">Cuca, Luis Enrique</mods:namePart>
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2.4. Isolation of biomarkers from bark of 
<taxonomicName id="27DC4D02FFFD2A13FB22FAB44D3BC371" ID-CoL="5CWQH" authority="(J. F. Gmel.) Reynel" box="[1198,1402,1386,1405]" class="Magnoliopsida" family="Rutaceae" genus="Zanthoxylum" kingdom="Plantae" order="Sapindales" pageId="7" pageNumber="8" phylum="Tracheophyta" rank="species" species="schreberi">Zanthoxylum schreberi</taxonomicName>
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According to our OPLS-DA results and comparing the exact masses and fragmentation patterns, we concluded that the variables 419POS and 232POS could correspond to putative berberine and chelerythrine. Since, these alkaloids were previously reported as cholinesterase inhibitors (
<bibRefCitation id="844D4B70FFFD2A13FC06F9CC4C20C029" author="Brunhofer, G. &amp; Fallarero, A. &amp; Karlsson, D. &amp; Batista-Gonzalez, A. &amp; Shinde, P. &amp; Gopi, C. &amp; Vuorela, P." box="[906,1121,1554,1573]" pageId="7" pageNumber="8" pagination="6669 - 6679" refId="ref11198" refString="Brunhofer, G., Fallarero, A., Karlsson, D., Batista-Gonzalez, A., Shinde, P., Gopi, C., Vuorela, P., 2012. Exploration of natural compounds as sources of new bifunctional scaffolds targeting cholinesterase and beta amyloid aggregation: the case of chelerythrine. Bioorg. Med. Chem. 20 (22), 6669 - 6679. https: // doi. org / 10.1016 / j. bmc. 2012.09. 040." type="journal article" year="2012">Brunhofer et al., 2012</bibRefCitation>
; 
<bibRefCitation id="844D4B70FFFD2A13FBFCF9CC4D6AC029" author="Ahmed, T. &amp; Gilani, A. &amp; Abdollahi, M. &amp; Daglia, M. &amp; Nabavi, S." box="[1136,1323,1554,1573]" pageId="7" pageNumber="8" pagination="970 - 979" refId="ref11008" refString="Ahmed, T., Gilani, A., Abdollahi, M., Daglia, M., Nabavi, S., 2015. Berberine and neurodegeneration: a review of literature. Pharmacol. Rep. 67 (5), 970 - 979. https: // doi. org / 10.1016 / j. pharep. 2015.03.002." type="journal article" year="2015">Ahmed et al., 2015</bibRefCitation>
), we selected an extract with high occurrence of these compounds for isolation and further evaluation of their activity against both enzymes, which would allow us to validate our statistical model. Thus, from the bark of 
<taxonomicName id="27DC4D02FFFD2A13FA31F9B84BC0C098" class="Magnoliopsida" family="Rutaceae" genus="Zanthoxylum" kingdom="Plantae" order="Sapindales" pageId="7" pageNumber="8" phylum="Tracheophyta" rank="species" species="schreberi">
<emphasis id="D2A8EA93FFFD2A13FA31F9B84BC0C098" bold="true" italics="true" pageId="7" pageNumber="8">Z. schreberi</emphasis>
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by bio-guided three compounds were isolated, which presented positive reaction with Dragendorff and showed AChE inhibition in TLC bioautography. The isolated alkaloids were analyzed by LC-MS in the same conditions of the metabolomic profiling and as well as by 
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H-NMR. The alkaloid A2 (
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: 18.42 min), with exact mass 336.1229 and molecular formula C 
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H 
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NO 
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showed the same ions in the MS 
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spectrum as the variable 419POS (
<emphasis id="D2A8EA93FFFD2A13FBF4F8F74CDCC130" bold="true" box="[1144,1181,1833,1852]" italics="true" pageId="7" pageNumber="8">m/z</emphasis>
321.0940, 320.0920, 306.0724, 292.0968 and 278.078). In the 
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H-NMR, A2 showed characteristic signals of the protoberberine scaffold and by comparison with data from previous reports this compound was fully identified as berberine (
<bibRefCitation id="844D4B70FFFD2A13FCB6F8474B9EC1A0" author="Jeon, Y. W. &amp; Jung, J. W. &amp; Kang, M. &amp; Chung, I. K. &amp; Lee, W. T." box="[826,991,1944,1964]" pageId="7" pageNumber="8" pagination="391 - 394" refId="ref12592" refString="Jeon, Y. W., Jung, J. W., Kang, M., Chung, I. K., Lee, W. T., 2002. NMR studies on antitumor drug candidates, berberine and berberrubine. Bull. Korean Chem. Soc. 23 (3), 391 - 394. https: // doi. org / 10.5012 / bkcs. 2002.23.3.391." type="journal article" year="2002">Jeon et al., 2002</bibRefCitation>
). The alkaloid A3 (
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: 17.89 min), presented a 
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H-NMR profile similar to berberine, but without the signal in ?? 6.1 ppm (s, 2H) that corresponds to methylenedioxy moiety and three signals in ?? 4,20, 4,10 and 4,02 ppm (s, 3H, each one), which indicate the presence of three methoxy substituents. In the HRMS analysis, A3 showed an exact mass of 338.1387 with molecular formula C 
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H 
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NO 
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and the same fragmentation ions in MS 
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as the variable 123POS (
<tableCitation id="AD5E033AFFF22A1CFD21FB7B4AB5C2B4" box="[685,756,1189,1208]" captionStart="Table 2" captionStartId="3.[100,150,161,177]" captionTargetPageId="3" captionText="Table 2 Tentative identification of potential anti-cholinesterase alkaloids selected from S-plot applying a cut-off for correlation (p[corr]) and covariance (w*)." httpUri="http://table.plazi.org/id/B4A36609FFF92A17FFE8FF7F4D26C6C7" pageId="8" pageNumber="9" tableUuid="B4A36609FFF92A17FFE8FF7F4D26C6C7">Table 2</tableCitation>
). By comparing the spectral data, A3 was identified as columbamine. Likewise, the alkaloid A1 (
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: 18.66 min), with exact mass 348.1299 and molecular formula C 
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H 
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NO 
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, showed the same profile in the MS 
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spectrum as the variable 232POS, with ions 
<emphasis id="D2A8EA93FFF22A1CFDFCFACB4AD4C324" bold="true" box="[624,661,1301,1320]" italics="true" pageId="8" pageNumber="9">m/z</emphasis>
321.0940, 320.0920, 306.0724, 292.0968 and 278.0780, and based on the 
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H-NMR results, this compound was successfully identified as chelerythrine.
</paragraph>
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<emphasis id="D2A8EA93FFF22A1CFEBFFC29492DC204" bold="true" box="[307,364,1015,1032]" pageId="8" pageNumber="9">Fig. 6.</emphasis>
HRMS biomarker 232POS: (A) MS spectrum, (B) MS 
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spectrum and, (C) proposed fragmentation pathway.
</paragraph>
</caption>
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<emphasis id="D2A8EA93FFF22A1CFCBEFBE64B34C245" bold="true" box="[818,885,1080,1097]" pageId="8" pageNumber="9">Table 3</emphasis>
IC 
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values (μM) of biomarker alkaloids isolated from 
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<emphasis id="D2A8EA93FFF22A1CFAAAFB8C4DC9C26E" bold="true" box="[1318,1416,1105,1122]" italics="true" pageId="8" pageNumber="9">Z. schreberi</emphasis>
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against Acetylcholinesterase and Butyrylcholinesterase.
</paragraph>
</caption>
<paragraph id="E0633681FFF22A1CFCCEFB4D4DFEC324" pageId="8" pageNumber="9">
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<th id="1D3D5DBFFFF2D5EBFCCEFB4D4B89C2AF" box="[834,968,1171,1187]" gridcol="0" gridrow="0" pageId="8" pageNumber="9">Compound</th>
<th id="1D3D5DBFFFF2D5EBFBC3FB4D4C82C2AF" box="[1103,1219,1171,1187]" gridcol="1" gridrow="0" pageId="8" pageNumber="9">IC50 (μM)</th>
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<td id="1D3D5DBFFFF2D5EBFBC3FB1E4C82C2C2" box="[1103,1219,1216,1230]" gridcol="1" gridrow="1" pageId="8" pageNumber="9">
<emphasis id="D2A8EA93FFF22A1CFBC3FB1E4C21C2C2" bold="true" box="[1103,1120,1216,1230]" italics="true" pageId="8" pageNumber="9">Ee</emphasis>
AChE
</td>
<td id="1D3D5DBFFFF2D5EBFAC7FB1E4DFEC2C2" box="[1355,1471,1216,1230]" gridcol="2" gridrow="1" pageId="8" pageNumber="9">
<emphasis id="D2A8EA93FFF22A1CFAC7FB1E4D1CC2C2" bold="true" box="[1355,1373,1216,1230]" italics="true" pageId="8" pageNumber="9">Eq</emphasis>
BChE
</td>
</tr>
<tr id="5EEC34C3FFF2D5EBFCCEFB324DFEC2F7" box="[834,1471,1260,1275]" gridrow="2" pageId="8" pageNumber="9">
<th id="1D3D5DBFFFF2D5EBFCCEFB324B89C2F7" box="[834,968,1260,1275]" gridcol="0" gridrow="2" pageId="8" pageNumber="9">A1- chelerythrine</th>
<td id="1D3D5DBFFFF2D5EBFBC3FB324C82C2F7" box="[1103,1219,1260,1275]" gridcol="1" gridrow="2" pageId="8" pageNumber="9">1.028 ± 0.110</td>
<td id="1D3D5DBFFFF2D5EBFAC7FB324DFEC2F7" box="[1355,1471,1260,1275]" gridcol="2" gridrow="2" pageId="8" pageNumber="9">3.554 ± 0.177</td>
</tr>
<tr id="5EEC34C3FFF2D5EBFCCEFADD4DFEC31D" box="[834,1471,1283,1297]" gridrow="3" pageId="8" pageNumber="9">
<th id="1D3D5DBFFFF2D5EBFCCEFADD4B89C31D" box="[834,968,1283,1297]" gridcol="0" gridrow="3" pageId="8" pageNumber="9">A2- berberine</th>
<td id="1D3D5DBFFFF2D5EBFBC3FADD4C82C31D" box="[1103,1219,1283,1297]" gridcol="1" gridrow="3" pageId="8" pageNumber="9">0.107 ± 0.020</td>
<td id="1D3D5DBFFFF2D5EBFAC7FADD4DFEC31D" box="[1355,1471,1283,1297]" gridcol="2" gridrow="3" pageId="8" pageNumber="9">6.404 ± 0.293</td>
</tr>
<tr id="5EEC34C3FFF2D5EBFCCEFAC44DFEC324" box="[834,1471,1306,1320]" gridrow="4" pageId="8" pageNumber="9">
<th id="1D3D5DBFFFF2D5EBFCCEFAC44B89C324" box="[834,968,1306,1320]" gridcol="0" gridrow="4" pageId="8" pageNumber="9">A3- columbamine</th>
<td id="1D3D5DBFFFF2D5EBFBC3FAC44C82C324" box="[1103,1219,1306,1320]" gridcol="1" gridrow="4" pageId="8" pageNumber="9">3.752 ± 0.160</td>
<td id="1D3D5DBFFFF2D5EBFAC7FAC44DFEC324" box="[1355,1471,1306,1320]" gridcol="2" gridrow="4" pageId="8" pageNumber="9">2.048 ± 0.088</td>
</tr>
</table>
</paragraph>
<paragraph id="E0633681FFF22A1CFF09FA5A4905C02F" blockId="8.[100,771,1078,1601]" pageId="8" pageNumber="9">
Although berberine and chelerythrine have been previously reported as cholinesterase inhibitors, in the present work it was also determinate the dual anti-cholinesterase activity in order to support our statistical results. The inhibition curves were determined at specified substrate concentration in the presence of different concentrations of A1, A2 and A3 (
<figureCitation id="78E72A04FFF22A1CFE8CF9CE4977C02F" box="[256,310,1552,1571]" captionStart="Fig" captionStartId="8.[100,130,1940,1957]" captionTargetBox="[340,1247,1642,1916]" captionTargetId="figure-293@8.[339,1249,1641,1917]" captionTargetPageId="8" captionText="Fig. 7. Dose-response inhibition curves of the isolated alkaloids (A1, A2 and A3) against EeAChE and EqBChE. The IC50 values were calculated after fitting the curves using non-linear regression function of GraphPad Prism. The curves are shown s mean of three independent experiments performed in triplicates." figureDoi="http://doi.org/10.5281/zenodo.10484105" httpUri="https://zenodo.org/record/10484105/files/figure.png" pageId="8" pageNumber="9">Fig. 7</figureCitation>
).
</paragraph>
<paragraph id="E0633681FFF22A1CFF09F9F24C29C37E" blockId="8.[100,771,1078,1601]" lastBlockId="8.[818,1488,1352,1394]" pageId="8" pageNumber="9">
The IC 
<subScript id="7C5834C4FFF22A1CFF47F9EB489CC04D" attach="left" box="[203,221,1589,1601]" fontSize="5" pageId="8" pageNumber="9">50</subScript>
values were calculated after fitting the curves using The IC 
<subScript id="7C5834C4FFF22A1CFCE7FA904B3AC356" attach="left" box="[875,891,1358,1370]" fontSize="5" pageId="8" pageNumber="9">50</subScript>
values are shown as the mean ± SEM of three independent experiments each performed in triplicates.
</paragraph>
<paragraph id="E0633681FFF22A1DFCBEFA494952C435" blockId="8.[818,1487,1431,1590]" lastBlockId="9.[100,771,159,1266]" lastPageId="9" lastPageNumber="10" pageId="8" pageNumber="9">
nonlinear regression function. The values are shown as mean of three independent experiments each performed by triplicates (
<tableCitation id="AD5E033AFFF22A1CFAC3FA6D4DD7C3CA" box="[1359,1430,1459,1478]" captionStart="Table 3" captionStartId="8.[818,868,1080,1096]" captionTargetPageId="8" captionText="Table 3 IC50 values (μM) of biomarker alkaloids isolated from Z. schreberi against Acetylcholinesterase and Butyrylcholinesterase." httpUri="http://table.plazi.org/id/B4A36609FFF22A1CFCBEFBE64C89C270" pageId="8" pageNumber="9" tableUuid="B4A36609FFF22A1CFCBEFBE64C89C270">Table 3</tableCitation>
). Our results are in accordance with those reported in literature for these compounds. Berberine is one of the isoquinoline alkaloids with the highest inhibition activity against AChE reporting IC 
<subScript id="7C5834C4FFF22A1CFAA9F9CE4D76C010" attach="left" box="[1317,1335,1552,1564]" fontSize="5" pageId="8" pageNumber="9">50</subScript>
values between 0.01 and 2.00 μM. However, berberine has been shown to be less active against BChE, the IC 
<subScript id="7C5834C4FFF32A1DFEB5FF76490AC6B8" attach="left" box="[313,331,168,180]" fontSize="5" pageId="9" pageNumber="10">50</subScript>
values reported for horse-serum BChE are highest than 3 μM (
<bibRefCitation id="844D4B70FFF32A1DFE91FF6549FCC6C2" author="Jung, H. &amp; Min, B. &amp; Yokozawa, T. &amp; Lee, J. &amp; Kim, Y. &amp; Choi, J." box="[285,445,187,206]" pageId="9" pageNumber="10" pagination="1433 - 1438" refId="ref12728" refString="Jung, H., Min, B., Yokozawa, T., Lee, J., Kim, Y., Choi, J., 2009. Anti-Alzheimer and antioxidant activities of Coptidis Rhizoma alkaloids. Biol. Pharm. Bull. 32 (8), 1433 - 1438. https: // doi. org / 10.1248 / bpb. 32.1433." type="journal article" year="2009">Jung et al., 2009</bibRefCitation>
; 
<bibRefCitation id="844D4B70FFF32A1DFE46FF654AC3C6C2" author="Murray, A. P. &amp; Faraoni, M. B. &amp; Castro, M. J. &amp; Alza, N. P. &amp; Cavallaro, V." box="[458,642,187,206]" pageId="9" pageNumber="10" pagination="388 - 413" refId="ref13623" refString="Murray, A. P., Faraoni, M. B., Castro, M. J., Alza, N. P., Cavallaro, V., 2013. Natural AChE inhibitors from plants and their contribution to alzheimer's disease therapy. Curr. Neuropharmacol. 11 (4), 388 - 413. http: // doi. org / 10.2174 / 1570159 X 11311040004." type="journal article" year="2013">Murray et al., 2013</bibRefCitation>
). In contrast, columbamine, another protoberberine alkaloid, is less potent than berberine against AChE with reported IC 
<subScript id="7C5834C4FFF32A1DFE6CFF2249B3C704" attach="left" box="[480,498,252,264]" fontSize="5" pageId="9" pageNumber="10">50</subScript>
value of 5 μM (
<bibRefCitation id="844D4B70FFF32A1DFD0BFF2D48D5C72E" author="Tsai, S. F. &amp; Lee, S. S." pageId="9" pageNumber="10" pagination="1632 - 1635" refId="ref14820" refString="Tsai, S. F., Lee, S. S., 2010. Characterization of acetylcholinesterase inhibitory constituents from Annona glabra assisted by HPLC microfractionation. J. Nat. Prod. 73, 1632 - 1635. https: // doi. org / 10.1021 / np 100247 r." type="journal article" year="2010">Tsai and Lee, 2010</bibRefCitation>
) which has not previous reports against BChE inhibition. With regards to our results, columbamine shows comparable activity against both enzymes 
<emphasis id="D2A8EA93FFF32A1DFF61FE994943C756" bold="true" box="[237,258,327,346]" italics="true" pageId="9" pageNumber="10">Ee</emphasis>
AChE and 
<emphasis id="D2A8EA93FFF32A1DFEEAFE99493CC756" bold="true" box="[358,381,327,346]" italics="true" pageId="9" pageNumber="10">Eq</emphasis>
BChE with IC 
<subScript id="7C5834C4FFF32A1DFE70FE914A4FC757" attach="left" box="[508,526,335,347]" fontSize="5" pageId="9" pageNumber="10">50</subScript>
values of 3.752 ± 0.160 and 2.048 ± 0.088 respectively. These results are an important finding in the search to multi-target compounds considering the role played by BChE in AD physiopathology. Although, AChE is the main responsible for the hydrolysis of the acetylcholine neurotransmitter, recent evidence (
<bibRefCitation id="844D4B70FFF32A1DFF26FE0C490BC7E9" author="Lane, R. &amp; Potkin, S. &amp; Enz, A." box="[170,330,466,485]" pageId="9" pageNumber="10" pagination="101 - 124" refId="ref13204" refString="Lane, R., Potkin, S., Enz, A., 2006. Targeting acetylcholinesterase and butyrylcholinesterase in dementia. Int. J. Neuropsychopharmacol. 9, 101 - 124. https: // doi. org / 10.1017 / S 1461145705005833." type="journal article" year="2006">Lane et al., 2006</bibRefCitation>
), has shown that BChE is also involved in the cholinergic regulation. In fact, during the progress of AD the activity of AChE decreases, while that of BChE increases gradually to keep the catalytic pathway.
</paragraph>
<caption id="B4A36609FFF22A1CFFE8F84A4D07C1B2" ID-DOI="http://doi.org/10.5281/zenodo.10484105" ID-Zenodo-Dep="10484105" httpUri="https://zenodo.org/record/10484105/files/figure.png" pageId="8" pageNumber="9" startId="8.[100,130,1940,1957]" targetBox="[340,1247,1642,1916]" targetPageId="8" targetType="figure">
<paragraph id="E0633681FFF22A1CFFE8F84A4D07C1B2" blockId="8.[100,1487,1940,1982]" pageId="8" pageNumber="9">
<emphasis id="D2A8EA93FFF22A1CFFE8F84A48DCC1A8" bold="true" box="[100,157,1940,1957]" pageId="8" pageNumber="9">Fig. 7.</emphasis>
Dose-response inhibition curves of the isolated alkaloids (A1, A2 and A3) against 
<emphasis id="D2A8EA93FFF22A1CFCC2F84A4B20C1A8" bold="true" box="[846,865,1940,1956]" italics="true" pageId="8" pageNumber="9">Ee</emphasis>
AChE and 
<emphasis id="D2A8EA93FFF22A1CFC35F84A4B8FC1A8" bold="true" box="[953,974,1940,1956]" italics="true" pageId="8" pageNumber="9">Eq</emphasis>
BChE. The IC 
<subScript id="7C5834C4FFF22A1CFBB0F8454C0DC1AB" attach="left" box="[1084,1100,1947,1959]" fontSize="5" pageId="8" pageNumber="9">50</subScript>
values were calculated after fitting the curves using non-linear regression function of GraphPad Prism. The curves are shown s mean of three independent experiments performed in triplicates.
</paragraph>
</caption>
<paragraph id="E0633681FFF32A1DFF09FD9F497CC58B" blockId="9.[100,771,159,1266]" pageId="9" pageNumber="10">
Furthermore, chelerythrine has been reported as less active than berberine against both enzymes with IC 
<subScript id="7C5834C4FFF32A1DFE70FDB84A4FC47E" attach="left" box="[508,526,614,626]" fontSize="5" pageId="9" pageNumber="10">50</subScript>
values in the range of 1–10 μM, which is in concordance to our results (
<bibRefCitation id="844D4B70FFF32A1DFDBBFDA74AB5C480" author="Konrath, L. &amp; dos Santos, C. &amp; Klein, L. &amp; Henriques, A." box="[567,756,633,653]" pageId="9" pageNumber="10" pagination="1701 - 1725" refId="ref13087" refString="Konrath, L., dos Santos, C., Klein, L., Henriques, A., 2013. Alkaloids as a source of potential anticholinesterase inhibitors for the treatment of Alzheimer's disease. J. Pharm. Pharmacol. 65 (12), 1701 - 1725. https: // doi. org / 10.1111 / jphp. 12090." type="journal article" year="2013">Konrath et al., 2013</bibRefCitation>
). However, kinetic studies showed that chelerythrine presents a mixed inhibition mechanism against 
<emphasis id="D2A8EA93FFF32A1DFE00FD6F49E0C4C8" bold="true" box="[396,417,689,708]" italics="true" pageId="9" pageNumber="10">Ee</emphasis>
AChE. Moreover, molecular docking models suggest that this alkaloid binds both the catalytic (CAS) and peripheral anionic sites (PAS) of the TcAChE (
<taxonomicName id="27DC4D02FFF32A1DFDADFD364A8EC4F7" authorityName="Ayres" authorityYear="1855" box="[545,719,744,763]" class="Elasmobranchii" family="Torpedinidae" genus="Torpedo" kingdom="Animalia" order="Torpediniformes" pageId="9" pageNumber="10" phylum="Chordata" rank="species" species="californica">
<emphasis id="D2A8EA93FFF32A1DFDADFD364A8EC4F7" bold="true" box="[545,719,744,763]" italics="true" pageId="9" pageNumber="10">Torpedo californica</emphasis>
</taxonomicName>
) and this characteristic has been linked with the inhibition of Aβ fibrillogenesis induced by AChE (
<bibRefCitation id="844D4B70FFF32A1DFE03FCFF4A1CC538" author="Brunhofer, G. &amp; Fallarero, A. &amp; Karlsson, D. &amp; Batista-Gonzalez, A. &amp; Shinde, P. &amp; Gopi, C. &amp; Vuorela, P." box="[399,605,801,820]" pageId="9" pageNumber="10" pagination="6669 - 6679" refId="ref11198" refString="Brunhofer, G., Fallarero, A., Karlsson, D., Batista-Gonzalez, A., Shinde, P., Gopi, C., Vuorela, P., 2012. Exploration of natural compounds as sources of new bifunctional scaffolds targeting cholinesterase and beta amyloid aggregation: the case of chelerythrine. Bioorg. Med. Chem. 20 (22), 6669 - 6679. https: // doi. org / 10.1016 / j. bmc. 2012.09. 040." type="journal article" year="2012">Brunhofer et al., 2012</bibRefCitation>
). Therefore, if all the benzophenanthridine alkaloids present this characteristic binding mode, they would present some high multimodal potential as amyloid aggregation inhibitors.
</paragraph>
<paragraph id="E0633681FFF32A1DFF09FC4E4B42C2FE" blockId="9.[100,771,159,1266]" pageId="9" pageNumber="10">
Considering that chelerythrine (232POS), berberine (419POS) and columbamine (123POS) were recognized as potential cholinesterase inhibitors, in the S-plot, the isolation and identification of these alkaloids from the bark of 
<taxonomicName id="27DC4D02FFF32A1DFEB1FC3A49E4C5FB" box="[317,421,996,1015]" class="Magnoliopsida" family="Rutaceae" genus="Zanthoxylum" kingdom="Plantae" order="Sapindales" pageId="9" pageNumber="10" phylum="Tracheophyta" rank="species" species="schreberi">
<emphasis id="D2A8EA93FFF32A1DFEB1FC3A49E4C5FB" bold="true" box="[317,421,996,1015]" italics="true" pageId="9" pageNumber="10">Z. schreberi</emphasis>
</taxonomicName>
, as well as, their dual inhibitory activity allowed us to validate our statistical model. Thus, our metabolomic profiling allows a rapid correlation of the chemical composition and the anticholinesterase activity of extracts, therefore decreasing the time spent in the isolation procedure, including the isolation of inactive compounds. Moreover, predictive statistical models were built in order to uncover the anticholinesterase potential of non-tested extracts. Therefore, our results constitute an important contribution to the search of naturally occurring anticholinesterase alkaloids using a targeted screening strategy based on the chemical profile of crude extracts.
</paragraph>
</subSubSection>
</treatment>
</document>