Screening of inhibitors of porcine dipeptidyl peptidase IV activity in aqueous extracts from marine organisms

Pascual, Isel; López, A. O.; Gómez, Hansel; Chappe, Mae; Saroyán, Angélika; González, Yoel Martínez; Cisneros, Miguel; Charli, Jean Louis; Chávez, María Á.

doi:10.1016/j.enzmictec.2006.07.012

Cited by 29 publications

(11 citation statements)

References 20 publications

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“…This extract also exhibited very poor values of SIA (specific inhibitory activity) and IC 50 . In a previous screening experiment against DPPIV, carried out with some of the marine invertebrate extracts analyzed in this study and others, it was observed that only three extracts showed inhibitory activity against the enzyme [46]. The study included the extract of B. granulifera with an IC 50 value in the same order (1.2 mg/mL) similar to that obtained in the current study, which also confirms a rather poor value.…”

Section: Discussionsupporting

confidence: 62%

Screening of Protease Inhibitory Activity in Aqueous Extracts of Marine Invertebrates from Cuban Coast

González¹,

Sánchez²,

Rojas³

et al. 2016

AJAC

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Protease inhibitors have been isolated from many variable sources; however, the need to identify and characterize new molecules has increased with the discovery of new therapeutic targets and the lack of specificity of already identified compounds with inhibitory activity. The goal of this work was to search for inhibitory activity against four proteolytic enzymes already recognized as therapeutic targets: human neutrophil elastase, dipeptidyl peptidase IV, subtilisin from Bacillus licheniformis and cathepsin K in selected marine invertebrates from the Caribbean Sea. A systematic screening was carried out with selected aqueous extracts belonging to 20 species from seven different phyla: Annelida, Bryozoa, Chordata, Cnidaria, Equinodermata, Mollusca and Porifera, all collected at the coast of Havana (Cuba). All extracts showing initial inhibitory activity were characterized in terms of IC50 values and specific inhibitory activity (SIA). Model enzymes were used in the case of human neutrophil elastase (porcine pancreatic elastase) and cathepsin K (papain) for the screening and all positive results were confirmed by testing toward the therapeutic targets. Ten extracts were identified showing inhibitory activity against human neutrophil elastase, for which the most promising values were obtained for Nerita peloronta. Only one extract, Bunodosoma granulifera, showed inhibitory activity against dipeptidyl peptidase IV with rather poor values of IC50 and SIA. Seven extracts showed inhibitory activity against B. licheniformis subtilisin with very good IC50 and SIA values for Lissodendorix isodyctialis, Cenchritis muricatus, and N. peloronta. Finally, eight extracts were positive for cathepsin K with almost similar parameters values among them. All these results confirmed the richness and potential of the marine invertebrate's fauna and indicated new promising sources for the identification of natural compounds with potential application in therapeutics.

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Section: Discussionsupporting

confidence: 62%

Screening of Protease Inhibitory Activity in Aqueous Extracts of Marine Invertebrates from Cuban Coast

González¹,

Sánchez²,

Rojas³

et al. 2016

AJAC

Self Cite

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“…Although IC 50 values are not the best parameters for evaluating the strength of the protease-inhibitor interaction, it is frequently used in the screening of protease inhibitors [44,45]. Furthermore, this strategy is very useful for the comparison of two or more inhibitors, in the interaction with their target proteases.…”

Section: Accepted Manuscriptmentioning

confidence: 99%

Heterologous expression of Cenchritis muricatus protease inhibitor II (CmPI-II) in Pichia pastoris system: Purification, isotopic labeling and preliminary characterization

Cabrera-Muñoz

Rojas

Gil

et al. 2016

Protein Expression and Purification

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Cenchritis muricatus protease inhibitor II (CmPI-II) is a tight-binding serine protease inhibitor of the Kazal family with an atypical broad specificity, being active against several proteases such as bovine pancreatic trypsin, human neutrophil elastase and subtilisin A. CmPI-II 3D structures are necessary for understanding the molecular basis of its activity. In the present work, we describe an efficient and straightforward recombinant expression strategy, as well as a cost-effective procedure for isotope labeling for NMR structure determination purposes. The vector pCM101 containing the CmPI-II gene, under the control of Pichia pastoris AOX1 promoter was constructed. Methylotrophic Pichia pastoris strain KM71H was then transformed with the plasmid and the recombinant protein (rCmPI-II) was expressed in benchtop fermenter in unlabeled or (15)N-labeled forms using ammonium chloride ((15)N, 99%) as the sole nitrogen source. Protein purification was accomplished by sequential cation exchange chromatography in STREAMLINE DirectHST, anion exchange chromatography on Hitrap Q-Sepharose FF and gel filtration on Superdex 75 10/30, yielding high quantities of pure rCmPI-II and (15)N rCmPI-II. Recombinant proteins displayed similar functional features as compared to the natural inhibitor and NMR spectra indicated folded and homogeneously labeled samples, suitable for further studies of structure and protease-inhibitor interactions.

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“…The activity was measured as described, 13,14) using the substrate Gly-Pro-p-nitroanilide, which can be decomposed to Gly-Pro and p-nitroaniline by DPP-4. Compounds H1-H45 were dissolved in 1% DMSO aqueous solution and were tested at a fixed level of 10 µg/mL.…”

Section: Resultsmentioning

confidence: 99%

Synthesis and Biological Evaluation of Xanthine Derivatives on Dipeptidyl Peptidase 4

Lin

Cai

Wang

et al. 2013

CHEMICAL & PHARMACEUTICAL BULLETIN

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A series of xanthine derivatives in which a methylene was inserted at position 8 of xanthine scaffold was synthesized and evaluated as inhibitors of dipeptidyl peptidase 4 (DPP-4) for the treatment of type 2 diabetes. As the results of structure-activity relationship (SAR) study of the series, the compounds with 4-methyl-quinazoline-2-yl-methyl group at N-1 position and 2-aminoethylaminomethyl group gave better activities. Compounds H4 and H9 showed good DPP-4 inhibition and more than 100-fold selectivity over DPP-7 and DPP-8.

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Screening of inhibitors of porcine dipeptidyl peptidase IV activity in aqueous extracts from marine organisms

Cited by 29 publications

References 20 publications

Screening of Protease Inhibitory Activity in Aqueous Extracts of Marine Invertebrates from Cuban Coast

Screening of Protease Inhibitory Activity in Aqueous Extracts of Marine Invertebrates from Cuban Coast

Heterologous expression of Cenchritis muricatus protease inhibitor II (CmPI-II) in Pichia pastoris system: Purification, isotopic labeling and preliminary characterization

Synthesis and Biological Evaluation of Xanthine Derivatives on Dipeptidyl Peptidase 4

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