Novel Regioselective Synthesis of 1,3,4,5-Tetrasubstituted Pyrazoles and Biochemical Valuation on F<sub>1</sub>F<sub>O</sub>-ATPase and Mitochondrial Permeability Transition Pore Formation
An efficient, eco-compatible, and very cheap method for the construction of fully substituted pyrazoles (Pzs) via eliminative nitrilimine-alkene 1,3-dipolar cycloaddition (ENAC) reaction was developed in excellent yield and high regioselectivity. Enaminones and nitrilimines generated in situ were se...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-02-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/15/2/498 |
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| author | Vincenzo Algieri Cristina Algieri Paola Costanzo Giulia Fiorani Antonio Jiritano Fabrizio Olivito Matteo Antonio Tallarida Fabiana Trombetti Loredana Maiuolo Antonio De Nino Salvatore Nesci |
| author_facet | Vincenzo Algieri Cristina Algieri Paola Costanzo Giulia Fiorani Antonio Jiritano Fabrizio Olivito Matteo Antonio Tallarida Fabiana Trombetti Loredana Maiuolo Antonio De Nino Salvatore Nesci |
| author_sort | Vincenzo Algieri |
| collection | DOAJ |
| description | An efficient, eco-compatible, and very cheap method for the construction of fully substituted pyrazoles (Pzs) via eliminative nitrilimine-alkene 1,3-dipolar cycloaddition (ENAC) reaction was developed in excellent yield and high regioselectivity. Enaminones and nitrilimines generated in situ were selected as dipolarophiles and dipoles, respectively. A deep screening of the employed base, solvent, and temperature was carried out to optimize reaction conditions. Recycling tests of ionic liquid were performed, furnishing efficient performance until six cycles. Finally, a plausible mechanism of cycloaddition was proposed. Then, the effect of three different structures of Pzs was evaluated on the F<sub>1</sub>F<sub>O</sub>-ATPase activity and mitochondrial permeability transition pore (mPTP) opening. The Pz derivatives’ titration curves of <b>6a</b>, <b>6h</b>, and <b>6o</b> on the F<sub>1</sub>F<sub>O</sub>-ATPase showed a reduced activity of 86%, 35%, and 31%, respectively. Enzyme inhibition analysis depicted an uncompetitive mechanism with the typical formation of the tertiary complex enzyme-substrate-inhibitor (<i>ESI</i>). The dissociation constant of the <i>ESI</i> complex (<i>K</i><sub>i</sub>’) in the presence of the <b>6a</b> had a lower order of magnitude than other Pzs. The pyrazole core might set the specific mechanism of inhibition with the F<sub>1</sub>F<sub>O</sub>-ATPase, whereas specific functional groups of Pzs might modulate the binding affinity. The mPTP opening decreased in Pz-treated mitochondria and the Pzs’ inhibitory effect on the mPTP was concentration-dependent with <b>6a</b> and <b>6o</b>. Indeed, the mPTP was more efficiently blocked with 0.1 mM <b>6a</b> than with 1 mM <b>6a</b>. On the contrary, 1 mM <b>6o</b> had stronger desensitization of mPTP formation than 0.1 mM <b>6o</b>. The F<sub>1</sub>F<sub>O</sub>-ATPase is a target of Pzs blocking mPTP formation. |
| first_indexed | 2024-03-11T08:17:00Z |
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| id | doaj.art-7abf87217f564943adb97d40f357b610 |
| institution | Directory Open Access Journal |
| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-11T08:17:00Z |
| publishDate | 2023-02-01 |
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| spelling | doaj.art-7abf87217f564943adb97d40f357b6102023-11-16T22:40:46ZengMDPI AGPharmaceutics1999-49232023-02-0115249810.3390/pharmaceutics15020498Novel Regioselective Synthesis of 1,3,4,5-Tetrasubstituted Pyrazoles and Biochemical Valuation on F<sub>1</sub>F<sub>O</sub>-ATPase and Mitochondrial Permeability Transition Pore FormationVincenzo Algieri0Cristina Algieri1Paola Costanzo2Giulia Fiorani3Antonio Jiritano4Fabrizio Olivito5Matteo Antonio Tallarida6Fabiana Trombetti7Loredana Maiuolo8Antonio De Nino9Salvatore Nesci10Department of Chemistry and Chemical Technologies, University of Calabria, Via P. Bucci, Cubo 12C, 87036 Rende, CS, ItalyDepartment of Veterinary Medical Sciences, Mitochondrial Biochemistry Lab, Via Tolara di Sopra, 50, 40064 Ozzano Emilia, BO, ItalyDepartment of Chemistry and Chemical Technologies, University of Calabria, Via P. Bucci, Cubo 12C, 87036 Rende, CS, ItalyDepartment Molecular Sciences and Nanosystems, University Ca’ Foscari Venezia, Via Torino 155, 30172 Venezia Mestre, VE, ItalyDepartment of Chemistry and Chemical Technologies, University of Calabria, Via P. Bucci, Cubo 12C, 87036 Rende, CS, ItalyDepartment of Chemistry and Chemical Technologies, University of Calabria, Via P. Bucci, Cubo 12C, 87036 Rende, CS, ItalyDepartment of Chemistry and Chemical Technologies, University of Calabria, Via P. Bucci, Cubo 12C, 87036 Rende, CS, ItalyDepartment of Veterinary Medical Sciences, Mitochondrial Biochemistry Lab, Via Tolara di Sopra, 50, 40064 Ozzano Emilia, BO, ItalyDepartment of Chemistry and Chemical Technologies, University of Calabria, Via P. Bucci, Cubo 12C, 87036 Rende, CS, ItalyDepartment of Chemistry and Chemical Technologies, University of Calabria, Via P. Bucci, Cubo 12C, 87036 Rende, CS, ItalyDepartment of Veterinary Medical Sciences, Mitochondrial Biochemistry Lab, Via Tolara di Sopra, 50, 40064 Ozzano Emilia, BO, ItalyAn efficient, eco-compatible, and very cheap method for the construction of fully substituted pyrazoles (Pzs) via eliminative nitrilimine-alkene 1,3-dipolar cycloaddition (ENAC) reaction was developed in excellent yield and high regioselectivity. Enaminones and nitrilimines generated in situ were selected as dipolarophiles and dipoles, respectively. A deep screening of the employed base, solvent, and temperature was carried out to optimize reaction conditions. Recycling tests of ionic liquid were performed, furnishing efficient performance until six cycles. Finally, a plausible mechanism of cycloaddition was proposed. Then, the effect of three different structures of Pzs was evaluated on the F<sub>1</sub>F<sub>O</sub>-ATPase activity and mitochondrial permeability transition pore (mPTP) opening. The Pz derivatives’ titration curves of <b>6a</b>, <b>6h</b>, and <b>6o</b> on the F<sub>1</sub>F<sub>O</sub>-ATPase showed a reduced activity of 86%, 35%, and 31%, respectively. Enzyme inhibition analysis depicted an uncompetitive mechanism with the typical formation of the tertiary complex enzyme-substrate-inhibitor (<i>ESI</i>). The dissociation constant of the <i>ESI</i> complex (<i>K</i><sub>i</sub>’) in the presence of the <b>6a</b> had a lower order of magnitude than other Pzs. The pyrazole core might set the specific mechanism of inhibition with the F<sub>1</sub>F<sub>O</sub>-ATPase, whereas specific functional groups of Pzs might modulate the binding affinity. The mPTP opening decreased in Pz-treated mitochondria and the Pzs’ inhibitory effect on the mPTP was concentration-dependent with <b>6a</b> and <b>6o</b>. Indeed, the mPTP was more efficiently blocked with 0.1 mM <b>6a</b> than with 1 mM <b>6a</b>. On the contrary, 1 mM <b>6o</b> had stronger desensitization of mPTP formation than 0.1 mM <b>6o</b>. The F<sub>1</sub>F<sub>O</sub>-ATPase is a target of Pzs blocking mPTP formation.https://www.mdpi.com/1999-4923/15/2/498pyrazoles1,3-dipolar cycloadditionhydrazonyl chloridesmitochondrial permeability transition poreF<sub>1</sub>F<sub>O</sub>-ATPase |
| spellingShingle | Vincenzo Algieri Cristina Algieri Paola Costanzo Giulia Fiorani Antonio Jiritano Fabrizio Olivito Matteo Antonio Tallarida Fabiana Trombetti Loredana Maiuolo Antonio De Nino Salvatore Nesci Novel Regioselective Synthesis of 1,3,4,5-Tetrasubstituted Pyrazoles and Biochemical Valuation on F<sub>1</sub>F<sub>O</sub>-ATPase and Mitochondrial Permeability Transition Pore Formation Pharmaceutics pyrazoles 1,3-dipolar cycloaddition hydrazonyl chlorides mitochondrial permeability transition pore F<sub>1</sub>F<sub>O</sub>-ATPase |
| title | Novel Regioselective Synthesis of 1,3,4,5-Tetrasubstituted Pyrazoles and Biochemical Valuation on F<sub>1</sub>F<sub>O</sub>-ATPase and Mitochondrial Permeability Transition Pore Formation |
| title_full | Novel Regioselective Synthesis of 1,3,4,5-Tetrasubstituted Pyrazoles and Biochemical Valuation on F<sub>1</sub>F<sub>O</sub>-ATPase and Mitochondrial Permeability Transition Pore Formation |
| title_fullStr | Novel Regioselective Synthesis of 1,3,4,5-Tetrasubstituted Pyrazoles and Biochemical Valuation on F<sub>1</sub>F<sub>O</sub>-ATPase and Mitochondrial Permeability Transition Pore Formation |
| title_full_unstemmed | Novel Regioselective Synthesis of 1,3,4,5-Tetrasubstituted Pyrazoles and Biochemical Valuation on F<sub>1</sub>F<sub>O</sub>-ATPase and Mitochondrial Permeability Transition Pore Formation |
| title_short | Novel Regioselective Synthesis of 1,3,4,5-Tetrasubstituted Pyrazoles and Biochemical Valuation on F<sub>1</sub>F<sub>O</sub>-ATPase and Mitochondrial Permeability Transition Pore Formation |
| title_sort | novel regioselective synthesis of 1 3 4 5 tetrasubstituted pyrazoles and biochemical valuation on f sub 1 sub f sub o sub atpase and mitochondrial permeability transition pore formation |
| topic | pyrazoles 1,3-dipolar cycloaddition hydrazonyl chlorides mitochondrial permeability transition pore F<sub>1</sub>F<sub>O</sub>-ATPase |
| url | https://www.mdpi.com/1999-4923/15/2/498 |
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