N-(3-{4-[3-(trifluoromethyl)phenyl]piperazin-1-yl}propyl)-1H-indazole-3-carboxamide (D2AAK3) as a potential antipsychotic: In vitro, in silico and in vivo evaluation of a multi-target ligand

Kaczor, Agnieszka A.; Targowska-Duda, Katarzyna M.; Stępnicki, Piotr; Silva, Andrea G.; Koszła, Oliwia; Kędzierska, Ewa; Grudzińska, Angelika; Kruk-Słomka, Marta; Biała, Grażyna; Castro Pérez, María de los Ángeles

doi:10.1016/j.neuint.2021.105016

N-(3-{4-[3-(trifluoromethyl)phenyl]piperazin-1-yl}propyl)-1H-indazole-3-carboxamide (D2AAK3) as a potential antipsychotic: In vitro, in silico and in vivo evaluation of a multi-target ligand

Kaczor, Agnieszka A.; Targowska-Duda, Katarzyna M.; Stępnicki, Piotr; Silva, Andrea G.; Koszła, Oliwia; Kędzierska, Ewa; Grudzińska, Angelika; Kruk-Słomka, Marta; Biała, Grażyna; Castro Pérez, María de los Ángeles

Published: 2021-03-01
Publisher: Elsevier Ltd.
DOI: 10.1016/j.neuint.2021.105016
Repository: minerva.usc.es
Language: en

Abstract

Schizophrenia is a mental illness of not adequately understood causes that is not satisfactorily enough treated by current antipsychotics. In search for novel potential antipsychotics we performed structure-based virtual screening aimed to identify new dopamine D2 receptor antagonists. We found compound D2AAK3 with affinity to dopamine D2 receptor of 115 nM. D2AAK3 possesses additional nanomolar or low micromolar affinity to D1, D3, 5-HT1A, 5-HT2A and 5-HT7 receptors, which makes it a good hit for further development as a multifunctional ligand. The compound has also some affinity to M1 and H1 receptors. We used homology modeling, molecular docking and molecular dynamics to study interactions of D2AAK3 with its molecular targets at the molecular level. In behavioral studies D2AAK3 decreases amphetamine-induced hyperactivity (when compared to the amphetamine-treated group) measured as spontaneous locomotor activity in mice. In addition, passive avoidance test demonstrated that D2AAK3 improves memory consolidation after acute treatment in mice. Elevated plus maze tests indicated that D2AAK3 induces anxiogenic activity 30 min after acute treatment, whereas this effect has no longer been observed 60 min after administration of the studied compound in mice.

Full text

Supplementary material N-(3-{4-[3-(trifluoromethyl)phenyl]piperazin-1-yl}propyl)-1H-indazole-3-carboxamide (D2AAK3) as a potential antipsychotic: in vitro, in silico and in vivo evaluation of a multi-target ligand Agnieszka A. Kaczora,b*, Katarzyna M. Targowska-Dudac, Piotr Stępnickia, Andrea G. Silvad, Oliwia Koszłaa, Ewa Kędzierskae, Angelika Grudzińskaa, Marta Kruk-Słomkae, Grażyna Białae, Marián Castrod a Department of Synthesis and Chemical Technology of Pharmaceutical Substances with Computer Modeling Laboratory, Faculty of Pharmacy, Medical University of Lublin, 4A Chodźki St., PL-20093 Lublin, Poland b School of Pharmacy, University of Eastern Finland, P.O. Box 1627, FI-70211 Kuopio, Finland c Department of Biopharmacy, Faculty of Pharmacy, Medical University of Lublin, 4A Chodźki St., PL-20093 Lublin, Poland d Department of Pharmacology, Universidade de Santiago de Compostela, Center for Research in Molecular Medicine and Chronic Diseases (CIMUS), Avda de Barcelona, E15782 Santiago de Compostela, Spain e Department of Pharmacology and Pharmacodynamics, Faculty of Pharmacy, Medical University of Lublin, 4A Chodźki St., PL-20093 Lublin, Poland; * Correspondence: [email protected]; Tel.: +48 81 448 7273 Figures S1-S4 Fig. S1. Competition binding curves of D2AAK3 in radioligand binding assays at the indicated receptors. Concentration-dependent displacement of the specific radioligand binding by compounds D2AAK3 and corresponding reference compound at the human cloned receptors indicated. The graphs show the data (media ± SEM) of a representative experiment out of 2-3 independent experiments performed in duplicate. D2 receptor -10 -9 -8 -7 -6 -5 -4 0 25 50 75 100 125 Haloperidol D2AAK3 log [compound], M % [ 3H]-Spiperone specific binding D1 receptor -10 -9 -8 -7 -6 -5 -4 0 25 50 75 100 125 D2AAK3 Haloperidol log [compound], M % [ 3H]-SCH-23390 specific binding D3 receptor -10 -9 -8 -7 -6 -5 -4 0 25 50 75 100 125 D2AAK3 Haloperidol log [compound], M % [ 3H]-Spiperone specific binding 5-HT 1A receptor -11 -10 -9 -8 -7 -6 -5 -4 0 25 50 75 100 125 D2AAK3 5-CT log [compound], M % [ 3H]-8-OH-DPAT specific binding 5-HT 2A receptor -11 -10 -9 -8 -7 -6 -5 -4 0 25 50 75 100 125 D2AAK3 Methysergide log [compound], M % [ 3H]-Ketanserin specific binding 5-HT 7 receptor -10 -9 -8 -7 -6 -5 -4 0 25 50 75 100 125 D2AAK3 C loza pine log [compound], M % [ 3H]-SB269970 specific binding H1 receptor -11 -10 -9 -8 -7 -6 -5 -4 0 25 50 75 100 125 D2AAK3 Doxepin log [compound], M % [ 3H]-Pyrilamine specific binding M1 receptor -11 -10 -9 -8 -7 -6 -5 -4 0 25 50 75 100 125 Ipratropium D2AAK3 log [compound], M % [ 3H]-Pirenzepine specific binding Fig. S2. Ligand RMSD values during 200 ns molecular dynamics simulations for D2AAK3 in complex with dopamine D1 (A), D2 (B), D3 (C) and serotonin 5-HT1A (D), 5-HT2A (E) and 5-HT7 (F) receptors. Fig. S3. Molecular interactions of D2AAK3 with human dopamine D1 (A), D2 (B) and D3 (C) receptor during 200 ns molecular dynamics simulations: summary of contacts. Interactions that occur more than 30% of the simulation time in the selected trajectory (0 through 200 ns) are shown. Fig. S4. Molecular interactions of D2AAK3with human serotonin 5-HT1A (A), 5-HT2A (B) and 5-HT7 (C) receptor during 200 ns molecular dynamics simulations: summary of contacts. Interactions that occur more than 30% of the simulation time in the selected trajectory (0 through 200 ns) are shown.