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. 2018 Dec;33(1):1181-1193.
doi: 10.1080/14756366.2018.1490732.

Synthesis and pharmacological evaluation of novel isoquinoline N-sulphonylhydrazones designed as ROCK inhibitors

Ramon Guerra de Oliveira  1   2 , Fabiana Sélos Guerra  2   3 , Cláudia Dos Santos Mermelstein  4 , Patrícia Dias Fernandes  2   3 , Isadora Tairinne de Sena Bastos  5 , Fanny Nascimento Costa  6 , Regina Cely Rodrigues Barroso  5 , Fabio Furlan Ferreira  6 , Carlos Alberto Manssour Fraga  1   2
Affiliations

Synthesis and pharmacological evaluation of novel isoquinoline N-sulphonylhydrazones designed as ROCK inhibitors

Ramon Guerra de Oliveira et al. J Enzyme Inhib Med Chem. 2018 Dec.

Abstract

In this study, we synthesized a new congener series of N-sulphonylhydrazones designed as candidate ROCK inhibitors using the molecular hybridization of the clinically approved drug fasudil (1) and the IKK-β inhibitor LASSBio-1524 (2). Among the synthesized compounds, the N-methylated derivative 11 (LASSBio-2065) showed the best inhibitory profile for both ROCK isoforms, with IC50 values of 3.1 and 3.8 µM for ROCK1 and ROCK2, respectively. Moreover, these compounds were also active in the scratch assay performed in human breast cancer MDA-MB 231 cells and did not display toxicity in MTT and LDH assays. Molecular modelling studies provided insights into the possible binding modes of these N-sulphonylhydrazones, which present a new molecular architecture capable of being optimized and developed as therapeutically useful ROCK inhibitors.

Keywords: LASSBio-1524; Molecular hybridization; N-sulphonylhydrazone; ROCK inhibitor; Rho kinase; fasudil.

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Figures

Figure 1.
Figure 1.
Design concept used to generate a novel class of N-sulphonylhydrazones as potential ROCK inhibitors.
Figure 2.
Figure 2.
Representation of ROCK (PDB code: 2ESM) with the co-crystallized isoquinoline inhibitor fasudil (1) (magenta) and best interaction mode of LASSBio-1524 (2) (green) obtained through docking studies.
Scheme 1.
Scheme 1.
Synthetic route exploited to prepare the N-sulphonylhydrazones (5a–h). a) NaHCO3 aqueous, dichloromethane; b) N2H4.H2O, dichloromethane, 0 °C, 4 h, 80%; c) EtOH, HCI (cat), r.t., 24 h, 70–90%.
Scheme 2.
Scheme 2.
Synthesis of compound 5h (LASSBio-2055) following the removal of N-Boc from derivative 6.
Figure 3.
Figure 3.
Unit cell representation of compound 5f (LASSBio-2024) along the a-axis.
Figure 4.
Figure 4.
Crystal structure of compound 5f (LASSBio-2024) showing the labels for all non-hydrogen atoms.
Scheme 3.
Scheme 3.
Synthetic route exploited to prepare the N-acylhydrazone derivative 10 (LASSBio-2064). a) KOH, I2, MeOH, 0 °C, 4h, 80%; b) N2H4.H2O, EtOH, reflux, overnight, 80%; c) EtOH, benzaldehyde, HCl (cat), overnight, 75%.
Figure 5.
Figure 5.
1H NMR shift of the amide proton of compound 10 (LASSBio-2064). (A) Experiment performed at 25 °C, where the duplication of the amide hydrogen was observed. (B) Experiment performed at 90 °C, where the coalescence of the signal was observed, indicating a conformational effect.
Scheme 4.
Scheme 4.
Synthesis of the N-methyl-N-sulphonylhydrazone derivative 11 (LASSBio-2065).
Figure 6.
Figure 6.
Predicted binding mode of compound 11 (LASSBio-2064) in complex with ROCK. Docking studies were performed using the program GOLD 5.4.1.
Figure 7.
Figure 7.
(A) Predicted binding mode of compound 5b (LASSBio-2020) in complex with ROCK. (B) Predicted binding mode of compound 11 (LASSBio-2065) in complex with ROCK. (C) Ligplot 2D representation of compound 5b (LASSBio-2020). (D) Ligplot 2D representation of compound 11 (LASBio-2065).
Figure 8.
Figure 8.
(A) Viability of MDA-MB-231 cells in the presence of compounds 5b (LASSBio-2020) and (B) 11 (LASSBio-2065).
Figure 9.
Figure 9.
Measurements the serum lactate dehydrogenase (LDH) levels in the culture medium of MDA-MB-231 cells treated with compounds 5b (LASSBio-2020) (A) and 11 (LASSBio- 2065) (B).
Figure 10.
Figure 10.
(A) Effects of compounds 5b (LASSBio-2020) and (B) 11 (LASSBio-2065) on the migration of MDA-MB 231 cells. Images were obtained using phase-contrast microscopy. Filled areas represented migrating cells were calculated using ImageJ software. Scale bars represent 50 μm.
Figure 11.
Figure 11.
Effects of compounds 5b (LASSBio-2020) and 11 (LASSBio-2065) on the migration of MDA-MB 231 cells. Filled areas representing migrating cells were calculated using ImageJ software. The results are presented as the medians ± standard deviations (n = 3) of independent experiments. Statistical analyses were performed using analysis of variance followed by the Newman-Keuls post-test. *p < 0.05 compared with the control group.
Figure 12.
Figure 12.
Chemical stability of compounds 5b (LASSBio-2020) and 11 (LASSBio-2065). (A) Recovery (%) of compounds at pH 2. (B) Recovery (%) of compounds at pH 7.4. The experiments were conducted in triplicate, and the values represent the averages from the experiments.
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References

    1. Riento K, Ridley AJ. Rocks: multifunctional kinases in cell behaviour. Nat Rev Mol Cell Biol 2003;4:446–56. - PubMed
    1. Julian L, Olson MF. Rho-associated coiled-coil containing kinases (ROCK): structure, regulation, and functions). Small GTPases 2014;5:e29846. - PMC - PubMed
    1. Feng Y, Lograsso PV, Defert O. Rho kinase (ROCK) inhibitors and their therapeutic potential. J Med Chem 2016;59:2269–300. - PubMed
    1. Mueller BK, Mack H, Teusch N. Rho kinase, a promising drug target for neurological disorders. Nat Rev Drug Discov 2005;4:387–98. - PubMed
    1. Glyn M. A Rho-associated kinase mitigates reperfusion-induced change in the shape of cardiac capillary endothelial cells in situ. Cardiovasc Res 2003;57:195–206. - PubMed

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