Research paper
Structure-activity relationship with pyrazoline-based aromatic sulfamates as carbonic anhydrase isoforms I, II, IX and XII inhibitors: Synthesis and biological evaluation

https://doi.org/10.1016/j.ejmech.2019.111638Get rights and content

Highlights

  • Pyrazoline aryl sulfamates were designed and synthesized.

  • Sulfamates were assayed in vitro as human Carbonic Anhydrase (CA) isoforms inhibitors.

  • Synthesized compound inhibited selectively CAII, CAII, CAIX and CAXII isoforms.

  • Sulfamates 8h, 9e, 9i, 17d, 18e show CAIX and CAXII Ki in the nanomolar to sub-nanomolar range.

  • Docking revealed CA isoform selective inhibition related to distinguish interactions.

Abstract

Four new series of aromatic sulfamates were synthesized and investigated for the inhibition of four human (h) isoforms of zinc enzyme carbonic anhydrase (CA, EC 4.2.1.1), hCA I, II, IX, and XII. The reported derivatives, obtained by a sulfamoylation reaction of the corresponding phenolic precursors, bear 3,5-diarylpyrazoline moieties as spacers between the benzenesulfamate fragment which binds the zinc ion from the active site, and the tail of the inhibitor. Pyrazolines are biologically privileged scaffolds, endowed with versatile biological activity, such as an anti-proliferative action. The derivatives were tested for the inhibition of the cytosolic, hCA I and II (off target isoforms) and the trans-membrane, tumor-associated hCA IX and XII enzymes (anticancer drug targets). Generally, hCA I was not effectively inhibited, whereas many low nanomolar inhibitors were evidenced against hCA II (KIs in the range of 0.42–90.1 nM), IX (KIs in the range of 0.72–63.6 nM), and XII (KIs in the range of 0.88–85.2 nM). The best substitution fragments at the pyrazoline ring included for CA II a 4-sulfamic group on the 3-aryl and halogens on the 5-aryl or a methoxy group on the 3-aryl and a 4-sulfamate group on the 5-aryl; for CA IX and CA XII they included the sulfamic group on the 3- or 4-position of the 5-aryl and an electronwithdrawing group on the 4-postion of the 3-aryl ring.

Introduction

Pyrazoline ring is a privileged structure in medicinal chemistry because of its wide spectrum of pharmacological activities. A number of pyrazoline derivatives have been reported for their antibacterial [1], antimalarial [2,3], anti-inflammatory [4], MAO inhibitory [5,6], antioxidant [7,8], neuroprotective [9], antidepressant [10] and anticancer activity [[11], [12], [13], [14], [15], [16], [17], [18]]. Furthermore, several series of pyrazoline derivatives displayed carbonic anhydrase (CA, EC 4.2.1.1) inhibitory activity [[19], [20], [21], [22], [23], [24], [25]]. CAs are a superfamily of metalloenzymes that catalyze the CO2 hydration/dehydration reaction, and are classified into seven genetically distinct families, named α-, β-, γ-, δ-, ζ-, η-, and ɵ-CAs [[26], [27], [28]]. All human (h) CAs are α-class enzymes [28]. Fifteen different human isoforms have been identified and characterized to date, among which twelve are catalytically active (hCAs I-IV, VA, VB, VI, VII, IX, XII-XIV). Human CAs can be further categorized into four different subsets depending on their subcellular localization. Among those identified, hCA I, II, III, VII, VIII, X, XI, XIII are cytosolic proteins, hCA VA and VB are present in the mitochondrial matrix, hCA VI is a secreted enzyme, hCA IV is a glycosylphosphatidylinositol (GPI)-anchored protein and hCA IX, XII and XIV are trans-membrane isoforms [26,28]. These enzymes are widely distributed in many tissues and organs where they are implicated in a wealth of pivotal physiological processes. Dysregulated expression and/or abnormal activity of hCAs can result into severe pathological conditions [26,[29], [30], [31], [32]]. hCA IX and XII have been validated as markers of disease progression in many hypoxic tumors and their targeted inhibition has been associated with a significant reduction of the growth of both primary tumors and metastases [26]. In contrast, ubiquitous isoforms hCA I and II are the main off-target isoforms because their promiscuous inhibition might lead to undesired side effects [26]. CA inhibitors (CAIs) can be clustered into several different groups considering their binding mode to the enzyme active site, among which the zinc-binders are the most effective and thus most investigated for drug-design purposes [33]. Within this subset, sulfonamides are the ideal zinc-binding group (ZBG) owing to a peculiar combination of interactions that this moiety can solely establish with the zinc ion and the residues nearby [33]. Pyrazoline sulfonamides were recently reported as CA inhibitors [34] and as dual CA and AchE inhibitors [[35], [36], [37], [38], [39]]. Bioisosteric replacement of the sulfonamide with the sulfamate group, one of sulfonamide most related congeners, produced interesting examples of CAs selective inhibitors [[40], [41], [42], [43]]. These considerations led us to report a small library of pyrazoline sulfamates displaying interesting profile and selectivity as CA inhibitors [44]. Here, we have extended these studies to have full SAR of this class of compounds.

Section snippets

Chemistry

The synthetic pathway to obtain N1-acetyl-3,5-diaryl-4,5-dihydropyrazole bearing a sulfamate group at 4- or 3-position on 3-aryl ring (compounds 8a-m and 9a-m, Scheme 1) and on 5-aryl ring (compounds 17a-h and 18 a-h, Scheme 2) started with the preparation of chalcones (4, 5, 14 and 15) through the Claisen-Schmidt condensation [[45], [46], [47]] between substituted acetophenone (1, 2 and 10) and substituted benzaldehydes (3, 11 and 12) in methanol in the presence 50% aqueous NaOH. Chalcones

Conclusions

Four new series of aromatic sulfamates were synthesized by a sulfamoylation reaction of the corresponding phenolic precursors. The reported derivatives bearing 3,5-diarylpyrazoline moieties as spacers between the benzenesulfamate fragment which binds the zinc ion from the active site, and the tail of the inhibitor were investigated for the inhibition of four human hCAs, namely the cytosolic hCA I and II (off target isoforms) and the trans-membrane, tumor-associated hCA IX and XII enzymes

General methods

All commercially available solvents and reagents were used without further purification. NMR spectra were recorded on an Inova 500 spectrometer (Varian, Palo Alto, CA, USA). The chemical shifts (δ) are reported in part per million downfield from tetramethylsilane (TMS), which was used as internal standard. The spectra were recorded in hexadeuteriodimethylsulphoxide (DMSO‑d6). Infrared spectra were recorded on a Vector 22 spectrometer (Bruker, Bremen, Germany) in Nujol mulls. The main bands are

Acknowledgements

This study was supported in part by FIR grants from the University of Cagliari, Italy (to V.O. and G.B.).

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