Investigation of the inhibitory effects of isoindoline-1,3-dion derivatives on hCA-I and hCA-II enzyme activities

doi:10.1016/j.molstruc.2019.07.070

Journal of Molecular Structure

Volume 1197, 5 December 2019, Pages 386-392

https://doi.org/10.1016/j.molstruc.2019.07.070 Get rights and content

Highlights

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The therapeutic potential of isoindoline-1,3-diones were synthesized.
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Synthesized compounds were evaluated for their hCA I and hCA II enzymes inhibitory activities.
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Compounds 3a and 3c were found to be the most effective compound on hCA I and hCA II enzymes, respectively.
•
A good Absorption, Distribution, Metabolism and Excretion and Blood, Brain, Barier profile was predicted for all compounds.
•
Molecular docking studies were carried out for all compounds and acetazolamide (AAZ), the reference agent.

Abstract

Inhibition of carbonic anhydrase (CA) has emerged as a promising approach for the treatment of a variety of diseases such as glaucoma, epilepsy, obesity and cancer. As a result, the design of CA inhibitors (CAIs) is an outstanding field of medicinal chemistry. Due to the therapeutic potential of isoindoline-1,3-diones as CAIs, herein hCA I and hCA II isozymes were purified from human erythrocytes using affinity chromatography and the inhibitory effects of a series of isoindoline-1,3-diones on hydratase activities of these isozymes were investigated. Among these compounds, compound 3a was found to be the most effective compound on hCA I with an IC₅₀ value of 7.02 μM, whereas compound 3c was the most potent compound on hCA II with an IC₅₀ value of 6.39 μM. Moreover, molecular docking studies were carried out for all compounds and acetazolamide (AAZ), the reference agent, in the active sites of hCA I and hCA II. Generally, the compounds showed high affinity through salt bridge formation and metal coordination with Zn²⁺ ion and π-stacking interaction with His94 residue. According to in silico Absorption, Distribution, Metabolism and Excretion (ADME) studies, the pharmacokinetic parameters of all compounds were within the acceptable range.

Introduction

Carbonic anhydrases (CAs; EC 4.2.1.1) are ubiquitous zinc-containing metalloenzymes and they play a pivotal role in all living metabolism. The CA enzyme is present in 16 isoforms in mammals [1]. These isoforms are found to be specialized in different tissues of the organism. Although CAs are involved in many different reactions, the removal of waste carbon dioxide in the metabolism is their main task (Fig. 1) [2]. Due to their role in the conversion of carbon dioxide to bicarbonate, CAs participate in key biosynthetic reactions such as lipogenesis, gluconeogenesis and ureagenesis. Some CA isoenzymes contain zinc in their active regions, while others catalyze the reaction in the active site without zinc [3].

The regulation and control of enzyme activity is important for all living things. There are many regulations that increase the activity of enzymes, as well as decreasing regulations. In addition, enzyme activity is regulated by covalent modifications, phosphorylation and allosteric effects. One of the most important approaches for the reduction of this type of enzyme activity is the inhibition of CAs [4,5].

Over the past 60 years, heterocyclic and aromatic sulfonamides have been used as CA inhibitors in the treatment of many diseases such as glaucoma, obesity, epilepsy, cancer, altitude sickness. Moreover, in the last years CA inhibitors have emerged as anti-infective agents [6].

In addition to the sulfonamide derivatives such as AAZ, methazolamide, ethoxzolamide, aromatic/heteroaromatic sulfonamide-based Schiff bases have also been identified as CA inhibitors [6,7].

Isoindoline-1,3-dione is a prominent compound in organic synthesis for the preparation of diverse biologically active molecules [[8], [9], [10], [11], [12]]. Isoindoline-1,3-dione derivatives exhibit various biological activities, like anticancer [13], antimicrobial [14], antioxidant [15], anti-inflammatory [16] and analgesic [17] activities. The hydrophobic character of isoindoline-1,3-diones increases their potential to cross different biological membranes in vivo [18].

In this paper, we synthesized a series of N-substituted isoindoline-1,3-diones and investigated their inhibitory effects on hCA I and hCA II isoenzymes. All compounds were docked to the active sites of hCA I and hCA II with AAZ to explore their possible binding modes in the active sites of these enzymes. A computational study for the prediction of ADME properties of all compounds was also performed.

Section snippets

Chemistry

All chemicals were commercially available and were used without purification or after distillation and treatment with drying agents. Melting points (M.p.) were determined on a capillary melting apparatus (Buechi 530) and are uncorrected. IR spectra were obtained from solutions in 0.1 mm cells with a PerkinElmer spectrophotometer. The ¹H (400 MHz) and ¹³C NMR (100 MHz) spectra were recorded on Varian and Bruker spectrometers; δ in ppm. Elemental analyses were performed on a Leco CHNS-932

Chemistry

In this study, isoindoline-1,3-dione derivatives were obtained by the reaction of 1,3-dioxo-1,3-dihydroisobenzofuran-5-carboxylic acid (1) with methyl, ethyl, phenyl amine (2a, 2b, 2c), then converted to the methyl ester derivatives (3a, 3b, 3c) (Scheme 1). The structures of these compounds were approved on the basis of IR, ¹H NMR, ¹³C NMR spectral data and elemental analysis. All spectral data were in good agreement with their structures.

Biochemistry

hCA I and hCA II isoenzymes were purified in one step

Conclusion

In the present paper, 1,3-dioxoisoindoline derivatives were found to inhibit hCA I and hCA II at high levels. Accordingly, it has been found that 1,3-dioxoisoindoline derivatives have the potential to be used as hCA inhibitors for the development of drugs against various diseases. Furthermore, molecular docking studies showed that these compounds presented proper interactions with Zn²⁺ ion, His94 and Thr199 residues in the active sites of hCA I and hCA II. According to in silico ADME studies,

Acknowledge

All studies related to the kinetics and inhibition studies of hCA I and hCA II enzyme in this study was financially supported by a the project numbered FAD-2018-6321 from Research Development Center of Ataturk University, Turkey.

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Investigation of the inhibitory effects of isoindoline-1,3-dion derivatives on hCA-I and hCA-II enzyme activities

Highlights

Abstract

Introduction

Section snippets

Chemistry

Chemistry

Biochemistry

Conclusion

Acknowledge

Eur. J. Med. Chem.

J. Mol. Struct.

Bioorg. Med. Chem.

Molecules

Bioorg. Med. Chem.

Farmaco

Eur. J. Med. Chem.

J. Biol. Chem.

Purification and characterization of carbonic anhydrase from bovine erythrocyte plasma membrane

Prep. Biochem. Biotechnol.

Structure-based drug discovery of carbonic anhydrase inhibitors

J. Enzym. Inhib. Med. Chem.

Regulation of human carbonic anhydrase I (hCA I) activity by using a photochromic inhibitor

Angew. Chem. Int. Ed.