Ömer İrfan KÜFREVİOĞLU, Orhan ERDOĞAN, Saltuk Buğrahan CEYHUN, Ramazan DEMİRDAĞ, Deryanur ERDEM, Murat ŞENTÜRK, Emrah YERLİKAYA

Inhibition effects of some sulfonamides on α-carbonic anhydrase from european seabass (Dicentrarchus labrax)

Alfa-karbonik anhidraz (EC: 4.2.1.1; CA) levrek solungaç ve karaciğerden saflaştırıldı. Saflaştırma prosedürü homojenat hazırlama ve Sepharose 4B-tirozin-sülfanilamid afinite kramatografisinden oluştu. Bazı sülfonamidlerin enzim aktivitesi üzerine in vitro inhibisyon etkisi incelendi. Bu maddeler için Ki sabitleri ve IC50 değerleri Lineweaver-Burk grafikleri ve % aktivite-[I] grafikleri kullanılarak belirlendi. Sülfanilamid, mafenid, asetazolamid, 2-amino-1,3,5-tiyadiazol-5 sülfonamid için IC50 değerleri sırası ile solungaç karbonik anhidraz (GCA) için 980, 142, 20 ve 34 μM, karaciğer karbonik anhidraz (LCA) için 126, 23, 14 ve 2.58 μM, olarak belirlendi. Bazı sülfonamidlerin diğer CA izoenzimleri ile Ki sabitleri karşılaştırıldığında düşük mikromolar derişimlerde 0.21’den 76.0 μM’a kadar GCA ve LCA’ya karşı çok güçlü inhibitör aktiviteleri olduğu belirlendi.

Levrek (Dicentrarchus labrax) alfa-karbonik anhidraz üzerine bazı sülfonamidlerin inhibisyon etkisi

Alpha-carbonic anhydrase (EC: 4.2.1.1; CA) was purified from European seabass gill and liver. The purification procedure was composed of preparation of homogenate (or hemolysate) and affinity chromatography on Sepharose 4B-tyrosine-sulfanilamide. Some sulfonamides exhibited in vitro inhibitory effects on the enzyme activity. Ki constants and IC50 values for these drugs were determined by Lineweaver-Burk graphs and plotting activity % vs. [I], respectively. IC50 values of sulfanilamide, mafenide, acetazolamide, 2-amino-1,3,5-tiyadiazol-5 sulfonamide were 980 μM, 142 μM, 20 μM and 34 μM for gill carbonic anhydrase (GCA), 126 μM, 23 μM, 14 μM and 2.58 μM for liver carbonic anhydrase (LCA), respectively. Some sulfonamides exhibited much stronger inhibitory activity against GCA and LCA at low micromolar concentrations with the Ki values ranging from 0.21 to 76.0 μM as compared with other CAs.

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1. C.T. Supuran, Carbonic anhydrases: novel therapeutic applications for inhibitors and activators. Nat. Rev. Drug Discov., 7 (2008) 168.
2. M. Senturk, O.I. Kufrevioglu, In vitro effects of some nucleoside analogue drugs on enzyme activities of carbonic anhydrase isozymes I and II from human erythrocytes. Hacettepe J. Biol. Chem., 37 (2009) 289.
3. C.T. Supuran, A. Scozzafava, Carbonic anhydrase inhibitors and their therapeutic potential. Exp. Opin. Ther. Patents, 10 (2000) 575.
4. D. Hewett-Emmett, Evolution and distribution of the carbonic anhydrase gene families. In: W.R. Chegwidden, N.D. Carter, Y.H. Edwards (Eds). The Carbonic Anhydrases. New Horizons. Basel: Birkhauser Verlag, (2000) pp 29.
5. T. Stams, D.W. Christianson, X-ray crystallographic studies of mammalian carbonic anhydrase isozymes. In: W.R. Chegwidden, N.D. Carter, Y.H. Edwards, (Eds.), The Carbonic Anhydrases: New Horizons. Birkhauser Verlag, Boston (2000) pp. 159.
6. E. Bayram, M. Senturk, O.I. Kufrevioglu, C.T. Supuran, In vitro inhibition of salicylic acid derivatives on human cytosolic carbonic anhydrase isozymes I and II, Bioorg. Med. Chem, 16 (2008) 9101.
7. S.B. Ceyhun, M. Senturk, O. Erdogan, O.I. Kufrevioglu, In vitro and in vivo effects of some pesticides on carbonic anhydrase enzyme from rainbow trout (Oncorhynchus mykiss) gills. Pesticide Biochem. Physiol., 97 (2010) 177.
8. ARENA (Applied Research Ethics National Association) Institutional Animal Care and Use Committee Guidebook, second ed., Boston, (2002) pp. 121.
9. H. Soyut, S. Beydemir, Purification and Some Kinetic Properties of Carbonic Anhydrase from Rainbow Trout (Oncorhynchus mykiss) Liver and Metal Inhibition. Protein peptide lett. 15 (2008) 528.
10. S. Durdagi, M. Senturk, D. Ekinci, H.T. Balaydin, S. Goksu, O.I. Kufrevioglu, A. Innocenti, A. Scozzafava, C.T. Supuran, Kinetic and docking studies of phenolbased inhibitors of carbonic anhydrase isoforms I, II, IX and XII evidence a new binding mode within the enzyme active site. Bioorg. Med. Chem, 19 (2011) 1381.
11. D. Ekinci, S.B. Ceyhun, M. Senturk, D. Erdem, O.I. Kufrevioglu, C.T. Supuran, Characterization and anions inhibition studies of an α-carbonic anhydrase from the teleost fish Dicentrarchus labrax. Bioorg. Med. Chem, 19 (2011) 744.
12. M. Senturk, O. Talaz, D. Ekinci, H. Cavdar, O.I. Kufrevioglu, In vitro inhibition of human erythrocyte glutathione reductase by some new organic nitrates. Bioorg. Med. Chem. Lett, 19 (2009) 3661.
13. C.T. Supuran, A. Scozzafava, A. Casini, Carbonic Anhydrase Inhibitors. Med. Res. Rev, 23 (2003) 146.
14. K.M. Wilbur, N.G. Anderson, Electrometric and colorimetric determination of carbonic anhydrase J. Biol. Chem, 176 (1948) 147.
15. J.A. Verpoorte, S. Mehta, J.T. Edsall, Esterase activities of human carbonic anhydrase. J. Biol. Chem, 242 (1967) 4221.
16. H. Lineweaver, D. Burk, The determination of enzyme dissocation constants. J. Am. Chem. Soc, 57 (1934) 685.
17. M.M. Bradford, A rapid and sensitive method for the quantition of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem, 72 (1976) 248.
18. D.K. Laemmli, Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature, 227 (1970) 680.
19. D. Ekinci, S. Beydemir, Risk assessment of pesticides and fungicides for acid-base regulation and salt transport in rainbow trout tissues. Pestic. Biochem. Phys, 97 (2010) 66.
20. S. Dogan, The in vitro effects of some pesticides on carbonic anhydrase activity of Oncorhynchus mykiss and Cyprinus carpio carpio fish. J. Hazard. Mater, 132 (2006) 171.
21. I.J. Morgan, R.P. Henry, C.M. Wood, The mechanism of acute silver nitrate toxicity in freshwater rainbow trout (Oncorhynchus mykiss) is inhibition of gill Na+ and Cl– transport. Aquat. Toxicol, 38 (1997) 145.
22. Ekinci, D., Beydemir, S., Kufrevioglu, O.I., In vitro inhibitory effects of some heavy metals on human erythrocyte carbonic anhydrases. J. Enzyme Inhib. Med. Chem, 22 (2007) 745.
23. G.K. Bielmyer, K.V. Brix, M. Grosell, Is Cl- protection against silver toxicity due to chemical speciation? Aquat. Toxicol, 87 (2008) 81.