Alzheimer Hastalığı Tedavisinde Kullanılan Rivastigmin ve Galantamin İlaç Etken Maddelerinin CA I ve II İzoenzimleri Üzerine in vitro Etkilerinin İncelenmesi

Öz

Karbonik anhidraz (CA, EC.4.2.1.1), CO₂’in, HCO₃^- ve H⁺ iyonlarına iki basamakta dönüşümlü hidratasyonunu katalizleyen, çinko (Zn²⁺) iyonlu metallo enzimlerden biridir. Fizyolojik ve patolojik birçok proseste önemli rol oynar. Bu çalışmanın amacı, Alzhaimer hastalığının tedavisinde kullanılan bazı ilaç etken maddelerinin insan eritrosit CA I ve II izoenzimleri üzerine in vitro etkilerini araştırmaktır. İnsan kanından CA-I ve II izoenzimleri Sefaroz-4B-L-tirozin-sülfamid afinite kromotografisi yöntemi ile saflaştırıldı. CA I izoenzimi: yaklaşık % 61.7 verimle 122.2 kat; CA II izoenzimi: % 60 verimle 807.5 kat saflaştırılmıştır. CA I için rivastigmin yarışmalı inhibisyon, galantamin ise yarışmasız inhibisyon gösterdi. CA II için ise her iki etken madde de yarışmasız inhibisyon gösterdi. K_i değerleri ise rivastigmin için CA I: 0.79 µM; CA II: 1.08 µM, galantamin için CA I: 0.41µM; CA II: 0.40 µM olarak hesaplandı. Elde edilen sonuçlara bakıldığında en kuvvetli inhibitör galantamin olarak tespit edildi.

Anahtar Kelimeler

• Rivastigmin,galantamin,inhibisyon,karbonik anhidraz

Investigation in vitro Effects of Rivastigmine and Galantamine Used to Treatment of Alzheimer's Disease on CA Isozymes I and II

Öz

The carbonic anhydrases (CA, EC. 4.2.1.1) are an expanding family of zinc-containing enzymes catalyzing the reversible hydration of CO₂ in a two-step reaction to yield HCO₃^-and H⁺. These enzymes play important roles in several physiological/pathological processes. The aim of this study is to evaluate in vitro the effects of these drug active substances which use which use for treatment of Alzheimer disease on CA I and II isoenzyme. CA I and II isoenzymes from human blood have been purified using Sepharose-4B-l-tyrosine-sulfanilamide affinity chromatography method. The CA I isoenzyme was purified ~122.2-fold with a yield of 61.7%. The CA II isoenzyme was purified ~807.5-fold with a yield of 60%. rivastigmine was showed competitive inhibition and galantamine was showed uncompetitive inhibition for CAI. Both of these drug active substances were showed uncompetitive inhibition for CAII. K_i values were determined for rivastigmine CA I: 0.79 µM; CA II: 1.08 µM and for galantamine CA I: 0.41µM; CA II: 0.40 µM. The results obtained in this study galantamine was identified as the most potent inhibitor.

Anahtar Kelimeler

• Rivastigmine,galantamine,inhibition,carbonic anhydrases

Kaynakça

1. Bartolucci, C., Perola, E., Pilger, C., Fels, G., Lamba, D. 2001. Three-dimensional structure of a complex of galanthamine (Nivalin) with acetylcholinesterase from Torpedo californica: Implications for the design of new anti-Alzheimer drugs. Proteins, 42, 182–191.
2. Bayram, E., Şentürk, M., Küfrevioğlu, Ö.İ. 2008. Supuran C.S. In vitro inhibition of salicylic acid derivatives on human cytosolic carbonic anhydrase isozymes I and II. Bioorganic and Medicinal Chemistry, 16, 9101-9105.
3. Beydemir, Ş., Çiftçi, M., Küfrevioğlu, Ö.İ., Büyükokuroğlu, M.E. 2002, Effects of gentamicine sulfate on enzyme activities of carbonic anhydrase from human erythrocytes in vitro and from rat erythrocytes in vivo. Biological and Pharmaceutical Bulletin, 25, 966-969.
4. Birks, J. 2006. Cholinesterase inhibitors for Alzheimer's disease. The Cochrane Database of Systematic Reviews, 1, CD005593.
5. Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of micro gram quantities of protein utilizing the principle of protein-dyebinding. Analytical Biochemistry, 72, 248-254.
6. Cankaya, M., Aktas, M., Kuzucu, M., Gülçin, I., Coban, T.A. 2012 Effects of some drugs on human cord blood erythrocyte carbonic anhydrases I and II: an in vitro study. Journal of Enzyme Inhibition and Medicinal Chemistry, 27(5), 641-645.
7. Ciftçi, M., Küfrevioglu, O.I., Gündogdu, M., Ozmen, I. 2000. Effects of some antibiotics on enzyme activity of glucose-6-phosphate dehydrogenase from human erythrocytes. Pharmacological Research, 41,109-113.
8. Coban, T.A., Beydemir, S., Gülcin, I., Gücin, I., Ekinci, D., Innocenti, A. 2009. Sildenafil is a strong activator of mammalian carbonic anhydrase isoforms I-XIV. Bioorganic and Medicinal Chemistry, 17, 5791-5795.

9. Colovic, M.B., Lazarevic-Pasti, T.D., Bondzic, A.M., Vasic, V.M. 2013. Acetylcholin esterase Inhibitors: Pharmacology and Toxicology. ‎Current Neuropharmacology, 11(3), 315–335.
10. Desai, A.K., Grossberg, G.T. 2005. Rivastigmine for Alzheimer's disease. Expert Review of Neurotherapeutics, 5, 563–580.
11. Dilek, E., Caglar, S. 2015. Effects of mono and dinuclear copper (II) complexes derived from non-steroidal anti-inflammatory drug naproxen on human serum paraoxanase1 (PON1) activity. International Journal of Chemistry and Pharmaceutical Sciences, 5(6), 189-195.
12. Dilek, E. Polat, M.F. 2016. In vitro ınhibition of three different drugs used ın rheumatoid arthritis treatment on human serum paraoxanase 1 enzyme activity. Protein and Peptide Letters, 23, 3-8.
13. Feldstein, J.B., Silverman, D.N. 1984. Purification and characterization of carbonic anhydrase from the saliva of therat. Journal of Biological Chemistry, 259: 5447-5453.
14. Gocer, H., Akincioglu, A., Goksu, S., Gulcin, I., Supuran. C.T. 2016. Carbonic anhydrase and acetyl cholin esterase inhibitory effects of carbamates and sulfamoyl carbamates. Journal of Enzyme Inhibition and Medicinal Chemistry, 30(2), 316–320.
15. Gocer, H., Topal, F., Topal, M., Kucuk, M., Teke, D., Gulcin, I., Alwase S.H., Supuran C.T. 2016. Acetyl cholin esterase and carbonic anhydrase isoenzymes I and II inhibition profiles of taxifolin. Journal of Enzyme Inhibition and Medicinal Chemistry, 31(3), 441–447.
16. Keha, E.E., Küfrevioğlu, Ö.İ. 2014, Biyokimya, Aktif Yayınevi.Kitisripanya, N., Saparpakorn, P., Wolschann, P., Hannongbua, S. 2011. Binding of huperzine A and galanthamine to acetylcholinesterase, based on ONIOM method. Nanomedicine: Nanotechnology, 7, 60–68.
17. Krungkrai, S.R., Suraveratum, N., Rochanakij, S., Krungkrai, J. 2001. Characterization of carbonic anhydrase in plasmodium falciparum. International Journal of Parasitology, 31, 661-668.
18. Laemmli, D.K. 1970. Cleavage of structural proteins during in assembly of the head of Bacteriophage T4. Nature 227, 680-685.
19. Lineweaver, H., Burk, D.J. 1934. The Determination of Enzyme Dissociation Constants. Journal of the American Chemical Society, 56, 658-66.
20. Nelson, D.L., Cox, M.M. 2004. Lehninger Biyokimyanın İlkeleri, 3.Baskı, Palme Yayıncılık, S. 259-261
21. Özgeris, B., Göksu, S., Köse, L.P., Gülçin, I., Salmas, R.E., Durdagi, S., Tümer, F., Supuran. C.T. 2016. Acetylcholin esterase and carbonic anhydrase inhibitory properties of novel urea and sulfamide derivatives in corporating dopaminergic 2-aminotetralin scaffolds. Bioorganic and Medicinal Chemistry, 24(10), 2318-29.
22. Pilger, C, Bartolucci, C., Lamba, D., Tropsha, A., Fels, G. 2001. Accurate prediction of the bound conformation of galanthamine in the active site of torpedo californica acetylcholinesterase using molecular docking. Journal of Molecular Graphics and Modelling, 19, 288–296. Sarıkaya, S.B.O., Topal, F., Şentürk, M., Gülçin, I., Supuran, C.T. 2001. In vitro inhibition of α-carbonic anhydrase isozymes by some phenolic compounds. Bioorganic and Medicinal Chemistry Letters, 5, 4259-4262.
23. Sarikaya, S.B.O., Sisecioglu, M., Cankaya, M., Gulcin, I., Ozdemir, H. 2014. Inhibition profile of a series of phenolic acids on bovine lactoperoxidase enzyme. Journal of Enzyme Inhibition and Medicinal Chemistry, 29, 1-5.
24. Scozzafava, A., Kalın, P., Supuran, C.T., Gülcin İ., Alwasel, S.H. 2015. The impact of hydroquinone on acetyl choline esterase and certain human carbonic anhydrase isoenzymes (hCA I, II, IX, and XII). Journal of Enzyme Inhibition and Medicinal Chemistry, 30(6), 941–946.
25. Telefoncu, A. 1986. Temel ve Uygulamalı Enzimoloji. Ege Üniversitesi, Fen-Edebiyat Fak. Yayını; S. 59, İzmir. Verpoorte, J.A., Mehta, S., Edsall, J.T. 1967. Esterase activities of human carbonic anhydrase. Journal of Biological Chemistry, 242, 4221-4230.