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Comparison of two structurally related thienothiopyran-2sulfonamides as carbonic anhydrase II inhibitors

Arunima, Kushant & Zain

Carbonic anhydrase - Introduction

Zinc-metalloenzymes involved in catalysing the hydration of CO2 CO2 + H2O HCO3- + H+ 7 isoenzymes found in human tissues CA II expressed in erythrocytes and in the eye CA II contains a single polypeptide chain containing 259 amino acids and 1 essential zinc ion bound by 3 histidine residues.

Carbonic anhydrase II in the eye

Plays an important role in maintaining Intraocular Pressure (IOP) Increased Intraocular pressure leads to Glaucoma
Glaucoma :

Caused by trabecular blockage Can be treated using carbonic anhydrase inhibitors

Treating Glaucoma
Inhibition of Carbonic anhydrase II (in the ciliary process)

Slows down HCO3- (bicarbonate) production

Reduction in sodium and fluid transport

Reduction in aqueous humor secretion

Lowering of IOP

Carbonic anhydrase inhibitors

Acetazolamide Brinzolamide Methazolamide
ETS (4-ethylamino-5,6-dihydro-6-methyl-4Hthienothiopyran-2-sulfonamide or Dorzolamide) PTS (4-amino-5,6-dihydro-6-methyl-4Hthienothiopyran-2-sulfonamide)

Development of thienothiopyran-2sulfonamides
Dorzolamide was developed from a structure-based design. Structure-based drug design: Involves knowledge of the drug target to guide drug discovery Structurally ETS and PTS only vary in the length of the substituent on the 4-amino group. ETS has an ethyl group whereas PTS has a proton.

Mechanism of inhibitor action on CAII

Inhibitors target the active site of the enzyme. His-64 side chain observed in the native position for the PTS inhibited form of CA II. His-64 side chain forced in an alternate position by the size of the 4-amino ethyl group of ETS with displacement of the adjacent water molecule.

Figure : Shows two superimposed crystal structures of complexes ETSCA II (Cyan) and PTS-CA II (Green)

Thermodynamics - Revision
Gibbs equation:

G = H - T S
Free Energy Change


Entropy Temperature

Enthalpy (H) Energy of making and breaking bonds. Entropy (S) - Disorder of a system. Free Energy Change (G) Spontaneity of a reaction.

Thermodynamics of inhibitor-enzyme binding

Structural change from PTS to ETS involves replacing a proton on the primary amine with an ethyl group. This resulted in:
- greater conformational flexibility - increase in entropic cost - additional surface area - gain in desolvation energy.

Net effect:
- ETS forces the His-64 side chain in an alternate position - displacement of an ordered water molecule

Kinetics of CA II inhibition
Inhibition constant (KI ): Dissociation constant for the complex between an enzyme and the inhibitor. KI of ETS 4 fold smaller than the KI of PTS. Smaller KI Tighter Binding ETSCA II binding is 4 fold tighter than PTS-CA II binding. ETS or Dorzolamide more effective drug than PTS in inhibiting carbonic anhydrase II

In Conclusion:
Benefits of Dorzolamide Topical Carbonic anhydrase inhibitor
Dorzolamide inhibits

CO2 + H2O HCO3- + H+ Dorzolamide blocks the transformation of CO2 to bicarbonate Increased CO2 in the eye improves ocular blood flow Less bicarbonate in the eye leads to reduced aqueous humor production, thus lowering IOP.


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