Secondary structure of cyclosporine in a spray-dried liquid crystal by FTIR.
The conformational state of cyclosporine in liquid crystalline spray-dried powders and the solution structure of cyclosporine, in a series of organic solvents where solvent dipole and hydrogen bonding ability varied, were determined. Fourier transformed infrared spectra (FTIR) were obtained on cyclosporine powders, and cyclosporine solutions in a series of organic solvents. Tetragonal crystalline cyclosporine revealed an intermolecular aggregate band at 1614 cm(-1), a beta-sheet band at 1627 cm(-1), a gamma-loop band at 1648 cm(-1), a gamma-turn band at 1658 cm(-1) (formed from a hydrogen bond between D-Ala(8)NH and MeLeu(6)Cdbond;O) and a Type II beta-turn band at 1673 cm(-1) (centered at the hydrogen bond betweenVal(5)NH to Abu(2)Cdbond;O). A similar conformation was observed in chloroform or octanol (apolar), where a second beta-sheet band emerged at 1638 cm(-1) and a turn structure associated with the beta-OH on MeBmt(1) appeared at 1685 cm(-1). However, the spray dried liquid crystal structure resembled the solution conformation in acetone or acetonitrile (hydrogen bond acceptor). The conformation in acetone suggested that the beta-sheet, gamma-loop, Type II beta-turn and MeBmt(1) turn remained intact. Interestingly, the spray-dried powder conformations did not resemble the solution structure of the solvent (ethanol) from which they had been obtained. The conformation in ethanol and methanol (hydrogen bond donor) showed only beta-sheet, gamma-turn, MeBmt(1) turn structure. Only a small population of molecules retained the Type II beta-turn. Finally, cyclosporine is essentially insoluble in water, so the water conformation has never been elucidated; however, a conformation resembling the active structure was obtained in a cosolvent solution containing both hydrogen bond donors and acceptors. This conformation is in good agreement with molecular modelling studies where cyclosporine is docked in the active site of cyclophilin. Spray-dried cyclosporine formed a liquid crystal that can be described as maintaining the Type II beta-turn, beta-sheet, and gamma-loop structures seen in crystalline material. However, the hydrogen bond between D-Ala(8)NH and MeLeu(6)Cdbond;O was disrupted.
Stevenson
Cynthia L
CL
Nektar Therapeutics, 150 Industrial Road, San Carlos, California 94070, USA. cstevenson@ca.nektar.com
Tan
Mandy M
MM
Lechuga-Ballesteros
David
D
D016572:Cyclosporine / Q000737:chemistry*; D015394:Molecular Structure; D019999:Pharmaceutical Solutions; D011487:Protein Conformation; D012997:Solvents; D017550:Spectroscopy, Fourier Transform Infrared
D019999:Pharmaceutical Solutions; D012997:Solvents; D016572:Cyclosporine