Characterization and application of single-isomer cyclodextrin derivatives

Transcription

1 Characterization and application of single-isomer cyclodextrin derivatives Gábor Benkovics CycloLab, Cyclodextrin R&D Ltd., Budapest, Hungary Faculty of Science, Charles University in Prague, Czech Republic 1

2 Cyclodextrins primary side hydrophobic cavity 6 OH 3 OH 2 OH CD Number of glucose units α 6 β 7 γ 8 secondary side hydrophilic outer surface 2

3 Isomerism in cyclodextrin derivatization Random/statistical substitution position 2-O position 3-O position 6-O Monosubstitution Persubstitution Disubstitution Top view for β-cd pseudoenantiomers 3

4 CD-Screen Inclusion-assisted liquid chromatography CD-Screen columns H-acceptor hydrophilic hydrophobic H-donors dipole-dipole AD/AE CD-Screen 50 AC/AF 40 AB/AG 30 mau Benkovics et al. Int. J. Pharm 2017, Ahead of print min 4

5 mv mv CD-Screen-IEC Inclusion-assisted ion-exchange chromatography (CD-Screen-IEC columns) H-acceptor hydrophobic hydrophilic ionic H-donor a DS=6 mv Randomly carboxymethylated γ-cd Single-isomer carboxymethyl-γ-cd In process control DS=5 DS=7 ODMCM-γ-CD DS= DS= b Purity assesment ODMCM-γ-CD Purity: 95.5 Area % t/min t/min t/min HPLC conditions: CD-Screen-IEC stationary phase, ammonium acetate (ph 4.0), gradient with ACN, ELS detection 5

6 Single-isomer carboxymethyl cyclodextrins HDMCM ODMCM Preparation: heptakis-(2,3-di-o-methyl-6-o-carboxymethyl)-β-cyclodextrin (HDMCM) octakis-(2,3-di-o-methyl-6-o-carboxymethyl)-γ-cyclodextrin (ODMCM) 4-step synthesis from native CDs in 52 % (HDMCM) and 48 % (ODMCM) total yield Characterization: HPLC - inclusion-assisted ion-exchange chromatography (CD-Screen-IEC columns) with ELS detection Capillary electrophoresis with indirect UV detection 1 H NMR titration 1. Fejős et al. Electrophoresis 2017, Ahead of print 2. Benkovics et al. J. Chromatogr. A. 2016, 1467,

7 Single-isomer carboxymethyl cyclodextrins (application) ph tunable ionic interactions on a well-defined position electroneutral * * octaanionic CE electropherograms applying ODMCM as chiral selector applicability in aqueous as well as in non-aqueous capillary electrophoresis (NACE) NACE electropherograms applying HDMCM as chiral selector 7

8 Single-isomer sulfobutyl-β-cyclodextrin Sulfobutylated β-cd (DS=6.4) (Dexolve TM, Captisol TM ) Heptakis-(6-O-sulfobutyl)-β-CD (DS=7.0) 6-(SB) 7 -β-cd HPLC conditions: CD-Screen-IEC stationary phase, solvent gradient of 1.5% triethylamine-formate (ph 4.5) with ACN, ELS detection 8

9 Single-isomer sulfobutyl β-cyclodextrin Racemic dapoxetin * CE electropherograms applying 6-(SB) 7 -β-cd as chiral selector Racemic dapoxetine : 6-(SB) 7 -β-cd HO OH H C 2 H C 3 C 1 H O O C 6 H O H N + H - O S -O H H 7 9 S C 4 C 5 C 6 O H R 8S 5S 5R Racemic dapoxetine : 6-(SB) 7 -β-cd system spiked with S-dapoxetine 3S 3R 2R 2S Malanga et al. J. Chromatogr. A. 2017, 1514,

10 Beyond enantiomer separation: Methylated cyclodextrins randomly-methylated β-cyclodextrins (RAMEB, CRYSMEB) heptakis-(6-o-methyl)-β-cyclodextrin (SIXMEB) heptakis-(2-o-methyl)-β-cyclodextrin (TWOMEB) heptakis-(2,6-di-o-methyl)-β-cyclodextrin (2,6-DIMEB) heptakis-(2,3-di-o-methyl)-β-cyclodextrin (2,3-DIMEB) heptakis-(2,3,6-tri-o-methyl)-β-cyclodextrin (TRIMEB) 10

11 Beyond enantiomer separation Nanocarriers for the delivery of antimicrobials to fight resistance mechanisms Cyclodextrin-penicillin interactions ODMCM, HDMCM and TRIMEB are able to protect the β-lactam ring from catalytic degradation Popielec et al. Pharmazie 2016, 71, Popielec et al. Int. J. Pharm 2017, Ahead of print 11

12 Summary New negatively charged single-isomer CDs have been synthesized: ODMCM, HDMCM (carboxymethylated) 6-SB 7 -β-cd (sulfoalkylated) Chiral resolution properties of these derivatives have been studied with a wide set of test racemates applying aqueous and non-aqueous capillary electrophoresis The synthesized library of single-isomer carboxymethylated and methylated CD derivatives helped us to understand the inclusion phenomenon between CDs and β-lactam antibiotics ODMCM, HDMCM and TRIMEB are the first CD derivatives showing stabilization effect of the β-lactam ring 12

13 Acknowledgement Milo Malanga Ph.D., Julianna Szemán Ph.D., Tamás Sohajda Ph.D., Éva Fenyvesi Ph.D., Erzsébet Varga, Lajos Szente Ph.D.,D.Sc. CycloLab Cyclodextrin R&D, Ltd., Budapest, Hungary Assoc. Prof. Jindřich Jindřich Faculty of Science, Charles University in Prague, Czech Republic András Darcsi Ph.D., Ida Fejős Ph.D., Assoc. Prof. Szabolcs Béni Department of Pharmacognosy, Semmelweis University, Budapest, Hungary Agnieszka Popielec, Prof. Thorsteinn Loftsson, Faculty of Pharmacy, University of Iceland, Iceland European Comission, FP7 PEOPLE-ITN project n CycloN Hit Prof. Gyula Vígh 13

14 Thank you for your kind attention! 14