title: pH-dependent complexation of hydroxypropyl-beta-cyclodextrin with chlorin e6: effect on solubility and aggregation in relation to photodynamic efficacy.
abstract: OBJECTIVE
  The activity of chlorin e6 (Ce6) in photodynamic therapy of cancers is significantly reduced by its propensity to form aggregates. It was postulated that disaggregation of Ce6 could be achieved with the use of hydroxypropyl-beta-cyclodextrin (HP-β-CD) through solubility enhancement.
  
  
  METHODS
  An initial phase solubility study of Ce6 was conducted with various concentrations of HP-β-CD at three different pH conditions, i.e. pH 3, pH 5 and pH 7. Solubility-induced disaggregation of Ce6 was illustrated by fluorescence spectroscopy and singlet oxygen generation studies. Interaction between Ce6 and HP-β-CD was further demonstrated by solid-state characterization techniques. Inclusion complex formulations were tested for improved efficacy on squamous cancer cell lines.
  
  
  RESULTS
  Increase in Ce6 solubility was observed, especially at pH 7, indicating the formation of inclusion complex between Ce6 and HP-β-CD. This resulted in disaggregation of Ce6 aggregates illustrated by fluorescence spectroscopy. The mode of binding was predominated by H-bonding supported by temperature-dependent binding studies and molecular simulation work. The inclusion complex demonstrated improved photodynamic efficacy through enhanced singlet oxygen generation and phototoxicity on human oral squamous carcinoma cells.
  
  
  CONCLUSIONS
  pH-dependent complexation between Ce6- and HP-β-CD-induced disaggregation of Ce6 aggregates and the resultant formulations facilitated improved PDT efficacy on tested cancer cell lines.
authors.affiliation: Department of Pharmaceutics, University of Minnesota, Minneapolis, MN, USA.
authors.forename: Shubhajit
authors.initials: S
authors.lastname: Paul
authors.affiliation: Department of Pharmacy, National University of Singapore, Singapore.
authors.forename: Paul Wan Sia
authors.initials: PW
authors.lastname: Heng
authors.affiliation: Department of Pharmacy, National University of Singapore, Singapore.
authors.forename: Lai Wah
authors.initials: LW
authors.lastname: Chan
mesh_terms: D000073738:2-Hydroxypropyl-beta-cyclodextrin; D002152:Calorimetry, Differential Scanning; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002735:Chlorophyllides; D018360:Crystallography, X-Ray; D004305:Dose-Response Relationship, Drug; D004339:Drug Compounding; D006258:Head and Neck Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D009062:Mouth Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D010778:Photochemotherapy / Q000379:methods*; D017319:Photosensitizing Agents / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D011166:Porphyrins / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D033362:Powder Diffraction; D026082:Singlet Oxygen / Q000737:chemistry; D012995:Solubility; D013050:Spectrometry, Fluorescence; D017550:Spectroscopy, Fourier Transform Infrared; D000077195:Squamous Cell Carcinoma of Head and Neck; D013816:Thermodynamics; D013997:Time Factors; D047392:beta-Cyclodextrins / Q000737:chemistry*
chemical_list: D002735:Chlorophyllides; D017319:Photosensitizing Agents; D011166:Porphyrins; D047392:beta-Cyclodextrins; D026082:Singlet Oxygen; D000073738:2-Hydroxypropyl-beta-cyclodextrin; C062985:phytochlorin
keywords: drug delivery to specific tissues; pharmaceutical analysis; pharmaceutics and drug delivery, drug characterisation studies