title: A systematic evaluation of solubility enhancing excipients to enable the generation of permeability data for poorly soluble compounds in Caco-2 model.
abstract: The study presented here identified and utilized a panel of solubility enhancing excipients to enable the generation of flux data in the Human colon carcinoma (Caco-2) system for compounds with poor solubility. Solubility enhancing excipients Dimethyl acetamide (DMA) 1 % v/v, polyethylene glycol (PEG) 400 1% v/v, povidone 1% w/v, poloxamer 188 2.5% w/v and bovine serum albumin (BSA) 4% w/v did not compromise Caco-2 monolayer integrity as assessed by trans-epithelial resistance measurement (TEER) and Lucifer yellow (LY) permeation. Further, these excipients did not affect P-glycoprotein (P-gp) mediated bidirectional transport of digoxin, permeabilities of high (propranolol) or low permeability (atenolol) compounds, and were found to be inert to Breast cancer resistant protein (BCRP) mediated transport of cladribine. This approach was validated further using poorly soluble tool compounds, atazanavir (poloxamer 188 2.5% w/v) and cyclosporine A (BSA 4% w/v) and also applied to new chemical entity (NCE) BMS-A in BSA 4% w/v, for which Caco-2 data could not be generated using the traditional methodology due to poor solubility (<1 µM) in conventional Hanks balanced salt solution (HBSS). Poloxamer 188 2.5% w/v increased solubility of atazanavir by >8 fold whereas BSA 4% w/v increased the solubility of cyclosporine A and BMS-A by >2-4 fold thereby enabling permeability as well as efflux liability estimation in the Caco-2 model with reasonable recovery values. To conclude, addition of excipients such as poloxamer 188 2.5% w/v and BSA 4% w/v to HBSS leads to a significant improvement in the solubility of the poorly soluble compounds resulting in enhanced recoveries without modulating transporter-mediated efflux, expanding the applicability of Caco-2 assays to poorly soluble compounds.
authors.forename: Devang
authors.initials: D
authors.lastname: Shah
authors.forename: Sundeep
authors.initials: S
authors.lastname: Paruchury
authors.forename: Muralikrishna
authors.initials: M
authors.lastname: Matta
authors.forename: Gajendra
authors.initials: G
authors.lastname: Chowan
authors.forename: Murali
authors.initials: M
authors.lastname: Subramanian
authors.forename: Ajay
authors.initials: A
authors.lastname: Saxena
authors.forename: Matthew G
authors.initials: MG
authors.lastname: Soars
authors.forename: John
authors.initials: J
authors.lastname: Herbst
authors.forename: Roy
authors.initials: R
authors.lastname: Haskell
authors.forename: Punit
authors.initials: P
authors.lastname: Marathe
authors.affiliation: Pharmaceutical candidate optimization, Biocon Bristol-Myers Squibb R&D Center, Syngene International Limited, Biocon Park Plot 2 & 3, Bommasandra IV Phase, Bangalore - 560 099, India. devang.shah@syngeneintl.com.
authors.forename: Sandhya
authors.initials: S
authors.lastname: Mandlekar
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism*; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D001692:Biological Transport / Q000187:drug effects; D018938:Caco-2 Cells; D005079:Excipients / Q000494:pharmacology*; D006801:Humans; D007546:Isoquinolines / Q000378:metabolism; D009363:Neoplasm Proteins / Q000378:metabolism*; D010539:Permeability; D004364:Pharmaceutical Preparations / Q000737:chemistry / Q000378:metabolism*; D012995:Solubility
chemical_list: C116614:ABCG2 protein, human; D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D005079:Excipients; D007546:Isoquinolines; D009363:Neoplasm Proteins; D004364:Pharmaceutical Preparations; C017475:lucifer yellow