1. Development and characterisation of novel fentanyl-delta opioid receptor antagonist based bivalent ligands 
M.F. Bird, R.S. Vardanyan, V.J. Hruby, G. Calò, R. Guerrini, S. Salvadori, C. Trapella, J. McDonald, D.J. Rowbotham, D.G. Lambert 
British Journal of Anaesthesia 
Volume 114, Issue 4, Pages (April 2015)
DOI: /bja/aeu454
Copyright © 2015 The Author(s) Terms and Conditions

2. Fig 1
(a) Ligand stimulated GTPγ[35S] binding for fentanyl and compounds #9, #10 and #11 in CHOhMOP cell membranes. (b) Bivalent compounds were co-incubated, at a concentration of 1 µM, with fentanyl. Data are shown as mean (sem) for 5 experiments.
British Journal of Anaesthesia  , DOI: ( /bja/aeu454)
Copyright © 2015 The Author(s) Terms and Conditions

3. Fig 2
(a) DPDPE stimulated GTPγ[35S] binding. Compounds (#9, #10, and #11) showed no activity in CHOhDOP cell membranes. (b) DPDPE stimulated binding in the absence and presence of 100 nM of bivalent compounds (#9, #10, and #11). All data are mean (sem) of 5 experiments. Reference ligand, DPDPE; [D-Pen2, D-Pen5 Enkephalin].
British Journal of Anaesthesia  , DOI: ( /bja/aeu454)
Copyright © 2015 The Author(s) Terms and Conditions

4. Fig 3
(a) Dynorphin-A stimulated GTPγ[35S] binding. Compounds (#9, #10, and #11) showed no activity in CHOhKOP cell membranes. (b) N/OFQ stimulated GTPγ[35S] binding. Bivalent compounds (#9, #10, and #11) showed no activity in CHOhNOP cell membranes. Data are shown as the mean (sem) for 3 experiments.
British Journal of Anaesthesia  , DOI: ( /bja/aeu454)
Copyright © 2015 The Author(s) Terms and Conditions

5. Fig 4
(a) Fentanyl stimulated GTPγ[35S] binding for #11 and the Gly-extended compounds (#12, #13), is shown in CHOhMOP cell membranes. (b) Fentanyl stimulated binding in the absence and presence of 300 nM of bivalents #12 and #13. Reference ligand, fentanyl. Data are shown as mean (sem) for 5 experiments.
British Journal of Anaesthesia  , DOI: ( /bja/aeu454)
Copyright © 2015 The Author(s) Terms and Conditions

6. Fig 5
(a) DPDPE, #11 and the Gly-extended compound (#12, #13)-stimulated GTPγ[35S] binding in CHOhDOP membranes. (b) DPDPE stimulated binding in the absence and presence of 30nM of bivalents #12 and #13. Reference ligand, DPDPE; [D-Pen2, D-Pen5 Enkephalin]. Data are shown as mean (sem) for 5 experiments.
British Journal of Anaesthesia  , DOI: ( /bja/aeu454)
Copyright © 2015 The Author(s) Terms and Conditions

7. Fig 6
(a) Ligand stimulated GTPγ[35S] in CHOhKOP membranes. (b) Dynorphin-A stimulated binding in the absence and presence of #12 (300nM) and #13 (1 µM). Reference ligand; Dynorphin-A. Data are shown as mean (sem) for 5 experiments.
British Journal of Anaesthesia  , DOI: ( /bja/aeu454)
Copyright © 2015 The Author(s) Terms and Conditions

8. Fig 7
(a) Fentanyl and morphine concentration response curves for β-arrestin recruitment. (b) shows the recruitment of β-arrestin in the presence of 10 µM of fentanyl, morphine and the various bivalent ligands in CHOhDOP cells. The activation factor is a ratio relative to basal β-arrestin activity. Data were significant (anova) and *P< compared with fentanyl post hoc Bonferroni test. Data are shown as mean (sem) for n≥5.
British Journal of Anaesthesia  , DOI: ( /bja/aeu454)
Copyright © 2015 The Author(s) Terms and Conditions

9. Fig 8
(a) Depicts a concentration response curve in the absence, and presence, of 10 nM Dmt-Tic. (b) Shows the inhibition caused by selected concentrations of the various bivalent ligands in CHOhDOP cells. Data were significant (anova) and *P<0.0005; **P< compared with DPDPE post hoc Bonferroni test. Data are shown as mean (sem) for n≥5.
British Journal of Anaesthesia  , DOI: ( /bja/aeu454)
Copyright © 2015 The Author(s) Terms and Conditions