Journal: eLife

Article Title: A peptide-neurotensin conjugate that crosses the blood-brain barrier induces pharmacological hypothermia associated with anticonvulsant, neuroprotective, and anti-inflammatory properties following status epilepticus in mice

doi: 10.7554/eLife.100527

Figure Lengend Snippet: ( A, C, and E ) Chemical structure and molecular weight of the VH-N21, VH-N41, and VH-N412 conjugates, containing the eight amino acid cyclic brain penetrating peptide that recognizes the LDLR (VH445 for VH-N21 and VH4129 for VH-N41 and VH-N412), and either the neurotensin (NT) tridecapeptide (VH-N21 and VH-N41) or its C-terminal NT(8–13) fragment (VH-N412). ( B, D, and F ) Hypothermic response to VH-N21, VH-N41, and VH-N412 conjugates in mice after single i.v. (bolus) injection at increasing dose levels. Core body (rectal) temperature was measured before (baseline) and at indicated times after injection. Data are presented as means ± SEM, n=4–8 per group. ( G ) Dose-response curves of VH-N21, VH-N41, and VH-N412 hypothermic response. ED 50 values for each conjugate were estimated by plotting the response vs log[dose(mg/kg eq. NT)] followed by nonlinear regression (three parameters) using GraphPad Prism software.

Article Snippet: Recombinant human LDLR (His-tagged) was purchased from Sino Biological (Beijing, China).

Techniques: Molecular Weight, Injection, Software

Journal: eLife

Article Title: A peptide-neurotensin conjugate that crosses the blood-brain barrier induces pharmacological hypothermia associated with anticonvulsant, neuroprotective, and anti-inflammatory properties following status epilepticus in mice

doi: 10.7554/eLife.100527

Figure Lengend Snippet: ( A ) Surface plasmon resonance (SPR) sensorgrams of the free VH4129 and the VH-N412 compound on immobilized human LDLR (hLDLR). Red lines show the specific binding of molecules obtained after double subtraction of the signal measured on the control flow cell (without immobilized LDLR) and a blank run. Black lines show fit curves of the experimental data with a 1:1 binding model. The illustrated data are representative of two to five independent experiments. ( B ) Dose-response inhibition curves of tritiated NT, bound on hNTSR-1 or rNTSR-1 membrane extracts, in the presence of indicated concentrations of NT or VH-N412. Indicated Ki values were estimated from mean IC50 values obtained by logarithmic regression of experimental data. Data were plotted as means ± SD of biological duplicates. ( C and D ) Comparison of degradation rates for NT or peptide-NT conjugates in mouse blood. NT or peptide-NT conjugates were incubated in freshly collected mouse ( C ) or human ( D ) blood and analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) at indicated times in the plasma fraction. Data were plotted as means ± SD of n=3 biological replicates. T 1/2 values were estimated from nonlinear regression (one-phase decay) of experimental data. ( E ) Blood-brain barrier (BBB) transport of tritium-labeled NT or peptide-NT conjugates using in situ brain perfusion in mice. Data were presented as mean ± SEM for three to six animals. Student’s t-test vs NT: *p<0.05, **p<0.01.

Article Snippet: Recombinant human LDLR (His-tagged) was purchased from Sino Biological (Beijing, China).

Techniques: SPR Assay, Binding Assay, Control, Inhibition, Membrane, Comparison, Incubation, Liquid Chromatography, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Labeling, In Situ

Journal: eLife

Article Title: A peptide-neurotensin conjugate that crosses the blood-brain barrier induces pharmacological hypothermia associated with anticonvulsant, neuroprotective, and anti-inflammatory properties following status epilepticus in mice

doi: 10.7554/eLife.100527

Figure Lengend Snippet:

Article Snippet: Recombinant human LDLR (His-tagged) was purchased from Sino Biological (Beijing, China).

Techniques: Transfection, Construct, Plasmid Preparation, Membrane, Binding Assay, Sequencing, RNA Binding Assay, Recombinant, DC Protein Assay, Software