The anti-inflammatory action of the analgesic kyotorphin neuropeptide derivatives: insights of a lipid-mediated mechanism.
Recently, a designed class of efficient analgesic drugs derived from an endogenous neuropeptide, kyotorphin (KTP, Tyr-Arg) combining C-terminal amidation (KTP-NH2) and N-terminal conjugation to ibuprofen (Ib), IbKTP-NH2, was developed. The Ib moiety is an enhancer of KTP-NH2 analgesic action. In the present study, we have tested the hypothesis that KTP-NH2 is an enhancer of the Ib anti-inflammatory action. Moreover, the impact of the IbKTP-NH2 conjugation on microcirculation was also evaluated by a unified approach based on intravital microscopy in the murine cremasteric muscle. Our data show that KTP-NH2 and conjugates do not cause damage on microcirculatory environment and efficiently decrease the number of leukocyte rolling induced by lipopolysaccharide (LPS). Isothermal titration calorimetry showed that the drugs bind to LPS directly thus contributing to LPS aggregation and subsequent elimination. In a parallel study, molecular dynamics simulations and NMR data showed that the IbKTP-NH2 tandem adopts a preferential "stretched" conformation in lipid bilayers and micelles, with the simulations indicating that the Ib moiety is anchored in the hydrophobic core, which explains the improved partition of IbKTP-NH2 to membranes and the permeability of lipid bilayers to this conjugate relative to KTP-NH2. The ability to bind glycolipids concomitant to the anchoring in the lipid membranes through the Ib residue explains the analgesic potency of IbKTP-NH2 given the enriched glycocalyx of the blood-brain barrier cells. Accumulation of IbKTP-NH2 in the membrane favors both direct permeation and local interaction with putative receptors as the location of the KTP-NH2 residue of IbKTP-NH2 and free KTP-NH2 in lipid membranes is the same.
Faculdade de Medicina de Lisboa, Instituto de Medicina Molecular, Av. Professor Egas Moniz, 1649-028, Lisbon, Portugal. katia.conceicao@unifesp.br.|Departamento de Ciência e Tecnologia, Universidade Federal de São Paulo, UNIFESP, Rua Talim, 330, São José dos Campos, Brazil. katia.conceicao@unifesp.br.
Katia
K
Conceição
Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, 2780-157, Oeiras, Portugal.
Pedro R
PR
Magalhães
Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, 2780-157, Oeiras, Portugal.
Sara R R
SR
Campos
Faculdade de Medicina de Lisboa, Instituto de Medicina Molecular, Av. Professor Egas Moniz, 1649-028, Lisbon, Portugal.
Marco M
MM
Domingues
Laboratori d'Innovació en Processos i Productes de Síntesi Orgànica (LIPPSO), Departament de Química, Universitat de Girona, Campus Montilivi, 17071, Girona, Spain.
Vasanthakumar G
VG
Ramu
UMR7177, Institut de chimie de Strasbourg, CNRS, University of Strasbourg, 67000, Strasbourg, France.
Matthias
M
Michalek
UMR7177, Institut de chimie de Strasbourg, CNRS, University of Strasbourg, 67000, Strasbourg, France.
Philippe
P
Bertani
Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, 2780-157, Oeiras, Portugal.
António M
AM
Baptista
Laboratori d'Innovació en Processos i Productes de Síntesi Orgànica (LIPPSO), Departament de Química, Universitat de Girona, Campus Montilivi, 17071, Girona, Spain.
Montserrat
M
Heras
Laboratori d'Innovació en Processos i Productes de Síntesi Orgànica (LIPPSO), Departament de Química, Universitat de Girona, Campus Montilivi, 17071, Girona, Spain.
Eduard R
ER
Bardaji
UMR7177, Institut de chimie de Strasbourg, CNRS, University of Strasbourg, 67000, Strasbourg, France.
Burkhard
B
Bechinger
Unidade de Imunorregulação, Laboratório Especial de Toxinologia Aplicada, Instituto Butantan, Av. Vital Brasil, São Paulo, 1500, Brazil.
Mônica Lopes
ML
Ferreira
Faculdade de Medicina de Lisboa, Instituto de Medicina Molecular, Av. Professor Egas Moniz, 1649-028, Lisbon, Portugal. macastanho@medicina.ulisboa.pt.
Miguel A R B
MA
Castanho
D000700:Analgesics / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000737:chemistry* / Q000378:metabolism; D004723:Endorphins / Q000737:chemistry / Q000378:metabolism*; D005260:Female; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000737:chemistry / Q000378:metabolism; D008297:Male; D051379:Mice; D056004:Molecular Dynamics Simulation; D015394:Molecular Structure
D000700:Analgesics; D000893:Anti-Inflammatory Agents; D004723:Endorphins; D008051:Lipid Bilayers; D008070:Lipopolysaccharides; C019448:kyotorphin
Analgesia; Ibuprofen; Kyotorphin; Kyotorphin amide; Microcirculation