Low-molecular mimetics of brain-derived neurotrophic factor (BDNF) loops 1 and 4 representing to monomeric and dimeric amides of N-acyldipeptides were constructed and synthesized. The sequence of these dipeptides coinside with the central regions of beta-turns of corresponding loops of neurotrophine sequence, and acyl groups are bioisosters of preceding amino acid residues. Hexa- and heptamethylenediamine were used as spacers linking C-terminus ofdipeptides in BDNF dimeric mimetics. These substances were synthesized by classic peptide synthesis methods in solution and got laboratory codes GSB-104 (HO-Suc-Ser-Lys-NH2), GSB-106 ([HO-Suc-Ser-Lys-NH-(CH2)3-]2), GSB-207 (HO-Suc-Met-Ser-NH2) and GSB-214 ([HO-Suc-Met-Ser-NH-(CH2)7/2-]2). By using the culture of immortalized hippocampal cell line HT-22 on the oxidative stress conditions it was shown that dimeric mimetics of both loops demonstrated neuroprotective activity in the concentration rage of 10(-5)-10(-8) M. Monomeric loop 1 mimetic GSB-207 was inactive in the same concentrations and monomeric loop 4 mimetic GSB-104 in a concentration of 10(-7) M decreased survival of neurons. Presence of neuroprotective activity only for dimeric mimetics correlates with the data that BDNF is active only in homodimeric form. As opposed to dimeric mimetic of loop 1 GSB-214, dimeric mimetic of loop 4 GSB-106 demonstrates specific for BDNF antidepressive activity in Porsolt test on rats in doses 0.1 and 1 mg/kg i.p. It is suggested that antidepressive activity of BDNF is associated with its loop 4. We consider that compounds obtained will be useful for investigation of BDNF action mechanism and can lead to creation of a new group of medicinal substances with antidepressive and neuroprotective activities.
BackgroundTwo dimeric dipeptides, bis-(N-monosuccinyl-l-seryl-l-lysine)hexamethylenediamide (GSB-106) and bis-(N-monosuccinyl-l-methionyl-l-serine) heptamethylenediamide (GSB-214), were designed based on the brain-derived neurotrophic factor (BDNF) loop 4 and loop 1 β-turn sequences, respectively. Earlier, both of these dipeptides were shown to exhibit neuroprotective activity in vitro (10−5–10−8 M). The present study aimed to investigate the mechanisms of action of these peptides and their neuroprotective activity in an experimental stroke model.MethodsWe used western blot and HT-22 hippocampal neuronal cell line to investigate whether these peptides induced phosphorylation of the TrkB receptor and the AKT and ERK kinases. Rat middle cerebral artery occlusion (MCAO) was used as a stroke model. GSB-106 and GSB-214 were administered intraperitoneally (0.1 mg (1.3×10−7 mol)/kg) 4 hours after MCAO and daily for 7 days. The cerebral infarct volumes were measured with 2,3,5-triphenyltetrazolium chloride staining 21 days after MCAO.ResultsBoth compounds were shown to elevate the TrkB phosphorylation level while having different post-receptor signaling patterns. GSB-106 activated the PI3K/AKT and MAPK/ERK pathways simultaneously, whereas GSB-214 activated the PI3K/AKT only. In experimental stroke, the reduction of cerebral infarct volume by GSB-106 (∼66%) was significantly greater than that of GSB-214 (∼28% reduction), which could be explained by the fundamental role of the MAPK/ERK pathway in neurogenesis and neuroplasticity. Notably, between these two dipeptides, only GSB-106 exhibited antidepressant activity, as was found previously.ConclusionThe results provided support for the beneficial pharmacological properties of BDNF loop 4 mimetic GSB-106, thereby suggesting a potential role for this dipeptide as a therapeutic agent.