Neurosteroids are endogenous steroidal compounds that can modulate neuronal receptors. N-Methyl-D-aspartate receptors (NMDARs) are glutamate-gated, calcium-permeable ion channels that are of particular interest, as they participate in synaptic transmission and are implicated in various processes, such as learning, memory, or long-term neuronal potentiation. Positive allosteric modulators that increase the activity of NMDARs may provide a therapeutic aid for patients suffering from neuropsychiatric disorders where NMDAR hypofunction is thought to be involved, such as intellectual disability, autism spectrum disorder, or schizophrenia. We recently described a new class of pregn-5-ene and androst-5-ene 3beta-dicarboxylic acid hemiesters (2-24) as potent positive modulators of NMDARs. Considering the recommended guidelines for the early stage development of new, potent compounds, we conducted an in vitro safety assessment and plasma stability screening to evaluate their druglikeness. First, compounds were screened for their hepatotoxicity and mitochondrial toxicity in a HepG2 cell line. Second, toxicity in primary rat postnatal neurons was estimated. Next, the ability of compounds 2-24 to cross a Caco-2 monolayer was also studied. Finally, rat and human plasma stability screening revealed an unforeseen high stability of the C-3 hemiester moiety. In summary, by using potency/efficacy towards NMDARs data along with toxicity profile, Caco-2 permeability and plasma stability, compounds 14 and 15 were selected for further in vivo animal studies.
- MeSH
- androstenoly chemie farmakologie krev MeSH
- buňky Hep G2 MeSH
- cholesterol chemie farmakologie krev MeSH
- estery chemie farmakologie krev MeSH
- kyseliny dikarboxylové chemie farmakologie krev MeSH
- lidé MeSH
- poruchy autistického spektra farmakoterapie metabolismus MeSH
- stabilita léku MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
Neurosteroids are endogenous steroidal compounds that can modulate neuronal receptors. N-Methyl-D-aspartate receptors (NMDARs) are glutamate-gated, calcium-permeable ion channels that are of particular interest, as they participate in synaptic transmission and are implicated in various processes, such as learning, memory, or long-term neuronal potentiation. Positive allosteric modulators that increase the activity of NMDARs may provide a therapeutic aid for patients suffering from neuropsychiatric disorders where NMDAR hypofunction is thought to be involved, such as intellectual disability, autism spectrum disorder, or schizophrenia. We recently described a new class of pregn-5-ene and androst-5-ene 3β-dicarboxylic acid hemiesters (2-24) as potent positive modulators of NMDARs. Considering the recommended guidelines for the early stage development of new, potent compounds, we conducted an in vitro safety assessment and plasma stability screening to evaluate their druglikeness. First, compounds were screened for their hepatotoxicity and mitochondrial toxicity in a HepG2 cell line. Second, toxicity in primary rat postnatal neurons was estimated. Next, the ability of compounds 2-24 to cross a Caco-2 monolayer was also studied. Finally, rat and human plasma stability screening revealed an unforeseen high stability of the C-3 hemiester moiety. In summary, by using potency/efficacy towards NMDARs data along with toxicity profile, Caco-2 permeability and plasma stability, compounds 14 and 15 were selected for further in vivo animal studies.
- MeSH
- androstenoly krev chemie farmakologie MeSH
- buňky Hep G2 MeSH
- cholesterol krev chemie farmakologie MeSH
- estery krev chemie farmakologie MeSH
- krysa rodu rattus MeSH
- kyseliny dikarboxylové krev chemie farmakologie MeSH
- lidé MeSH
- mentální retardace farmakoterapie metabolismus MeSH
- mitochondrie účinky léků metabolismus MeSH
- molekulární struktura MeSH
- nádorové buňky kultivované MeSH
- neurony účinky léků metabolismus MeSH
- neuroprotektivní látky krev chemie farmakologie MeSH
- poruchy autistického spektra farmakoterapie metabolismus MeSH
- potkani Wistar MeSH
- pregnenolon analogy a deriváty krev farmakologie MeSH
- receptory N-methyl-D-aspartátu antagonisté a inhibitory metabolismus MeSH
- schizofrenie farmakoterapie metabolismus MeSH
- stabilita léku MeSH
- viabilita buněk účinky léků MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A broad variety of central nervous system diseases have been associated with glutamate induced excitotoxicity under pathological conditions. The neuroprotective effects of neurosteroids can combat this excitotoxicity. Herein, we have demonstrated the neuroprotective effect of novel steroidal N-methyl-D-aspartate receptor inhibitors against glutamate- or NMDA- induced excitotoxicity. Pretreatment with neurosteroids significantly reduced acute L-glutamic acid or NMDA excitotoxicity mediated by Ca2+ entry and consequent ROS (reactive oxygen species) release and caspase-3 activation. Compounds 6 (IC50 = 5.8 μM), 7 (IC50 = 12.2 μM), 9 (IC50 = 7.8 μM), 13 (IC50 = 1.1 μM) and 16 (IC50 = 8.2 μM) attenuated glutamate-induced Ca2+ entry more effectively than memantine (IC50 = 18.9 μM). Moreover, compound 13 shows comparable effect with MK-801 (IC50 = 1.2 μM) and also afforded significant protection without any adverse effect upon prolonged exposure. This drop in Ca2+ level resulted in corresponding ROS suppression and prevented glutamate-induced caspase-3 activation. Therefore, compound 13 has great potential for development into a therapeutic agent for improving glutamate-related nervous system diseases.
- MeSH
- kultivované buňky MeSH
- kyselina glutamová škodlivé účinky MeSH
- N-methylaspartát škodlivé účinky MeSH
- neurony cytologie účinky léků metabolismus MeSH
- neuroprotekce účinky léků MeSH
- neuroprotektivní látky chemie farmakologie MeSH
- neurotransmiterové látky chemie farmakologie MeSH
- potkani Wistar MeSH
- reaktivní formy kyslíku metabolismus MeSH
- receptory N-methyl-D-aspartátu antagonisté a inhibitory metabolismus MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
