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ABSTRACT: PURPOSE: In vivo and ex vivo inhibition of ectopic activity of clinically used and newly developed sodium channel (NaV) blockers were quantified in the rat spinal nerve ligation (SNL) model using a pharmacokinetic-pharmacodynamic (PKPD) approach and correlated to in vitro NaV1.7 channel inhibition and clinical effective concentrations. METHODS: In vivo, drug exposure and inhibition of ectopic activity were assessed in anaesthetized SNL rats at two dose levels. Ex vivo, compounds were applied at increasing concentrations to dorsal root ganglias isolated from SNL rats. The inhibitory potency (IC ( 50 )) was estimated using PKPD analysis. In vitro IC ( 50 ) was estimated using an electrophysiology-based assay using recombinant rat and human NaV1.7 expressing HEK293 cells. RESULTS: In vivo and ex vivo inhibition of ectopic activity correlated well with the in vitro inhibition on the rat NaV1.7 channel. The estimated IC ( 50s ) for inhibition of ectopic activity in the SNL model occurred at similar unbound concentrations as clinical effective concentrations in humans. CONCLUSIONS: Inhibition of ectopic activity in the SNL model could be useful in predicting clinical effective concentrations for novel sodium channel blockers. In addition, in vitro potency could be used for screening, characterization and selection of compounds, thereby reducing the need for in vivo testing.
Pharmaceutical Research 02/2013; · 4.09 Impact Factor
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Inger Kers,
Istvan Macsari,
Gabor Csjernyik,
Martin Nylöf,
Karin Skogholm,
Lars Sandberg,
Alexander Minidis,
Tjerk Bueters,
Jonas Malmborg, Anders B Eriksson,
Per-Eric Lund,
Elisabet Venyike,
Lei Luo,
Jan-Erik Nyström,
Yevgeni Besidski
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ABSTRACT: The Na(V)1.7 ion channel is an attractive target for development of potential analgesic drugs based on strong genetic links between mutations in the gene coding for the channel protein and inheritable pain conditions. The (S)-N-chroman-3-ylcarboxamide series, exemplified by 1, was used as a starting point for development of new channel blockers, resulting in the phenethyl nicotinamide series. The structure and activity relationship for this series was established and the metabolic issues of early analogues were addressed by appropriate substitutions. Compound 33 displayed acceptable overall in vitro properties and in vivo rat PK profile.
Bioorganic & medicinal chemistry letters 08/2012; 22(19):6108-15. · 2.65 Impact Factor
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Istvan Macsari,
Yevgeni Besidski,
Gabor Csjernyik,
Linda I Nilsson,
Lars Sandberg,
Ulrika Yngve,
Kristofer Ahlin,
Tjerk Bueters, Anders B Eriksson,
Per-Eric Lund,
Elisabet Venyike,
Sandra Oerther,
Karin Hygge Blakeman,
Lei Luo,
Per I Arvidsson
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ABSTRACT: The voltage-gated sodium channel Na(V)1.7 is believed to be a critical mediator of pain sensation based on clinical genetic studies and pharmacological results. Clinical utility of nonselective sodium channel blockers is limited due to serious adverse drug effects. Here, we present the optimization, structure-activity relationships, and in vitro and in vivo characterization of a novel series of Na(V)1.7 inhibitors based on the oxoisoindoline core. Extensive studies with focus on optimization of Na(V)1.7 potency, selectivity over Na(V)1.5, and metabolic stability properties produced several interesting oxoisoindoline carboxamides (16A, 26B, 28, 51, 60, and 62) that were further characterized. The oxoisoindoline carboxamides interacted with the local anesthetics binding site. In spite of this, several compounds showed functional selectivity versus Na(V)1.5 of more than 100-fold. This appeared to be a combination of subtype and state-dependent selectivity. Compound 28 showed concentration-dependent inhibition of nerve injury-induced ectopic in an ex vivo DRG preparation from SNL rats. Compounds 16A and 26B demonstrated concentration-dependent efficacy in preclinical behavioral pain models. The oxoisoindoline carboxamides series described here may be valuable for further investigations for pain therapeutics.
Journal of Medicinal Chemistry 07/2012; 55(15):6866-80. · 4.80 Impact Factor
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Inger Kers,
Gabor Csjernyik,
Istvan Macsari,
Martin Nylöf,
Lars Sandberg,
Karin Skogholm,
Tjerk Bueters, Anders B Eriksson,
Sandra Oerther,
Per-Eric Lund,
Elisabet Venyike,
Jan-Erik Nyström,
Yevgeni Besidski
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ABSTRACT: Recent findings showing a relation between mutations in the Na(V)1.7 channel in humans and altered pain sensation has contributed to increase the attractiveness of this ion channel as target for development of potential analgesics. Amido chromanes 1 and 2 were identified as blockers of the Na(V)1.7 channel and analogues with modifications of the 5-substituent and the carboxamide part of the molecule were prepared to establish the structure-activity relationship. Compounds 13 and 29 with good overall in vitro and in vivo rat PK profile were identified. Furthermore, 29 showed in vivo efficacy in a nociceptive pain model.
Bioorganic & medicinal chemistry letters 07/2012; 22(17):5618-24. · 2.65 Impact Factor
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Istvan Macsari,
Lars Sandberg,
Yevgeni Besidski,
Ylva Gravenfors,
Tobias Ginman,
Johan Bylund,
Tjerk Bueters, Anders B Eriksson,
Per-Eric Lund,
Elisabet Venyike,
Per I Arvidsson
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ABSTRACT: Blocking of certain sodium channels is considered to be an attractive mechanism to treat chronic pain conditions. Phenyl isoxazole carbamate 1 was identified as a potent and selective Na(V)1.7 blocker. Structural analogues of 1, both carbamates, ureas and amides, were proven to be useful in establishing the structure-activity relationship and improving ADME related properties. Amide 24 showed a good overall in vitro profile, that translated well to rat in vivo PK.
Bioorganic & medicinal chemistry letters 07/2011; 21(13):3871-6. · 2.65 Impact Factor
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ABSTRACT: Because ion channel function is a fundamental element of any nociceptive signalling, it is not surprising that numerous channelopathies have recently emerged as likely causes of several inherited clinical pain conditions. For example, numerous missense mutations in the Na(v)1.7 gene SCN9A have recently been linked to a congenital inability to sense pain. Establishing the link between a clinical pain phenotype to an inherited molecular dysfunction of a specific protein has its challenges and requires the collaboration between many specialists. However, once established, such a linkage offers the promise of a powerful and elegant way to mechanistically explain the aspects of the disease studied.
Methods in molecular biology (Clifton, N.J.) 01/2010; 617:309-25.
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Sultan Ahmad,
Leif Dahllund, Anders B Eriksson,
Dennis Hellgren,
Urban Karlsson,
Per-Eric Lund,
Inge A Meijer,
Luc Meury,
Tracy Mills,
Adrian Moody,
Anne Morinville,
John Morten,
Dajan O'donnell,
Carina Raynoschek,
Hugh Salter,
Guy A Rouleau,
Johannes J Krupp
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ABSTRACT: The general lack of pain experience is a rare occurrence in humans, and the molecular causes for this phenotype are not well understood. Here we have studied a Canadian family from Newfoundland with members who exhibit a congenital inability to experience pain. We have mapped the locus to a 13.7 Mb region on chromosome 2q (2q24.3-2q31.1). Screening of candidate genes in this region identified a protein-truncating mutation in SCN9A, which encodes for the voltage-gated sodium channel Na(v)1.7. The mutation is a C-A transversion at nucleotide 984 transforming the codon for tyrosine 328 to a stop codon. The predicted product lacks all pore-forming regions of Na(v)1.7. Indeed, expression of this altered gene in a cell line did not produce functional responses, nor did it cause compensatory effects on endogenous voltage-gated sodium currents when expressed in ND7/23 cells. Because a homozygous knockout of Na(v)1.7 in mice has been shown to be lethal, we explored why a deficiency of Na(v)1.7 is non-lethal in humans. Expression studies in monkey, human, mouse and rat tissue indicated species-differences in the Na(v)1.7 expression profile. Whereas in rodents the channel was strongly expressed in hypothalamic nuclei, only weak mRNA levels were detected in this area in primates. Furthermore, primate pituitary and adrenal glands were devoid of signal, whereas these two glands were mRNA-positive in rodents. This species difference may explain the non-lethality of the observed mutation in humans. Our data further establish Na(v)1.7 as a critical element of peripheral nociception in humans.
Human Molecular Genetics 10/2007; 16(17):2114-21. · 7.64 Impact Factor
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ABSTRACT: We describe a novel three-dimensional (3-D) imaging tool for analysis of protein conformation of in situ samples. Sidec (Sidec Technologies AB, Stockholm, Sweden) electron tomography (SET) uses low-dose electron tomography and a refinement algorithm to reconstruct individual proteins and macromolecular complexes. The approach has successfully reconstructed therapeutic proteins in solution. In this study, we investigate the use of SET to visualize ion channels in cells and tissue samples. SET successfully resolved the volume and structural features of the target complex, showing that it was a tetrameric channel with a central pore. The technology could distinguish and provide 3-D images of the intra- and extracellular domains in the ion channel. In addition, SET was able to show that the channel associates in the form of a tetramer with the four subunits preorganized into dimers. While additional studies using smaller antibody markers are needed to resolve the subunit assembly further, this study demonstrates that SET is a valuable tool for visualization of in situ specimens and can provide important information on the subunit assembly of these macromolecular complexes, and thereby aid in the screening assay process in drug development.
Assay and Drug Development Technologies 11/2004; 2(5):561-7. · 1.73 Impact Factor
