I need to inject SKF-96365 (1-[-(3-(4-methoxyphenyl)propoxy)-4-methoxyphenethyl]-1H-imidazole hydrochloride) , an inhibitor of TRPC channels, in rat brain stem by cannula injections, but i have not been able to find a paper where this drug has been used in-vivo in rat, especially in brain. Can any body suggest me how to find the appropriate concentration of the doze , although in cell cultures it has been used at 20-50 micromolar concentrations to inhibit TRPC channels.
Dear Rehaan Fayaz
I would suggest to use the same concentrations that are effective in cell culture, in the high range since dilution might quickly decrease concentration in situ.
I was not aware of this drug and I'm curious about its properties. Do you know if it crosses the blood-brain barrier? Do you know if it is very toxic? Do you have any reference I can read to study this?
I've not been very helpful, I'm afraid. Instead I'm asking questions to you! Many thanks in advance.
Good luck
Fernando
Dear Rehaan Fayaz, here again Fernando
I've looked for literature on the in vivo effects of this drug. It has been used in rats for analysing its properties in the control of pain through its effects on nociception induced by injection of the bee venom melittin.
Neurosci Bull. 2012 Apr;28(2):173-81. doi: 10.1007/s12264-012-1213-y.
Effects of a non-selective TRPC channel blocker, SKF-96365, on melittin-induced spontaneous persistent nociception and inflammatory pain hypersensitivity.
Ding J1, Zhang JR, Wang Y, Li CL, Lu D, Guan SM, Chen J.
The authors inject melittin subcutaneously, in the forepaw of a rat, and (either before or after venom injection) SKF-96365 at three concentrations: 100, 10 or 1 μg in 20 μL saline. Their figures 2A an d 2B indicate that when applied at 10 or 100 μg in 20 μL saline, the drug had an analgesic effect, as revealed by a reduction of the paw-flinch reflex. If the analgesic effects of this drug, as the authors suggest, are related to its unspecific antagonistic effects on TRPC channels, I imagine that the same concentrations would be effective in other tissues, e.g. the brain.
Good luck again
All the best
Fernando
Thank you for your valuable inputs . I am planning to inject this drug in rat brainstem (exactly in Locus Coeruleus) with the help of cannula implantation with stereotaxic surgery. I have read an article where in mice they have given 0.2 and 002 nano moles per 2 micro liter but still i am apprehensive about the final dosage. Hereby i am attaching that article please go through it if you don't mind.
Thank you very much Fernando for your immense valuable inputs, we have decided to proceed with 100 micro molar concentration as some studies in mice have been carried out with intra-cerebro-ventricular injections.
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