Is it possible to metabolize Buprenorphine into Norbuprenorphine in vitro?

[Animoe]

The title kind of says it all.. Is there a way to metabolize Buprenorphine into Norbuprenorphine in vitro? I'm no expert in biology/pharmacology so please excuse me if this is a dumb question or I'm using incorrect terminology..
I was wondering if it would be possible to take the enzyme (Cyp3A4, I think..) that metabolizes Buprenorphine into Norbuprenorphine, and introduce some Buprenorphine to the mix and end up with Norbuprenorphine?

Obviously it would probably (I'm guessing) be a bit more complicated than just mixing the two up in a beaker and have Norbuprenorphine form, but you guys get the gist of what I'm saying...

Soooooo, what do you guys think?

 

[G_Chem]

Theoretically one could spit into a cup, mix up some buprenorphine strips and let it “react.” CYP3A4 is found in saliva. Not sure what kind of microbial life would flourish as well in such conditions.

I’d bet though a simple A/B, assuming buprenorphine can handle it, the NorBupe could be cleaned up enough to be safe to use.

One could send the product to a lab after the fact mentioning it may contain NorBupe and to look for it as well as Bupe.

-GC

 

[Animoe]

Ooh, interesting! Thank you for the reply!

BTW, what does A/B mean?

 

[plumbus-nine]

BTW, what does A/B mean?

Acid/base

Recrystallization and Acid/Base Extraction - The Basics - [www.rhodium.ws]

 

[Animoe]

Acid/base

Recrystallization and Acid/Base Extraction - The Basics - [www.rhodium.ws]

Ah! Thank you very much!

 

[DeathIndustrial88]

I thought the issue was norbuprenorphine's poor ability to pass the blood brain barrier?



Would the spit thing really work? Would one need to really worry about microbial life if they were to just sublingual it? How long would it need to sit & by 'react', does that mean doing more than just mixing up the two?

 

[Skorpio]

Theoretically one could spit into a cup, mix up some buprenorphine strips and let it “react.” CYP3A4 is found in saliva. Not sure what kind of microbial life would flourish as well in such conditions.

I’d bet though a simple A/B, assuming buprenorphine can handle it, the NorBupe could be cleaned up enough to be safe to use.

One could send the product to a lab after the fact mentioning it may contain NorBupe and to look for it as well as Bupe.

-GC

I'm interested in this, do you have the source you got this info from? Even better would be a functional comparison with hepatocytes/microsomes from whole liver to compare activity.

Edit:

This paper was the only one i found about cyps in the salivary glands.

Expression of two drug-metabolizing cytochrome P450-enzymes in human salivary glands - PubMed

The results indicate a xenobiotic metabolizing capability of salivary glands. This may have implications for development of oral mucosal disease as a result of mucosal exposure to metabolites originating from internal sources (blood) as well as from saliva.

pubmed.ncbi.nlm.nih.gov

They did staining to visualize protein and in situ hybridization to measure mRNA expression.

To believe this, i would also like to see cytochrome b reductase colocalization (this is required for cyp activity because).

Either way, i highly doubt that functional cyp enzymes are secreted into the saliva. They are situated in membranes and require other proteins in complex to function.

You could probably do homemade microsomes from fresh calf liver to metabolize bupe in vitro. Wouldn't be too easy but seems possible to adapt a lab protocol. Hardest part would be getting all the cofactors in the solution.

 

[G_Chem]

I'm interested in this, do you have the source you got this info from? Even better would be a functional comparison with hepatocytes/microsomes from whole liver to compare activity.

Edit:

This paper was the only one i found about cyps in the salivary glands.

Expression of two drug-metabolizing cytochrome P450-enzymes in human salivary glands - PubMed

The results indicate a xenobiotic metabolizing capability of salivary glands. This may have implications for development of oral mucosal disease as a result of mucosal exposure to metabolites originating from internal sources (blood) as well as from saliva.

pubmed.ncbi.nlm.nih.gov

They did staining to visualize protein and in situ hybridization to measure mRNA expression.

To believe this, i would also like to see cytochrome b reductase colocalization (this is required for cyp activity because).

Either way, i highly doubt that functional cyp enzymes are secreted into the saliva. They are situated in membranes and require other proteins in complex to function.

You could probably do homemade microsomes from fresh calf liver to metabolize bupe in vitro. Wouldn't be too easy but seems possible to adapt a lab protocol. Hardest part would be getting all the cofactors in the solution.

Thanks @Skorpio yea that was my article I was basing that info off of, admittedly you know much more about these things than I

So what if I puked all over some buprenorphine, would we get some NorBupe then? Lol. One could eat a bunch of hot pepper, induce CYP3A4 then brutally yak it all up crying as the pepper burns your sinuses. This synthesis would need to be done large scale so as to avoid ever doing it again.

That last paragraph has me interested, what about other animal livers? Blend it like a smoothie, then go about extracting the mess, sounds like a project.

-GC

 

[Skorpio]

Thanks @Skorpio yea that was my article I was basing that info off of, admittedly you know much more about these things than I

So what if I puked all over some buprenorphine, would we get some NorBupe then? Lol. One could eat a bunch of hot pepper, induce CYP3A4 then brutally yak it all up crying as the pepper burns your sinuses. This synthesis would need to be done large scale so as to avoid ever doing it again.

That last paragraph has me interested, what about other animal livers? Blend it like a smoothie, then go about extracting the mess, sounds like a project.

-GC

Yeah other mammal livers should do the trick. Just need to get all the cofactors in order and blend it up in a way that keeps the cyps embedded in membranes.. Look up microsome preparation for studies in drug metabolism, it was a very common technique before people got into expressing cyps in cell lines.

Ive been seeing a lot about capsaicin inducing cyp3a4. There's honestly something missing there - I feel like it would be contraindicated like st johns wort, rifampicin, and barbs if it was potent enough to do that at physiological doses. I'll pursue this tonight when i get home or tomorrow when i have more bandwidth.

 

[G_Chem]

Yeah other mammal livers should do the trick. Just need to get all the cofactors in order and blend it up in a way that keeps the cyps embedded in membranes.. Look up microsome preparation for studies in drug metabolism, it was a very common technique before people got into expressing cyps in cell lines.

Ive been seeing a lot about capsaicin inducing cyp3a4. There's honestly something missing there - I feel like it would be contraindicated like st johns wort, rifampicin, and barbs if it was potent enough to do that at physiological doses. I'll pursue this tonight when i get home or tomorrow when i have more bandwidth.

I don’t think capsaicin is potent enough to be contraindicative, I do believe based on my experimentation there’s something there but the amounts I eat to get there are far from normal. Amounts that only true hot spice lovers can handle and enjoy. To get there I also use preparations that are fat heavy so a lot more capsaicin can be ingested.

Compared to most other substances known to mess with enzymes it’s relatively weak. Also there’s kind of less risks/problems with inducing this enzyme than inhibiting for most drugs. It’s one thing if it just metabolizes in a different way then excreted, it’s another if a build up occurs because of improper metabolism.

That’s my take on it at least. On top of that the research around capsaicin inducing CYP3A4 is weak from what I see, more needs to be done.

-GC

 

[Feretile]

Yes - I'm sure it's metabolism was elucidated before it ever received a licence.

 

[DeathIndustrial88]

I eat a lot of jalapenos & peppers on bupe, but have never felt a subjective difference.

I've always wondered if norbupe did become active, if it's receptor profile would make it easy to overdose on or possibly a dysphoric or strange experience.
Though it is a full agonist apparently. But I dunno. I assume buprenorphine probably blocks it's own metabolite.

Also if it were easy to get norbupe into the brain, there'd most likely be more reports of buprenorphine deaths, which are rare (not impossible tho I guess).
Atleast through unsuspected metabolism changes from eating certain foods or taking certain drugs I mean.

 

[Animoe]

Theoretically one could spit into a cup, mix up some buprenorphine strips and let it “react.” CYP3A4 is found in saliva. Not sure what kind of microbial life would flourish as well in such conditions.

I’d bet though a simple A/B, assuming buprenorphine can handle it, the NorBupe could be cleaned up enough to be safe to use.

One could send the product to a lab after the fact mentioning it may contain NorBupe and to look for it as well as Bupe.

-GC

I decided to give this method a go.
I've got 2mg Buprenorphine mixed up with saliva in an oral syringe that I'm going to let sit and (maybe/hopefully) metabolize into Norbuprenorphine for 8 or so hours, then I'm going to SL it like normal and see if there's any noticeable effect.
I'll report back with the results.

 

[Animoe]

I decided to give this method a go.
I've got 2mg Buprenorphine mixed up with saliva in an oral syringe that I'm going to let sit and (maybe/hopefully) metabolize into Norbuprenorphine for 8 or so hours, then I'm going to SL it like normal and see if there's any noticeable effect.
I'll report back with the results.

Ok, so I did it.. I'm not sure if it's just the placebo effect or not but it does feel a bit better than normal.. I'd say about 10% better feeling, also a little stimulating feeling, which isn't what I'd expect from a full agonist like Norbuprenorphine.. But again, it could be completely chalked up to the placebo effect.
Someone else should give it a try and either verify/refute my results.
I'm kinda leaning toward it being a placebo, but if anyone wants to give it a go, please let me know your thoughts.