Potential Suramin Alternatives - Sytrinol and Kudzu (Anti Purinergic Therapy)

[necessary8]

I'm still very crashed, but I just wanted to say to be very careful with conclusions like these, @nandixon If you watch Naviaux's presentations, be it about ME/CFS or about autism, he clearly says that the therapeutic effect of suramin is about blocking the channels through which ATP is released. And if you look close at his slides, the channels are labeled Panx1/P2X7. He might have lied for the sake of simplicity, but I doubt it.

The way I see it, the decrease in P2X7 expression in autism is probably compensatory, because of their constant activation. I hadnt seen any indication from Naviaux that it is the cause of autism. (if you had, show me?)

As for suramin not blocking P2X7 in the quantities used in the study... First, can you show us your math and sources for that?

And second, there is another thing to consider, which is that the suramin might have blocked Panx1 directly, even without affecting P2X7 activation. There are substances known to do that, such as Brilliant Blue FCF. You can try calculating this as well if you want, I'm too crashed for that, and would like to see it. Because you might still be right, I just doubt it at this moment.

There is also the issue that P2X7 activity downregulation being the cause of ME/CFS contradicts like half of all the observations we have in this illness, which I talked about in detail in my essays about purinergic signaling.

 

[Sushi]

So far, just one-half of a 1.34 mg tablet taken sometime between dinner and bedtime.

I've been trying clemastine for a few days. 1/2 of a 1.34 tablet made me really sedated the whole of the day after. I tried 1/4 tab and that may help. I do notice more energy and a better antihistamine effect than 2nd generation antihistamines that I have tried.

@nandixon are you still trying it?

 

[necessary8]

And for all of you guys, I'm gonna spell it out in simple terms: I discourage taking clemastine or any other P2X7 upregulator, until we get more light on this issue. Of course the choice is yours, but you should understand that if nandixon's speculation is wrong, and I think it is, clemastine can make you worse, possibly permanently. Please be careful.


In other news, I tried emodin. According to Hip's math, for my weight I would need 9g of rheum palmatum for IC50. I took about 5-10g (my scale is not very accurate). No change in my ME/CFS, but since it triggered my digestive symptoms, I won't be trying it again. As for why it didnt work, it might be that it wont work in general, but it also might be the issue of dosage or purity. We're dealing with a dried plant after all, not a synthethized drug.
There might be a dransdermal option though, which would be easier on the gut. There is something called the Liu He Dan ointment, which has emodin, and it does get into the blood. @Hip, could you look into this, and do the math?
I also found something that looks like a pharmacodynamic study of Brilliant Blue FCF. Do you think we can calculate the dosage more accurately with this?

 

[nandixon]

I've been trying clemastine for a few days. 1/2 of a 1.34 tablet made me really sedated the whole of the day after. I tried 1/4 tab and that may help. I do notice more energy and a better antihistamine effect than 2nd generation antihistamines that I have tried.

@nandixon are you still trying it?

Yes, it still seems to be working consistently well, and I'm still taking ½ tablet at night between dinner and bedtime (the earlier the better to help avoid the next day sedation).

It's given me what feels like about a 50-60% increase in energy, which is one of the largest I've experienced, but the reality is that it's taken me from bordering on severe (i.e., nearly bedbound) to maybe the middle to milder end of moderate (i.e., housebound to at least some degree).

It's a little difficult to compare to previous successes because I have a progressive form of ME/CFS that has slowly but steadily worsened over nearly 20 years. (I started out mild.) So this improvement with clemastine is probably not even quite putting me back to where cimetidine (Tagamet) - my previous “big” success - put me 2 or 3 years ago, and that felt like maybe a 30% subjective improvement at the time. (I'm not sure whether the cimetidine has stopped working. It may just be that it’s no longer able to compensate for a further deterioration. I'm still taking it for gastritis, though.)

It's been interesting with the clemastine. It's the very first thing I’ve tried that’s actually been able to seemingly help reset my circadian rhythm since developing ME/CFS.

(It's also working fairly well as a sleep aid all by itself, at least enough so that I don't have to take and rotate combinations of other meds and supplements to be able to sleep.)

Like @Sushi, I've also found it to be a better antihistamine for hayfever purposes than any other I've tried. (The antihistamine effect carries over well into the next day.) So I no longer need to take chlorpheniramine during the day or diphenhydramine at night (the latter I'd use only once or twice a week for sleep).

I haven't noticed any downsides so far. Clemastine is one of the “cleanest” drugs in the antihistamine class, except for the sedation effect, of course.

I did go for one night without taking it (inadvertently) and promptly seemed to revert back to near bedridden the next day, but it also could have been PEM from overexerting myself 2 days prior.

(@necessary8, I have some thoughts you may be interested in, but they'll have to wait until sometime after Thanksgiving.)

 

[Jesse2233]

@necessary8 why do you think Clemastine might make someone permanently worse

 

[necessary8]

It's given me what feels like about a 50-60% increase in energy, which is one of the largest I've experienced, but the reality is that it's taken me from bordering on severe (i.e., nearly bedbound) to maybe the middle to milder end of moderate (i.e., housebound to at least some degree).

Interesting, how long have you been taking it?

If you say it helps from experience, then that is a better argument that it might be worth trying, than any biochemical speculations, tbh. Keep us posted.

@necessary8 why do you think Clemastine might make someone permanently worse

Should be obvious, no? Because it increases P2X7 activation, which nandixon proposes might be a good thing, but I disagree. Literally all of my theories are built around the notion that P2X7 is overactivated in ME/CFS, and that's what Naviaux is suggesting as well.

 

[Hip]

In other news, I tried emodin. According to Hip's math, for my weight I would need 9g of rheum palmatum for IC50. I took about 5-10g (my scale is not very accurate). No change in my ME/CFS, but since it triggered my digestive symptoms, I won't be trying it again. As for why it didnt work, it might be that it wont work in general, but it also might be the issue of dosage or purity. We're dealing with a dried plant after all, not a synthethized drug.
There might be a dransdermal option though, which would be easier on the gut. There is something called the Liu He Dan ointment, which has emodin, and it does get into the blood. @Hip, could you look into this, and do the math?
I also found something that looks like a pharmacodynamic study of Brilliant Blue FCF. Do you think we can calculate the dosage more accurately with this?

Emodin is a laxative, so that's likely why you experienced digestive symptoms. It may be that if you use transdermal emodin in the form of Liu He Dan ointment, you will not get these symptoms.

If you put Liu He Dan ointment on your skin, though, it's not really going to be possible to calculate blood levels of emodin accurately, unless you can find data detailing what percentage of the emodin is transdermally absorbed by the skin.

If we assume that the amount of emodin is absorbed by the skin is roughly the same amount that is absorbed in the gut, then we can use my original emodin calculation earlier on this thread, which found that and oral dose of 83 mg of emodin achieved the IC50 concentration in the blood. So with this assumption, you would want to put enough Liu He Dan ointment on your skin to provide 83 mg of emodin. I could not find the percentage of emodin in Liu He Dan, though.



That pharmacokinetic study on Brilliant Blue FCF in rats provides a figure of for the percentage absorption in the gut of only 2%. This is lower that the 10% figure I originally used in my earlier Brilliant Blue FCF pharmacokinetic calculation. So on the basis of this new 2% figure, we can revise my earlier calculation for the Brilliant Blue FCF dose, to the following:

Brilliant Blue FCF oral dosage that achieves the IC50 concentration in the blood plasma = 855 mg

However, remember that this dose calculation is based on a guess, because we don't have any data for the plasma protein binding of Brilliant Blue FCF, so I had to guess a plasma protein binding of 50%.

 

[frozenborderline]

Yes, it still seems to be working consistently well, and I'm still taking ½ tablet at night between dinner and bedtime (the earlier the better to help avoid the next day sedation).

It's given me what feels like about a 50-60% increase in energy, which is one of the largest I've experienced, but the reality is that it's taken me from bordering on severe (i.e., nearly bedbound) to maybe the middle to milder end of moderate (i.e., housebound to at least some degree).

It's a little difficult to compare to previous successes because I have a progressive form of ME/CFS that has slowly but steadily worsened over nearly 20 years. (I started out mild.) So this improvement with clemastine is probably not even quite putting me back to where cimetidine (Tagamet) - my previous “big” success - put me 2 or 3 years ago, and that felt like maybe a 30% subjective improvement at the time. (I'm not sure whether the cimetidine has stopped working. It may just be that it’s no longer able to compensate for a further deterioration. I'm still taking it for gastritis, though.)

It's been interesting with the clemastine. It's the very first thing I’ve tried that’s actually been able to seemingly help reset my circadian rhythm since developing ME/CFS.

(It's also working fairly well as a sleep aid all by itself, at least enough so that I don't have to take and rotate combinations of other meds and supplements to be able to sleep.)

Like @Sushi, I've also found it to be a better antihistamine for hayfever purposes than any other I've tried. (The antihistamine effect carries over well into the next day.) So I no longer need to take chlorpheniramine during the day or diphenhydramine at night (the latter I'd use only once or twice a week for sleep).

I haven't noticed any downsides so far. Clemastine is one of the “cleanest” drugs in the antihistamine class, except for the sedation effect, of course.

I did go for one night without taking it (inadvertently) and promptly seemed to revert back to near bedridden the next day, but it also could have been PEM from overexerting myself 2 days prior.

(@necessary8, I have some thoughts you may be interested in, but they'll have to wait until sometime after Thanksgiving.)

could it be that the antihistamine effects, not the effects on purinergic signalling, are why clemastine is effective? it seems to be an antihistamine with serotonin receptor antagonism, which could help with inflammation and fatigue

 

[necessary8]

If you put Liu He Dan ointment on your skin, though, it's not really going to be possible to calculate blood levels of emodin accurately, unless you can find data detailing what percentage of the emodin is transdermally absorbed by the skin.

It has proper studies done on it:
http://www.sciencedirect.com/science/article/pii/S0378874112007337
https://www.hindawi.com/journals/ecam/2014/389576/abs/

As for BB FCF, I hoped that this study maybe would allow you to not have to guess the protein binding. But I guess the study doesnt list plasma levels?

 

[Hip]

As for BB FCF, I hoped that this study maybe would allow you to not have to guess the protein binding. But I guess the study doesnt list plasma levels?

That's right, it does not give plasma levels, so it's not a very useful pharmacokinetic study.

The most useful pharmacokinetic studies are those that give humans a drug orally, and then measure the peak drug concentration (called the Cmax) achieved in the blood plasma by taking blood samples.

But even these studies do not give the plasma protein binding, because when you take a blood sample and measure how much of the drug you have in the plasma, that measurement includes both the free drug (dissolved in the plasma), and the bound drug (bound to blood plasma proteins). So these pharmacokinetic studies usually only measure the total plasma concentration = free drug concentration + bound drug concentration.

So in order to know how much free drug is in the plasma, you need data for the percentage plasma protein binding. It is usually only the free drug that is active in the body, so that's why you need to know the free drug concentration.

It has proper studies done on it:
http://www.sciencedirect.com/science/article/pii/S0378874112007337
https://www.hindawi.com/journals/ecam/2014/389576/abs/

OK, well the first study found that putting Liu He Dan ointment on the belly of rats achieved a peak blood plasma concentration (Cmax) of 3.52 ng/ml, which if we convert to molar concentrations is 0.013 μM. However, remember that my estimate for emodin plasma protein binding is 90%, meaning that only 10% of the emodin is free in the plasma, so in fact the free emodin concentration in the plasma will be 10 times lower than the 0.013 μM figure, at only 0.0013 μM.

That does not look very promising, because the IC50 concentration we want to achieve is 0.5 μM, and you can see that the 0.0013 μM is much lower than this target.

 

[frozenborderline]

are there studies that compare the therapeutic and toxic doses of emodin?

 

[Hip]

are there studies that compare the therapeutic and toxic doses of emodin?

I've been looking at lots of in vitro antiviral studies recently, including antiviral studies on emodin. In these studies, they infect a human cell line culture in a test tube with the virus, and then see what concentration of the antiviral compound is able to inhibit viral replication by 50% in these cells. The concentration that achieves this 50% inhibition of viral replication is called the EC50 or IC50.

But at the same time, these in vitro studies will also see what concentration of the compound leads to cell damage (cytotoxicity). So in the study they work out what concentration causes cell death in 50% of the human cells in your test tube. That concentration is called the CC50.

For a good antiviral, you would want the CC50 concentration to be much higher than the EC50 concentration, so that you can get a useful antiviral effect, without causing any cell damage. Usually for a good antiviral drug, you can expect the CC50 concentration to be around 20 to 1000 times higher than the EC50.

The higher the CC50, the higher the dose of the antiviral you can safely give.

However, in the case of emodin, its CC50 is not much higher than its EC50: in a cell line composed of MRC-5 cells, one antiviral study I looked at recently found that for cytomegalovirus, the EC50 was 4.9 μM, and the CC50 was 9.8 μM. So that makes emodin a poor antiviral for cytomegalovirus, because the CC50 is too close to the EC50.

So that's how they look at toxic dose levels in in vitro studies.


You can also examine toxic dose levels via in vivo studies in animals, where you see what sort of oral doses lead to ill effects. The most crude measure of ill effects is the LD50 (lethal dose 50), which is the oral dose in milligrams that causes 50% of your batch of test animals to die. Once you know the LD50, you would want to make sure that any oral dose you take is well below the LD50 oral dose level.

 

[frozenborderline]

I've been looking at lots of in vitro antiviral studies recently, including antiviral studies on emodin. In these studies, they infect a human cell line culture in a test tube with the virus, and then see what concentration of the antiviral compound is able to inhibit viral replication by 50% in these cells. The concentration that achieves this 50% inhibition of viral replication is called the EC50 or IC50.

But at the same time, these in vitro studies will also see what concentration of the compound leads to cell damage (cytotoxicity). So in the study they work out what concentration causes cell death in 50% of the human cells in your test tube. That concentration is called the CC50.

For a good antiviral, you would want the CC50 concentration to be much higher than the EC50 concentration, so that you can get a useful antiviral effect, without causing any cell damage. Usually for a good antiviral drug, you can expect the CC50 to be around 20 to 1000 times higher than the EC50.

The higher the CC50, the higher the dose of the antiviral you can safely give.

However, in the case of emodin, its CC50 is not much higher than its EC50: in a cell line composed of MRC-5 cells, one antiviral study I looked at recently found that for cytomegalovirus, the EC50 was 4.9 μM, and the CC50 was 9.8 μM. So that makes emodin a poor antiviral, because the CC50 is too close to the EC50.

So that's how they look at toxic dose levels in in vitro studies.


You can also examine toxic dose levels via in vivo studies in animals, where you see what sort of oral doses lead to ill effects. The most crude measure of ill effects is the LD50 (lethal dose 50), which is the oral dose in milligrams that causes 50% of your batch of test animals to die. Once you know the LD50, you would want to make sure that any oral dose you take is well below the LD50 oral dose level.

so this sounds Bad--but is it possible that the toxic to therapeutic dose ratio would be different in vivo? I just tend to avoid things that don't have a good toxic to therapeutic dose ratio as I don't have the equipment to monitor my serum levels or anything.

 

[Hip]

is it possible that the toxic to therapeutic dose ratio would be different in vivo?

There should not be much difference with in vitro and in vivo, but what does make a difference is the type of cells in your human cell line. Human cell lines can be made from: kidney epithelial cells, skeletal muscle cells, lung cells, and various other types of cell.

Each type of cell will have a different CC50 value for the same substance.

But cell toxicity is not the only way a drug or compound can be dangerous; for example, if a drug dangerously lowers or increases blood pressure, or increases the risk of stomach perforation, etc, that's also a problem.


But if you are taking a drug or supplement, you don't usually get into these sort of details. You just look at the normal dosing guidelines, and be aware of the maximum safe dose, and be aware of the known side effects.

 

[frozenborderline]

There should not be much difference with in vitro and in vivo, but what does make a difference is the type of cells in your human cell line. Human cell lines can be made from: kidney epithelial cells, skeletal muscle cells, lung cells, and various other types of cell.

Each type of cell will have a different CC50 value for the same substance.

But cell toxicity is not the only way a drug or compound can be dangerous; for example, if a drug dangerously lowers or increases blood pressure, or increases the risk of stomach perforation, etc, that's also a problem.


But if you are taking a drug or supplement, you don't usually get into these sort of details. You just look at the normal dosing guidelines, and be aware of the maximum safe dose, and be aware of the known side effects.

so I imagine you'd go with whatever the lowest CC50 value is, right?

 

[Hip]

so I imagine you'd go with whatever the lowest CC50 value is, right?

Yes, I would guess so.

In the emodin antiviral studies I looked at, the CC50 was 9.8 μM in MRC-5 cells, but a higher 76 μM in HEp-2 cells in a different study.

Presumably pharmaceutical companies will test their drugs in development on a whole range of human cell lines in vitro, as well as performing animal studies, before the first human tests are conducted.

 

[necessary8]

Thanks so much @Hip, you're a great help doing all those calculations for us, I really appreciate it.

Whenever you feel like, you can also look at other antipurinergic herbs, and see if there is maybe something better than emodin to take. Idk, puerarin?

I'm also trying to find more Panx1 inhibitors (because thats the main thing I would want to shoot for - closing of Panx1 channels), but no luck so far.

 

[Hip]

Whenever you feel like, you can also look at other antipurinergic herbs, and see if there is maybe something better than emodin to take. Idk, puerarin?

All the Panx1, P2X7 and P2Y2 antagonists listed in this post that I could get data on, I calculated their oral dose in this post (click on the spoiler button to see the calculations).

The puerarin study does not give any IC50 value, so I cannot calculate the oral dose. The only thing you can do is take a safe dose of puerarin from Kudzu herb, and see if there are any benefits. There is a Kudzu root 20:1 extract for sale here.

The Rheedia longifolia study uses the methanol extract of this herb, which will contain a number of compounds, but without knowing which compounds, it's not possible to accurately calculate an oral dose. As a very rough guess calculation (assuming bioavailability = 50% and plasma protein binding = 50%), to attain a blood concentration of IC50 of 1.31 μg/ml of the methanol extract, you would need a oral dose of the methanol extract of:

Dosage in milligrams = 400,000 x C / ( B x (100 - P)) = 400,000 x 1.31 / ( 50 x (100 - 50)) = 210 mg

This uses the equation detailed in this post.

If we assume that the methanol extract of Rheedia longifolia is around half the weight of the raw herb, then that would be an oral dose of 2 x 210 = 420 mg of the Rheedia longifolia raw herb to obtain the IC50 concentration in the blood. I am not sure if this herb is available to buy though.

 

[Sushi]

Yes, it still seems to be working consistently well, and I'm still taking ½ tablet at night between dinner and bedtime (the earlier the better to help avoid the next day sedation).

I've been taking it too late--at bedtime. I'll try taking it earlier. How long after you take it do you begin to feel the sedation effect?

It's given me what feels like about a 50-60% increase in energy, which is one of the largest I've experienced,

I've also noticed a significant increase in energy in the few days I've been taking it. I really hope that the effect doesn't fizzle out.

 

[dreampop]

"Most gap junction blockers also attenuate the function of Panx1, including carbenoxolone, mefloquine and flufenamic acid. However, in contrast to connexin based gap junction channels, Panx1 channel activity can be attenuated by several groups of drugs hitherto considered very specific for other proteins. The drugs affecting Panx1 channels include several transport inhibitors, chloride channel blockers, mitochondrial inhibitors, P2X7 receptor ligands, inflammasome inhibitors and malaria drugs."

The title of this paper it the "Bizarre Pharmacology of the ATP Release Channel Pannexin".

Also, Probeneicd & Arachidonic Acid.

Which I guess makes a good point for things that can be tested in the impedence test and may be more or less effective than suramin.