Anti-purinergic therapy restores itaconate/IFNa (Dr. Phair's theory) and Dr. Prusty's proposed abnormalities in ME/CFS

[serg1942]

@serg1942 and all,



We know that the inflammation/cytokines lead to the negative symptoms and so first thought was to reduce them via inhibiting the NLRP3 inflammasome at minimum in earlier phases which would therefore stop all the subsequent downstream cascade of effects by blocking the cytokines. That is where palmitoylethanolamide, doxycycline, and colchicine which all inhibit the NLRP3 inflammasome that have shown benefits may be effective before all the major downstream issues set in. Suramin happens to be another NLRP3 inhibitor and why its helpful in inflammatory conditions including autism and likely ME/CFS in earlier phases. I have lists of dozens of them, but those also happen to have potential additional effects on other areas known to be related to ME/CFS and some have reported benefits.

Extracellular ATP can cause P2X receptors to activate the NOD-like receptor 3 (NLRP3) inflammasome and cause IL-1β and IL-18 maturation and release. https://pubmed.ncbi.nlm.nih.gov/23434541/ The NLRP3 inflammasome is present primarily in immune and inflammatory cells, including mast cells, neutrophils, and macrophages, following activation by inflammatory stimuli and is a good target to stop downstream effects that can lead to ME/CFS.

Consistently, apyrase (extracellular ATP scavenger), suramin (P2 receptor inhibitor), TNP-ATP (P2X receptor antagonist), and 5-BDBD (P2X4 inhibitor) downregulate NLRP3 expression https://www.nature.com/articles/s41401-022-00886-7

Viruses can trigger a biochemical pathway, known as the immune complement system...Complement activation then elicits secretion of both IL-1β and IL-18 via activation of the NLRP3 inflammasome https://pubmed.ncbi.nlm.nih.gov/23817414/

These articles were discussing the activation of the NLRP3 inflammasome in regard to COVID-19 which is not the same
https://www.sciencedirect.com/science/article/pii/S0024320520308651
Palmitoylethanolamide inhibits NLRP3 inflammasome expression (such as mentioned here expressed by sars-cov-2 spike protein). However it also helped with the gut barrier, restructuring the gut bacteria and short chain fatty acid profile, and inhibits STING so that was my first thought over some of the other options that appear to help from their NLRP3 inhibition which I can provide a list I started awhile ago.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8472716/ And of course sodium butyrate in later stages should help with the pem and energy as another thought I've mentioned so far in addition to inflammation and perhaps helping to restore TH1.

Regarding Dr Phairs latest video about IL-1a, which they suggested as a potential target:

This article confirms IL-1 can induce production and release of more IL-1, a process described as an autoinflammatory loop. Anakinra is a bio-engineered form of the naturally occurring interleukin-1 receptor antagonist (IL-1ra) that blocks the action of interleukin-1. It is routinely used in patients with autoimmune and inflammatory disorders and MAS. Within the IL-1 family of cytokines, several inhibitory mechanisms are in place to prevent runaway inflammation induced by IL-1α or IL-1β. The main mechanism is the IL-1 receptor antagonist (IL-1Ra), which blocks the IL-1R1 and prevents binding of IL-1α and IL-1β. In contrast to JAKinibs, anakinra will not directly block the IFN-STAT1/STAT2 pathway critical for host defense against viral infections. Third, in contrast to tocilizumab (an IL-6 inhibitor), it targets and inhibits the core mechanism in the pathogenesis of MAS, namely the hyperactive inflammasome loop https://ccforum.biomedcentral.com/articles/10.1186/s13054-020-03166-0

This one metions neutralization of IL-1α inhibits the antiviral activity of IFN-γ by 90%, whereas no inhibition of type I IFN activity was observed. Indeed, the antiviral activity of IFN-γ depends largely on the basal level of NF-κB, which is maintained by constitutively expressed IL-1α. https://www.sciencedirect.com/science/article/pii/S1568997221000227#s0070

Thanks @bob800 The paper says Several studies in inflammatory and noninflammatory illnesses have assessed the effect of IL-1 inhibition on fatigue severity, and most have found positive effects. Studies using daily anakinra injections (100 mg) found decreased fatigue within 4 weeks of treatment. To investigate the role of proinflammatory activity in CFS, we conducted a randomized, placebo controlled trial in female patients with CFS using the IL-1 receptor antagonist anakinra. All patients had severe fatigue leading to functional impairment. a decision was made to exclude patients with illness lasting more than 10 years - Median illness duration (range), mo 44 (7–109).

So its possible here that this treatment can work on earlier states of ME/CFS where the inflammation level is still currently high as would things like NLRP3 inflammation inhibition to a greater degree which would help prevent further deterioration. However in the later phase it does not appear that IL-1 inhibition alone at that point ( at least via anakinra that may not be able to reach the brain) can recover the subsequent lasting alterations that happen from the chronic inflammatory state and requires a different treatment approach as expected, which is important to note the two phases patients can be in.

Hi @datadragon,

Thank you for your message, it's really interesting.

Perhaps the IL-1 receptor is too downstream to reverse the cell danger response cascade. I prefer to inhibit the purinergic system from the very top, so that not only the inflammasome is inhibited, but also the rest of the inflammatory response, including IFNs I secretion and Janus Kinase activation. And most importantly, it does so without suppressing the immune system (This fits with the review you've quoted where it's explained that IL-1a neutralization inhibits IFN gamma activity, but not IFNI).

I have actually been taking an anti-purinergic combo for a month, including lidocaine (anti-p2x7), probenecid (anti-pannexin-1), glycyrrhizic acid (anti-pannexin-1 and anti-connexin 43) and brilliant blue FCF (anti-pannexin-1), and my IL1beta secreted by PBMCs ex vivo, after stimulation with LPS and ATP has decreased to half of the baseline level.

Another problem with the study is that it is too short. One month won't be enough to heal a long term body with CFS. I have actually experienced a hard flare from the anti-purinergic drugs for about 20 days or so, and now that I have recovered my baseline, I don't think I'll be seeing any improvement for the next months. This happens with most successful treatments for ME/CFS, such as GcMAF, LDN, antibiotics, antivirals, etc. There are cross-reactive "inflammation-producing" antibodies and memory cells which will live for months. Also, there are many intracellular infections and accumulated toxins which will have to be dealt with once the immune system starts to connect better with itsself and the CNS.

I had read about palmitoylethanolamide time ago, but I didn't remember it to inhibit the inflammasome assembly or STING expresson. I will take a look at it!

As for butyrate, yes, I like it, and that's why I am following a very strict ketogenic diet, because I think that beta-hydroxy-butyrate can behave identically to luminal butyrate in the gut.

Thanks again,
Sergio

 

[Violeta]

Just some notes I found when looking up adenosine and glutamate.


Adenosine has several functions in the central nervous system (CNS) that are critical for proper brain function. As a neuromodulator, one of the main functions of adenosine is to inhibit glutamate release via presynaptic A1 receptors in the nucleus accumbens.

Adenosine inhibits the action potential-dependent excitatory inputs to large Ih GABAergic neurons via A1 receptors and a presynaptic mechanism, but has no effect on their inhibitory inputs.

Studies have also shown that adenosine is released [3] from cortical slices and inhibits acetylcholine (ACh) release from cholinergic terminals evoked by axonal stimulation via activation of theophylline-sensitive receptors.

Adenosine and glutamate signaling are interrelated, with adenosine decreasing glutamate neurotransmission but glutamate, and receptor agonists, increasing cellular release of adenosine.

Adenosine and Glutamate Signaling in Neuron-Glial interactions: Implications in Alcoholism and Sleep Disorders​

https://www.ncbi.nlm.nih.gov/pmc/ar... are,increasing cellular release of adenosine.

 

[Thebirdman333]

Just some notes I found when looking up adenosine and glutamate.


Adenosine has several functions in the central nervous system (CNS) that are critical for proper brain function. As a neuromodulator, one of the main functions of adenosine is to inhibit glutamate release via presynaptic A1 receptors in the nucleus accumbens.

Adenosine inhibits the action potential-dependent excitatory inputs to large Ih GABAergic neurons via A1 receptors and a presynaptic mechanism, but has no effect on their inhibitory inputs.

Studies have also shown that adenosine is released [3] from cortical slices and inhibits acetylcholine (ACh) release from cholinergic terminals evoked by axonal stimulation via activation of theophylline-sensitive receptors.

Adenosine and glutamate signaling are interrelated, with adenosine decreasing glutamate neurotransmission but glutamate, and receptor agonists, increasing cellular release of adenosine.

Adenosine and Glutamate Signaling in Neuron-Glial interactions: Implications in Alcoholism and Sleep Disorders​

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3349794/#:~:text=Adenosine and glutamate signaling are,increasing cellular release of adenosine.

Probably why GABA drugs like Pregabalin, Ativan, etc. Have been known to help a large portion of us sick people.

 

[Violeta]

Hi @datadragon,

Thank you for your message, it's really interesting.

Perhaps the IL-1 receptor is too downstream to reverse the cell danger response cascade. I prefer to inhibit the purinergic system from the very top, so that not only the inflammasome is inhibited, but also the rest of the inflammatory response, including IFNs I secretion and Janus Kinase activation. And most importantly, it does so without suppressing the immune system (This fits with the review you've quoted where it's explained that IL-1a neutralization inhibits IFN gamma activity, but not IFNI).

I have actually been taking an anti-purinergic combo for a month, including lidocaine (anti-p2x7), probenecid (anti-pannexin-1), glycyrrhizic acid (anti-pannexin-1 and anti-connexin 43) and brilliant blue FCF (anti-pannexin-1), and my IL1beta secreted by PBMCs ex vivo, after stimulation with LPS and ATP has decreased to half of the baseline level.

Another problem with the study is that it is too short. One month won't be enough to heal a long term body with CFS. I have actually experienced a hard flare from the anti-purinergic drugs for about 20 days or so, and now that I have recovered my baseline, I don't think I'll be seeing any improvement for the next months. This happens with most successful treatments for ME/CFS, such as GcMAF, LDN, antibiotics, antivirals, etc. There are cross-reactive "inflammation-producing" antibodies and memory cells which will live for months. Also, there are many intracellular infections and accumulated toxins which will have to be dealt with once the immune system starts to connect better with itsself and the CNS.

I had read about palmitoylethanolamide time ago, but I didn't remember it to inhibit the inflammasome assembly or STING expresson. I will take a look at it!

As for butyrate, yes, I like it, and that's why I am following a very strict ketogenic diet, because I think that beta-hydroxy-butyrate can behave identically to luminal butyrate in the gut.

Thanks again,
Sergio

Sergio, are you still on anti-purinergics? If so, how is it going?

 

[serg1942]

Sergio, are you still on anti-purinergics? If so, how is it going?

Hello Violeta,

Apologies for the delay in my response. Yes, I've been on several anti-purinergic agents for a year now, inhibiting the Panexin I and P2X7 receptors. Symptomatically, I’m slightly better, perhaps around 20% improved. However, the most significant progress I’ve noticed is in my immune system, as for the first time, my NKCD56+ levels have increased significantly, and my NKCD57+ levels, which were always very low, are now within a more normalized range.

Additionally, I have tested negative for Lyme disease for the first time in 20 years, confirmed by three different tests: PCR Phagos, and lymphocyte transformation test. I didn't achieve this with years of ABX.

I believe the treatment is doing what I anticipated it would do—helping to lift me out of immunosuppression. However, it’s clearly not enough, as the improvement is mild.

It’s possible that I'm not inhibiting enough purinergic receptors, as we don’t fully understand which ones are most critical. Alternatively, we might need to address other aspects of the CDR that, together with the purinergic receptors, contribute to a chronic inflammatory cellular state. This could involve targeting mechanisms like the Integrated Stress Response or the Unfolded Protein Response which I am planing to address as well.

 

[SqueakyPScott]

Hello Violeta,

Apologies for the delay in my response. Yes, I've been on several anti-purinergic agents for a year now, inhibiting the Panexin I and P2X7 receptors. Symptomatically, I’m slightly better, perhaps around 20% improved. However, the most significant progress I’ve noticed is in my immune system, as for the first time, my NKCD56+ levels have increased significantly, and my NKCD57+ levels, which were always very low, are now within a more normalized range.

Additionally, I have tested negative for Lyme disease for the first time in 20 years, confirmed by three different tests: PCR Phagos, and lymphocyte transformation test. I didn't achieve this with years of ABX.

I believe the treatment is doing what I anticipated it would do—helping to lift me out of immunosuppression. However, it’s clearly not enough, as the improvement is mild.

It’s possible that I'm not inhibiting enough purinergic receptors, as we don’t fully understand which ones are most critical. Alternatively, we might need to address other aspects of the CDR that, together with the purinergic receptors, contribute to a chronic inflammatory cellular state. This could involve targeting mechanisms like the Integrated Stress Response or the Unfolded Protein Response which I am planing to address as well.

Probably going to try and use just Probenecid to inhibit Pannexin-1. Will update if anything interesting happens. I will probably be aiming at 2-3g daily depending on how I tolerate it. Things Ive tried recently are Metformin, Rapamycin, and Prodrome glia. All of these things seem to intervene somewhere down stream of CDR and have not proved useful. Hopefully Pannexin Therapeutics actually gets through their human studies soon.

 

[SqueakyPScott]

Probably going to try and use just Probenecid to inhibit Pannexin-1. Will update if anything interesting happens. I will probably be aiming at 2-3g daily depending on how I tolerate it. Things Ive tried recently are Metformin, Rapamycin, and Prodrome glia. All of these things seem to intervene somewhere down stream of CDR and have not proved useful. Hopefully Pannexin Therapeutics actually gets through their human studies soon.

Actually, I might try Spironolactone instead as I already have treatment resistant hypertension https://pmc.ncbi.nlm.nih.gov/articles/PMC5815904/

 

[serg1942]

Probably going to try and use just Probenecid to inhibit Pannexin-1. Will update if anything interesting happens. I will probably be aiming at 2-3g daily depending on how I tolerate it. Things Ive tried recently are Metformin, Rapamycin, and Prodrome glia. All of these things seem to intervene somewhere down stream of CDR and have not proved useful. Hopefully Pannexin Therapeutics actually gets through their human studies soon.

If you’ll allow me a piece of advice, at least based on my experience and that of my partner, starting with 2 or 3 grams of probenecid is too high, and I would personally recommend starting much slower. Typically, with probenecid, you should notice a relaxation effect, more fatigue, as if the parasympathetic nervous system is becoming more active, deeper and longer sleep, and you can experience this starting from 500 mg without any issues.

Probenecid is a very interesting drug as it is quite potent. It indeed inhibits pannexin and, on its own, is capable of reducing IL-1-beta levels in the blood, which I have measured myself. The main issue with it is that it significantly interferes with the detoxification of other endogenous substances and drugs, so you need to be very careful with potential interactions, as it could increase the half-life of any other medication.

Additionally, it will greatly lower your uric acid levels, and this poses a problem since uric acid is one of the body’s main antioxidants. It is very likely that this could raise your homocysteine levels, which is dangerous. To address this, I recommend following the Konynenburg's protocol, at least by taking hydroxy or methylcobalamin sublingually along with 5MTHF and folinic acid. This way, you can lower homocysteine levels, as my partner and I managed to do while taking probenecid.

In my opinion, rapamycin does indeed address a very important part of the CDR, unlike metformin. The latter mainly works by inhibiting mitochondrial complexes to elicit a response from healthy mitochondria, such as mitochondrial biogenesis, and so on. However, with our mitochondria so exhausted and in CDR, I don’t think it’s the best approach, and I don’t see it as part of the CDR strategy, at least from my humble perspective.

Best of luck with it and please tag me if you don't mind when you write an update on how probenecid works for you!

 

[serg1942]

Actually, I might try Spironolactone instead as I already have treatment resistant hypertension https://pmc.ncbi.nlm.nih.gov/articles/PMC5815904/

Sorry, I didn’t read this text. Look, I ruled out spironolactone mainly because of its antiandrogenic effect. I realized that at doses sufficient to inhibit pannexin-1, the effects were quite terrible in studies. But in any case, if you’re going to try it, please let me know as well, because I’m very curious.

Probenecid and brilliant blue FCF have a much safer profile.

 

[SqueakyPScott]

Sorry, I didn’t read this text. Look, I ruled out spironolactone mainly because of its antiandrogenic effect. I realized that at doses sufficient to inhibit pannexin-1, the effects were quite terrible in studies. But in any case, if you’re going to try it, please let me know as well, because I’m very curious.

Probenecid and brilliant blue FCF have a much safer profile

How did you come to the calculation for spironolactone? And what were the doses roughly?

 

[SqueakyPScott]

If you’ll allow me a piece of advice, at least based on my experience and that of my partner, starting with 2 or 3 grams of probenecid is too high, and I would personally recommend starting much slower. Typically, with probenecid, you should notice a relaxation effect, more fatigue, as if the parasympathetic nervous system is becoming more active, deeper and longer sleep, and you can experience this starting from 500 mg without any issues.

Probenecid is a very interesting drug as it is quite potent. It indeed inhibits pannexin and, on its own, is capable of reducing IL-1-beta levels in the blood, which I have measured myself. The main issue with it is that it significantly interferes with the detoxification of other endogenous substances and drugs, so you need to be very careful with potential interactions, as it could increase the half-life of any other medication.

Additionally, it will greatly lower your uric acid levels, and this poses a problem since uric acid is one of the body’s main antioxidants. It is very likely that this could raise your homocysteine levels, which is dangerous. To address this, I recommend following the Konynenburg's protocol, at least by taking hydroxy or methylcobalamin sublingually along with 5MTHF and folinic acid. This way, you can lower homocysteine levels, as my partner and I managed to do while taking probenecid.

In my opinion, rapamycin does indeed address a very important part of the CDR, unlike metformin. The latter mainly works by inhibiting mitochondrial complexes to elicit a response from healthy mitochondria, such as mitochondrial biogenesis, and so on. However, with our mitochondria so exhausted and in CDR, I don’t think it’s the best approach, and I don’t see it as part of the CDR strategy, at least from my humble perspective.

Best of luck with it and please tag me if you don't mind when you write an update on how probenecid works for you!

My reasoning for taking Metformin was for this study which found that eATP was limited by Metformin.
https://researchfeatures.com/metfor...ct-diabetic-glucose-levels-finally-uncovered/
I also have treatment resistant bipolar type 1 with rare mania and this study found a large benefit measured by GAF with those who have insulin resistance who responded
https://www.psychiatrist.com/jcp/tr...t-resistant-bipolar-depression-trio-bd-study/
Unfortunately I was not a responder.

I messed up my calculations for Probenecid. https://pubmed.ncbi.nlm.nih.gov/7175716/ this study claims to reach a much high ug/ml than I had thought previously.

I was under the impression that Spironolactone is a more potent inhibitor of pannexin-1 am I mistaken? However it comes with its own downsides such as lowering androgens but might help me considering I have bipolar.
https://pmc.ncbi.nlm.nih.gov/articles/PMC5815904/
https://onlinelibrary.wiley.com/doi/pdfdirect/10.1002/brb3.3313.

Please do correct me where I am wrong I'm pretty new to this. (looking at studies and research not cfs).

 

[serg1942]

Any substance that reduces inflammation or free radicals will secondarily reduce extracellular ATP, as they always go hand in hand. There is a misconception: many antioxidant herbs are considered antipurinergic because they either decrease extracellular ATP or reduce the expression of purinergic receptors. However, this does not mean they are truly antipurinergic, and in general, there will not be a significant clinical effect. For this reason, inhibiting mitochondrial complexes with metmorfine, for instance, does not have an antipurinergic effect per se.

Regarding bipolar disorder, I strongly recommend that you listen to and read Dr. Georgia Ede, who, with a highly ketogenic carnivore diet, manages to eliminate symptoms in 50–70% of people with bipolar disorder and significantly reduce medication and symptoms in the rest. She has books, a blog with study reviews, and I have personally seen this with my aunt, who also suffers from this disorder.

If I remember correctly, neither Probenecid nor spironolactone have a very potent antipurinergic effect, with EC50 values around 100 micromolar. (This is from memory; if you're interested, I can look up the source.) However, given that Probenecid is a fairly safe drug and can be taken at doses between 500 mg and 2 g, antipurinergic doses are easily achieved. As for spironolactone, it is also not very potent, and reaching antipurinergic doses would involve significant side effects.

In the study you sent me, I do not interpret that inhibiting mineralcorticoides or androgen receptors helps with bipolar disorder. Rather, the benefit seems to come from reducing inflammation, regulating the HPA axis, and increasing brain-derived neurotrophic factor (BDNF), which I attribute to the antipurinergic effect. You can find similar results in many other studies.

On the other hand, let me suggest two more options that you might consider trying, as they are relatively simple. First, there is the food dye Brilliant Blue FCF, which you can purchase at 99% purity if you live in the U.S. Although oral absorption in mice is low, around 5%, as we're talking about EC50 of only a few nanograms, so the small amount absorbed is usually enough for a clinical effect. In fact, the maximum recommended dose is around 500 mg per day, and with just 50–100 mg, I noticed a strong clinical effect. I stopped using it because it caused mild MCAS in my case, but it is a very inexpensive and effective treatment.

Alternatively, you could make a lidocaine cream. If you'd like, I can teach you how to prepare it. I developed it based on a COVID study where subcutaneous lidocaine infusions were used for 24 hours. This lidocaine was directly absorbed by the lymph nodes, reaching sufficient concentrations to stimulate the immune system and subsequently travel throughout the body. It’s a relatively easy approach to implement at home with a blender and the necessary ingredients.

I wish you the best of luck on your journey with antipurinergics.

 

[SqueakyPScott]

Any substance that reduces inflammation or free radicals will secondarily reduce extracellular ATP, as they always go hand in hand. There is a misconception: many antioxidant herbs are considered antipurinergic because they either decrease extracellular ATP or reduce the expression of purinergic receptors. However, this does not mean they are truly antipurinergic, and in general, there will not be a significant clinical effect. For this reason, inhibiting mitochondrial complexes with metmorfine, for instance, does not have an antipurinergic effect per se.

Regarding bipolar disorder, I strongly recommend that you listen to and read Dr. Georgia Ede, who, with a highly ketogenic carnivore diet, manages to eliminate symptoms in 50–70% of people with bipolar disorder and significantly reduce medication and symptoms in the rest. She has books, a blog with study reviews, and I have personally seen this with my aunt, who also suffers from this disorder.

If I remember correctly, neither Probenecid nor spironolactone have a very potent antipurinergic effect, with EC50 values around 100 micromolar. (This is from memory; if you're interested, I can look up the source.) However, given that Probenecid is a fairly safe drug and can be taken at doses between 500 mg and 2 g, antipurinergic doses are easily achieved. As for spironolactone, it is also not very potent, and reaching antipurinergic doses would involve significant side effects.

In the study you sent me, I do not interpret that inhibiting mineralcorticoides or androgen receptors helps with bipolar disorder. Rather, the benefit seems to come from reducing inflammation, regulating the HPA axis, and increasing brain-derived neurotrophic factor (BDNF), which I attribute to the antipurinergic effect. You can find similar results in many other studies.

On the other hand, let me suggest two more options that you might consider trying, as they are relatively simple. First, there is the food dye Brilliant Blue FCF, which you can purchase at 99% purity if you live in the U.S. Although oral absorption in mice is low, around 5%, as we're talking about EC50 of only a few nanograms, so the small amount absorbed is usually enough for a clinical effect. In fact, the maximum recommended dose is around 500 mg per day, and with just 50–100 mg, I noticed a strong clinical effect. I stopped using it because it caused mild MCAS in my case, but it is a very inexpensive and effective treatment.

Alternatively, you could make a lidocaine cream. If you'd like, I can teach you how to prepare it. I developed it based on a COVID study where subcutaneous lidocaine infusions were used for 24 hours. This lidocaine was directly absorbed by the lymph nodes, reaching sufficient concentrations to stimulate the immune system and subsequently travel throughout the body. It’s a relatively easy approach to implement at home with a blender and the necessary ingredients.

I wish you the best of luck on your journey with antipurinergics.

I have tried strict keto and carnivore diets multiple times without significant benefits. Benefits I got lasted a couple weeks and after I was back to where I was pre-keto unfortunately.

 

[SqueakyPScott]

I have tried strict keto and carnivore diets multiple times without significant benefits. Benefits I got lasted a couple weeks and after I was back to where I was pre-keto unfortunately.

I should also mention that I did them for more than a few months. The longest keto I did was 15 mo.