Experiences with Succinic Acid ( Amber Acid )

[joshua.leisk]

The first line of this article explains that the Mitochondrial Dicarboxylate Carrier operates on the inner mitochondrial membrane.

Succinate cannot reach this transporter if it is not cell-permeable.

The paper I cited previously shows that succinate is not cell-permeable with experimental evidence: https://www.nature.com/articles/ncomms12317

As Hip explained, it's possible that succinate has other actions throughout the body due to the widespread presence of succinic acid receptors. I don't think it's fruitful to continue discussions about succinic acid entering cells in significant quantities when all literature indicates otherwise.

You've cited one study. Can it be reliably repeated?
Here's a contrasting study:
https://www.hindawi.com/journals/sci/2020/2016809/
and another:
https://www.haematologica.org/article/view/8615

SLC-13 appears to be the transporter:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1866268/
https://pubmed.ncbi.nlm.nih.gov/23506872/
https://link.springer.com/article/10.1007/s00424-013-1369-y
https://link.springer.com/article/10.1007/s00424-003-1128-6

As succinate can be released into the extracellular space from the cytoplasm, succinate may also enter into the cytosol. We then determined levels of intracellular and extracellular succinate after treating hPDLCs with succinate. Succinate concentration in the cytosol climbed in an early phase of succinate stimulation, peaked at 6 h, and then declined later on (Figure 4(a)). In contrast, succinate concentration in the extracellular space decreased over time in succinate-treated cells (Figure 4(b)). In line with elevated succinate in the cytosol, the protein level of HIF-1α was increased, whereas the level of PHD was reduced (Figure 4(c)). Then, changes in glycolysis-related genes were analyzed. Both HK2 and PFKFB3 were elevated with the succinate supplement (Figures 4(d) and 4(e)). Furthermore, it was observed that SDH transcription elevated at 4 h while the SDH protein level diminished in succinate-supplemented hPDLCs (Figures 4(d) and 4(e)). Additionally, reduced SDH enzyme activity was found in 5 mM succinate-treated cells (Figure 4(f)). Therefore, our present data suggest that exogenous succinate can be transferred into the cytosol, where it induced PHD suppression, leading to a pseudohypoxia condition and HIF-1α stabilization. Therefore, such HIF-1α pathway activation was crucial in succinate supplement-induced stem cell proliferation.

We've seen exogenous succinate reliably resolve hypoxia numerous times and presented as additional supporting evidence, has been successfully used to "rescue" 2 people who performed the GDH-knockdown fast and then (ignoring advice) consumed food during the GDH-knockdown period, leading to the state described in the third paper.
One other person who also ignored the advice on not eating was unable to reach his succinate due to repeatedly blacking out every few seconds due to loss of mitochondrial energy... for hours... until the EGCG wore off. This is why I put the large disclaimers in the third paper - if someone chose to ignore this advice and eg. drive a car in this state, while eating food, I could see this easily being fatal.

I have no doubts that further lab experiments will explain any gaps in the limited published literature on this.

50g of glucose contains 200 calories.

50mg of succinic acid is certainly not equivalent to 200 calories, and my 500mg dose was not equivalent to 2000 calories.
This is a physically impossible claim. No substance can be that calorie-dense.

I was talking subjectively and really this observation was based around PEM load.

50mg of succinic acid can induce roughly the same amount of PEM as about 50g of glucose. The a-KGDH bypass effect is likely a confounding factor in this observation and overall, it needs to be properly studied. The anaplerosis would also likely skew the lactateyruvate ratio, in the same way the glutaminolysis does.

As a dietary input, I don't believe anyone has quantified the calorific density of these organic acids in the literature. I haven't looked into the specifics yet either, this is only by general indications / observations.

I suppose it'll need to be done at some stage, so I guess I'll give it a brief spit-ball now.. Now this is currently straying slightly outside of my normal comfort zone, however I'll go out on a limb briefly, at least until I've better studied, digested and understood this. Maybe some input from others will help:

At the basic level, we're talking about chemical reactions that involve electrons, electron chain transport, hydrogen and the formation of ATP.. Great.. so let's look into the thermochemistry..

What's the energy density of hydrogen?
...It's probably the most energy dense thing I'm aware of, at >120-140kJ per g.
https://www.sciencedirect.com/science/article/pii/S1002007116303240
https://en.wikipedia.org/wiki/Energy_density

Succinic acid is C4H6O4
https://pubchem.ncbi.nlm.nih.gov/compound/Succinate_1#section=Structures

By weight, succinic acid is very close to 5% hydrogen, 40% carbon and 55% oxygen.
https://www.lenntech.com/calculators/molecular/molecular-weight-calculator.htm

So at this point, seems like a suitably energy dense compound.
https://www.sciencedirect.com/science/article/abs/pii/0021961472900754
https://royalsocietypublishing.org/doi/abs/10.1098/rsta.1955.0008

So, as a direct calorific measurement, from these musings, it appears to be less than glucose. Measured this way, it's roughly 3 Kcal/gram, however this really doesn't describe the relationship to avoiding all of the processes involved in glucose metabolism, glycolysis and fatty acid oxidation before it reaches the citric acid cycle.

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3345970/

Chemically, succinic acid sits directly before/after the "primary" reversible reaction which generates ATP:
http://vmh.uni.lu/minerva/index.xht...earchCoordinates=34269,30094,5974,10&zoom=10&

There is one reversible reaction to/from succinyl-coa:
https://en.wikipedia.org/wiki/Succinyl_coenzyme_A_synthetase

Overall, I believe there is a significantly underappreciated value to succinic acid, which I'll be detailing further in future papers.

 

[joshua.leisk]

Going back to what was said about it not being able to get into the cells, it obviously can do otherwise it would not be used as a treatment for mitochondrial disease https://www.medicinesforchildren.or...content-type/leaflet/pdf/20111209192733_0.pdf

I have decided not to take any more of this substance, it is very powerful ! it seems a bit like our equivalent of cocaine or meth amphetamine, gets you going but probably not good for the body in the long run. ( never took those illegal drugs just guessing what its like ).

It has specific uses, but care definitely needs to be taken. You'd consume a more reasonable / appropriate dose in Apple Cider Vinegar.

 

[joshua.leisk]

That website is the only one I could find which suggests succinate may help mitochondrial diseases. I cannot find anything else online which promotes succinate supplementation for mito diseases. It may be just wishful thinking that this supplement can into mitochondria.

Obviously it would good for those with mitochondrial diseases if we could get succinate into mitochondria, and that's why researchers are attempting to create pro-drug forms of succinate, such as:

NV118 = diacetoxymethyl succinate
NV189 = bis-(1-acetoxy-ethyl) succinate
NV241 = 1-acetoxyethyl acetoxymethyl succinate

@mitoMAN may be interested in these prodrug forms of succinate. Whether they might help ME/CFS though is another question.

see the study I linked here

 

[joshua.leisk]

They don't join a patients forum and provide step by step details on what substances to take, because they know that would be irresponsible.

I'd argue that it'd be irresponsible not to.

Chronically sick people are going to jump in and try anything that may/may not help/hurt them. Clarifying any questions to prevent someone from misinterpreting the information and doing something avoidable does take time away from my other work, but I see this as extremely helpful to everyone.

 

[joshua.leisk]

I also wonder, why is the only available amber acid from this Russian brand? Is it common in Russia to take it?

It's readily available as a raw food supplement and as a lab reagent. eg. I bought 2kg for about $20.

 

[joshua.leisk]

Sorry to be iff topic, but would you tell me how much Acetic acid is in a 1200 mg tablet of ACV?

Depends on the apples, brand, etc.
This inconsistency and lack of clear labeling / product information was why we later shifted to NAC.

 

[joshua.leisk]

@Reading_Steiner what dose of succinic acid did you trial?

Succinic acid is found in significant quantities in apple cider drinks and wine, among other things. Wine typically contains 0.5g/L to 1.5g/L, though some red wines have been observed with up to 3.0g/L. (Source https://scholar.sun.ac.za/handle/10019.1/4228 )

A typical glass of wine is 5oz, which translates to between 74mg and 222mg of succinic acid in an average glass of wine, or up to 444mg in certain red wines.

As much as some people would enjoy learning that CFS can be put into remission by drinking red wine, it doesn't appear that red wine has any such effects on CFS despite the relatively high succinic acid content. Similarly, commonly used doses of Apple Cider Vinegar contain small amounts of succinic acid in the range that supposedly produces these effects, yet none of the previous Apple Cider Vinegar reports on this forum or elsewhere on the internet suggest such dramatic results.



I trialed everything from small doses (1/4 of a 100mg tablet) up to the standard 500mg dose with no noticeable effects, and no reduction of delayed PEM after an exercise challenge. Previous reports on this forum also did not notice any effects from succinic acid. I would be interested to hear experiences from others so we can collect some more data points, but given the prevalence of low level succinic acid consumption in regular diet I'm skeptical that these 25mg doses provide such profound results.

If you read the other 2 papers I've written, you'll understand what succinic acid actually does in HHV infections, why it cannot put CFS into remission by itself (it merely provides specific symptomatic relief by bypassing the a-KGDH impairment, resolving hypoxia, HIF), why small doses are useful and conversely why high doses create CFS symptoms instead of resolving them.

I can't offer specific medical advice here, however according to our model and experiences, if someone with hepatic infection of HHV simultaneously resolves their ROS issues using eg. R-ALA and then "pulses" succinic acid to alter HIF factors, rather than overdosing on it, they'd be expected to put their symptoms into remission, in just the same way that some of the v2.x self-testers found.

 

[pattismith]

Depends on the apples, brand, etc.
This inconsistency and lack of clear labeling / product information was why we later shifted to NAC.

do you mean that you use NAC for some other thing than bringing cysteine for glutathione repletion?

 

[joshua.leisk]

Some very important context for this discussion: Joshua Leisk has filed for 4 patents on his protocols which he intends to commercialize. He mentions this in his first document and also on this forum. The patent filings can be found through the Australian Patent Search portal by searching for his name, although the contents of the patent filings are currently hidden.

Without seeing the patents we cannot know what is claimed. However, this does introduce a potential notable conflict of interest in this discussion: If he publicly acknowledges that any of his specific claims in the patents are flawed or incorrect, that could put the value of his patents in jeopardy.

All of my research agrees with everything Hip has explained in great detail already: It does not appear feasible that the low dose succinic acid in these protocols would enter mitochondria in sufficient quantities to make a difference. We have found multiple documents with experimental evidence that contradict the claim that succinic acid is cell permeable. Unfortunately, acknowledging this contradictory evidence would put the author's patent filings and eventual profits at risk.

I suggest we all keep this potential conflict of interest in mind when considering the author's claims.

I'd really appreciate it if you stopped "attacking the messenger" and instead put your time into reading my 3 papers. Perhaps you'll have a better appreciation for what I'm actually doing here, as it's exceedingly clear that you're basing your comments on having somewhat read the first paper, only.

Always happy to take any/all constructive criticism for any of my work presented, but your continuing obsession with who I am is bordering on breaking Rule 1.

I'm here to help people understand my research and I'm happy to do this, however it also takes time away from family and further research. This time is valuable.

What I have patented doesn't affect researchers, or anyone here - it affects any pharmaceutical companies or supplement companies looking to take my work and generate revenue from it, which has happened in the past.

Given the scope of application and the potential impact to existing pharmaceutical revenues, it seems like something that you'd regret not patenting.

 

[joshua.leisk]

do you mean that you use NAC for some other thing than bringing cysteine for glutathione repletion?

Oh definitely!
It's to replenish the acetyl-CoA being depleted by excretion as phenylacetylglutamine.

This is discussed in the third paper - https://www.researchgate.net/publication/350956432_CFSME_A_New_Hope

I've tried to elaborate further on this here - https://forums.phoenixrising.me/thr...toimmune-spectrum-disorder.83371/post-2335821

 

[godlovesatrier]

Alcohols always made me worse not better. It makes my neurological symptoms worse which is one of my primary symptoms after PEM. Dizziness basically. Comes and goes when I've got it under control. I was primarily a red wine drinker before ME.

 

[godlovesatrier]

Josh. How does putting yourself in remission but not treating the illnesss make the disease worse? In my experience when I do nothing is when things decline but when I actively stop the viral replication with something like andrographis paniculata or ginseng or a combination of alcohol tinctures. Things usually stop after a few weeks of taking them 3x a day. I say stop this isn't remission it just stops me getting worse.

Thoughts?

 

[joshua.leisk]

Josh. How does putting yourself in remission but not treating the illnesss make the disease worse? In my experience when I do nothing is when things decline but when I actively stop the viral replication with something like andrographis paniculata or ginseng or a combination of alcohol tinctures. Things usually stop after a few weeks of taking them 3x a day. I say stop this isn't remission it just stops me getting worse.

Thoughts?

So in the third paper, we described how anything you do to put energy into the mitochondrial reactions, or fix any of the metabolites being lost in urine will give energy and resources to the protein synthesis tasks being carried out by the infected cells.

These tasks can be somewhat managed by a late-stage replication intervention (spironolactone, tenofovir).. in fact, our early data showed a reduction of latent cell burden, over a few months, when combining this combination with a HDAC inhibitor to provoke latent cells into lytic phase, allowing a functioning immune system to detect and replace that cell.

(25% reduction as inferred by EBNA IgG)

This was great progress, but appears much slower than what we're seeing now with the v3.x protocol.

 

[joshua.leisk]

Alcohols always made me worse not better. It makes my neurological symptoms worse which is one of my primary symptoms after PEM. Dizziness basically. Comes and goes when I've got it under control. I was primarily a red wine drinker before ME.

I've seen alcohol free red wine provide temporary remission in 1 person.. It was a really cheap one and somewhat closer to 'vinegar' than wine.

 

[Reading_Steiner]

Josh. How does putting yourself in remission but not treating the illnesss make the disease worse? In my experience when I do nothing is when things decline but when I actively stop the viral replication with something like andrographis paniculata or ginseng or a combination of alcohol tinctures. Things usually stop after a few weeks of taking them 3x a day. I say stop this isn't remission it just stops me getting worse.

Thoughts?

same reason we don't catch cold I guess ? metabolic downshift. no need to go into the complicated science at first if one can reduce a problem to its simplest form, I speculate this could be why Joshua is succeeding in ways that more famous scientists have not.

One other person who also ignored the advice on not eating was unable to reach his succinate due to repeatedly blacking out every few seconds due to loss of mitochondrial energy... for hours... until the EGCG wore off. This is why I put the large disclaimers in the third paper - if someone chose to ignore this advice and eg. drive a car in this state, while eating food, I could see this easily being fatal.

That gave me some mad scientist vibes, probably not what you are going for but you have potential to be a good 'mad scientist'.

 

[joshua.leisk]

same reason we don't catch cold I guess ? metabolic downshift. no need to go into the complicated science at first if one can reduce a problem to its simplest form, I speculate this could be why Joshua is succeeding in ways that more famous scientists have not.


That gave me some mad scientist vibes, probably not what you are going for but you have potential to be a good 'mad scientist'.

View attachment 42766

Nikola Tesla is definitely someone I’ve enjoyed trying to understand. I actually have hard-copies of all his .. available.. written works, patents.

I think I have a reasonable grip on many of his ideas. Biefeld-Brown, Townsend have done some interesting work around this as well.

Funnily enough, on the “mad scientist” topic, there are some outstanding projects up my sleeve around this from my electronics engineering focused days that I intend to get back to one day. Electrogravitics, using the resonance + capacitance of the Earth for large scale applications and transmission purposes still seems like quite a fun area to apply myself again one day.

 

[godlovesatrier]

Hmm you see I have a down regulated immune system, mine has never been hyper active. I just don't fit that commonly well known ME subset. I've always been able to get sick, in fact the incidence of illness post ME went up. In the first few years every illness just ran one into the next. It was a never ending barrage of colds and viruses that my body never fully recovered from. Over the years I am now in a place where a cold (usually it's a cold much less a virus) takes 2-4 weeks for my immune system to recover from without help. I know this because my wound healing in my mouth drops for about 2-3 weeks and my general energy levels are very low + air hunger and general malaise is worse.

So in my case I am not sure that answers the question.

 

[joshua.leisk]

Hmm you see I have a down regulated immune system, mine has never been hyper active. I just don't fit that commonly well known ME subset. I've always been able to get sick, in fact the incidence of illness post ME went up. In the first few years every illness just ran one into the next. It was a never ending barrage of colds and viruses that my body never fully recovered from. Over the years I am now in a place where a cold (usually it's a cold much less a virus) takes 2-4 weeks for my immune system to recover from without help. I know this because my wound healing in my mouth drops for about 2-3 weeks and my general energy levels are very low + air hunger and general malaise is worse.

So in my case I am not sure that answers the question.

When I help people, we usually include (Labcorp):

T- and B-Lymphocyte and Natural Killer Cell Profile

TEST: 505370

Test number copied​

CPT: 86355; 86357; 86359; 86360

 

[godlovesatrier]

I was looking into this last week, do you know of anyone in the UK that does it?

 

[Dubitat]

You've cited one study. Can it be reliably repeated?
Here's a contrasting study:
https://www.hindawi.com/journals/sci/2020/2016809/
and another:
https://www.haematologica.org/article/view/8615

SLC-13 appears to be the transporter:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1866268/
https://pubmed.ncbi.nlm.nih.gov/23506872/
https://link.springer.com/article/10.1007/s00424-013-1369-y
https://link.springer.com/article/10.1007/s00424-003-1128-6

I'm sorry to say that these studies don't support your argument, as both Hip and I have been carefully trying to explain.

The first paper you cited is an in vitro study which used succinic acid concentrations up to 25mM. Hip previously calculated the approximate succinic acid dose required to reach such concentrations on page 1 of this thread:

However, the concentration of succinate they used in vitro was extremely high, 6 to 24 mM (6 mM equates to 0.7 mg/ml). This concentration resulted in an increase in the oxygen consumption rate of mitochondria by 20%.

If you wanted to achieve the same concentration in the body, the oral succinate dose needed to attain this concentration of 0.7 mg/ml (in the 40,000 ml of body fluids) we can calculate as 57 grams! That's assuming 100% bioavailability and zero plasma protein binding.

If 6mM concentration requires 57,000mg doses of succinic acid, then the 25mM concentration used in the paper you cited would require 237,500mg doses. These doses are thousands of times higher than the doses used in your protocol, and far higher than any safe dose that a person could take.

Many compounds can overwhelm natural balance and enter cells when applied at unnaturally high concentrations in a petri dish. However, those studies cannot be assumed to apply to doses that are 4 orders of magnitude lower.

Multiple studies in humans show that the typical range of succinic acid in the blood is between 2uM and 20uM, which is nowhere near the up to 25,000uM concentrations used in the study you cited. This is noted in your second link above.



Second, your citations about the SLC-13 transporter also disagree with your claims. As we've been explaining, the transporters you're citing are used for moving succinate out of the mitochondria, not into them.

The second link you provided has a diagram and a section which explains this:

Mitochondria are the physiological source for succinate, but accumulated succinate may be transported to the cytosol through the dicarboxylic acid translocator in the mitochondrial inner membrane and the voltage-dependent anion channel in the outer membrane (Figure 1)

and

Interestingly, succinate connects intracellular metabolic status and intercellular communication, as it may be released to the extracellular space through plasma membrane transporters of the SLC13 family (Figure 1)

The SLC13 transporter transports succinate out of the cell, not into the cell. The dicarboxylic acid translocator transports succinate out of the mitochondria, not into the mitochondria. This is information I'm taking from the exact citations you provided above.



We've been patiently reading your writings, your linked studies, and your citations in multiple threads and responding with valid concerns. At this point even your own citations are not supporting your claims, as I've outlined above. I hope that others reading this thread can take the time to read some of the careful critiques of the claims being made here.