591 replies to this topic

[Daniel Cooper]

This test was in vitro.  They cultured opossum kidney cells in some sort of buffered isotonic saline and then introduced them to a 500nmol solution of MitoQ.

 

For this test to have any meaning, you'd have to know what sort of concentration taking a normal dose of MitoQ (250mg) would induce in vivo in a human. I don't know what that number is, but 500nmol sounds on the high side of what might be realistic.  I would guess that you might see something in the tens of nmol.  But that's just a guess.

 

The fact of the matter is that if you put cells in any sort of sufficiently concentrated polar solution, the mitochondria will swell and depolarize.  

 

The right question for Greg would be "what plasma concentration of MitoQ have you seen as a peak after a standard dose?".  

 

Perhaps someone could invite Greg over to this thread.

 

 

[Daniel Cooper]

FYI - Sent an invite to Greg.

[smithx]

Here is the entire abstract, bolding mine. The depolarization and swelling was identified by the researchers to be due to the structure of the MitoQ, and another mitochondrial antioxidant didn't cause the problem even at a very high dose.

Kidney proximal tubules (PTs) contain a high density of mitochondria, which are required to generate ATP to power solute transport. Mitochondrial dysfunction is implicated in the pathogenesis of numerous kidney diseases. Damaged mitochondria are thought to produce excess reactive oxygen species (ROS), which can lead to oxidative stress and activation of cell death pathways. MitoQ is a mitochondrial targeted anti‐oxidant that has shown promise in preclinical models of renal diseases. However, recent studies in nonkidney cells have suggested that MitoQ might also have adverse effects. Here, using a live imaging approach, and both in vitro and ex vivo models, we show that MitoQ induces rapid swelling and depolarization of mitochondria in PT cells, but these effects were not observed with SS‐31, another targeted anti‐oxidant. MitoQ consists of a lipophilic cation (Tetraphenylphosphonium [TPP]) joined to an anti‐oxidant component (quinone) by a 10‐carbon alkyl chain, which is thought to insert into the inner mitochondrial membrane (IMM). We found that mitochondrial swelling and depolarization was also induced by dodecyltriphenylphosphomium (DTPP), which consists of TPP and the alkyl chain, but not by TPP alone. Surprisingly, MitoQ‐induced mitochondrial swelling occurred in the absence of a decrease in oxygen consumption rate. We also found that DTPP directly increased the permeability of artificial liposomes with a cardiolipin content similar to that of the IMM. In summary, MitoQ causes mitochondrial swelling and depolarization in PT cells by a mechanism unrelated to anti‐oxidant activity, most likely because of increased IMM permeability due to insertion of the alkyl chain.

 
Plasma concentrations are not relevant, as pointed out by the researchers, because MitoQ concentrates in the mitochondria and stays there:
 

However, since MitoQ accumulates into mitochondria at very high concentrations, the possibility remains that it could have effects on mitochondrial function other than on ROS levels, not all of which might be beneficial. For example, a recent study has reported that MitoQ can actually increase ROS production in some cancer cells, and this is associated with a decrease in Δψ _m and mitochondrial DNA (mtDNA) copy number (Pokrzywinski et al. 2016).

 
The structure of the molecule seems to be at fault:
 

Since SS‐31 did not cause the same mitochondrial swelling as MitoQ, even at a very high dose, we considered that the toxic effects observed with the latter are probably unrelated to its anti‐oxidant activity. To investigate this, we assessed the individual effects of different structural components of MitoQ. We found that TPP (500 nmol/L), the mitochondrial targeting cation of MitoQ, did not induce any acute deleterious effects on mitochondria in OK cells (Fig. 2A–B). However, dodecyltriphenylphosphomium (DTPP) (500 nmol/L), which consists of TPP plus the carbon alkyl chain (but lacks the anti‐oxidant quinone), caused acute mitochondrial swelling and depolarization, identical to that induced by MitoQ (Fig. 2A–B). These findings suggest that the toxic effect of MitoQ on PT cell mitochondria is indeed unrelated to anti‐oxidant activity, and that the carbon alkyl chain plays an important role.

 

They used the same solutions for MitoQ, for SS-31 and for the structural analog of MitoQ and found the depolarization and swelling issue only with MitoQ and its structural analog. So there was clearly no problem which can be blamed on a concentrated polar solution.

 

To me, this looks very bad for MitoQ, and is a compelling argument for avoiding it entirely.

 

 

This test was in vitro.  They cultured opossum kidney cells in some sort of buffered isotonic saline and then introduced them to a 500nmol solution of MitoQ.
 
For this test to have any meaning, you'd have to know what sort of concentration taking a normal dose of MitoQ (250mg) would induce in vivo in a human. I don't know what that number is, but 500nmol sounds on the high side of what might be realistic.  I would guess that you might see something in the tens of nmol.  But that's just a guess.
 
The fact of the matter is that if you put cells in any sort of sufficiently concentrated polar solution, the mitochondria will swell and depolarize.  
 
The right question for Greg would be "what plasma concentration of MitoQ have you seen as a peak after a standard dose?".  
 
Perhaps someone could invite Greg over to this thread.

Edited by smithx, 13 June 2019 - 06:54 PM.

[Daniel Cooper]

You're making many leaps in your logic there.  Plasma concentrations are certainly relevant. That paper absolutely does not say that concentrations are irrelevant.  

 

Keep in mind, there are studies out there testing MitoQ in vivo in human subjects at doses of 1200mg/day and 3600mg/day that noted no deleterious effects.  You're touting a in vitro study using opossum kidney cells and neither one of us knows if the plasma molarities have any bearing whatsoever on what you'll actually see in a human kidney.  And you are concluding that this takes MitoQ completely off the table as a dangerous substance.  In spite of the fact that we've had humans taking 14x the standard dose and no one measured any damage to their kidneys in the labs.

 

Using your criteria I think I'd have to swear off salt as well.  That stuff will kill you if you get too much of it.

 

Let's find out what the pharmacokinetics of this substance is.  What plasma concentrations you might expect and for how long.  The benefits of MitoQ are too intriguing to dismiss it off this one study.  Unless you're a possum perhaps.

 

 

And btw, MitoQ doesn't enter the mitochondria and "stay there".  If it did it would be useless.  It mops up the ROS generated by energy production in the mitochondria and is thus being continually consumed.

 

 

[Benko]

  ***You're touting a in vitro study using opossum kidney cells***

 

[smithx]

I'm not making many leaps in logic. I'm quoting the logic of the researchers in the cited paper.

 

MitoQ is preferentially taken up by the mitochondria and attains orders of magnitude higher concentrations there than in plasma. So plasma levels are not particularly germane: intra-mitochondrial concentrations are what is germane.

 

Even for drugs like antibiotics, plasma concentrations are not solely important. For example, many drugs which treat kidney or urinary tract infections may not achieve a minimum inhibitory concentration in blood plasma but are preferentially concentrated in urine, which is what makes them effective in those applications.

 

The researchers showed that MitoQ and its structural analog caused mitochondrial depolarization and swelling, and the solutions they used did not, and neither did far higher concentrations of a different mitochondrial targeted antioxidant. This seems like a compelling result to me, and is notably not something you've acknowledged or referred to at all in your comments, which seem to be an attempt to trivialize these findings rather than address them seriously. I find your responses to be odd and disingenuous.

 

My reason for taking MitoQ was that it was potentially able to produce incremental benefits. This research suggests that it could, instead, cause harm. I'd rather be safe than sorry in this case. If the potential benefits were an order of magnitude higher, I might be more willing to take some bigger risks.

 

 

You're making many leaps in your logic there.  Plasma concentrations are certainly relevant. That paper absolutely does not say that concentrations are irrelevant.  

 

Keep in mind, there are studies out there testing MitoQ in vivo in human subjects at doses of 1200mg/day and 3600mg/day that noted no deleterious effects.  You're touting a in vitro study using opossum kidney cells and neither one of us knows if the plasma molarities have any bearing whatsoever on what you'll actually see in a human kidney.  And you are concluding that this takes MitoQ completely off the table as a dangerous substance.  In spite of the fact that we've had humans taking 14x the standard dose and no one measured any damage to their kidneys in the labs.

 

Using your criteria I think I'd have to swear off salt as well.  That stuff will kill you if you get too much of it.

 

Let's find out what the pharmacokinetics of this substance is.  What plasma concentrations you might expect and for how long.  The benefits of MitoQ are too intriguing to dismiss it off this one study.  Unless you're a possum perhaps.

 

 

And btw, MitoQ doesn't enter the mitochondria and "stay there".  If it did it would be useless.  It mops up the ROS generated by energy production in the mitochondria and is thus being continually consumed.

 

Edited by smithx, 14 June 2019 - 07:55 AM.

[Daniel Cooper]

Plasma concentrations are not solely important, but they are relevant (you'll recall your saying they are not relevant)

 

MitoQ is being consumed in the mitochondria mopping up ROSs.  If the concentration is such that it is entering faster than it is being consumed, obviously you will see a buildup as time goes forward.  How fast it is accumulating and for how long is important.  Obviously there is an integration occurring here. 

 

Since we put in MitoQ as essentially a pulse the peak plasma concentration and the duration will determine to what extent it accumulates. 

 

And I keep coming back to the fact that we have in vivo human subjects that have taken substantially high doses (up to 3,600mg) of this compound in real phase 1 and 2 trials that looked fairly intensely at safety and did not see anything concerning.  Recall that MitoQ originally went down a drug development path, not a supplement path.  It failed to pass muster not because of any safety issues, but because they could't show efficacy for their target disease (at that time, Parkinson's). 

 

I'm not saying to discount this in vitro paper at all.  This issue should obviously be looked at.  But in vitro will frequently give different results from in vivo both in the positive and negative direction.  Ultimately though, it's how the compound performs in the target organism that matters. I'm just suggesting that we look at this issue coolly and not be alarmist over this.

 

 

[aribadabar]

Guys,

 

I also wondered how meaningful 500 nmol concentration is when the study first came out and here is QfL's calculation for your consideration.

Based on it, the observed deleterious effect looks fairly plausible possibility to also occur to some extent in humans, in vivo.

Edited by aribadabar, 16 June 2019 - 03:50 AM.

[Daniel Cooper]

Guys,

 

I also wondered how meaningful 500 nmol concentration is when the study first came out and here is QfL's calculation for your consideration.

Based on it, the observed deleterious effect looks fairly plausible possibility to also occur to some extent in humans, in vivo.

 

 

I did the same calculation and got something like 7.7uM, so a little higher.

 

The problem with that sort of analysis is that it doesn't consider time.  It assumes that 100% of the oral dose makes it to the blood plasma instantly.  You'll get something like that with an IV (however even that will have to drip in over 45 minutes to an hour).  With an oral dose you won't have that.  100% won't be absorbed, and that pulse of oral dose will be spread over time as it is absorbed in the gut.  The question is what peak concentration will you see in the kidneys?  We don't know the answer to that.

 

The other thing we don't know is whether opossum kidney cells floating in normal saline with a little glucose thrown in function like human kidney cells in a kidney in a human.  

 

As I keep reminding everyone, MitoQ originally went down a FDA drug approval process and at least got through phase I and phase II clinical trials.  That certainly doesn't mean it's completely safe, but it does mean that a reasonable amount of diligence has been done with respect to safety.  And we have other trials with humans consuming up to 3600 mg.  That does probably mean that MitoQ at least isn't acutely damaging, which is interesting since the researchers saw immediate damage.

 

So we have a conundrum here.   I can certainly understand someone electing to avoid MitoQ based on this paper.  But, it is premature to call MitoQ dangerous.

[aribadabar]

I did the same calculation and got something like 7.7uM, so a little higher.

 

The problem with that sort of analysis is that it doesn't consider time.  It assumes that 100% of the oral dose makes it to the blood plasma instantly.  You'll get something like that with an IV (however even that will have to drip in over 45 minutes to an hour).  With an oral dose you won't have that.  100% won't be absorbed, and that pulse of oral dose will be spread over time as it is absorbed in the gut.  The question is what peak concentration will you see in the kidneys?  We don't know the answer to that.

 

The other thing we don't know is whether opossum kidney cells floating in normal saline with a little glucose thrown in function like human kidney cells in a kidney in a human.  

 

As I keep reminding everyone, MitoQ originally went down a FDA drug approval process and at least got through phase I and phase II clinical trials.  That certainly doesn't mean it's completely safe, but it does mean that a reasonable amount of diligence has been done with respect to safety.  And we have other trials with humans consuming up to 3600 mg.  That does probably mean that MitoQ at least isn't acutely damaging, which is interesting since the researchers saw immediate damage.

 

So we have a conundrum here.   I can certainly understand someone electing to avoid MitoQ based on this paper.  But, it is premature to call MitoQ dangerous.

 

IV is certainly the best way to achieve maximum bio-availability.

I can say from a personal experience that I cannot "feel" MitoQ taken orally at the recommended 10mg dose.

OTOH, sublingual administration of the same 2 caps worth(10mg) leads to some slight head rush about 15-20 min later which seems to show that it is absorbed sufficiently in relatively short time otherwise it won't exert such/any perceptible effect.

 

Since it is a mito-targeted compound it won't preferentially go to the brain and skip reaching other organs' cells which leads me to believe that it is achieving significant concentrations in kidneys (as well as other organs) too in a relative short timeframe.

Even if we don't reach the 100% absorption sublingually in vivo we can conclude that we can allow a significant margin of error (in concentration levels) and still "hit the danger zone" as it requires mere 10% (at 20mg) to 20% (at 10mg) to achieve it. I understand that this is purely anecdotal and theoretical at this point but seems logical at face value.

Erring on the side of caution until more information is available seems the prudent approach IMO.

 

On the subject of clinical trials - at what dosage levels were those Phase I and II studies conducted?

I can't seem to find the 1000 and 3600mg trials you referred to?

Would you point me to the relevant references?

 

Thanks!

[smithx]

The question is what peak concentration will you see in the kidneys?  We don't know the answer to that.

 
This is actually not the question. The question is how much MitoQ ends up embedded in the mitochondria and for how long? If the affinity is high enough, and the retention long enough, a low plasma concentration over time may still result in a very high concentration in the mitochondria. 
 

As I keep reminding everyone, MitoQ originally went down a FDA drug approval process and at least got through phase I and phase II clinical trials.  That certainly doesn't mean it's completely safe, but it does mean that a reasonable amount of diligence has been done with respect to safety.

 
Phase I and II typically don't run for long periods of time. I don't know how long these ran, or what ended up causing the FDA to reject the compound as a drug, but short term trials may not reveal long term deletereous effects. 

Edited by smithx, 19 June 2019 - 05:58 PM.

[smithx]

Getting back to the topic at hand, an anecdotal report:

 

- My grandfather and father both died of heart attacks

- In my father's case it was due to an almost 100% coronary artery blockage (calcification)

- I am only 5 years younger than the age at which my grandfather died

- I have been taking Vitamin K-2 (4) 5mg / day for about 3 years

- I also take Vitamin D25 5000 IU most days, and 400mg of Magnesium (citrate).

 

- I had a calcium score in 2017 via electron beam tomography. It found 0% calcium in my coronary arteries (or anywhere).

- I had a chest CT scan 2 weeks ago (for an unrelated matter, thankfully negative), and they reported on my coronary artery as being "no evidence of calcification"

 

So a bit more anecdotal, evidence for K-2 (4) and Vitamin D. And maybe Magnesium.

 

[pamojja]

- My grandfather and father both died of heart attacks

- In my father's case it was due to an almost 100% coronary artery blockage (calcification)

- I am only 5 years younger than the age at which my grandfather died

 

Plaque forms in each of as, given only enough advanced age, or enough toxic exposures at early ages, as response to a damaged endothelium. Though it might seem hereditary, maybe you haven't had the same toxic exposures like your forefathers, and therefore no plaque to begin with. So not even anecdotal.
 

Please everyone understand how important it is to get it tested, only that way any intervention can be verified. Please don't waste an other 6 years, as this thread is already running, without any testing. Do yourself a favor.

[Advocatus Diaboli]

smithx, re: post 432.

 

Did you start your vitamin K2 (4), vitamin D, magnesium, regimen before, at the same time as, or perhaps a little bit after your 2017 calcium-score test? The "about 3 years" in your post 432 might fit any of those 3 scenarios depending upon what you consider "about" to mean, as well as when in 2017 the test was performed. For the pertinent case, what is your "best-guess estimate" of the time differential relative to your 2017 calcium-score test date? Possible choices might include--"the regimen was started": "at the same time as the test", "3 months before the test", or "2 weeks after the test", for example. I realize that trying to pin down the differential might involve more "about" statements, but I'd appreciate any attempt on your part to more closely define the time differential between your 2017 calcium-score-test-results date with your regimen-onset date. Thanks.

 

 

[smithx]

Yes, I should have been more specific. I was on the vitamin D25 and K-2(4) at those doses for somewhat over a year before the 1st calcium score.

 

 

smithx, re: post 432.

 

Did you start your vitamin K2 (4), vitamin D, magnesium, regimen before, at the same time as, or perhaps a little bit after your 2017 calcium-score test? The "about 3 years" in your post 432 might fit any of those 3 scenarios depending upon what you consider "about" to mean, as well as when in 2017 the test was performed. For the pertinent case, what is your "best-guess estimate" of the time differential relative to your 2017 calcium-score test date? Possible choices might include--"the regimen was started": "at the same time as the test", "3 months before the test", or "2 weeks after the test", for example. I realize that trying to pin down the differential might involve more "about" statements, but I'd appreciate any attempt on your part to more closely define the time differential between your 2017 calcium-score-test-results date with your regimen-onset date. Thanks.

 

[Advocatus Diaboli]

smithx, thanks for the info (post 435). Have you, by any chance, had a carotid-artery ultrasound?

[smithx]

I did have one, just prior to the first calcium score. It was negative, but I then read that for a more conclusive reading a calcium score was preferred.

smithx, thanks for the info (post 435). Have you, by any chance, had a carotid-artery ultrasound?

[Andey]

I did have one, just prior to the first calcium score. It was negative, but I then read that for a more conclusive reading a calcium score was preferred.
 

 

   I see how another argument could be made. Calcium score shows calcium deposits on repair sites where arteries are injured/ruptured.

What if interfering with calcium metabolism doesnt prevent injuries (why should it?) but simply limit the calcium deposition on injury sites? Formulated this way, it doesn't seem like there is much of benefit from it. 

[smithx]

Before developing hypotheses it's good to do sufficient research.
 
Far from "interfering with calcium metabolism", supplementing D and K support it. Vitamin D25 controls the amount of dietary calcium which can be absorbed, while K-2(4) is required to enable the osteoblasts to remove calcium from the blood and use it for bone formation.
 
Also, with regard to "... sites where arteries are injured..." this sort of injury happens in many disease conditions. See the table in this paper for example: https://www.scienced...735109714003283
 
With regard to whether Coronary Artery Calcification is anything to worry about or not, see this article:
https://annals.org/a...omatic-patients
 
Relevant quote:

In Cox models adjusted for risk factors for coronary artery disease, the CAC score was highly predictive of all-cause mortality (P < 0.001). Overall 15-year mortality rates ranged from 3% to 28% for CAC scores from 0 to 1000 or greater (P < 0.001). The relative hazard for all-cause mortality ranged from 1.68 for a CAC score of 1 to 10 (P < 0.001) to 6.26 for a score of 1000 or greater (P < 0.001).

 

t   I see how another argument could be made. Calcium score shows calcium deposits on repair sites where arteries are injured/ruptured.
What if interfering with calcium metabolism doesnt prevent injuries (why should it?) but simply limit the calcium deposition on injury sites? Formulated this way, it doesn't seem like there is much of benefit from it.

Edited by smithx, 21 June 2019 - 09:47 PM.

[Rocket]

I am going to apologize in advance if this is thread hijacking, but what happened to people taking trehalose? These threads are becoming too long and numerous to read everything. 

 

Mods maybe we need a sticky on this? This is a very important subject as EVERYONE will eventually have buildup in our arteries.

 

I have a bulk source for trehalose that I used for other chemicals and want to know if its worth taking it. How to take it. 

 

Feel free to move this post if needed.

[Rocket]

I am going to apologize in advance if this is thread hijacking, but what happened to people taking trehalose? These threads are becoming too long and numerous to read everything.

Mods maybe we need a sticky on this? This is a very important subject as EVERYONE will eventually have buildup in our arteries.

I have a bulk source for trehalose that I used for other chemicals and want to know if its worth taking it. How to take it.

Feel free to move this post if needed.

Bumpity bump

[Daniel Cooper]

Trehalose ingested orally is almost completely broken down in the gut. 

 

IV trehalose will definitely work (work in the sense of getting it into the bloodstream).  But getting some properly sterile and vetted trehalose solution in quantity and administering it over several months probably isn't going to happen. That leaves two other potential routes to administer - 

 

Liposomal and rectal. 

 

Our group buy of liposomal trehalose petered out, but you could make up some liposomal trehalose yourself and we can hope that we get enough encapsulated and that it makes it into the bloodstream.  If you do get a good encapsulation I think that liposomal might actually have a few benefits over IV.  But this is all highly speculative.  

 

For rectal, if you can make up some suppositories I suspect that will work as well.  There is however the "yuck" factor.  The trehalase enzyme doesn't seem to be present in the lower gut so if you could make a 3g trehalose suppository and administer daily you'd probably in the same ballpark of the dose that has been used in the weekly IV trehalose studies.

 

I think this is all in the separate trehalose thread we did.  If you can't find it let me know and I'll look it up.

 

 

 

 

 

[mikey]

In rats with CKD in might be 'preventing the build-up of calcium' - but it most definitely wouldn't have lifted the prediction of earlier death by reversing the build-up in the 3 cases detailed just above, without a 'patented' technology. We could long be gone by now.

 

 

If it works for preventing calcium build-up also in humans, it sure would become the next blog-buster drug. Imaging the whole population (which all are risk of calcification with age) on a antibiotic for life, a great business model.

 

 

A concern is taking and antibiotic, esp long-term and its effects on the microbiome.

[pamojja]

 

Imaging the whole population (which all are risk of calcification with age) on a antibiotic for life, a great business model.

 

A concern is taking and antibiotic, esp long-term and its effects on the microbiome.

 

Necessitating more prescriptions, not a concern for the business mode. On the contrary.

[unbreakable]

I couldn't read the whole thread, but my choice would probably be rosuvastatin (low dose, maybe every other day) in combination with pomegranate and GliSODin + natural cocoa (extract) or highly pure dark chocolate + garlic (extract), eventually diosmin/hesperidin.

Edited by unbreakable, 06 July 2019 - 11:24 AM.

[mikey]

I couldn't read the whole thread, but my choice would probably be rosuvastatin (low dose, maybe every other day) in combination with pomegranate and GliSODin + natural cocoa (extract) or highly pure dark chocolate + garlic (extract), eventually diosmin/hesperidin.

 

Aren't you concerned that rosuvastatin, like all statins, damages mitochodria, perhaps permanently?
 

See: The Dark Side of Statins, by Graveline. Then see Rosuvastatin Side Effects on Drugs.com, which lists side effects well, but I think may be understating how many people experience the side effects, because it is a "conventional" medical information (propaganda) site.

Rosuvastatin doesn't seen to be appropriate for life extension.
 

[unbreakable]

If I had significant arteriosclerosis / coronary artery disease I would likely take a statin.

[experimenting]

Vitamin K complex completely nukes my dental plaques. Dental hygienist doesn't know what to do.

Is it flippant to think it will not just stop further, but actually REVERSE existing arterial calcification?

I'm 30, so not really looking at this just yet, but just musing.

[ryukenden]

http://www.nutrition...ih-animal-study

Any of you taking Omega 7?

[smithx]

http://www.nutrition...ih-animal-study

Any of you taking Omega 7?

From the linked article
 

Researchers from the National Institutes of Health recently published a study¹ exploring palmitoleic acid’s effects on atherosclerosis in LDL-receptor knockout mice fed a high-fat Western diet.

 

In their study, alongside a Western diet, the researchers administered the mice either 1) palmitoleic acid, 2) oleate-rich olive oil (which is another monounsaturated fatty acid), or 3) placebo. Organic Technologies (Coshocton, OH) supplied both the palmitoleic acid (its AlaskOmega Omega-7 700 ingredient), as well as the olive oil.

 

At the end of a 12-week feeding period, the researchers found significant reductions in the palmitoleic acid group, especially in the areas of atherosclerotic lesions (approximately 45% reduction) and circulating triglycerides (approximately 30% reduction), compared to the olive oil and control groups. They noted that “our data showed that [the] palmitoleate-rich diet reduced atherosclerotic plaque areas and hyperlipidemia in LDLR-KO mice, although such effect was not observed in mice fed oleate-rich diet.” They also noted that mice in the palmitoleic acid group exhibited improved glucose metabolism compared to control.

 

The NIH researchers said that, following this study, they are beginning a clinical trial on palmitoleic acid on cardiometabolic health.