#264

That is a pretty high dose for a long cycle. 120mg over 10 days!!

The most I’ve seen is 2x 25mg doses in a 3 day cycle (on, off, on) per year or bi-annually.

Do you mind sharing your rational for this dose and cycle?

#265

I recently read that FOX04-DRI works well in combination with quercetin. I can’t find the paper now, but you’ve probably seen it.

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#266

There are many other closed forums doing self researching with FOXO4 and the protocols are very much the ‘Wild West’. One of the original mouse studies converted to human dosing would be 0.42 mg/kg 3x every other day. For me that would be 38mg x 3 = 114mg. In my research experiment, I just spread it out over 10 days.
Currently, no effects at all to report. Others reported flu like symptoms during their cycle.

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#267

The mouse studies have provided interesting results :slight_smile:

I’ve not experienced any negative effects at 3mg per dose for 6 doses (1 day on, 1 off) 18mg total cycle. Going to wait a few months to see if there are any measurable or physically noticeable results.

But I’ve been actively killing my zombie cells for 4 years now with another intervention. Although that one has limited “reach” on the cell types that it can clear from.

I have seen consistent positive changes in my epigenetic tests (5 in 4 years) and will be doing #6 in March.

What types of tests are others doing in these private groups?? Do you have a test plan?

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#268

According to ‘The Peptide Handbook,’ by James B. LaVelle and others, dosing FOX04 with MOTS-c or humanin can improve its ability to eliminate senescent cells. FOX04 is most effective against cells expressing high levels of SASP factors, which are increased by MOTS-c or humanin.

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#269

In this study they used navitoclax

Senolytics restore hematopoietic stem cell function in sickle cell disease

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#270

@Steve_Combi What do you think is an effective basic senolytics regimen that can be performed monthly? I’d like to clean out my senescent cells at this interval.

Thanks in advance.

#271

Near as I can tell there is no “basic” way to kill zombie cells. Go hard or don’t bother, cause they are immortal for a reason… as in hard to kill.

A couple questions;

  1. budget
  2. age

Those 2 basics help determine method.

But basically it can be done with a combination of quercetin and fisetin and EGCG. The doses are high to make it work though.

For anyone under 50 I would not be terribly concerned and might consider starting 1 cycle a year. Details on the attached on age, frequency and how to cycle it.

Attached is our formula and protocol. Feel free to copy it. I went a bit over board on the bio-availability enhancers due to the poor availability of polyphenol flavinoids, i.e. Q and F. Everything in the formula is there for a reason. Only took me 18 months to sort this out :slight_smile:

The volume is high for a reason, if not, there is no proof it would work. The volume of Q is based on 2 successful clinical studies, the volume of F is based on 2 planned studies, all by the Mayo Clinic.

My wife and I did this for 4 years, 4 times a year. until this year when we switched to FOX04-DRI a more “universal” senolytic. While this one may seem more expensive at retail, at wholesale it’s actually the same or less cost. It’s also a LOT easier to do.

Senolytic_formula_and_protocol.pdf (195.7 KB)

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#272

What would you measure before and after a cycle to confirm that it had the desired impact?

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#273

A question I’ve been asking since I started killing zombie cells 6 years ago.

It’s a difficult thing to measure directly. In clinical studies they use multiple measurements, one being tissue/cell counts. (biopsy or autopsy) Senescent cells have a very specific “look” and can be counted. SASP (senescence associated secretory phenotype) encompasses over 500 cytokines, chemokines and other secretions. These are not all exclusively markers of senescence and have other contributing factors but it appears a few may be exclusively related to senescence.

If you would like to know how difficult this is, check out this paper.

Further up this thread there is reference to a soon to be release test that is specific to senescence and this would be the first.

Theoretically at a high level, one would look for reduction in inflammation BUT many things can contribute to inflammation. Not just senescent cells increase inflammation so hsCRP is not great for identifying senescence, specifically. Other markers have multiple sources so it’s not easy to measure a level of senescence. It would be amazing to have a test so one could iterate and maintain a low healthy level.

β-galactosidase (β-Gal) assay is typically used in clinical studies BUT not on it’s own as there are, again, other things that can affect this marker besides senescence so it’s not the best marker either. And the reason multiple tests are done in senescence related studies.

Another issue is that SNC’s are typically tissue specific and not “mobile”. By tissue specific I mean “injury site” specific. Sun burns generate a lot of SNC’s in your skin… Injure your back and you will have them in your damaged vertebral disks. Get a concussion and you will have them in your brain, cut yourself, break a bone, bruise a muscle, have a heart attack, get a lung infection like Covid, etc. and there will be tissue specific senescent cells, localized to the injury.

I settled on epigenetic markers 5 years ago when I did my first DNAm test through Bioviva (Liz Parrish) and found my bio-age was 13 years younger than my chrono-age but they stopped offering that service so I switched to Trudiagnostic and have done 5 tests over 4 years with them. Every one a bit better, except the one shortly after getting covid. My results since that infection have continued to improve and are better then before covid.

I also used this test when I ran a very small evaluation with 4 strangers I met on the internet LoL! Over a 7 month, 3 cycle evaluation every participant had an improvement in their markers. A few were quite spectacular.

Combilytics_Corp_Product_Evaluation_Results.pdf (217.0 KB)

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#274

I am not a believer in senolytics.

If you listen to the most recent Attia AMA, there’s a roundtable with Prof Miller (from ITP) and he summarizes nicely. He points out that senolytic drugs failed the ITP, at two different dosing methods (Astaxanthin and meclizine extend lifespan in UM-HET3 male mice; fisetin, SG1002 (hydrogen sulfide donor), dimethyl fumarate, mycophenolic acid, and 4-phenylbutyrate do not significantly affect lifespan in either sex at the doses and schedules used - PubMed).

Worse yet, when they sent tissue samples to the investigators who recommended fisetin, blinded, there was no difference in p16, SA-B-GAL, or any of the markers, in liver, brain etc. So it’s questionable whether they really work in the real world.

Additionally, “senescence” as a whole is insanely complicated, and seems to be a word like “cancer” where it encompasses hundreds of different phenotypes. In the lab, we sometimes induce senescence by treating cells with H2O2, or stressing them with UV. But I’m not exactly sure what that phenotype represents in human aging… IMO, not much.

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#275

Here are 2 studies in humans. Both of which had interesting results.

Senolytics decrease senescent cells in humans: Preliminary report from a clinical trial of Dasatinib plus Quercetin in individuals with diabetic kidney disease.

https://www.thelancet.com/journals/ebiom/article/PIIS2352-3964(19)30591-2/fulltext

Senolytics in idiopathic pulmonary fibrosis: Results from a first-in-human, open-label, pilot study

https://www.thelancet.com/journals/ebiom/article/PIIS2352-3964(18)30629-7/fulltext

And some news from the Mayo Clinic.
https://newsnetwork.mayoclinic.org/discussion/senolytic-drugs-boost-key-protective-protein/

Orally-active, clinically-translatable senolytics restore α-Klotho in mice and humans

https://www.thelancet.com/journals/ebiom/article/PIIS2352-3964(22)00096-2/fulltext

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#276

They’re definitely interesting, but both are pretty flawed studies to my eyes.

The first is open-label, no proper control group (i.e. it’s a before/after). And no mention of blinding the investigators. Counting positive cells in tissue slices is very subjective, and easily affected by bias, selecting whichever areas to count etc. I note that the only truly quantitative data (gene expression and plasma factors) is kinda hidden by using a heatmap and ranking things into percentiles. IME, that usually means when they simply plotted the numbers, there was no difference. Combine that with the conflicts of interest that the authors have patents on Senolytics drugs, and I become even more sceptical.

The pulmonary fibrosis paper is also very preliminary and again the results are kinda “buried”. 23/50 measured cytokines had correlations. Improvement on physical tests is quite interesting, but again the before/after nature of the experiment means there are many explanations. For example, at the “after” test, the subjects had a much better idea of what was expected, as they’ve done it before. This is common in sports literature, where placebo groups gain strength simple due to familiarity with the tests. A proper trial design can account for that, but obviously requires more resources.

I just have to come back to the ITP, if those mice were given senolytics for their entire life and the blinded samples from multiple organs were indistinguishable… I have to stay sceptical that a few weeks of these compounds would move the needle in humans. However, I will love to be proven wrong by a “proper” trial or a senolytic passing the ITP.

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#277

And that “for entire life” thing is going to prevent any measurable difference in any study. Why? if senescent cells are constantly eliminated, there is no change to measure. That is the basis of the Threshold Theory of Aging.

threshold_theory_of_senescence

The “problem” with senescence is that it does not apply to the young. It’s when you hit 60+ that it’s going to present as a contributor to a wide variety of conditions.

When one looks up various conditions with the word “senescence” added to the search string there are quite a few interesting results. > heart disease senescence <> dementia senescence <> cancer senescence <> just pick your favourite condition… :slight_smile:

If so many human conditions were not affected by senescence, I’d agree with you.

There is more to research than the ITP… as if that program was all that mattered in human science… sure mice are an interesting species but many things that work in mice do not work in humans and vice versa. So I’ll keep an open mind on senescence and the overall science that indicates it is harmful as we humans age.

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#278

Hi Steve, thanks for the thoughtful response. I don’t quite understand this comment. In the ITP there is a control group of mice - thus, if the old mice without drug and the old mice with drug have the same amount of P16+ cells, it implies the drug didn’t work. (Note that old mice in both groups had higher P16 and SA-B-GAL than young mice).

I do agree with you that this is just one drug (Iresin, in this case). SO there are two possibilities, I think:

  1. This specific drug didn’t actually kill senescence cells, and that’s why no lifespan extension. Maybe other drugs would work.

  2. The whole approach just doesn’t work, and/or doesn’t translate to lifespan

I absolutely agree that interesting findings come up when looking at certain organs. My own lab has done some of that research ourselves, on senescence and cardiovascular diseases. I believe there is something going on. As you say, this phenomenon does appear to be involved in many disease; but as I mentioned before, to me, “senescence” is a word a bit like “cancer” where it’s very challenging to define, let alone target.

For example, in the heart, cardiomyocytes in older mice have higher P16 expression, and knocking this down does seem to lower inflammation and result in better cardiac function after injury. A potential good drug is this one: Navitoclax - Wikipedia

However, that’s an acute injury setting, and I’m not sure about prophylactic use for longevity.

I agree that there’s more to research than the ITP, but it’s a damn good model using heterozygous mice, multiple sites etc. The problem is that most small studies are rife with bias. James Kirkland is the author of the two D+Q papers you mentioned, and he’s the one who suggested Iresin to the ITP. But his whole reputation, financial future etc is staked on showing that senolytics work. I’m not picking on him specifically, because this is a very human thing, but that’s why I value the ITP and the total unbiased approach it takes. I absolutely hope they test more senolytic drugs, and if they can actually show reduction of signature (P16, SA-B-GAL etc) cells, and extension of lifespan, I’ll be delighted to be wrong, and I’ll get on board with taking them!

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#279

When it comes to mice, I get that they are a decent screening process but how many times have mice and rats passed the test (what ever test that may be) and the ensuing human trials failed miserably?? Not only that but they are a poor reference system for toxicity.

Even here on this forum we often see “BUT this is in mice, lets see what it does in humans”
99% failure in AD research
Typically 80% or worse in other areas.
https://journals.sagepub.com/doi/10.1177/02611929231157756

When I reference “senescence” I’m more specifically talking about senescent cells, not the entire process of “aging” that is often referred to as “senescence” For me, it’s not like the generic “cancer” but more specifically the cells that become senescent and hang around past their due date :slight_smile:

I appreciate your position and commentary on this topic.

I also get Kirklands commitment to this area of research. One thing about committed researchers, quite often that is what it takes for breakthroughs. Dogged determination against the headwinds, Katalin Karikó, PhD is one recent example of that.

I’ll keep doing my zombie killing processes as there does not seem to be a downside. So far :slight_smile: Unlike our fav intervention here with respect to Rapamycin, which appears to have more risk and yet I continue to take 6mg every 2 weeks :slight_smile:

#280

Hedging my bets by taking rapamycin, which is already senomorphic. But otherwise, I’ve become more and more skeptical of the current crop of senolytics, especially after listening to the Attia roundtable.

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#281

Mice die primarily from cancer - 85-95%. Humans die primarily from heart disease.

I think you have to ponder the question “How much do senescent cells impact cancer?” If the answer is “A lot” then senolytics (as shown by the ITP) probably are not very useful. If the answer is “Not much” but very useful for cardiovascular health or maintaining healthspan in old age, then senolytics may be very useful to humans who do not die primarily from cancer.

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#282

Yours too. Thanks for the civil discussion. And absolutely, I support guys like Kirkland pursuing things, and hopefully we can have bigger, better, longer studies to really answer these things.

Honestly, a lot of this could be answered if we had a better method to measure whether the treatment is actually working. For example, I see your protocol mentioned earlier. If there was something you could track or measure that would be super exciting, even in the absence of lifespan data.

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#283

There is supposed to be a new test that is only available to health care practitioners.

As you noted, we did use a DNAm test from Trudignositc in an attempt to validate our little eval. Plus it is an easy to do at home test. My hope was those markers might be a proxy for overall SASP reduction, but I can’t prove that. Of the 4 that completed the program (not counting myself and my wife, we both improved over a 4 year span) all showed an improvement. What does that mean/indicate? who knows for sure…

Once I saw it “work” for strangers with no skin in the game, that was enough to give me a bit of hope :slight_smile: regardless of what was actually happening LoL! I’d really love to know;

  1. is it actually clearing senescent cells
  2. are the improvements due to this clearance

There are so many mouse studies on this that indicate a benefit but those are mice and I don’t always trust rodents :slight_smile: although they do “naturally” experience cellular senescence as opposed to the need to be genetically manipulated as is the case with AD research.

And then if a level of healthy senescence can be determined, and then maintained/iterated with judicious use of senolytics and testing to evaluate levels, my guess is probably not easily maintained as we age, due to the varied types and locations of SNC’s

The D+Q combo with our addition of Fisetin is limited in “reach”. That combo only appears to reduce senescent cells in adipose and endothelial tissues. One of those limitations.

Which is why we started doing cycles of FOX04-DRI in Dec.

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