An Example of Senolytic Self-Experimentation with FOXO4-DRI

Senolytic drug candidates, those demonstrated to selectively remove senescent cells to some degree in animal studies, are fairly easy to obtain. They are not enormously expensive, considered in the grand scheme of things, even those that are not yet mass-manufactured. Removal of senescent cells is a form of rejuvenation, shown to extend life in mice and reverse a number of specific measures of aging and age-related disease. These cells cause harm through the signals they generate, generating inflammation, fibrosis, and many other harmful secondary effects. Given the potential benefits, people are starting to experiment, though so far without the sort of rigor that it would be useful to see. You really have to be measuring appropriate metrics, otherwise it is all too easy to generate no useful information about the effects.

In the example noted here, I'm pleased that someone is making the effort to self-experiment in a public way - something I'd like to see more of, as this is how more organized efforts get underway. He is using the drug candidate FOXO4-DRI recently shown to interfere in the FOXO4-p53 signaling that only takes place in senescent cells. However, he isn't picking useful endpoints to measure, I think, which means that the only evidence gathered here is that this isn't horribly dangerous - always assuming that the supplier is providing what they say they are, which should be checked for compounds that are not presently mass-manufactured and widely used. Bad batches are possible, even with the best of intentions.

Useful or possibly useful items to measure might include the Osiris Green DNA methylation biomarker, bloodwork focused on markers of inflammation, kidney function, and liver function, and CT scans focused on assessing calcification of arteries. If you are not in much later life, however, the changes might be small enough to be hard to detect reliably in easily available tests such as those above, or swamped by normal day to day variation, even if the treatment is useful. Thus the best measure is to take a biopsy and have it stained using the standard research assay for senescent cell presence, but that is custom lab work and harder to arrange for most people.

A senolytic (from the words "senescence" and "lytic" - destroying) is among the class of senotherapeutics, and refers to small molecules that can selectively induce death of senescent cells. Senescence is a potent tumor suppressive mechanism. It however drives both degenerative and hyperplastic pathologies, most likely by promoting chronic inflammation. Senescent cells accumulate in aging bodies and accelerate the aging process. Eliminating senescent cells increases the amount of time that mice are free of disease. The goal of those working to develop senolytic agents is to delay, prevent, alleviate, or reverse age-related diseases. Targeting premalignant senescent cells could also be a preventive and therapeutic strategy against late-life cancer given the deteriorated efficacy of the senescence response in stopping cancer.

Senolytics are arguably the best rejuvenation therapy currently available, and though costly, FOXO4-DRI is the most effective senolytic. This site is a repository for the first human experiences with this exciting new substance. And, though anecdotal, the hope is this information will prove valuable to early adopters and science. I'm a lifelong experimenter, a member of AAAS, and proud supporter of SENS. I'm hoping the risks I'm taking will benefit many people, and advance the science. I know, I know, this is not a controlled, double-blind experiment. I am patient zero in an n=1 study. But, is there something that can be learned here? Yes, especially if I have a serious reaction or die. Alternatively, if a remarkable rejuvenation becomes evident credibility will be lent to this therapy.

Link: http://foxo4dri.com/

Comments

The effort to log his experience is appreciated. But some critique: I assume this is a guy, but would like a short bio stating age, sex, whatever. He uses Units, mg, and ml, at alternate times and it is too confusing to follow, should stick with one consistent metric. I am skeptical that positive effects would be seen short term from taking any senolytic, it would seem to me that if any short term effects were experienced they would be negative due to the body having to process the apoptosis of the senolytic cells.

Posted by: JohnD at May 15th, 2017 8:35 AM

@JohnD actually some previous animal studies show pretty quick improvements if you review the study data. I agree with you comments about consistency though and it is interesting despite it being an n=1 which is not overly useful in general terms.

Posted by: Steve Hill at May 15th, 2017 9:58 AM

I've been supplementing 100 mg/day of fisetin for about a month and also adding moderate amounts of Indian long pepper to my food since last January. I've ordered the Osiris Green DNA methylation biomarker this week and will retake it next year. For what it's worth I did the Aging.AI test using last December's blood work (it said 30 but I'm actually 43) and will retake that as well. If there are any changes I'll post them here.

Posted by: Corbin at May 15th, 2017 1:32 PM

And the math does not make sense. The cheaper of the two sources I found for FOXO4-DRI is $1,072 for 15mg. His log says he is injecting 30mg-50mg at a time.

Posted by: JohnD at May 15th, 2017 1:59 PM

If some experts in the field would develop a meaningful framework for self-experimentation, I would definitely participate.

Posted by: bardu at May 15th, 2017 4:44 PM

@Matthias - good spot that this is self promotion for his online peptide business. Unfortunately this now makes any results he self reports suspect. He is obviously going for the hairloss market. I found it a bit strange that he has pictures of his hair apparently growing back as he has male pattern baldness, which as far as I know is unrelated to hair thinning with aging which does not occur in spots but all over the entire scalp and in women as well as men. Now it makes sense as he is hoping the hairloss crowd will buy his product.

I misread the site and thought it was $1,072 for 15 grams not 15 milligrams. Obviously if he is injecting 30mg-50mg at a time a few times per week he is not selling this peptide at close to manufacturing cost. Either that or he is a deca millionaire.

I wonder how hard it is to manufacture peptides like this and sell them to your friends at close to manufacturing cost plus mailing?

Posted by: Jim at May 15th, 2017 8:45 PM

OK, so he's got a profit motive and it's for the flimiest of flim-flam type treatments--hair loss. I guess we can watch and see what happens. I'm normally pretty open to observing such one-off experiments, but this one will need some seriously impressive results repeated my multiple people before it has any value at all.

Hair loss treatments have a long and illustrious history we all know well.

Posted by: Paul at May 15th, 2017 9:15 PM

@Jim

If he tests out the dosage, makes some money selling that stuff and sends half of it to the SENS foundation I'm completely fine with that. Applause for him for that stunt.

Hair grows about 2-3mm per week and I wouldn't expect much in that regard after just 2 weeks, so this claim is clearly exaggerated.

And there are many labs selling peptides on demand online. You can call one and ask for an offer if you're really curious.

Posted by: Matthias F at May 16th, 2017 12:56 AM

male pattern baldness may prove the most unstoppable force in the universe, youth drug or not. If he gets some hair back, that will be interesting, but if he doesn't, it probably doesn't mean much. The best question to ask is whether he sees an improvement to any regular exercise that he does. And of course, "how do you feel?" But we'll have to take his word on that.

Posted by: Paul Rattner at May 16th, 2017 5:03 AM

As much as I'd like to see experimental results and verification that this works and is safe in humans, this guy's reports fall woefully short. Proper experimental design is hard, and I don't see any attempt to do that here.

If you want an example of what "reasonably correct" self experimentation looks like, compare the above linked blog to this:

https://www.gwern.net/LSD%20microdosing

Posted by: Dennis Towne at May 16th, 2017 9:11 AM

That explains everything, thank you Matthias

Posted by: JohnD at May 16th, 2017 10:16 AM

@Matthias - I'd buy some FOXO4-DRI from a lab online on a few conditions, but a quick google just turned up a few sites here in Australia that seem to be just be selling growth hormone peptides for athletic enhancement.

The main condition would be the cost. If it would be thousands per month, I cannot afford that. It would be good if someone would do this and used the DNA methylation analysis by Osiris Green and a skin biospy. I think submitting facial photos with only the skin and not hair visible in highly controlled lighting for age appearance estimation before and after could be useful.

Here is where I saw this done in a BBC documentary (The Truth About Looking Young):

https://youtu.be/n94IPA9bj9g?t=36m32s

A good bit of the documentary on fibroblasts in skin aging:
https://youtu.be/n94IPA9bj9g?t=24m27s

Any help on sourcing cheaper and real FOXO4-DRI would be appreciated, as I'd probably just end up buying a fake bottle of saline from somewhere.

Posted by: Jim at May 16th, 2017 9:07 PM

@Jim

These labs I'm thinking of deliver custom made stuff based on ones
specifications. It's very likely that they can help one out with the
recipe, too.

https://www.google.de/?gws_rd=ssl#q=peptide+synthesis
https://www.google.de/?gws_rd=ssl#q=peptide+custom
https://www.google.de/?gws_rd=ssl#q=peptide+buy

One will get good quality this way but if you want to buy cheap for yourself alone
it won't work. That's because the pricing is mainly determined by what one wants,
not by how much he buys. And the initial costs are high - the FOXO4 protein has
somewhere around 2400 amino acids if Wikipedia is right.

OTOH - gather up enough like minded people (like 20 or so) and it's affordable.

Posted by: Matthias F at May 17th, 2017 1:44 PM

So this peptide is pretty much as complex as an antibody to make. And you don't really read about people getting knock off antibody drugs made, even though some of them sell for exhorbitant prices.

I did see something on experiment.com where a group was trying to manufacturer edible insulin in algae. Maybe you could also make this larger protein in algae? Or even a gen therapy like the guy at http://www.bf-sci.com performed on himself, except with the protein production being indictable by tetracycline or some agent.

Posted by: Jim at May 18th, 2017 2:55 AM

Guess what, the peptide is actually patented. Here's the recipe - and the important sequence is only 34 amino acids long:
https://www.google.com/patents/WO2016118014A2?cl=en
That means it's basically free for scientific and private use. Any volunteers for cooking that up in their basement?

Posted by: Matthias F at May 19th, 2017 11:54 AM

How hard is synthesising short chain peptides in your basement?

Posted by: Jim at May 20th, 2017 7:52 AM

@Jim

Depends on your expertise. With some experience in chemistry it's possible using basic lab equipment. It's some work, but it's possible to create kg-batches this way. Here's a YouTube video of someone doing it:

Solid Phase Peptide Synthesis Hands On Procedures from swelling resin to HPLC & MS analysis
https://www.youtube.com/watch?v=jVXUfC2pLh4

Or for a simpler procedure there are automatic peptide synthesizers. Works basically like a 3D printer - fill in the raw material, insert the recipe and let it run over night. They cost as much as a car, but there are some places where they sell used ones, just like in cars.

The last step should be purification and quality control which can both be done with e.g. an HPLC.

Posted by: Matthias F at May 20th, 2017 9:19 AM

I would consider participating if someone were to undertake to form a group so as to try to reduce cost. I'm 84 and am taking a variety of things in the hope of enhancing longevity but without any formal guidance by somebody who knows what they are talking about. I'd love to have said guidance!!

Posted by: John Moe, MD, MPH at May 21st, 2017 10:25 AM

@John

If you want formal guidance for using this untested drug and some safety measures (I suppose) then what you're really talking about is participating in a clinical trial. At least that's the only formalized way I know of. You could try to contact the inventor of the peptide p.dekeizer@erasmusmc.nl and ask him. I'm pretty sure he's already planning something in that direction.

Posted by: Matthias F at May 21st, 2017 11:20 AM

You've got to figure yields and upfront costs.
If there are 34 amino acids in the sequence and let's say each link in the sequence yields 90% (as an example), then your total peptide yield is going to be 0.9^34 which equals about a 2.8% yield.
Not great, but doable.
However, your huge upfront cost will be the fact that you're not using "L type" amino acids for this peptide, but "D type" which are not readily found in nature and typically have to be totally synthesized to extreme purity. That's a big upfront cost.
You can use Fmoc or Boc as synthesis protocols. You'll need the basic reagents, but you could do it binge watching Family Guy, then send your results out for purification.

Posted by: Nyles at May 27th, 2017 10:53 PM

The idea of the peptide is to disable FOX 04, by preventing it from interacting with p53.
If you model FOX 04 as a protein, and work out what part of FOX 04 binds to p53, then you could possibly discover a much shorter sequence of L amino acids that could block p53.

Then lab synthesis would be much easier and cheaper.

Alternatively, you might use something that cuts the Fox 04 in specific places, based on its unique sequence, so that it can no longer bind to p53.

So it is good to research cheaper ways of disabling FOX 04

Posted by: Craig Paardekooper at March 4th, 2018 2:27 PM

An experience I had myself self-experimenting. I'm a 67 year old woman and I've ordered 100 mg from Breakthrough Biolabs about 2 months ago, because I read good reviews of them online and because of the pricing. I had some pins and needles the day after injecting, having injected 25mg 3 times in 5 day intervals. The effects for me personally are amazing: I feel a decade younger in my overall well-being, vitality and energy levels. I feel like I'm alive again after using this compound. I don't know how long this will last, but it's been 3 weeks and I feel amazing!

Posted by: Suzannah at March 24th, 2019 8:55 PM

What we are talking about here is a peptide.
Fox04-dri. It can be obtained. What is referenced is a mice study. This peptide can be & is being reengineered to be more effective & safer for humans. It isn't there yet. By taking this peptide you are killing off senescent cells, as well as some healthy cells. This is why this version is not approved. There is no dosage. There is no awareness of side effects. There has been no trials. So no doctor would recommend.
That being said, are there any results besides I feel 10 years younger..?

Posted by: Jay Rich at December 27th, 2019 7:59 PM
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