Senolytic Effects of FOXO4-DRI

Thanks.   Do you know how frequently you have to take it?

This is very interesting. Check my thread on fisetin. I have been taking it and could swear my testosterone levels were being elevated. This looks to be exactly what was causing it. I can testify to the fact if you take fisetin for the senylotic effect it will raise testosterone levels.

I swear I think this the exact mechanism that is doing it.

I take fisetin and quercetin with black tea extract but haven't noticed anything.  I'm 69.  I did just started taking Sam-e as I have genetics on both sides that effect meth-elation.  I've always had to deal with a lack of energy.  It is starting to give me some energy unless it's the placbo effect.   I also started taking Ipamorelin for the growth hormone releasing effect so that might be having an effect.  Just seem to notice a little more get up and go.  Hope spring eternal as they say.

How are you taking fisetin? I do three large doses once a month. 3g three days In a row. I use swansons, and that means 30 tablets or a whole bottle each day for 3 days.

For me the results were unmistakable. Vastly more energy, more dense and stronger muscle definition, some increased aggression, mental clarity...and so on. I swear by this stuff. 

What Happened to Senolytics with FOXO4-DRI?

     This topic discussion was started several years ago, when the big news in senolytics was that a new treatment, FOXO4-DRI, had been developed that promised to provide a powerful new way of clearing senescent cells.  Unlike most alternatives, its senolytic effects should be relatively independent of cell type.  On March 5th, 2018, Reason in his Fight Aging! Blog provided a lengthy description of how one might obtain FOXO4-DRI and do carefully monitored senolytic self-experiments with it.  This writeup included an estimate of the total cost of a session of self-experiments, which added up to over $7,000.  About the same time, the web site FOXO4-DRI appeared, created by Darren Moore, 52, who describes himself as a lifelong self-experimenter and who was injecting himself with FOXO4-DRI.

     However, over the past three years interest in the topic seems to have fallen off precipitously, and I have been wondering why.  Has anyone on this Forum tried FOXO4-DRI?  Does anyone understand why it seems to have dropped out of sight?

     Let me summarize just what FOXO4-DRI is and how it is supposed to work.   First, FOXO4 is a human peptide (i.e., short protein) that is a member of the forkhead family transcription factors O subclass, which is characterized by a winged helix domain used for DNA binding.  FOXO4 is a peptide, a chain of 34 amino acids with the sequence LTLRKEPASEIAQSILEAYSQNGWANRRSGGKRP.  It has a molecular weight of 54 kDa.  It has many biological functions, among which is that in senescent cells and cancer cells it binds with the protein p53, preventing p53 from triggering apoptosis.  The protein p53, if released from the cell nucleus into the cytosol will trigger apoptosis and cell death.  However, if it is bound by FOXO4, it remains in the nucleus and apoptosis is avoided.

     Researchers have reasoned that if this FOXO4-p53 link could be broken, the p53 would not be bound, apoptosis could proceed in senescent and malignant cells, and they would be cleared.  To achieve this end, they synthesized a form of FOXO4 constructed with the same sequence but made of right-handed (D) amino acids instead to the natural left-handed (L) amino acids that are the basis of all cell molecular biology.  They labelled this form FOXO4 D-Retro-Inverso or FOXO4-DRI.  Cells seem to treat FOXO4 and FOXO4-DRI more or less interchangeably, but FOXO4-DRI is unable to form the binding of p53 that suppresses apoptosis.  Experiments with human cell cultures and with aging mice were performed and showed the expected senolytic effects.  Fast-aging mice with patches of missing fur began to recover their coats.  In three weeks fitness benefits began to show, with older mice running double the distance of their counterparts who did not receive the peptide.  In a month, aged mice showed an increase in markers indicating healthy kidney function.  Curiously, the experimenters did not report measurement of senescent-cell counts for treated and untreated mice after the trial was completed.   One of the researchers, Dr. Peter de Keizer, created a startup in early 2018 in the Netherlands called Cleara Biotech.  It is intended to pursue FOXO4-DRI applications in senolytics and cancer therapy.  However, its website seems to be in permanent “maintenance mode”, and there are no reports of recent corporate activity.

     At his FOXO4-DRI website, Darren Moore reports the results of three senolytic sessions performed between April 18, 2017 and February 12, 2018, in which he injected himself subcutaneously with FOXO4-DRI.  There’s a Belgian documentary on YouTube about his self-experiment.  He did before-and-after bloodwork monitoring telomere length, epigenetic DNA age, and a variety of other biomarkers, and he also timed his endurance on a treadmill.  He reports his treatments produced increased endurance, some head-hair regrowth, somewhat improved biomarkers, and a few-year drop in his bio-age estimates from telomere length and epigenetic DNA analysis.

     I would have to say that I do not find any evidence of dramatic anti-aging effects from FOXO4-DRI.  The observed improvements, while positive, are not markedly better than reports of senolytics with D+Q or Fisetin, and the monetary expense and investment of time and energy is far greater.

    Further, I have a few worries about the mechanics of the senolytics.  The injected FOXO4-DRI in the bloodstream must cross two barriers, the cell wall and the nuclear wall, and reach the nuclei of all cells that it is to affect with a sufficient concentration that it frees enough p53 for those molecules to exit the nucleus and initiate apoptosis.  One virtue of FOXO4-DRI is supposed to be that it induces apoptosis in senescent cells of all cell types, but the probability of this double-wall-penetration seems unlikely to be 100% for all cell types.

     Further, the natural peptide FOXO4 has other functions in cell biology besides binding p53, and it isn’t clear to me what the consequences of interfering with those functions might be.  The residence half-life of FOXO4-DRI is unknown, so once injected there could be lasting side effects.  The absence of apparent negative side effects in mouse models and in Darren Moore is encouraging but not definitive.  For the moment, for senolytics I plan to stick with sessions of D+Q+F+…

     Do other contributors to this Forum have information on FOXO4-DRI?  Do you know of any other tests on human or animal subjects?

FOXO4-DRI  and the Blood-Brain Barrier

    Darren Moore mentions that in injecting the peptide FOXO4-DRI, he did not experience the hangover-like symptoms (mild headache, nausea, ...) that he had experienced after the senolytic sessions done with Dasatinib.  That suggests that the blood-brain barrier actively is actively blocking the passage of FOXO4-DRI (molecular weight: ~54,000 Da), a fairly large molecule formed as a folded chain of 34 amino acids, while it passes small-molecule senolytics like Dasatinib (488.01 Da), Quercetin (302.2 Da), and Fisetin (286.24 Da).  The probable reason that D+Q+... senolytics produces mild headaches, etc. is that senescent brain cells are being cleared.

    Thus, the claim that FOXO4-DRI may be a universal senolytic senolytic that clears senescent cells of all cell types is probably misleading.  Clearing senescent cells in the brain is likely to be important, and FOXO4-DRI injected into the bloodstream can't reach those.