20 replies to this topic

[Chemically_Charmed]

Anti-Aging Effects Observed for New 2-Compound Cocktail

 

 

A promising new study published in BioRxiv reports that a simple cocktail containing just two small molecule compounds (TCP, an LSD1 inhibitor, and Repsox, a TGF-β inhibitor) were able to reverse multiple aspects of cellular aging in human cells and extend lifespan in worms.

 

Previous studies have  used  a 7 drug combo, which if ever used in humans or animals, would probably cause significant side effects. This study demonstrated that using just a LSD1 inhibitor and a TGF-β inhibitor alone was able to achieve similar results as the 7 drug cocktail.

 

The two drugs used in the study were Tranylcypromine and LSD1, an irreversible  LSD1 inhibitor, and Repsox, a TGF-beta inhibitor. TCP isn’t very selective in its effects and Repsox hasn’t been tested as safe for humans.

 

The best drugs to be used in humans right now would be Pulrodemstat, a potent selective reversible LSD1 inhibitor and Galunisertib for the  TGF-beta inhibitor. 

 

A brief overview of results:

 

Researchers treated cultured human fibroblasts from elderly donors with the 2 chemical cocktail for just  6 days and observed significant improvements across several key biomarkers associated with youthfulness and vitality. They also provided the drug combination to Caenorhabditis elegans nematodes throughout adulthood and recorded an impressive 42% increase in median lifespan.

 

DNA damage (as measured by γH2AX levels):

7-chemical cocktail reduced γH2AX levels by ~60%

2-chemical cocktail reduced γH2AX levels by ~75%

 

Heterochromatin marks:

7-chemical cocktail increased H3K9me3 by ~2-fold and H3K27me3 by ~3-fold

2-chemical cocktail increased H3K9me3 by ~2-fold and H3K27me3 by ~5-fold

 

Cellular senescence:

2-chemical cocktail decreased senescence-associated β-galactosidase levels by ~60% in stress-induced senescent cells and ~50% in replicative senescent cells

Additionally, the 2-chemical cocktail decreased p21, p53 and IL6 gene expression levels linked to senescence and aging.

Oxidative Stress: Imbalance between free radical production and antioxidant defenses that damages macromolecules. The cocktail in fact decreased oxidative stress levels, implying improvement of metabolic homeostasis.

 

AKG exerts some of its anti aging effects though DNA/histone demethylation so that I think could be used in combination with these. AKG was an add on compound they used in another cellular reprogramming study.

 

 

γH2AX refers to the phosphorylated form of the H2A histone protein variant H2AX. It is an important marker of DNA damage.

 

A reduction in γH2AX signal indicates the chemical cocktails can ameliorate this aging-associated hallmark and improve genomic integrity. Lowering DNA damage is thought to be an important mechanism through which reprogramming interventions may promote longevity.

H3K9me3 and H3K27me3 are histone marks associated with heterochromatin and gene silencing. Levels of these repressive marks tend to decrease with aging. Therefore, increasing H3K9me3 and H3K27me3 with chemical treatment, as was observed in this study, suggests a rejuvenation effect on the epigenome.

There are a few reasons why declining H3K9me3 and H3K27me3 is considered an aging hallmark:

• Loss of these marks leads to heterochromatin deficits and derepression of normally silenced regions, contributing to genomic instability
• Removal of the marks occurs in an age-dependent manner due to altered histone modifier activity
• Declining levels correlate with conditions like premature aging disorders and senescence
• Experimental interventions that maintain or increase H3K9me3 & H3K27me3 can promote longevity and healthspan

Senescence-associated β-galactosidase serves as an enzyme marker indicating the burden of senescent cells. The percentage of cells displaying SA-β-Gal activity in cultures or tissues directly correlates with the degree of senescence. During the aging process, there is an accumulation of senescent cells in tissues, leading to an observable increase in measured SA-β-Gal activity.

 

The dose of Pulrodemstat is about 10-15 mg a day and Galunisertib 300mg per day each divided doses probably makes the most sense.

 

Please let me know if you are interested in a group buy as of now I have a chemist making Pulrodemstat as it is very expensive to buy. Galunisertib I have a source for a reasonable price.

 

For now Pulrodemstat would be 360 per gram which would be enough for about 10, 1 week cycles

 

Galunisertib would be 60 a gram if we get enough people.

 

Link to paper: https://www.biorxiv.....08.29.505222v1

 

 

Edited by Daniel Cooper, 11 January 2024 - 07:11 PM.

[BieraK]

Please, change the font type... it makes difficult to read

[Daniel Cooper]

Not saying that I'm not interested, but have you any idea of how long the list is of things that extend the lives of C. elegans? Or how short the list is of those things that subsequently pan out to be useful in humans?

 

Also, agree on the font issue.

[Chemically_Charmed]

It wont let me edit it. Can you do it?  The font looked different on my computer vs phone. Yes I am aware of that but it is possibly to make a cell into a pluripotent stem cell by using a drug combo that include these two types of drugs. The Yamanaka factors also do the same and Sinclair and his team are reversing aging with those. Sinclair also is planning on testing a drug combo to reverse aging. He has recently published a paper about it. https://www.aging-us...cle/204896/text

 

"A hallmark of eukaryotic aging is a loss of epigenetic information, a process that can be reversed. We have previously shown that the ectopic induction of the Yamanaka factors OCT4, SOX2, and KLF4 (OSK) in mammals can restore youthful DNA methylation patterns, transcript profiles, and tissue function, without erasing cellular identity, a process that requires active DNA demethylation. To screen for molecules that reverse cellular aging and rejuvenate human cells without altering the genome, we developed high-throughput cell-based assays that distinguish young from old and senescent cells, including transcription-based aging clocks and a real-time nucleocytoplasmic compartmentalization (NCC) assay. We identify six chemical cocktails, which, in less than a week and without compromising cellular identity, restore a youthful genome-wide transcript profile and reverse transcriptomic age. Thus, rejuvenation by age reversal can be achieved, not only by genetic, but also chemical means."

 

I think this has a good chance of age reversing effects in vivo.

[Daniel Cooper]

Fixed the font.

 

Yes, this is very interesting. But it would be very nice to see an experiment in a higher order animal like a mouse. Surely that is underway or planned.

 

I may be interested in your buy. Need to do some reading.

[Chemically_Charmed]

I'm surprised that on a longevity forum no one is interested in something that actually has the potential to reverse hallmarks of aging.

[Marcus Gitterle]

I am interested. Excellent project. I will join a group buy.

Edited by Marcus Gitterle, 04 February 2024 - 03:30 PM.

[sub7]

I'm surprised that on a longevity forum no one is interested in something that actually has the potential to reverse hallmarks of aging.

 

You have compared the functioning of this combination to the use of a small amount of Yamaka factors?

Do you happen to know what happens when one overdoses the Yamanake Factors? Yes cancer with 100 percent certainty

 

How on earth will you dose these things?
If you were using the exact same drugs as used in worms, even then doing the dose conversion would be extremely imprecise. Here you are substituting totally different things instead and not in a million years will you get the dosing right...

 

very very bad idea IMO

[Chemically_Charmed]

I am interested. Excellent project. I will join a group buy.

 

Send me a DM I'm starting a telegram group.

[LOOKINGFORTIME]

I am also interested in the group buy    john_faiola@yahoo.com

[Chemically_Charmed]

You have compared the functioning of this combination to the use of a small amount of Yamaka factors?

Do you happen to know what happens when one overdoses the Yamanake Factors? Yes cancer with 100 percent certainty

 

How on earth will you dose these things?
If you were using the exact same drugs as used in worms, even then doing the dose conversion would be extremely imprecise. Here you are substituting totally different things instead and not in a million years will you get the dosing right...

 

very very bad idea IMO

 

These are two anti cancer drugs and you would just be doing short bursts. I don't think the dosing has to be an exact amount to get a positive effect. Using 3/4 of the Yamanaka factors for short periods does not cause cancer.

[floret]

Galunisertib comes with considerable side effects and questionable results. There are reasons why its development was discontinued. I would take a deep dive into the papers published about the stuff--there are several.

 

Extending median lifespan is not the same as extending maximum lifespan

 

I would be very hesitant to try anything that did not have at least a robust mouse/other higher animal study backing up its safety and efficacy.  It's not just about whether it would be useless, but whether you'd do yourself irreparable harm.

[Chemically_Charmed]

Galunisertib comes with considerable side effects and questionable results. There are reasons why its development was discontinued. I would take a deep dive into the papers published about the stuff--there are several.

 

Extending median lifespan is not the same as extending maximum lifespan

 

I would be very hesitant to try anything that did not have at least a robust mouse/other higher animal study backing up its safety and efficacy.  It's not just about whether it would be useless, but whether you'd do yourself irreparable harm.

 

Sounds like Vactosertib would be better:

"Vactosertib is a highly selective and a potent small molecule inhibitor against Type 1 TGF-β Receptor (TGF-βR1), also known as activin receptor-like kinase 5 (ALK5), which binds at the ATP binding site on TGF-βR1. This leads to specific downstream down-regulation of the SMAD pathway. Vactosertib is orally available and has 10 times the potency (IC50=11nM) of another ALK5 inhibitor, Galunisertib (IC50=110nM) (23). Although high doses of Galunisertib over long durations caused cardiac toxicities (24), whereas Vactosertib showed limited toxicity without cardiac toxicity, and is well tolerated with minimal side effects in adults."
 

https://ascopubs.org....15_suppl.11557

[floret]

Nice find.

 

Were any ageing studies ever done with Vactosertib? I'm not finding any, although it appears to be a promising anti-cancer drug.

[Chemically_Charmed]

Nice find.

 

Were any ageing studies ever done with Vactosertib? I'm not finding any, although it appears to be a promising anti-cancer drug.

 

I don think so but I found this interesting one:

 

https://www.ncbi.nlm...les/PMC6186741/

 

"TEW-7197 (Vactosertib) induced significant regression of fibrotic plaque in PD rats in vivo through reduced infiltration of inflammatory cells and reduced expression of phospho-Smad2, which resulted in a recovery of erectile function. TEW-7197 also abrogated TGF-β1-induced enhancement in extracellular matrix production and hydroxyproline content in PD fibroblast in vitro by blocking TGF-β1-induced phosphorylation and nuclear translocation of Smad2 and Smad3, and by inhibiting TGF-β1-induced transdifferentiation of fibroblasts into myofibroblasts."

[floret]

Vactosertib does look more promising, but there's no evidence of it being an effective Yamaka-factor inducer (at least none I'm aware of).

 

What's curious here is how far this has strayed from the study OP quoted. The two substances used there were tranylcypromine and Repsox. Tranylcypromine is sold under the brand name of Parnate and is a proven safe/effective antidepressant (although as an MAOI inhibitor it can synergise dangerously with certain other substances). Repsox isn't a proven anything in any higher animals--yet (I'd be surprised if there weren't already experiments underway using 7c and 2c cocktails in mice or rats etc.) although there was some research done on its ability to help metabolise fat. Vactosertib does not appear to have been used anywhere in these molecular Yamanaka-factor experiments; neither has galunisertib. I wouldn't count on just being able to swap in substitutes for something so complex with so many unknowns.  That's a dangerous gamble especially when dealing with something that has a really high chance of giving you a lot of cancer really fast if you don't know exactly what you are doing. Until we see it done successfully in at least higher animals, we have no idea what we are doing.

 

That's not to say I'm not hopeful. I'm following this research and will happily give it a go once it is demonstrated to be safe and effective.

Edited by floret, 23 April 2024 - 07:23 AM.

[floret]

Update on this (I can't edit my other post so I'm double posting)  I did some more digging. Repsox DOES have some promising research done on rodents. Apparently it helps osteoporosis.

 

That is still not enough for me to feel safe risking comboing it up with Parnate (for which you have to avoid certain foods to take safely) until at least a good in vivo study on the 2c is done.

[Chemically_Charmed]

Vactosertib does look more promising, but there's no evidence of it being an effective Yamaka-factor inducer (at least none I'm aware of).

 

What's curious here is how far this has strayed from the study OP quoted. The two substances used there were tranylcypromine and Repsox. Tranylcypromine is sold under the brand name of Parnate and is a proven safe/effective antidepressant (although as an MAOI inhibitor it can synergise dangerously with certain other substances). Repsox isn't a proven anything in any higher animals--yet (I'd be surprised if there weren't already experiments underway using 7c and 2c cocktails in mice or rats etc.) although there was some research done on its ability to help metabolise fat. Vactosertib does not appear to have been used anywhere in these molecular Yamanaka-factor experiments; neither has galunisertib. I wouldn't count on just being able to swap in substitutes for something so complex with so many unknowns.  That's a dangerous gamble especially when dealing with something that has a really high chance of giving you a lot of cancer really fast if you don't know exactly what you are doing. Until we see it done successfully in at least higher animals, we have no idea what we are doing.

 

That's not to say I'm not hopeful. I'm following this research and will happily give it a go once it is demonstrated to be safe and effective.

 

The targets in the paper are LSD1 inhibition and TGF beta inhibition. Tranylcypromine is not selective at all and Repsox has not been tested in humans. I am using two drugs that selectively inhibit the targets that are also human safe.
 

[Chemically_Charmed]

https://www.youtube....h?v=PE5pTlvJPhg

 

At 29.17 David Sinclair is referring to the two chemical study results.

[floret]

It might be safe, but there's no evidence it'd do what 2c or 7c do. The chemical cocktail paper I read said they tested hundreds of compounds to come up with 7c. I think a lot of the usefulness of the cocktails has to do with how their components synergise.

 

Still, long as you guys stay safe, I say good luck!

[Chemically_Charmed]

It might be safe, but there's no evidence it'd do what 2c or 7c do. The chemical cocktail paper I read said they tested hundreds of compounds to come up with 7c. I think a lot of the usefulness of the cocktails has to do with how their components synergise.

 

Still, long as you guys stay safe, I say good luck!

 

Here is a Paper on the reprogramming effects of LSD1 inhibition.

https://www.ncbi.nlm...les/PMC4969595/

 

And here is one showing

Inhibition of transforming growth factor β (TGF-β) signaling can substitute for Oct4 protein in reprogramming and maintain pluripotency

https://pubmed.ncbi....h.gov/25548277/

Edited by Chemically_Charmed, Yesterday, 07:35 PM.