Senolytics

I've just finished my third Senolytic cycle.

Fisetin 1500 mgs on 2 consecutihve days repeated one month later

Dasatinab 180 mgs on 2 consecutive days + Quercitn 2250 mgs on the same days

 

I also take 4 mgs of Rapamycin once a week and 500 mgs of MetforminER twice a day.

65 years young.

128replies Oldest first
  • Oldest first
  • Newest first
  • Active threads
  • Popular
    • Dan Nave
    • Dan_Nave
    • 3 yrs ago
    • Reported - view

    A new wide spectrum senolytic SSK1 in development.  They imply that it works on all senescent cell types and works better than Dasatinib, Fisetin, and Quercetin.

     

    https://www.nature.c...1422-020-0314-9

     

    In this study, we designed a new prodrug based on the major senescence marker — the elevated β-gal — to selectively target senescent cells. This prodrug, SSK1, specifically killed both human and mouse senescent cells independent of senescent cell types and inducers. In a mouse lung injury model, SSK1 cleared stress-induced senescent cells in vivo and alleviated associated symptoms. Importantly, SSK1 also effectively reduced naturally occurring senescent cells in aged mice, decreased the senescence- and age-associated gene signatures, down-regulated SASP both locally and systemically, and restored physical functions. These results demonstrated the robustness and specificity of our prodrug in reducing senescent cells.

    We successfully developed a new prodrug strategy that directed gemcitabine to kill senescent cells in a highly selective manner. The foundation of our prodrug strategy was to select a highly specific senescence marker and identify an appropriate potent drug to efficiently kill senescent cells. First, the elevated enzymatic activity of β-gal, a universal senescence biomarker both in vitro and in vivo,18,52 was utilized to direct the prodrug to specifically target senescent cells. Second, through the screening of hundreds of FDA-approved drugs, we found gemcitabine to be one of the most potent agents in killing non-dividing senescent cells (Fig. 1a; Supplementary information, Fig. S1b, c, and Table S1). In addition, gemcitabine is a widely used FDA-approved drug with proven safety and short plasma circulation time.25,26 Our designed prodrug SSK1 is activated to release gemcitabine selectively in senescent cells (Fig. 1c), following which senescent cells were effectively eliminated (Fig. 1d). Our further study showed that gemcitabine activates p38 to induce apoptosis in non-dividing senescent cell (Supplementary information, Fig. S2a, b). As a nucleoside analog, gemcitabine could kill senescent cells through inducing mitochondrial DNA damage (Supplementary information, Fig. S2d), similar to a reported nucleoside analog, Ganciclovir.33 These results demonstrated that prodrug SSK1 is superior to current reported senolytics in terms of design strategy and specificity.

    Cellular senescence is a highly heterogeneous process due to the different cell origins and stimuli,6,53 whereas the key feature of SSK1 is the ability to efficiently clear senescent cells with a broad spectrum of cell types and senescence inducers, including replication, irradiation, oncogene and genotoxic stress (Fig. 2a, c). In this study, we compared SSK1 with other reported senolytics on HEFs, human preadipocytes and HUVECs, which were used to test senolytics. ABT263 (a classical anti-apoptosis BCL-2 inhibitor) eliminated senescent HEFs and HUVECs, but showed little effect on human preadipocytes (Supplementary information, Fig. S3b, f, j), which was also reported by other previous studies.12,13 The combination of dasatinib (a pan-tyrosine kinase inhibitor) and quercetin (a plant flavonoid) killed all three types of senescent cells in a dose-dependent manner (Supplementary information, Fig. S3c, g, k), but had a high toxic effect on non-senescent cells in consistence with others’ results.11,16,36 Another natural flavonoid fisetin, reported as a potential senotherapeutic agent,15 showed modest effect on eliminating senescent HEFs and preadipocytes even at higher concentrations (Supplementary information, Fig. S3d, h) which was comparable to other’s results.54 Most importantly, SSK1 could overcome these limitations, including cell-type dependency, high toxicity on non-senescent cells, and low efficiency on senescent cells (Supplementary information, Fig. S3a, e, i). Therefore, SSK1 possessed a better senolytic activity regarding of specificity and efficiency on a wider range of cell types, demonstrating the superiority of β-gal-based prodrug strategy to target senescence.

    Moreover, we found SSK1 exerted its elimination effect on senescent cells in vivo. SSK1 eliminated stress-induced senescent cells effectively and decreased different senescence markers in bleomycin-induced lung injury model, highlighting its effectiveness in vivo (Fig. 3). SSK1 treatment also relieved lung fibrosis (Fig. 3d; Supplementary information, Fig. S4c, d) and attenuated the impaired physical function as tested by treadmill assay (Supplementary information, Fig. S4f). In this study, we also treated naturally aged mice with SSK1 and tested its effects on senescent cells, chronic inflammation and physical function. First, SSK1 could remove senescent cells in multiple tissues and decrease the senescence-associated signatures as shown by the GSEA analysis (Fig. 4b–g). Second, SSK1 could decrease the expression of SASP-associated genes in aged livers and kidneys and reduce chronic low-grade inflammation in the blood (Fig. 5a, b, e–h). Third, SSK1 ameliorated the impaired motor function, balance, exhausted exercise, muscle strength, and spontaneous exploration in aged mice (Fig. 6a–e). Most importantly, the performance of rotarod and beam balance in the SSK1-treated group was improved compared with that in the initial pretreatment condition (Supplementary information, Fig. S8e, f). Collectively, our prodrug SSK1 targeting β-gal-positive cells exerted very significant biological effects in naturally aged mice.

    While SA-β-gal is widely used as a marker of cellular senescence,20,21,22 its elevated activity can be found in some other cells such as activated macrophages.48,55 These SA-β-gal-positive macrophages can be harmful and have been found to accumulate in injured and aged tissues contributing to chronic inflammation.44,45 Importantly, we have shown that SSK1 decreases the number of SA-β-gal-positive macrophages in injured lungs and aged livers (Supplementary information, Fig. S6g–j), which is consistent with our observation of reduced secretion of chronic inflammation-related cytokines. Therefore, eliminating macrophage accumulation by SSK1 might reduce chronic inflammation and benefit aged organisms. In addition, activated macrophages play crucial roles in acute inflammation and cytokine storm,56 especially those induced by virus infection. During virus-induced acute inflammation, macrophages produce pro-inflammatory factors and trigger initiation of cytokine storms.57 For instance, the depletion of macrophages could protect mice from coronavirus-induced lethal infection.58 Accordingly, activated macrophages could be potential targets for treating acute inflammation and cytokine storms via SSK1. The future potential for SSK1 in treatment of acute inflammation, injury, and age-induced chronic inflammation is promising.

    An understanding of the toxicological effects of SSK1 in vivo is critical to clinical applications. Importantly, our data showed that high concentration (100 mg/kg) and high frequency SSK1 treatment had no apparent systemic toxicities (Supplementary information, Fig. S9). This provides strong evidence for the in vivo safety of SSK1. This safety profile is further supported by comparison to gemcitabine, an SSK1 effector and approved clinical drug. First, our effective in vivo dosage of SSK1 was approximately 60-fold lower than the clinical dosage of gemcitabine (30 mg/kg) and implies a greatly reduced risk of in vivo toxicity of SSK1.24 Second, gemcitabine is a nucleoside analog that potently affects rapidly dividing cells and the off-target effects of gemcitabine could be shown in different proliferating cell types.59,60 SSK1, however, targets only β-gal-positive cells. Notably, in addition to macrophages, we found that the majority of non-senescent β-gal-positive cells are rapidly dividing epithelial cells (Supplementary information, Fig. S10a, b). As a result, the potential types of proliferating cells with SSK1 sensitivity are greatly narrowed relative to gemcitabine. Even though small numbers of β-gal-positive epithelial cells are targeted by SSK1 treatment, these cells have robust self-renewal ability.61 Consistent with this self-renewal property, we found that epithelial tubular cells in the kidney with elevated β-gal activity had a high percentage of Ki67-positive cells (Supplementary information, Fig. S10c). Following SSK1 treatment, we observed only a few cells of this tubular cell population undergoing apoptosis (Supplementary information, Fig. S10d), and such low-level impairment could be easily repaired by the rapid self-renewal of tubular cells during SSK1 treatment. Moreover, the short duration of SSK1 treatment might avoid significant impairment in related tissues and further minimize its side effects. In summary, our study demonstrates the superiority and safety of this prodrug strategy by targeting β-gal to selectively remove senescent cells of multiple cell types. These findings open a new avenue for the treatment of age-associated diseases and provide a clinical opportunity for intervention into the aging process.

    Like 2
    • Dan Nave That is fascinating news. I wonder how effective regular gemcitabine would be if we compare it to Dasatinib? great info thank you.

      Like 1
    • Staffan Olsson i don't understand, gemcitabine and ssk1 is the same? Or does it act with the same benefits without affecting other cells that are not senescent?

      Like 1
    • Pablo Reinaldos López As  I understand, they have created something that is better to deliver gemcitabin into senescent cells. "..designed prodrug SSK1 is activated to release gemcitabine selectively in senescent cells"

      Like 2
      • Karl
      • Karl.1
      • 3 yrs ago
      • Reported - view

      Dan Nave great find. Unfortunately sounds like this would become a patented drug and fetch a high price?

      Like 1
      • Dan Nave
      • Dan_Nave
      • 3 yrs ago
      • Reported - view

      Karl We can only hope for a reasonable price.  Who wouldn't want to use it. 

      Like 1
      • JGC
      • Retired Professor of Physics
      • JGC
      • 3 yrs ago
      • Reported - view

      Dan Nave 

          I don't think SSK1 could be made available here in the USA without having first a trial on humans. It would be very hard to get the FDA to approve such a trial on a drug that treats aging (because in the view of the FDA, aging is not a disease and therefore doesn't require treatment.)  Oisin Bio, with their plasmid/liposome treatment to clear senescent cells by inducing aptosis, has just that problem in getting a trial, and it took many years to get approval for the ongoing Mayo trial of metformin.

      Like 1
      • JGC
      • Retired Professor of Physics
      • JGC
      • 3 yrs ago
      • Reported - view

      Dan Nave 

          On the other hand, the Chinese paper gives explicit instructions for how to synthesize SSK1, and it's simple enough that most organic chemists in well-equipped labs could do the job.  I calculate that SSK1 should have a net molecular mass of about 925.9 Da.  That's not a very large molecule.  Vitamin B12, which is taken orally, has a mass of 1,355.4 Da.  I wonder what the water solubility of SSK1 is.

      Like 2
  • attacking senescent cells is evolving as the #1 treatment for aging

    Like 2
    • Paul Beauchemin 

      I just took my first senolytic cocktail yesterday and had a unusual reaction and wondering if anyone else had similar

      I am 70 yr old male (n=1) who has taken rapamycin for two years weekly.  Also metformin for years (nondiabetic).  Took the following after lunch yesterday :

      90mg dasatinib , 1200 mg quercetin, 250mg AZM, 300mg fisetin

      No reaction for 5 hours. Then had a bout of diarrhea for 30 minutes.Then OK.  Eat dinner. 2 hours later started having some body aches but not bad.  Went to bed 2 hours later had more aches and then the chills and shivering in bed and had pain in all my joints.  Lasted 2 hours and then went away.  Woke up fine.

      I was going to take 2 more days of this protocol but now I'm having second thoughts.

      Anyone have similar complaints.  Was it a reaction to dasatinib?  I think senolytic's is the future but maybe a different protocol.  Any ideas appreciated

      Like 2
      • Van
      • Van
      • 3 yrs ago
      • Reported - view

      Edward Meininger Wow, it is not necessary to take all of them at one time.  There are 4 different types of senolytics, and each supplement works best on one.  Please read Dr. Green's new website.  At the bottom he recommends the one's your taking and the doses and schedules.  I have also taken Rapa for 3 years, and do the senolytics quarterly.   https://senolyticstreatment.com/

      Like 1
      • Joe smith
      • Joe_smith
      • 3 yrs ago
      • Reported - view

      Edward Meininger please stop what you doing and see doctor ASAP!

      Like 1
      • Dan Nave
      • Dan_Nave
      • 3 yrs ago
      • Reported - view

      Edward Meininger Do you normally have joint problems, arthritis, rheumatism, etc?

      Like 1
    • Dan Nave I don't have any joint problems or pre existing conditions.  Read Dr Green's page re senolytics.  Seems some of studies used D + Q and others just D. I combined all 4 senolytics which may heve been a mistake.  Do most people use Singular senolytic?

      Like 1
      • Karl
      • Karl.1
      • 3 yrs ago
      • Reported - view

      Edward Meininger I have only taken D&Q once, but it kicked my ass.  Nausea started at 2 hrs and last for about 10 hrs with vomiting at the end. Body aches and fatigue for several hrs, and about 10 hrs post dose I had fever and rigors. I will definitely change dosing next time.

      Like 3
    • Karl Your reaction was certainly worse than mine.  I was able to do 3 days of AZM 250 mg with no real problem.  I will try fiesetin alone 1500 mg next quarter.  I think senolytics with weekly Rapamycin is a game changer

      Like 2
      • Joe smith
      • Joe_smith
      • 3 yrs ago
      • Reported - view

      Edward Meininger My concern with the senolitics is that big-time researchers like Judith Campisi and Dimitry Bulavin clearly state and demonstrate through their research that senolitics don’t extend the lifespan. Further, Dimity Bulavin research indicates that senolitics may actually very negatively affect quality of life. For instance liver is not very good at regenerating after senolitic treatment in older age resulting in fibrosis. The fibrosis is not just in liver. He actually has to kill poor mice under treatment as their bodies are clearly damaged and they are suffering due to experiments. He doesn’t care that they may live longer. He does the humane thing and terminates them. Based on this controversy, I question the value of senolitic treatment at this time.

      Like 1
      • Van
      • Van
      • 3 yrs ago
      • Reported - view

      Edward Meininger I really feel that this is important.  In Dr. Green's new website he talks about the 4 different kinds of senolytics and the different treatments for each one.  He recommends quarterly treatments for anti aging benefits, and monthly for people treating specific diseases.  Fistein 1500 mg X 3 days.   Azithromycin 500 mg x 3 in 1 week. (appx. every other day)  This is the same dose that has been used by MS patients for years to prolong there lives.   Dasatinib 100 mg x 3 days.  On his website he talks about the science behind using these drugs and dosages.  Either way, I would not take them together.  Rapamycin reduces the amount of senolytics, but cannot clear out the zombie cells like specific senolytics can.

      https://senolyticstreatment.com/

      Like 2
      • JGC
      • Retired Professor of Physics
      • JGC
      • 3 yrs ago
      • Reported - view

      Joe smith 

          You said "big-time researchers like Judith Campisi and Dimitry Bulavin clearly state and demonstrate through their research that senolitics don’t extend the lifespan."

          I am very suspicious of the lifespan vs. healthspan dichotomy.  I believe it is a cop-out employed by age-treatment researchers to convince the FDA and NIH that they are working to improve a patient's health (which the NIH funds) rather than making the patient live longer (which the NIH refuses to fund).   Why wouldn't a significant improvement in health make the subject live longer?  It makes no sense.

      Like 4
      • JOHN
      • JOHN.1
      • 3 yrs ago
      • Reported - view

      Edward Meininger The first time I took Dastinib I took 100mg plus 1500mg quercetin. I too had chills and shivering in bed. I also had a temperature and threw up constantly for a few hours. Next day I was fine. I waited a few more months to try this again. Just finished a 3 day protocol and was relatively fine. Bad headaches and loss of appetite at times but nothing serious. I too am a Dr Green patient and take rapa as well.

      Like 2
  • this is what Dr Green lists:

    Dasatinib 100 mg dose for 3 days.

    Quercetin 1000 mg for 3 days

    Fisetin 1500 mg for 3 days.

     

    I get diarrhea every time I take more than 500 mg of quercetin and I imagine the antibiotic would affect me the same

    I never had aches and pains but I read others report flu like symptoms the first time they take Dasatinib

    Like 2
    • Van
    • Van
    • 3 yrs ago
    • Reported - view

    Just received my order for Fisetin 50 grams and Dasatinib 5 grams from China.  No problems with Customs was shipped via Fedex.  Fisetin was $2.50 gram including $25 Fedex.  Dasatinib was $20 gram + $25 shipping.  I received exactly 50 grams of Fisetin, and 5.6 grams of Dasatinib.  Over paid for Fisetin, better to order online.  Dasatinib and Rapamycin also are such small amounts that they can ship in an envelope and less likely to have a problem with customs.  Dr. Green is now recommending to take Fisetin and Dasatinib together and you can skip or not quercetin because Fisetin works better.  Green recommends 1500 mg Fisetin x 3 day x quarterly for anti ageing.  I prefer to use Mayo clinic's formula of 20mg/kg.  On July 01, 2020, I will take Fisetin 1700 mg x 3(with olive oil) + Dasatinib 100 mg x 3.(quarterly)  The next week will  take 250 mg Zithromycin x 6 days (quarterly)  Will try taking Vitamin C, 500 mg with all of the above.  These 3 drugs target 3 of the 4 senescent putrid cells, (aka, zombie)  WIll be waiting for a treatment for the 4th. cell type.

    Like 2
    • JGC
    • Retired Professor of Physics
    • JGC
    • 3 yrs ago
    • Reported - view
    Paul Beauchemin said:
    attacking senescent cells is evolving as the #1 treatment for aging

         In my view, clearing senescent cells is the first step (but by no means the last) in treating aging.  The cell's internal programming has declared senescent cells to be too damaged to replicate, and that decision should be taken seriously.  One should clear them before any other intervention.  After that, if quasi-embryonic pluripotent stem cells are available, they should be provided to replace the cleared cells.  (Apparently, given the recent Unity Bio trial failure, this is important.)  Then, if an intervention is available to reset the epigenetic programming of the remaining cells (e.g., by AAV delivery of DNA or liposome delivery of mRNA to cause OSK Yamanaka factor expression), this should be done.  In principle, we know how to do all of these steps right now.  However, FDA roadblocks and the expense of human testing put access to these therapies far down the road, at least in the USA.

    Like 4
      • JOHN
      • JOHN.1
      • 3 yrs ago
      • Reported - view

      JGC In your opinion do you think Unity Bio has a shot with their next trial on vision?

      Like 1
    • JGC
    • Retired Professor of Physics
    • JGC
    • 3 yrs ago
    • Reported - view
    JOHN said:
    In your opinion do you think Unity Bio has a shot with their next trial on vision?

         I wouldn't put any money on it.  Essentially, the flawed approach of Unity Bio involves repurposing a highly toxic anti-cancer drug that they call UBX-0101 and have patented for senolytic applications.  UBX-0101 cannot be used as a general senolytic because of its toxicity.  To avoid whole-body toxic effects, they inject UBX-0101 into relatively isolated body systems like the knee capsule or the eye capsule.  The problem with this approach, in my opinion, is that the cleared senescent cells need to be replaced with healthy one, they are not clearing nearby senescence outside the capsule, and the SASP effects of these neighboring senescent cells are preventing replacement by stem cells.

        In my view, one needs to use a general non-toxic senolytic that clears all senescent cells.  Oisin Bio's DNA plasmid/liposome technique does this.  The recent Chinese paper describing a new synthetic small-molecule senolytic called SSK1 that targets the senescence-marker beta-galactosidase seems to promise to do the same.  However, the toxicity of SSK1 under all conditions has not been well established, it has not been tested on humans, and all senescent cells do not express beta-galactosidase.  Both such treatments should be followed up by providing a supply of pluripotent stem cells to replace the cleared senescent ones.

    Like 1
      • JOHN
      • JOHN.1
      • 3 yrs ago
      • Reported - view

      JGC That's interesting. I didn't realize they were repurposing an anti cancer drug.  I've seen you make a few posts on taking senolytics yourself. Do you think it's necessary to replace these senescent cells with stem cells?

      Like 2
Like8 Follow
  • 8 Likes
  • 2 yrs agoLast active
  • 128Replies
  • 4179Views
  • 27 Following