34 replies to this topic

[sub7]

This research is of course very important

https://www.ncbi.nlm...les/PMC6710051/

and has already been talked about

 

Both of the substances are available: Oxytocin can be obtained. ALK5 inhibitors are also available. Example: https://www.peprotec...nhibitor-repsox

 

Is anyone ready to try this combination?

Can you comment on the safety of ALK5 inhibitors in general and whether the above product is suited for that purpose?

[xEva]

where do you obtain oxytocin?

thanks

[adamh]

where do you obtain oxytocin?

thanks

 

I got some from ebay, look around

[sub7]

I am sorry I messed up the title, is that why there have been no responses?

 

I think this is a very critical piece of research that is definitely worth talking about.

 

any one...?

[Oakman]

I am sorry I messed up the title, is that why there have been no responses?

 

I think this is a very critical piece of research that is definitely worth talking about.

 

any one...?

 

There is so much research being done...with only some getting even a limited bit of acknowledgement. This is a problem of being overwhelmed with like information. Everyone wants to extend life, rejuvenate old tissues. Is this the 'one'? Possibly, but will it be lost in the flood of similar research? Possibly, or more likely yes. Oxytocin to me seems to be a very valuable molecule, overlooked for it's capabilities. But strangley, it's hard to procure at a reasonable price, or any price. Of course it is in various foods, but there is no way to know exactly how much or how much is even needed.

 

And currently, this forum has almost been turned in a one or a few thread forum, of course, COVID-19 being the central focus. There have been threads here on oxytocin, making it with yougurt, eating certain foods, etc. There were spurts of interest and effort, but then topics and people move on. Never underestimate the ability of people to ignore good advice and possible effective protocols to focus on something new, unproven and topical. Quite sad.

[Vitalist]

Thanks for posting sub7, the study looks credible. Excellent results (in mice). 

 

But as is often the case, these substances are not so easy to acquire. Even thought the ALK5 Inhibitor is carried by many of the chemical supply sites, they typically won't ship to home addresses, or deal with private individuals at all.

 

Too bad because I wouldn't mind giving this a try. 

 

[bosharpe]

I'd potentially be willing to give it a go. I've been eyeing an Oxytocin nasel spray from Khemcorp for a couple of months. How/where does one aquire Alk5?

[sub7]

Thanks a lot for helping me revive the thread. A moderator can come in and correct me here, but I do not think we are supposed to openly ask for sources here -not that I have a source which I am hiding, I am not... In my opinion, regardless of rules in place on this forum, a message board is not the right place to discuss sources.

 

Can anyone, who is proficient in mice-to-human dose conversion give us an idea about how much of the ALK5-inhibitor as well as Oxytocin we may have to use, so people can personally assess the feasibility / affordability here?

Of course we have to keep in mind that mice studies tend to be more "proof of concept" type analysis, where researchers are primarily interested in seeing if there is any significant effect at all. The safety of test animals as well as side effects are either a very minor concern or of no importance at all. We therefore need some help from members who know this stuff, so we can not only convert the dose, but also speculate on how we would adjust this "direct conversion" where we strike a balance between likely seeing some benefit while managing the associated danger.

[sub7]

There is so much research being done...with only some getting even a limited bit of acknowledgement. This is a problem of being overwhelmed with like information. Everyone wants to extend life, rejuvenate old tissues. Is this the 'one'? Possibly, but will it be lost in the flood of similar research? Possibly, or more likely yes. Oxytocin to me seems to be a very valuable molecule, overlooked for it's capabilities. But strangley, it's hard to procure at a reasonable price, or any price. Of course it is in various foods, but there is no way to know exactly how much or how much is even needed.

 

There have been threads here on oxytocin, making it with yougurt, eating certain foods, etc. There were spurts of interest and effort, but then topics and people move on. Never underestimate the ability of people to ignore good advice and possible effective protocols to focus on something new, unproven and topical. Quite sad.

 

Absolutely agree with every one of your eloquent comments. I definitely believe that Oxytocin is a very valuable molecule and am eating a high dose Reuteri yogurt myself almost every day.

 

You are totally correct in your assessment...  We are involved in a very tough endeavor, where a lot of good findings are ignored and the majority tend to bark at the wrong tree. There is also a large crowd, which doesn't want to take even a moderate amount of risk. As soon as a molecule is shown to have any effect at all, they engage in an extreme effort to identify and consume foods that have the said molecule but would never even topically apply a cream containing the substance -unless proven very safe in large studies. No disrespect to such people, but they are wasting their time. If you want to extend your lifespan without taking even a moderate risk, there is essentially nothing available for you at the moment. You will just have to lead a healthy lifestyle and nobody needs to browse the forums to identify what such a lifestyle involves.

 

Take the Reuteri yogurt for example; it turned into a back-and-forth about how to make the yogurt tastier and creamier. A small percentage of the discussion was focused on how to maximize the amount of the Reuteri strain concentration. All good and well, but we are not going to extend lifespan with such discussions.

 

OK, with all that out of the way, let's stay positive and concentrate our collective effort here on how we can use ALK5-i and Oxytocin.

[sub7]

Here is something very interesting. The referenced study I posted says this

OT and various Alk5 inhibitors [43] are already FDA approved for applications that are different from age-related degenerative pathologies, and hence repositioning this combination is facilitated. 

 

What the hell??? Really

When I follow the link next to this comment I get just this reference to the mentioned ALK5 inhibitors:
 

TGF-β1 Type I Receptor Kinase Alk5 inhibitor 2-(3-(6-Methylpyridin-2-yl)- 407 1H-pyrazol-4-yl)-1,5-naphthyridine

 

This is FDA approved??? Doesn't sound like it...

[Daniel Cooper]

Following this thread. 

 

I looked for a currently approved ALK5 inhibitor but so far have come up empty. But my search is hardly exhaustive.

 

 

[OP2040]

I've wanted to act on this study, in particular because my woman is experiencing pretty terrible menopausal symptoms and that's a second bird I'd like to kill.  But the oxytocin nasal spray doesn't seem like what we would want.  It is marketed specifically for one-time uses and it's super-expensive.  What we need is something that raises blood levels of oxytocin consistently over time, and thus it must be affordable for longer term use.  Outside of those parameters, it would be quite worthless.

 

I can also back up what Daniel is saying. A fairly extensive search for AlK5 inhibitors, etc. doesn't provide much to go on at this point.

 

It would be great if ALK5 was key to TGFb since that target is notoriously pleiotropic and hard to target but definitely involved in aging and regeneration in many ways.

[OP2040]

When Self-hacked has an entry it is usually a goldmine of consolidated information.  Their entry on oxytocin sort of confirms what we are saying.  There's a lot of enticing hints and suggestions but very little evidence.  Maybe someone with more time than myself can look up listed citations listed there for more clarity on the matter

 

https://selfhacked.c...-receptor-snps/

Edited by OP2040, 30 April 2020 - 03:21 PM.

[sub7]

When Self-hacked has an entry it is usually a goldmine of consolidated information.  Their entry on oxytocin sort of confirms what we are saying.  There's a lot of enticing hints and suggestions but very little evidence.  Maybe someone with more time than myself can look up listed citations listed there for more clarity on the matter

 

https://selfhacked.c...-receptor-snps/

 

Perhaps better to focus this thread on the combination of ALK5-i plus Oxytocin? Oxytocin alone is the safe and easy part of the equation. It is easier to obtain and use and has been used a lot without mush ill effect (if dose is moderate).

 

BTW, any ideas about the dosage (of each of the 2 substances) to be used in a human self-experiment?

[Hieon]

Here it goes again with "New Mice study, must buy this".

ALK5 inhibitor is nothing new. These researches are for restoring the optimum level of TGF-ß after overexpression of the signal associated with age degeneration or deregulation. It is not to reduce the signal below optimum level or overinhibition of ALK5 pathology. Deviating either direction from the optimum level may have an inverse negative effect. You make a knockout mouse of this pathology, I'm pretty sure it will instantly die.

Sure, if you clinically have indications of excessive TGF-ß, it may or may not help but you can try. But there are human vivo tested alternatives.

You really think mice model is directly translatable? And this in no way is a model for aging prevention for healthy individuals. It is rejuvenation for age-related degeneration.

[sub7]

1 - "there are human vivo tested alternatives."

2- " this in no way is a model for aging prevention for healthy individuals. It is rejuvenation for age-related degeneration."

 

1- Can you please explain what you  are referring to?

 

2- So if you are a 60 year old human with no specific disease, you will not benefit from this intervention? If I understand correctly and this is what you mean, can you please explain how you reached this conclusion?

 

Thank you 

[Hieon]

1- Can you please explain what you are referring to?

2- So if you are a 60 year old human with no specific disease, you will not benefit from this intervention? If I understand correctly and this is what you mean, can you please explain how you reached this conclusion?

Thank you

1. I'm just Bio MSc grad in med school currently and not a practitioner so I'm not gonna make personal recommendations or name all the alternative triggers of the same pathology, because I don't wanna be responsible for anyone's irresponsibility of simply buying or trying something just because "This guy said this". You can research yourself more or look for human vivo trialed drugs such as dasatinib of the similar modular inhibition of ALK5.

2. Everything in our biology has a mechanistic function. Even amyloid or tau tangles or TNF-alpha or cellular senescence all have their functions given their optimum level at homeostasis. Inflammatory response is also a defense mechanism, same applies to TGF-ß. These aspects are bad when they accumulate more than homeostatic level or artificially deregulated.

Therefore if TGF-ß signal is overexpressed particularly in aged mice, it makes sense to bring it down to optimum level through ALK5 inhibition, and this is the primary reason for investigation. But if you are at an optimum level already, and you downregulate it for whatever reason, then it will cause an inverse effect. That's why this is a rejuvenation tactic in the aged model, not a preventative tactic for healthy model.

Perhaps a contributing reason why metformin produced a lifespan increase in obese individuals but not proven in healthy individuals. Because their overall mechanisms are deregulated, metformin will bring them to homeostatic levels so improving overall healthspan. However in healthy folks, overactivating AMPK or overinhibition of mTOR will do no good or cause inverse effect because same reasoning applies; mTOR has its function as a growth regulator and should not be always inhibited, and only an idiot would think inhibition of any pathology is always good.

That's why if you make a knockout mouse of any of these will die instantly. All the investigative senolytics have other potentiating factors than just inhibition/activation, that is why it is still unclear how even metformin actually works.

So bottom line, there are alternative ways to induce ALK5 inhibition, but inhibition alone isn't what is giving off benefits from whatever compound. And if you think just inhibiting something will always be good, then you are no different than a druggy following hypes with zero understanding.

All that being said, I could be just as wrong as anyone. So do your own research and be skeptical please.

[sub7]

Thank you very much Hieon for your thorough response. I appreciate the time and thought you put into this; made even more meaningful by virtue of your background in the field. 

Please do not be offended by anything I express below, because I truly desire a productive and fruitful exchange on this site. We have nothing to gain from pure conflict and all that I write below is backed by a spirit of cooperation -even if I do a bad job of communicating that at times...

 

1. I'm just Bio MSc grad in med school currently and not a practitioner so I'm not gonna make personal recommendations or name all the alternative triggers of the same pathology, because I don't wanna be responsible for anyone's irresponsibility of simply buying or trying something just because "This guy said this". You can research yourself more or look for human vivo trialed drugs such as dasatinib of the similar modular inhibition of ALK5.

2. Everything in our biology has a mechanistic function. Even amyloid or tau tangles or TNF-alpha or cellular senescence all have their functions given their optimum level at homeostasis. Inflammatory response is also a defense mechanism, same applies to TGF-ß. These aspects are bad when they accumulate more than homeostatic level or artificially deregulated.

Therefore if TGF-ß signal is overexpressed particularly in aged mice, it makes sense to bring it down to optimum level through ALK5 inhibition, and this is the primary reason for investigation. But if you are at an optimum level already, and you downregulate it for whatever reason, then it will cause an inverse effect. That's why this is a rejuvenation tactic in the aged model, not a preventative tactic for healthy model.

Perhaps a contributing reason why metformin produced a lifespan increase in obese individuals but not proven in healthy individuals. Because their overall mechanisms are deregulated, metformin will bring them to homeostatic levels so improving overall healthspan. However in healthy folks, overactivating AMPK or overinhibition of mTOR will do no good or cause inverse effect because same reasoning applies; mTOR has its function as a growth regulator and should not be always inhibited, and only an idiot would think inhibition of any pathology is always good.

That's why if you make a knockout mouse of any of these will die instantly. All the investigative senolytics have other potentiating factors than just inhibition/activation, that is why it is still unclear how even metformin actually works.

So bottom line, there are alternative ways to induce ALK5 inhibition, but inhibition alone isn't what is giving off benefits from whatever compound. And if you think just inhibiting something will always be good, then you are no different than a druggy following hypes with zero understanding.

All that being said, I could be just as wrong as anyone. So do your own research and be skeptical please.

 

You are making so many assumptions; so to make this discussion more productive I kindly request that you refrain from assuming.

 

"I'm not gonna make personal recommendations"

Please; nothing at all that anybody wrote anywhere in this thread can be construed as asking for personal recommendations, who ever asked you for that?

 

" if you think just inhibiting something will always be good"

When did I ever express anything remotely resembling this? Not in any way, shape or form

 

"You really think mice model is directly translatable? "

Who ever said directly translatable. I even was the one who said in such experiments the safety of mice is a non-concern and we should take steps accordingly.

 

Every single time we take an action in life, we assume a risk. Hoping into your car to go to Starbucks constitutes a great risk. The number of people killed while driving and through no fault of their own is what... millions maybe since the invention of the automobile? However, if the risk is acceptable and we have already taken all possible precautions, we just have to move on

 

"because I don't wanna be responsible for anyone's irresponsibility of simply buying or trying something just because "This guy said this"."

If this is your mindset, this whole anti-aging thing is a non-starter really. If you go ahead and say, for example: "to accomplish the same end goal with fewer side effects, substance X makes more sense than ALK5-i. But this is not tested in humans and I do not deem self-experimentation a viable alternative" than you're good to go in my humble opinion.

If someone goes out and tries said substance based on such a harmless sentence that has been written on a discussion forum as part of an intellectual discussion, consequences are entirely and I mean entirely, their responsibility. If we are going to be this concerned with such individuals misusing the information herein, we really cannot talk on an open forum. People do wrong things and hurt themselves all the time. We cannot take every step in our lives with the primary goal of preventing such harm caused by others to themselves. We'd be betraying ourselves if we do so and would thereby allow such individuals to hurt us and society at large far more than they would (or will anyway despite our best efforts) hurt themselves.

 

"You can research yourself more or look for human vivo trialed drugs such as dasatinib of the similar modular inhibition of ALK5."

Sure I can research myself and I am. I can also produce a primitive shoe to wear and make a terrible bread to eat. However we chose to join our efforts to benefit from each other's accumulated wisdom. No matter how much I research, I will not become as wise in these said drugs as the combined wisdom of the people viewing this thread.

Not sharing our wisdom just because someone may do something extremely stupid and harm themselves as a result would be a shame.

If that will be our approach, why bother hanging out on a discussion board?

 

We simply cannot operate under the assumption that nature has optimized everything

"Everything in our biology has a mechanistic function. Even amyloid or tau tangles or TNF-alpha or cellular senescence all have their functions given their optimum level at homeostasis. Inflammatory response is also a defense mechanism, same applies to TGF-ß. These aspects are bad when they accumulate more than homeostatic level or artificially deregulated."

 

I disagree with the spirit expressed in the above comment and here is why:

The only way I can interpret the above is: "A 60 year old individual with no specific disease should not go in and do something to reverse or slow down aging". The very results we observe suggest -NOT THEY ARE NOT CONCLUSIVE AND YES THERE IS YET INSUFFICIENT HUMAN DATA, HENCE I SAY "SUGGEST" - that we can take a 60 year old who is in excellent shape for his age and has no specific disease, and remove some amount of the senescent cells in his body to make him function better. No, in the case of such an individual the amount of senescence in his body is not optimized by nature -we can do better by fine tuning it. Do we have the mechanism at hand for this? No. Do we know which cells to remove and how many of them to remove? No. Could it turn out that we are wrong and that removing even one single senescent cell by way or exogenous intervention will do more harm than good? Weeeellll, that last question is where it's at. One has to say Yes since we can never be certain and there is a very low probability here, but there is a very strong possibility that the more suited answer is No. It appears that probably removing some senescent cells will do more good than harm, but the "some" here is yet to be defined.

 

At the end some people like myself want to take such risks and others do not, and that is perfectly fine. We all have different priorities as well as risk profiles, which makes the world an interesting place...

 

To make matters all the more viable in this particular case, the very ALK5-i used in the mice study is referred to as FDA-approved. But, as you will see in the earlier posts, we could not figure out when / where etc.... However, at least it seems (SEEMS) that we are talking about a substance that at some dose won't outright kill you.

 

With all of that said, I would like to very respectfully submit the following questions:

 

1- What other more feasible alternatives may be worth intellectually discussing here other than the specific ALK5-i mentioned in the study? You did mention Dasatinib; anything else?

 

2- Would you speculate that the impact of those other ALK5-inhibitors too would possibly be boosted by adding Oxytocin?

 

Thank you very much for your input and, more crucially, your understanding.

 

[Hieon]

Thank you very much Hieon for your thorough response. I appreciate the time and thought you put into this; made even more meaningful by virtue of your background in the field.
Please do not be offended by anything I express below, because I truly desire a productive and fruitful exchange on this site. We have nothing to gain from pure conflict and all that I write below is backed by a spirit of cooperation -even if I do a bad job of communicating that at times...


You are making so many assumptions; so to make this discussion more productive I kindly request that you refrain from assuming.

"I'm not gonna make personal recommendations"
Please; nothing at all that anybody wrote anywhere in this thread can be construed as asking for personal recommendations, who ever asked you for that?

" if you think just inhibiting something will always be good"
When did I ever express anything remotely resembling this? Not in any way, shape or form

"You really think mice model is directly translatable? "
Who ever said directly translatable. I even was the one who said in such experiments the safety of mice is a non-concern and we should take steps accordingly.

Every single time we take an action in life, we assume a risk. Hoping into your car to go to Starbucks constitutes a great risk. The number of people killed while driving and through no fault of their own is what... millions maybe since the invention of the automobile? However, if the risk is acceptable and we have already taken all possible precautions, we just have to move on

"because I don't wanna be responsible for anyone's irresponsibility of simply buying or trying something just because "This guy said this"."
If this is your mindset, this whole anti-aging thing is a non-starter really. If you go ahead and say, for example: "to accomplish the same end goal with fewer side effects, substance X makes more sense than ALK5-i. But this is not tested in humans and I do not deem self-experimentation a viable alternative" than you're good to go in my humble opinion.
If someone goes out and tries said substance based on such a harmless sentence that has been written on a discussion forum as part of an intellectual discussion, consequences are entirely and I mean entirely, their responsibility. If we are going to be this concerned with such individuals misusing the information herein, we really cannot talk on an open forum. People do wrong things and hurt themselves all the time. We cannot take every step in our lives with the primary goal of preventing such harm caused by others to themselves. We'd be betraying ourselves if we do so and would thereby allow such individuals to hurt us and society at large far more than they would (or will anyway despite our best efforts) hurt themselves.

"You can research yourself more or look for human vivo trialed drugs such as dasatinib of the similar modular inhibition of ALK5."
Sure I can research myself and I am. I can also produce a primitive shoe to wear and make a terrible bread to eat. However we chose to join our efforts to benefit from each other's accumulated wisdom. No matter how much I research, I will not become as wise in these said drugs as the combined wisdom of the people viewing this thread.
Not sharing our wisdom just because someone may do something extremely stupid and harm themselves as a result would be a shame.
If that will be our approach, why bother hanging out on a discussion board?

We simply cannot operate under the assumption that nature has optimized everything
"Everything in our biology has a mechanistic function. Even amyloid or tau tangles or TNF-alpha or cellular senescence all have their functions given their optimum level at homeostasis. Inflammatory response is also a defense mechanism, same applies to TGF-ß. These aspects are bad when they accumulate more than homeostatic level or artificially deregulated."

I disagree with the spirit expressed in the above comment and here is why:
The only way I can interpret the above is: "A 60 year old individual with no specific disease should not go in and do something to reverse or slow down aging". The very results we observe suggest -NOT THEY ARE NOT CONCLUSIVE AND YES THERE IS YET INSUFFICIENT HUMAN DATA, HENCE I SAY "SUGGEST" - that we can take a 60 year old who is in excellent shape for his age and has no specific disease, and remove some amount of the senescent cells in his body to make him function better. No, in the case of such an individual the amount of senescence in his body is not optimized by nature -we can do better by fine tuning it. Do we have the mechanism at hand for this? No. Do we know which cells to remove and how many of them to remove? No. Could it turn out that we are wrong and that removing even one single senescent cell by way or exogenous intervention will do more harm than good? Weeeellll, that last question is where it's at. One has to say Yes since we can never be certain and there is a very low probability here, but there is a very strong possibility that the more suited answer is No. It appears that probably removing some senescent cells will do more good than harm, but the "some" here is yet to be defined.

At the end some people like myself want to take such risks and others do not, and that is perfectly fine. We all have different priorities as well as risk profiles, which makes the world an interesting place...

To make matters all the more viable in this particular case, the very ALK5-i used in the mice study is referred to as FDA-approved. But, as you will see in the earlier posts, we could not figure out when / where etc.... However, at least it seems (SEEMS) that we are talking about a substance that at some dose won't outright kill you.

With all of that said, I would like to very respectfully submit the following questions:

1- What other more feasible alternatives may be worth intellectually discussing here other than the specific ALK5-i mentioned in the study? You did mention Dasatinib; anything else?

2- Would you speculate that the impact of those other ALK5-inhibitors too would possibly be boosted by adding Oxytocin?

Thank you very much for your input and, more crucially, your understanding.

This is why I don't like this forum sometimes.

I respect anyone who admits not knowing something and asking to learn.

But I have no respect for someone who pretends to know something they don't from reading some headlines or abstracts with no actual understanding nor skepticism.

I'm dumb as hell as anyone. That's why I study so I don't end up being conservative or victim to false beliefs.

I self-experiment too. And there are world class researchers who know more than you ever could regarding aging. Even reversal is possible in labs, no doubt. But as much as we know what works, we know just as much about what not to do. That's why these folks aren't all taking whatever hundreds of substance that worked in labs.

Seems like you are conservative in your view and would probably deny whatever I assert and still asking me for info spoonfed to you only for what you want to know or listen. If you had enough time to write this long response to me, you got time to not be lazy as well; plethra of publically available information online. But why should I bother. You do you, and find whatever you seek yourself.

If biology is just as simple as turning things on and off as you suppose, we would've solved aging already. But no. Life is f-ing complex.

No further replies.

Good luck.

[sub7]

Take care Hieon, wish you the best of luck...

[aribadabar]

This is why I don't like this forum sometimes.

I respect anyone who admits not knowing something and asking to learn.

But I have no respect for someone who pretends to know something they don't from reading some headlines or abstracts with no actual understanding nor skepticism.

I'm dumb as hell as anyone. That's why I study so I don't end up being conservative or victim to false beliefs.

I self-experiment too. And there are world class researchers who know more than you ever could regarding aging. Even reversal is possible in labs, no doubt. But as much as we know what works, we know just as much about what not to do. That's why these folks aren't all taking whatever hundreds of substance that worked in labs.

Seems like you are conservative in your view and would probably deny whatever I assert and still asking me for info spoonfed to you only for what you want to know or listen. If you had enough time to write this long response to me, you got time to not be lazy as well; plethra of publically available information online. But why should I bother. You do you, and find whatever you seek yourself.

If biology is just as simple as turning things on and off as you suppose, we would've solved aging already. But no. Life is f-ing complex.

No further replies.

Good luck.

 

I don't think we need pontificators like yourself here wasting everyone's time.

If you have/want nothing useful to contribute/share, spare us the admonitions.

 

FYI most people on this forum are willing to risk with self-experimentation based on results from animal studies so you are in the wrong place with this attitude of yours. 

 

[sub7]

Ladies and Gentlemen,

Can anyone help convert the dose mentioned in the study into a human-equivalent

They said:

We selected a combination of 1/10 the previously published [14] dose of Alk5i (0.02 nmol/g-day), and OT dosed at 1 μg/g-day by subcutaneous administration into mice to provide systemic in vivo delivery of these molecules. 

 

(the above is so unclear; I think the numbers provided above are the doses used in the prior study. So they used 10% of the doses mentioned above. Please also do let me know if this is how you interpret the comment above)

[aribadabar]

I will give it a try:

 

Alk5 inhibitor (A5i)

TGF-β1 Type I Receptor Kinase Alk5 inhibitor 2-(3-(6-Methylpyridin-2-yl)- 407 1H-pyrazol-4-yl)-1,5-naphthyridine, was purchased from Enzo Life Sciences, and a 25 mM concentrated stock dissolved in DMSO. OT and/or A5i or control vehicle (HBSS) was injected subcutaneously to old male C57.B6 mice (23-24 month), daily for 7 days before sacrifice. 

 

 

A5i MW = 287.3g/mol

1nM=287300 mg/10^9=0.2873mcg

 

A C57BL/6J mouse is considered to weigh  ~25-30g at this age. 

 

 

 

dose of Alk5i (0.02 nmol/g-day), and OT dosed at 1 μg/g-day

 

So the daily doses were at 25g: 0.02nmol=0.5nmol=0.1437mcg/d  and OT 1mcg/g = 25mcg/d (or 5.746mcg/kg Alk5i and 1mg/kg OT)

 

HED in mg/kg = Mouse dose /12.3=0.467mcg/kg Akl5i and 0.0813mg/kg OT

 

I think the Oxytocin dose to be substantial for humans at about ~6.5mg for 80kg person.

[sub7]

Thank you so very much aribadabar

I really appreciate it

I see that you used the dose straight up, i.e. you used the dose they they have referenced to have been used in the prior study. Do you also read the statement in my prior message they way I am reading it and conclude that in the Conboy study one-tenth of this dose was used?

If that is the case, we'd be using something like 10 percent of the numbers, right? If such is the case, 

instead of the 0.467mcg/kg Akl5i and 0.0813mg/kg OT  you kindly calculated above, the ALK5i would be dosed at approximately 0.05 mcg/kg for a human.

Am I making sense?

[aribadabar]

You are right - that sentence is indeed confusing - if those figures in the brackets are the original or the 10% dose.

 

So I went to the referenced paper to compare the dosages to see if those 0.02nmol/g per day are already reduced 90% or not:

 

 

 

Aged (24 month old) C57BL6/J male mice were injected intraperitonially (IP) with TGF-β1 Type I Receptor Kinase Alk5 inhibitor 2-(3-(6-Methylpyridin-2-yl)-1H-pyrazol-4-yl)-1,5-naphthyridine (Enzo Life Sciences) diluted in sunflower seed oil to a concentration of 57.4 μM. Mice (n = 4) received 100 uL IP injections of the Alk5 inhibitor or vehicle control once daily for 11 days

 

57.4 μM=57.4 x 287 300 000 mcg/10^6 x 1 L/1000 mL= 16.49102 mcg/mL

100 μL = 0.1mL= 1.649mcg/d for a ~25g mouse

1.649 mcg x 40= 65.964 mcg/kg

 

Compared to 5.746mcg/kg in the latter study ( the slight difference is probably due to my taking 25g avg weight) - it looks like the 10% is already factored in and the figures in the brackets is the reduced dose.

 

[RWhigham]

I double checked the doses calculated by aribadabar. They are correct as shown below.

• In the earlier experiment the old mice were injected daily with (0.1mL)(57.4 uM/L) = 5.74 nM of ALK5i
• Old male lab mice weigh about 28.7g.  5.74 nM / 28.7g  = 0.2 nM/g-day
• The new experiment injected the mice with 0.02 nM/g-day  (1/10 of previous dose).
• 287.3 g/M is the molar weight of the ALK5i (from Enzio Life Sciences ALK5 Inhibitor II)
• (287.3 g/M)(0.02 nM)  = 5.746 ng  = 5.746 ng/g-day, for mice
• Mouse to human HED is 1/12  (5.746)(1/12) = 0.479 ng/g-day, for humans
• This is 0.479 ug/kg-day for humans
• 70kg x 0.479 ug/kg = 33.5 ug/day for a 70 kg human

1) I don't know if this is a safe dose to inject in humans for 11 days

2) I don't know if the HED of 1/12 works here.

3) It won't work without enough Oxytocin which has a short halflife, so multiple injections/day would work best

Edited by RWhigham, 11 October 2020 - 12:44 AM.

[sub7]

Thank you so much RWhigham,

 

If you don't mind me asking, would you be so kind as to also calculate the Oxytocin HED in ius? I am struggling with that.

 

Also, your post in another thread led me to believe that you have actually done an ALK5-i plus Oxytocin cycle before. Do I understand that correctly? If so, will you be sharing your experience with that? You would be doing a tremendous service to us all if you do.

 

Finally, let me know if you also have this general impression:
It appears to me that such items that can reverse aging when used in a  high-dose / short-term manner also have a strong anti-aging effect when used at low doses over an extended period. Do you think ALK5-i may be worth dosing at a very low dose, combined with Oxytocin  for a significantly longer duration? (the question then becomes whether such longer-term use may have a more pronounced adverse effect on the heart valves and other similar tissues that may be sensitive to it)

[RWhigham]

In this paper https://www.ncbi.nlm...les/PMC6710051/ 

• Old adult mice were injected subcutaneously daily for 7 days with a mix of ALK5i and OT
• Old adult mice weigh approximately 27.8 g
• .
• The daily dose of ALK5i given each mice was 0.02 ng/g
• Each mouse was injected daily with 27.8 g  x 0.02 ng/g = 0.556 ng of ALK5i
• The ALK5i was Enzo Life Sciences "ALK5 Inhibitor II"
• This is pure (not diluted) and is soluble in DMSO.
• A 25 mM "concentrated" stock was prepared in DMSO.
• ALK5 Inhibitor's molar weight is 287.3 g/M = 0.2873 g/mM
• ​A 25 mM stock has 25 mM x 0.2873 g/mM = 7.18 g of the Enzio product per Liter of DMSO
• The daily injection would require 0.556 ng /(7.18 ng/nL) = 0.077 nL of the 25 mM "concentrated" stock 
• This seems too small to measure. Any additional dilution is not specified in the article.
• When speaking of a molar concentration the per Liter is understood

• The daily dose of oxytocin (OT) given each mice was 1 ug/g.
• Each mouse was injected daily with 27.8 g x 1 ug/g = 27.8 ug of OT
• The OT was Bachem H-2510.
• This is pure (not diluted) oxytocin and is soluble in water.
•  A 30 mM stock was prepared in sterile water
• Oxytocin's molar weight is 1007.2 g/M = 1.007g/mM.
• A 30 mM stock has 30 mM x 1.007 g/mM = 30.21 g of the Bachem product per Liter of sterile water.
• The daily injection would require 27.8 ug/(30.21ug/uL) = 0.92 uL.

In the earlier experiment the mice were injected with 100 uL per day of sunflower seed oil containing ALK5i. The tiny amounts of the above "stock" solutions required per injection indicates that additional dilution/prep steps are missing from the article.

[sub7]

 

The daily dose of oxytocin (OT) given each mice was 1 ug/g.

 

So a human would get 1/12 of 1ug of OT per gram of bodyweight.

Which equals to 1/12 * 0.001 milligrams of OT per gram of bodyweight.

 

An 80 kg human would be getting the following amount of OT

 

80,000 * 1/12 * 0.001

 

= 6.66 milligrams of Oxytocin

 

Now, the part I am having a hard time with is the conversion of the above to iu. 

 

This link (https://www.who.int/...FINALJune10.pdf) says:

By convention, for the purpose of labelling oxytocin preparations, 1 mg of oxytocin peptide (C43H66N12O12S2) is equivalent to 600 IU of biological activity.

 

which equates the abpve mg dose to 4,000 iu of Oxytocin per day. This sounds way too high. Is the bolded statement above accurate at all?

[RWhigham]

By convention, for the purpose of labelling oxytocin preparations, 1 mg of oxytocin peptide (C43H66N12O12S2) is equivalent to 600 IU of biological activity.

For Oxytocin 1 iu = 1.68 ug, so 1 mg /(1.68 ug/iu) = 595 iu.  This conversion factor was hard to find. I don't have a reference.

 

The following patent should be studied. I'm not sure the 7.5-30 nMol(/L) oxytocin concentration specified has any adjustment for species (human vs mouse). It appears that mammalian cells were to be subjected to a concentration in this range. The oxytocin doses may be impractical.

 

Patent number: 10265372 "Molecular composition for enhancing and rejuvenating maintenance and repair of mammalian tissues"  - rejuvenation of tissues by oxytocin together with an ALK5 antagonist. 

Edited by RWhigham, 23 October 2020 - 09:51 PM.