Found The Patent Recipes For The Original And Modified Brotzu Lotions

[Georgie]

Hello all

Many of you may have seen this already, many may not have, but upon my usual googling frivolities I came across this page which gives the original and modified patent Brotzu lotion recipes - the first containing PGE1 and then the second with DGLA. I know we are all inclined now to be somewhat skeptical and pissed off about all the things we have heard regarding this “breakthrough” treatment, but regardless, I thought I’d share and perchance, we might look into a group buy and lab concoction, or that one of the smarter members might just whip the sh*t up full beast mode style, zero fucks given in their kitchen sink and save us all. Anyways. Here ya go:

Example 1


Three solutions are prepared as follows: 1 m! of equal is diluted in 1 ml of ethanol and the solution is brought to 10 ml with water;


1 mg of PGE1 in 1 ml of ethanol brought to 10 ml with water;


2 g of carnitine in 10 ml of water.


The above three solutions are placed in a sonicator together with 10 g of phosphatidylcholine (Lipid S75 Human-grade) and sonicated for a sufficient time to obtain liposomes with diameter of less than 100 nm.


The liposome solution thus obtained is poured into an aqueous solution of polylysine 40 - 100 MW (0.01 mg/10 ml) and constantly stirred for 30 minutes. The solution thus obtained can be used directly on the scalp at least once daily The results of the hair loss cessation can already be observed after just 7 days and the first new hair regrowth results between 45 and 90 days.


Example 2


Three solutions are prepared as follows:


1 ml of equol is diluted in 1 ml of ethanol;


1 mg of PGE1 in 1 ml ethanol;


2 g of carnitine in 2 ml of ethanol;


The above three solutions are placed in a sonicator together with 10 g of phosphatidylcholine (Lipid S75 Human-grade) and sonicated for a sufficient time to obtain liposomes with a diameter of less than 100 nm.


The ethanol is completely evaporated and the liposomes obtained are placed in contact with 5 ml of a polylysine 40-100 MW (0.01 / 10 ml) solution. The liposome solution thus obtained is poured into an aqueous solution of polylysine 40 - 100 MW (0.01 mg/10 ml) and constantly stirred for 30 minutes then adding 5 ml of buffered water or of saline solution (NaCl 0:9/100 )


Characterisation of liposomes


The diameter of the liposomes was found to be 60 nm on average with polydispersity index equal to 0.2


The amount of PGE 1 in the liposomes following purification is in the range of 30- 50 pg/ml while the amount of carnitine is between 0.05 and 0.2 mg/ml.


The liposomes were characterised in terms of size, polydispersity index (PI) and zeta potential respectively by means of Photon Correlation Spectroscopy (PCS) (dimensions and PI) and M3-PALS (Phase Analysis Light Scattering), which measures the electrophoretic mobility of the particles in a thermostated cell, (zeta potential) using the Zetasizer nano (Malvern Instrument, UK).

AND THE DGLA LOTION:

Example 1: preparation of a liposome mixture in the form of a lotion comprising 1% stearylamine (method A)

Phosphatidylcholine (Lipid S75 Humangrade) 1 g

DGLA 1.25 mg

S-Equol 7 mg

Ethanol 1 ml

L-propionylcarnitine 7 mg

Stearylamine 10 mg

Sterile water 5 ml

DGLA, S-equol and stearylamine are dissolved in the dose of ethanol and phosphatidylcholine is added to the solution. Propionylcarnitine is dissolved in 5 ml of water and the thus obtained solution is added to the previous one. The resulting mixture is placed in a sonicator (Sonipress 150 kw) and submitted to 25 sonication cycles. Each cycle is composed of 5 seconds of full power sonication alternating with 2 seconds of rest. The lotion thus obtained is divided into 5 vials and each one is brought to the volume of 7 ml with additional sterile water. The lotion thus obtained is ready for use.

Example 2: preparation of a liposome mixture in the form of a lotion comprising 1% stearylamine (method B)

Phosphatidylcholine (Lipid S75 Humangrade) 1 g

DGLA 1.25 mg

S-Equol 7 mg

Ethanol 1 ml

L-propionylcarnitine 7 mg

Stearylamine 10 mg

Sterile water 5 ml

DGLA, S-equol, stearylamine and L-propionylcarnitine are dissolved in the dose of ethanol and the solution obtained, after addition of phosphatidylcholine, is placed in a sonicator (Sonipress 150 kw) and submitted to 25 sonication cycles. Each cycle is composed of 5 seconds of full power sonication alternating with 2 seconds of rest. Ethanol is then completely evaporated and the liposomes obtained are brought into contact with 5 ml of sterile water. The lotion thus obtained is divided into 5 vials and each one is brought to the volume of 7 ml with additional sterile water. The lotion is ready for use. Example 3: preparation of a liposome mixture in the form of a lotion comprising 2% stearylamine (method A)

See example 1, with the only variation of the dose of stearylamine employed, equal to 20 mg.

Example 4: preparation of a liposome mixture in the form of a lotion comprising 3% stearylamine (method A)

See example 1, with the only variation of the dose of stearylamine employed, equal to 30 mg.

To increase their shelf-life, the lotions thus prepared can also be freeze-dried according to methods known by the expert in the field, and reconstituted with water at the time of application.

 

[d3nt3dsh0v3l]

Sorry man, you need to lurk a lot more. What you posted is not as interesting as it may seem.

Firstly these "recipes" are simply examples from the patent. Patents have these because they have to enable the technology in that field so that other people can benefit from your work also. You can't just say you made a lotion and that it works, but that you won't say what is in it, how to make it, and why it should work. So what you usually do is patent the technology so that you have exclusive rights to it for the next few decades and then you keep all the real processes a trade secret. The process would be the real, actual lotion recipe with all of the steps, which I am suggesting of course is a secret.

Sure you know that liposomes are used and you know vaguely the compositions/characteristics of the liposomes. But you don't know the exact secret sauce they have to use, so it's not like reading the patent makes you as good as Brotzu. It very likely does not.

Additionally, liposomes are somewhat inherently unstable; like bubbles in a liquid, they will tend to aggregate together so as to minimize surface area.

This phenomenon is one of the main things that needs to be addressed to have a reasonable shelf life for the product. Stability is what Fidia is claiming to have focused on from the beginning of last year and ongoing.

Finally people like Spato and Wei Wu claim to have made versions of the lotions of course starting with the examples in the patent (which are very basic preparation methods for making liposomes - someone working in the field wouldn't be surprised) have discussed these things and people a lot and so has IESON.

tl;dr: fake news, lurk moar, etc

 

[Georgie]

Sorry man, you need to lurk a lot more. What you posted is not as interesting as it may seem.

Firstly these "recipes" are simply examples from the patent. Patents have these because they have to enable the technology in that field so that other people can benefit from your work also. You can't just say you made a lotion and that it works, but that you won't say what is in it, how to make it, and why it should work. So what you usually do is patent the technology so that you have exclusive rights to it for the next few decades and then you keep all the real processes a trade secret. The process would be the real, actual lotion recipe with all of the steps, which I am suggesting of course is a secret.

Sure you know that liposomes are used and you know vaguely the compositions/characteristics of the liposomes. But you don't know the exact secretly sauce they have to use, so it's not like reading the patent makes you as good as Brotzu. It very likely does not.

Additionally, liposomes are somewhat inherently unstable; like bubbles in a liquid, they will tend to aggregate together so as to minimize surface area.
This phenomenon is one of the main things that needs to be addressed to have a reasonable shelf life for the product. Stability is what Fidia is claiming to have focused on from the beginning of last year and ongoing.

Finally people like Spato and Wei Wu claim to have made versions of the lotions of course starting with the examples in the patent (which are very basic preparation methods for making liposomes - someone working in the field wouldn't be surprised) have discussed these things and people a lot and so has IESON.

tl;dr: fake news, lurk moar, etc

Well f***.

 

[Georgie]

Don't be down on yourself man, Lord Brotzu will release it in 2016

Lol. That Italian c*** better release it soon or I will actually lose my sh*t

 

[d3nt3dsh0v3l]

Well f***.

Haha sorry. I tried to make a lotion also. I'm a chemical engineering graduate student, so I spoke to one of my buddies who researches liposomal drug delivery, amidst other things. I discussed the patent, preparation and principle of action with him and it seemed pretty straight forward.

I have access to sonicators (that's no big deal). I priced out equol, DGLA, carnatine, sterylamine, the lipids, etc. but s-equol can be a bit tricky to get. Also all of the suppliers will say something like "For laboratory use only. Not for human consumption or veterinary use" which of course they are obliged to say, but not knowing if the substances are safe for consumption made me uneasy. I called one of the suppliers and asked them what the impurities might be, but they said that they do not make the equol in house and so they do not know. They were also offering racemic equol (50/50 mixture of r-equol and s-equol) so I know that the equol was chemically synthesized in a fashion insensitive to stereochemistry and then purified. Direct synthesis of s-equol is tricky, and the other scalable way to get s-equol is to enslave some gut bacteria to make it in a reactor, because the bacteria only produce s-equol. How lucky. r-equol also supposedly binds to DHT, but not the estrogen receptor like s-equol, so maybe a 50/50 mix is not ideal but also not a huge deal.

I pussed out of buying chems from suppliers for my own use because I'm not a cool kid I guess. Bacteria-made, purified s-equol appeared to cost way too much.

Then there is the issue of characterizing your lotion so that you know wtf you made. Need dynamic light scattering for finding liposome size. Zeta potential would also be useful. I'd probably be able to ask that other student for a few favors so I wasn't too worried about this.

So that's a bunch of work I have to do, with inevitable unforeseen troubleshooting as well, all the while not knowing if it will even work.

So then the next thing that I did was that I found a Japanese supplier who sells s-equol supplements made from fermentation of soy (bacteria method), but not purified.

It seemed reasonably priced, and contained a fair amount of s-equol per tablet.

The tablets are a packed powder of fermented soy encased in a titanium dioxide (insoluble and inert diffusion barrier).

I bought a mortar and pestle, crushed the tablets, placed the contents in a flask, along with ethanol (everclear) and magnetically stirred the mixture overnight. One big problem here is that since the pills are mostly soy powder by weight, it is very difficult to get greater than ~ 1 mg/mL equol concentration. This is because if you don't add enough ethanol, the mixture will be a slurry; also the soy powder appears to absorb some of the solvent, but I wasn't worried about this as much because I planned to centrifuge it anyway; filter paper would consume most of what I made and I was making very small batches - ~50 ml. I considered using excess solvent and then subsequently evaporating excess solvent from the final solution, but drying at room temp was slow, I had a limited supply of tablets, and I did not want to risk oxiding the equol or something by heating the mixture to drive the solvent out. Alternatively I could have applied a vacuum, but I did not really strive for higher concentration of equol.

Brotzu uses much lower concentrations of equol anyway (~ 0.1 mg/ml). But then again, I wasn't using liposomal delivery and apparently some literature suggests that liposomes deposit preferentially in the hair follicle. Perhaps because they are too large to diffuse through the skin (as a rule of thumb, anything 500 daltons and above does not penetrate the skin very well) but they are small enough to enter through other means, such as shunting through the side of the hair shaft, straight to the follicle.

Source: http://doi.org.https.sci-hub.tw/10.1016/S0923-1811(96)00557-9

Anyway, I took whatever slurry I made, centrifuged it at some ridiculous RPM and isolated the supernatent and stored it in my fridge. I applied ~2 ml/day for like a month and noticed no changes at all, other than needing to keep making this damn thing and smelling like soy.

Perhaps I needed a much higher concentration.
Perhaps equol by itself doesn't work, as Brotzu suggests (I have no idea why. Equol should be the only thing in the mixture that can address DHT).
Perhaps my equol extraction was unsuccessful (I unfortunately couldn't find someone to help me run GC-MS to determine the actual equol concentration in the supernatent).

Perhaps the whole thing doesn't work at all. There are no human data and few studies on equol supplementation. Even within the supplementation studies, there seems to be a mild effect in alleviating DHT based symptoms.

So I decided to give it a break. If someone releases human data with liposomal equol stopping miniaturization, then I swear I'll drop whatever I am doing and take a crack at a true version of the lotion.

But unless I have some kind of assurance, it is a fair amount of work on my part with little return.

Don't worry! If it works, people will make it. It's not too hard But idk how one would address stability without doing a bunch of work, so perhaps the homebrew version would have to be made often and used soon after preparation.

But for now...I wait..

 

[Georgie]

Haha sorry. I tried to make a lotion also. I'm a chemical engineering graduate student, so I spoke to one of my buddies who researches liposomal drug delivery, amidst other things. I discussed the patent, preparation and principle of action with him and it seemed pretty straight forward.

I have access to sonicators (that's no big deal). I priced out equol, DGLA, carnatine, sterylamine, the lipids, etc. but s-equol can be a bit tricky to get. Also all of the chems will say something like "For laboratory use only. Not for human consumption or veterinary use" which of course they are obliged to say, but not knowing if the substances are safe for consumption made me uneasy. I called one of the suppliers and asked them what the impurities might be, but they said that they do not make the equol in house and so they do not know. They were also offering racemic equol (50/50 mixture of r-equol and s-equol) so I know that the equol was chemically synthesized in a fashion insensitive to stereochemistry and then purified. Direct synthesis of s-equol is tricky, and the other scalable way to get s-equol is to enslave some gut bacteria to make it in a reactor, because the bacteria only produce s-equol. How lucky. r-equol supposedly binds to DHT, but not the estrogen receptor, like s-equol, so maybe a 50/50 mix is not a huge deal.

I pussed out of buying chems from suppliers for my own use because I guess I'm not a cool kid I guess.

Then there is the issue of characterizing your lotion so that you know wtf you made. Need dynamic light scattering for finding liposome size. Zeta potential would also be useful. I'd probably be able to ask that other student for a few favors so I wasn't too worried about this.

So that's a bunch of work I have to do, with inevitable unforeseen troubleshooting as well, all the while not knowing if it will even work.

So then the next thing that I did was that I found a Japanese supplier who sells s-equol supplements made from fermentation of soy (bacteria method).

View attachment 76280
It seemed reasonably priced, and contained a fair amount of s-equol per tablet.

The tablets are a packed powder of fermented soy encased in a titanium dioxide (insoluble and inert diffusion barrier).

I bought a mortar and pestle, crushed the tablets, placed the contents in a flask, along with ethanol (everclear) and magnetically stirred the mixture overnight. One big problem here is that since the pills are mostly soy powder by weight, it is very difficult to get greater than ~ 1 mg/mL equol concentration. This is because if you don't add enough ethanol, the mixture will be a slurry; also the soy powder appears to absorb some of the solvent. But Brotzu uses much lower concentrations than that.

But then again, I wasn't using liposomal delivery and apparently some literature suggests that liposomes deposit preferrentially in the hair follicle. Perhaps because they are too large to diffuse through the skin (as a rule of thumb, anything 500 daltons and above does not penetrate the skin very well) but they are small enough to enter through other means.

View attachment 76276

View attachment 76277

Source: http://doi.org.https.sci-hub.tw/10.1016/S0923-1811(96)00557-9

Anyway, I took whatever slurry I made, centrifuged it at some ridiculous RPM and isolated the supernatent and stored it in my fridge. I applied ~2 ml/day for like a month and noticed no changes at all, other than needing to keep making this damn thing and smelling like soy.

Perhaps I needed a much higher concentration.
Perhaps equol by itself doesn't work.
Perhaps my equol extraction was unsuccessful (I unfortunately couldn't find someone to help me run GC-MS to determine the actual equol concentration in the supernatent).

Perhaps the whole thing doesn't work at all. There is no human data and few studies on equol supplementation. Even within the supplementation studies, there seems to be a mild effect in alleviating DHT based symptoms.

So I decided to give it a break. If someone releases human data with liposomes stoping miniaturizing, then I swear I'll drop whatever I am doing and take a crack at a true version of the lotion.

But unless I have some kind of assurance, it is a fair amount of work on my part with little return.

Don't worry! If it works, people will make it. It's not too hard But idk how one would address stability without doing a bunch of work, so perhaps the homebrew version would have to be made often and used soon after preparation.

But for now...I wait..

Man that sounds so complicated and annoying. You really did go to a lot of effort huh?
From what I’ve read, the consistency needs to come out as a lotion. The whole sonoification process needs to dissolve the ethanol leaving only the liposomal contents from the DGLA are left. Maybe the recipe is wrong, because it seems like you tried to replicate it as closely as possibly.
That’s disappointing.

 

[d3nt3dsh0v3l]

Man that sounds so complicated and annoying. You really did go to a lot of effort huh?
From what I’ve read, the consistency needs to come out as a lotion. The whole sonoification process needs to dissolve the ethanol leaving only the liposomal contents from the DGLA are left. Maybe the recipe is wrong, because it seems like you tried to replicate it as closely as possibly.
That’s disappointing.

No, I did not try to replicate a liposomal solution. Basically I made a much simplified version: s-equol + isoflavones in ethanol.

The actual method is much more annoying.
The way it works is that the lipids are organic, so they want to phase separate from aqueous solution. So you want to put your organic compounds and the lipids together in an organic solvent then dry off the solvent with rotovapping. If dried slowly, as the last of the solvent leaves, the residual liposomes deposit layer-wise at the bottom of the contained, along with whatever organic delivery compounds are trapped between the layers.

Now you can bring in an aqueous solution with all of your aqueous drugs dissolved in it. When you fill the thing up with an aqueous solution, the deposited layers at the bottom of the container break away and form multilamellar vesicles - large spheres with multiple layers of lipids for walls. Within the spheres is the aqueous solution with aqueous drugs. The organics reside near/inside the lipid walls, from before.

Now if you sonicate the mixture, the pressure waves break up the multilamellar vesicles into smaller liposomes of the desired size.

Finally you can add more water to get the desired concentration of liposomes.

Here is a video. It's like infinitely easier to understand if you just see what is happening. It makes sense.

That's just one way to make liposomal solutions. Good for small batches but not very scalable. Doesn't matter for the home enthusiast. As the video shows, there are like many standard processes one can use to make/manipulate liposomes. That's what I meant by the process being like well known to a person in the field. It's a standard recipe. It's just there to make the patent sufficiently enabling.

 

[Georgie]

No, I did not try to replicate a liposomal solution. Basically I made a much simplified version: s-equol + isoflavones in ethanol.

The actual method is much more annoying.
The way it works is that the lipids are organic, so they want to phase separate from aqueous solution. So you want to put your organic compounds and the lipids together in an organic solvent then dry off the solvent with rotovapping. If dried slowly, as the last of the solvent leaves, the residual liposomes deposit layer-wise at the bottom of the contained, along with whatever organic delivery compounds are trapped between the layers.

Now you can bring in an aqueous solution with all of your aqueous drugs dissolved in it. When you fill the thing up with an aqueous solution, the deposited layers at the bottom of the container break away and form multilamellar vesicles - large spheres with multiple layers of lipids for walls. Within the spheres is the aqueous solution with aqueous drugs. The organics reside near/inside the lipid walls, from before.

Now if you sonicate the mixture, the pressure waves break up the multilamellar vesicles into smaller liposomes of the desired size.

Finally you can add more water to get the desired concentration of liposomes.

Here is a video. It's like infinitely easier to understand if you just see what is happening. It makes sense.

That's just one way to make liposomal solutions. Good for small batches but not very scalable. Doesn't matter for the home enthusiast. As the video shows, there are like many standard processes one can use to make/manipulate liposomes. That's what I meant by the process being like well known to a person in the field. It's a standard recipe. It's just there to make the patent sufficiently enabling.

Oh ok so many roads lead to Rome sort of thing. Sorry I find it a little difficult to comprehend because I’m just a lay person haha. I often looked at the ingredients involved and wondered how they could lead to the kind of hair restoration that was claimed - “5 years worth of hairloss” or whatever it was. It may be that the dr himself holds the unknown answer to what makes it an effective treatment, which we have no access to. Maybe it’s a scam. Either way it breaks my heart every time I hear that there is some kind of a cure in the makings and all we get thereafter is ambiguity and disappointment.

 

[d3nt3dsh0v3l]

Oh ok so many roads lead to Rome sort of thing. Sorry I find it a little difficult to comprehend because I’m just a lay person haha. I often looked at the ingredients involved and wondered how they could lead to the kind of hair restoration that was claimed - “5 years worth of hairloss” or whatever it was. It may be that the dr himself holds the unknown answer to what makes it an effective treatment, which we have no access to. Maybe it’s a scam. Either way it breaks my heart every time I hear that there is some kind of a cure in the makings and all we get thereafter is ambiguity and disappointment.

Oh I see. It's hard to tell who is proficient in what on here. Some people know way more than I expected when I joined the site.

I think the cliffs are to keep your eyes on follica, seti, Shiseido and Tsuji. Between them, I think something real will pan out.

They all have bits and pieces of clinical trial data, with the exception of seti itself, but Cots has shown a strong link between PDG2 and hairloss, and seti addresses this. And by shown, I mean he has published in very prestigious journals and has presented his work at international conferences; pretty legit.

Systemic drug use can compromise function in other parts of the body as we know.
Drug delivery to the scalp only and nowhere else is hard.
Blocking the androgen receptor itself with an antagonist like RU may give results that change over time, since the body may react by upregulating the receptors since less of them are now producing a signal. So overtime, your sensitivity to androgens may increase, actually. Plus, some report that RU likely goes systemic, so there is that problem again. That's why I'm not crazy for Breezula but I mean I'd still try it.
The chemical cascade that leads to actual miniaturization is still not entirely clear. Thus, if there are proposed small molecule drugs that interrupt the chemical messengers along this pathway, I think it could go either way. There have been things that "should" have worked, but did not, such as Samumed's drug, so I won't really hold my breath for Seti or that work with CXXC5, etc. Then there is the problem of some treatments giving regrowth but not maintenance, or vice versa. Perhaps the small molecule approach is limited, or perhaps not.

The stem cell route is a much more elegant solution that circumvents a lot of this, and there are pieces of clinical data to show that it the odds of such an approach working (in principle) are fantastic.

Wounding as a general approach seems like an orthogonal approach that works as well, since there are human data showing efficacy, and none of the concerns listed above really apply to this method. A potential downside is that perhaps it is difficult to get high terminal hair counts with this method if you don't address DHT sensitivity in some way because the scalp environment is already very bad for hair growth. But combined with something like Seti, wounding might end up being extremely effective.

 

[Georgie]

Oh I see. It's hard to tell who is proficient in what on here. Some people know way more than I expected when I joined the site.

I think the cliffs are to keep your eyes on follica, seti, Shiseido and Tsuji. Between them, I think something real will pan out.

They all have bits and pieces of clinical trial data, with the exception of seti itself, but Cots has shown a strong link between PDG2 and hairloss, and seti addresses this. And by shown, I mean he has published in very prestigious journals and has presented his work at international conferences; pretty legit.

Systemic drug use can compromise function in other parts of the body as we know.
Drug delivery to the scalp only and nowhere else is hard.
Blocking the androgen receptor itself with an antagonist like RU may give results that change over time, since the body may react by upregulating the receptors since less of them are now producing a signal. So overtime, your sensitivity to androgens may increase, actually. Plus, some report that RU likely goes systemic, so there is that problem again. That's why I'm not crazy for Breezula but I mean I'd still try it.
The chemical cascade that leads to actual miniaturization is still not entirely clear. Thus, if there are proposed small molecule drugs that interrupt the chemical messengers along this pathway, I think it could go either way. There have been things that "should" have worked, but did not, such as Samumed's drug, so I won't really hold my breath for Seti or that work with CXXC5, etc. Then there is the problem of some treatments giving regrowth but not maintenance, or vice versa. Perhaps the small molecule approach is limited, or perhaps not.

The stem cell route is a much more elegant solution that circumvents a lot of this, and there are pieces of clinical data to show that it the odds of such an approach working (in principle) are fantastic.

Wounding as a general approach seems like an orthogonal approach that works as well, since there are human data showing efficacy, and none of the concerns listed above really apply to this method. A potential downside is that perhaps it is difficult to get high terminal hair counts with this method if you don't address DHT sensitivity in some way because the scalp environment is already very bad for hair growth. But combined with something like Seti, wounding might end up being extremely effective.

Have you seen the study on PDM threading for hair regrowth? There’s a thread about I on the forum, but you can also find it easily via google searche. This all comes back to wounding and it’s potential to cause follicle neogenesis. It’s very true interesting.
I honestly believe that stem cells will eventually be the way forward. Treating things by trying to patch up what it already damaged just doesn’t seem to work. What we need are new follicles. How to maintain growth in said follicles is a challenge when, like you said, the scalp environment is already not great for hair growth, but I suppose that’s where your dht blockers and anti androgens might help.

For me, it seems like very little works, and I have very damaged hair growth within the space of only 4 years. For people like myself who just don’t response well to anything, it’s the only hope that is left.

 

[BaldAsshole]

Haha sorry. I tried to make a lotion also. I'm a chemical engineering graduate student, so I spoke to one of my buddies who researches liposomal drug delivery, amidst other things. I discussed the patent, preparation and principle of action with him and it seemed pretty straight forward.

I have access to sonicators (that's no big deal). I priced out equol, DGLA, carnatine, sterylamine, the lipids, etc. but s-equol can be a bit tricky to get. Also all of the suppliers will say something like "For laboratory use only. Not for human consumption or veterinary use" which of course they are obliged to say, but not knowing if the substances are safe for consumption made me uneasy. I called one of the suppliers and asked them what the impurities might be, but they said that they do not make the equol in house and so they do not know. They were also offering racemic equol (50/50 mixture of r-equol and s-equol) so I know that the equol was chemically synthesized in a fashion insensitive to stereochemistry and then purified. Direct synthesis of s-equol is tricky, and the other scalable way to get s-equol is to enslave some gut bacteria to make it in a reactor, because the bacteria only produce s-equol. How lucky. r-equol supposedly binds to DHT, but not the estrogen receptor, like s-equol, so maybe a 50/50 mix is not a huge deal.

I pussed out of buying chems from suppliers for my own use because I'm not a cool kid I guess. Bacteria-made, purified s-equol appeared to cost way too much.

Then there is the issue of characterizing your lotion so that you know wtf you made. Need dynamic light scattering for finding liposome size. Zeta potential would also be useful. I'd probably be able to ask that other student for a few favors so I wasn't too worried about this.

So that's a bunch of work I have to do, with inevitable unforeseen troubleshooting as well, all the while not knowing if it will even work.

So then the next thing that I did was that I found a Japanese supplier who sells s-equol supplements made from fermentation of soy (bacteria method), but not purified.

View attachment 76280
It seemed reasonably priced, and contained a fair amount of s-equol per tablet.

The tablets are a packed powder of fermented soy encased in a titanium dioxide (insoluble and inert diffusion barrier).

I bought a mortar and pestle, crushed the tablets, placed the contents in a flask, along with ethanol (everclear) and magnetically stirred the mixture overnight. One big problem here is that since the pills are mostly soy powder by weight, it is very difficult to get greater than ~ 1 mg/mL equol concentration. This is because if you don't add enough ethanol, the mixture will be a slurry; also the soy powder appears to absorb some of the solvent, but I wasn't worried about this as much because I planned to centrifuge it anyway; filter paper would consume most of what I made and I was making very small batches - ~50 ml.

Brotzu uses much lower concentrations of equol anyway (~ 0.1 mg/ml). But then again, I wasn't using liposomal delivery and apparently some literature suggests that liposomes deposit preferentially in the hair follicle. Perhaps because they are too large to diffuse through the skin (as a rule of thumb, anything 500 daltons and above does not penetrate the skin very well) but they are small enough to enter through other means, such as shunting through the side of the hair shaft, straight to the follicle.

View attachment 76284

View attachment 76276

View attachment 76277

Source: http://doi.org.https.sci-hub.tw/10.1016/S0923-1811(96)00557-9

Anyway, I took whatever slurry I made, centrifuged it at some ridiculous RPM and isolated the supernatent and stored it in my fridge. I applied ~2 ml/day for like a month and noticed no changes at all, other than needing to keep making this damn thing and smelling like soy.

Perhaps I needed a much higher concentration.
Perhaps equol by itself doesn't work, as Brotzu suggests (I have no idea why. Equol should be the only thing in the mixture that can address DHT).
Perhaps my equol extraction was unsuccessful (I unfortunately couldn't find someone to help me run GC-MS to determine the actual equol concentration in the supernatent).

Perhaps the whole thing doesn't work at all. There are no human data and few studies on equol supplementation. Even within the supplementation studies, there seems to be a mild effect in alleviating DHT based symptoms.

So I decided to give it a break. If someone releases human data with liposomal equol stopping miniaturization, then I swear I'll drop whatever I am doing and take a crack at a true version of the lotion.

But unless I have some kind of assurance, it is a fair amount of work on my part with little return.

Don't worry! If it works, people will make it. It's not too hard But idk how one would address stability without doing a bunch of work, so perhaps the homebrew version would have to be made often and used soon after preparation.

But for now...I wait..

Are you God?

 

[whatevr]

'Man' is a she, in this case.

 

[d3nt3dsh0v3l]

Are you God?

Dog* actually.

'Man' is a she, in this case.

Right. I figured that out recently. Whoops.

 

[Georgie]

Dog* actually.


Right. I figured that out recently. Whoops.

Not for long with the rate at which I’m balding

 

[whatevr]

Not for long with the rate at which I’m balding

At least the cure for female pattern hair loss isn't testosterone, as estrogen is for the male version

 

[charlie76761]

Hello all

Many of you may have seen this already, many may not have, but upon my usual googling frivolities I came across this page which gives the original and modified patent Brotzu lotion recipes - the first containing PGE1 and then the second with DGLA. I know we are all inclined now to be somewhat skeptical and pissed off about all the things we have heard regarding this “breakthrough” treatment, but regardless, I thought I’d share and perchance, we might look into a group buy and lab concoction, or that one of the smarter members might just whip the sh*t up full beast mode style, zero fucks given in their kitchen sink and save us all. Anyways. Here ya go:

Example 1


Three solutions are prepared as follows: 1 m! of equal is diluted in 1 ml of ethanol and the solution is brought to 10 ml with water;


1 mg of PGE1 in 1 ml of ethanol brought to 10 ml with water;


2 g of carnitine in 10 ml of water.


The above three solutions are placed in a sonicator together with 10 g of phosphatidylcholine (Lipid S75 Human-grade) and sonicated for a sufficient time to obtain liposomes with diameter of less than 100 nm.


The liposome solution thus obtained is poured into an aqueous solution of polylysine 40 - 100 MW (0.01 mg/10 ml) and constantly stirred for 30 minutes. The solution thus obtained can be used directly on the scalp at least once daily The results of the hair loss cessation can already be observed after just 7 days and the first new hair regrowth results between 45 and 90 days.


Example 2


Three solutions are prepared as follows:


1 ml of equol is diluted in 1 ml of ethanol;


1 mg of PGE1 in 1 ml ethanol;


2 g of carnitine in 2 ml of ethanol;


The above three solutions are placed in a sonicator together with 10 g of phosphatidylcholine (Lipid S75 Human-grade) and sonicated for a sufficient time to obtain liposomes with a diameter of less than 100 nm.


The ethanol is completely evaporated and the liposomes obtained are placed in contact with 5 ml of a polylysine 40-100 MW (0.01 / 10 ml) solution. The liposome solution thus obtained is poured into an aqueous solution of polylysine 40 - 100 MW (0.01 mg/10 ml) and constantly stirred for 30 minutes then adding 5 ml of buffered water or of saline solution (NaCl 0:9/100 )


Characterisation of liposomes


The diameter of the liposomes was found to be 60 nm on average with polydispersity index equal to 0.2


The amount of PGE 1 in the liposomes following purification is in the range of 30- 50 pg/ml while the amount of carnitine is between 0.05 and 0.2 mg/ml.


The liposomes were characterised in terms of size, polydispersity index (PI) and zeta potential respectively by means of Photon Correlation Spectroscopy (PCS) (dimensions and PI) and M3-PALS (Phase Analysis Light Scattering), which measures the electrophoretic mobility of the particles in a thermostated cell, (zeta potential) using the Zetasizer nano (Malvern Instrument, UK).

AND THE DGLA LOTION:

Example 1: preparation of a liposome mixture in the form of a lotion comprising 1% stearylamine (method A)

Phosphatidylcholine (Lipid S75 Humangrade) 1 g

DGLA 1.25 mg

S-Equol 7 mg

Ethanol 1 ml

L-propionylcarnitine 7 mg

Stearylamine 10 mg

Sterile water 5 ml

DGLA, S-equol and stearylamine are dissolved in the dose of ethanol and phosphatidylcholine is added to the solution. Propionylcarnitine is dissolved in 5 ml of water and the thus obtained solution is added to the previous one. The resulting mixture is placed in a sonicator (Sonipress 150 kw) and submitted to 25 sonication cycles. Each cycle is composed of 5 seconds of full power sonication alternating with 2 seconds of rest. The lotion thus obtained is divided into 5 vials and each one is brought to the volume of 7 ml with additional sterile water. The lotion thus obtained is ready for use.

Example 2: preparation of a liposome mixture in the form of a lotion comprising 1% stearylamine (method B)

Phosphatidylcholine (Lipid S75 Humangrade) 1 g

DGLA 1.25 mg

S-Equol 7 mg

Ethanol 1 ml

L-propionylcarnitine 7 mg

Stearylamine 10 mg

Sterile water 5 ml

DGLA, S-equol, stearylamine and L-propionylcarnitine are dissolved in the dose of ethanol and the solution obtained, after addition of phosphatidylcholine, is placed in a sonicator (Sonipress 150 kw) and submitted to 25 sonication cycles. Each cycle is composed of 5 seconds of full power sonication alternating with 2 seconds of rest. Ethanol is then completely evaporated and the liposomes obtained are brought into contact with 5 ml of sterile water. The lotion thus obtained is divided into 5 vials and each one is brought to the volume of 7 ml with additional sterile water. The lotion is ready for use. Example 3: preparation of a liposome mixture in the form of a lotion comprising 2% stearylamine (method A)

See example 1, with the only variation of the dose of stearylamine employed, equal to 20 mg.

Example 4: preparation of a liposome mixture in the form of a lotion comprising 3% stearylamine (method A)

See example 1, with the only variation of the dose of stearylamine employed, equal to 30 mg.

To increase their shelf-life, the lotions thus prepared can also be freeze-dried according to methods known by the expert in the field, and reconstituted with water at the time of application.

Hi George or anyone else with knowledge of the Brotzu lotions ingredients / patents.

I'm procuring some PGE1 and interested in making something not too dissimilar to the Brotzu lotion (altho will be ethanol based not lipo - should still help... worth a try) although I'm bit confused as to the S-Equol dosage - seem v v low.

If I've understood the above correctly, Brotzu has 7mg of S-Equol within 35ml - thus 0.2mg of S-Equol per 1ml (which i'm assuming is the ml amount applied per day in line with other topicals).

However, if you look at threads such as the one pasted below, users are stating that 30mg of S-Equol orally per day had no effect and looking to use 50 mg per day i.e. x250 more than 0.2mg in Bortzu.

Further, i think it's safe to assume that topicals needs to be in greater dosage to have the same effect as the oral route e.g. minoxidil - users go for 5% twice a day (50mg x 2) topically to get a result, but 10mg orally is the normal dosage that for oral to get results (and further, these are normally far superior results to topical)

I know S-Equol works in synergy with the other active ingredients in the Brotzu lotion, but i can't for a moment think the synergy effect is x250.

Seems odd - S-Equol is not known to be powerful hence why classed as therapeutic - 0.2mg seems v v low

Thoughts?


https://www.hairlosstalk.com/interact/threads/s-equol-as-a-hair-loss-inhibitor.66755/

 

[Georgie]

Hi George or anyone else with knowledge of the Brotzu lotions ingredients / patents.

I'm procuring some PGE1 and interested in making something not too dissimilar to the Brotzu lotion (altho will be ethanol based not lipo - should still help... worth a try) although I'm bit confused as to the S-Equol dosage - seem v v low.

If I've understood the above correctly, Brotzu has 7mg of S-Equol within 35ml - thus 0.2mg of S-Equol per 1ml (which i'm assuming is the ml amount applied per day in line with other topicals).

However, if you look at threads such as the one pasted below, users are stating that 30mg of S-Equol orally per day had no effect and looking to use 50 mg per day i.e. x250 more than 0.2mg in Bortzu.

Further, i think it's safe to assume that topicals needs to be in greater dosage to have the same effect as the oral route e.g. minoxidil - users go for 5% twice a day (50mg x 2) topically to get a result, but 10mg orally is the normal dosage that for oral to get results (and further, these are normally far superior results to topical)

I know S-Equol works in synergy with the other active ingredients in the Brotzu lotion, but i can't for a moment think the synergy effect is x250.

Seems odd - S-Equol is not known to be powerful hence why classed as therapeutic - 0.2mg seems v v low

Thoughts?


https://www.hairlosstalk.com/interact/threads/s-equol-as-a-hair-loss-inhibitor.66755/

Hey

I am no a bio chemist so unfortunately I can’t answer your questions about vehicle even if I thought I knew, but we haven’t seen or heard anything from Brotzu/Fidia for some time now and question whether it would work at all. Indeed, others have attempted to recreate with no results, but this is only a rough recipe. The real thing is obviously far more detailed.

Hopefully one of the more scientifically knowledgeable members can chime in to answer some of your questions here

 

[d3nt3dsh0v3l]

Hi George or anyone else with knowledge of the Brotzu lotions ingredients / patents.

I'm procuring some PGE1 and interested in making something not too dissimilar to the Brotzu lotion (altho will be ethanol based not lipo - should still help... worth a try) although I'm bit confused as to the S-Equol dosage - seem v v low.

If I've understood the above correctly, Brotzu has 7mg of S-Equol within 35ml - thus 0.2mg of S-Equol per 1ml (which i'm assuming is the ml amount applied per day in line with other topicals).

However, if you look at threads such as the one pasted below, users are stating that 30mg of S-Equol orally per day had no effect and looking to use 50 mg per day i.e. x250 more than 0.2mg in Bortzu.

Further, i think it's safe to assume that topicals needs to be in greater dosage to have the same effect as the oral route e.g. minoxidil - users go for 5% twice a day (50mg x 2) topically to get a result, but 10mg orally is the normal dosage that for oral to get results (and further, these are normally far superior results to topical)

I know S-Equol works in synergy with the other active ingredients in the Brotzu lotion, but i can't for a moment think the synergy effect is x250.

Seems odd - S-Equol is not known to be powerful hence why classed as therapeutic - 0.2mg seems v v low

Thoughts?


https://www.hairlosstalk.com/interact/threads/s-equol-as-a-hair-loss-inhibitor.66755/

Hey bud, just F.Y.I. according to Brotzu himself, PGE1 oxidizes very easily and is therefore hard to deliver it within a topical; Brotzu first attempted to use PGE1 directly:
https://patents.google.com/patent/WO2013171668A1/en

The following is written in his second patent:
"Encapsulation in liposomes is not able to increase the very poor stability of PGE1, though; the findings of WO2013171668 in fact, must be freeze-dried or, if in suspension, stored at low temperature (about -20°C) up to the time of use, if one intends to prevent the almost complete oxidation of PGE1. This obviously represents a complication not only on the industrial production level, but mainly on the product safety level. To act against the oxidative process, in fact, the trend is to resort to PGE1 doses that might be dangerous for the body, considered the repeated employment of these products, which must be administered over very long periods."
https://patents.google.com/patent/WO2015170247A1/en

He claims to have switched to DGLA because 1) it is a precursor to PGE1; that is, your body can use it to produce PGE1 within your cells, 2) it does not easily oxidize and 3) unlike PGE1, it is not considered a drug and thus does not require clinical trials.

So I do think if you try to use PGE1, it won't work.

I posted earlier that liposomes can deliver drugs straight to the follicle; this improves drug delivery and so you can use less to get a greater effect. This is partially discussed in the patent. Large oral (i.e. systemic) doses of equol do little.

I cannot tell you why the doctor claims that s-equol only works in conjunction with DGLA. Or why this detail had to be amended on the patent. Or how, having started the work by investigating PGE1, he would have seen results without using s-equol or why he would ever think to use a plant phytoestrogen to begin with. I am equally befuddled, but that's what the man claims.

 

[Ollie]

@d3nt3dsh0v3l any chance of being able to utilise liposomes for other topical treatments ? Ru etc..

 

[d3nt3dsh0v3l]

@d3nt3dsh0v3l any chance of being able to utilise liposomes for other topical treatments ? Ru etc..

Yes definitely! As you saw from the post above, liposomal delivery improves absorption and may even deposit selectively within the hair follicle.

But my experience as a researcher tells me that you want to have some characterization tools before you play around, because most of the time, things go wrong for unforseen reasons and force you to troubleshoot; rarely does the thing work the first time. If you could measure the liposome size, it would be so helpful. Otherwise you're totally in the blind - maybe you made it right or maybe you didn't, or maybe it was perfect but degraded after 24 hours, etc.

Getting liposome size and measuring its composition may be annoying.