0:00
I would like to introduce to you Doctor Papas Andreas Papas.
0:04
Dr Papas is adjunct professor of the College of Medicine, East Tennessee State University, and the CEO and member of the board of directors of Antares Health Products.
0:14
A Fulbright Scholar, Doctor Papas is a graduate of the University of Illinois and author of the Vitamin E Factor paperback and editor of the scientific book Antioxidant State of his diet, Nutrition and Health don't.
0:25
Papas also founded Yasu Health and LED the company as president and chair of the board of directors.
0:30
He developed products, concepts and managed successful commercialization, including formulation, clinical evaluation supported by the National Institute of Health and the Cystic Fibrosis Foundation, stability, safety testing, pilot and commercial production.
0:47
Good afternoon and thank you very much for joining us.
0:50
Today's it's getting along there to be a long day and I'm sure you've been experienced to be well known speakers and experts.
1:00
I tried to find something to distinguish myself other than those things that I don’t, and I realised that probably the oldest person in the conference.
1:09
So I know that you heard about mRNA technologies and the therapeutics in terms of revolutionising the field of vaccines and in other application ranging from gene and cancer treatment and special diseases.
1:30
This all this excitement probably overshadowed some of the challenges in applying the mRNA technology and we are not involved in this field at all, but our customers ask for the use of our products to address some of the challenges.
1:48
So I thought I discussed those with you, some of those with you.
1:54
As you know, mRNA is a large molecule.
1:59
It's very difficult to administer it orally because it will be of course digested but also will be very difficult to absorb because of its size.
2:08
So most of the therapies have to be given parenterally and not orally.
2:15
Also the stability we to address the stability in the formulation but also after administration and in the formulation being in parenteral has to be sterilised to be exposed to temperature and of course the other components in the formulation and that after administration obviously to the enzymes and so on.
2:41
And of course there is challenges in the terms of the targeted delivery, uptake and efficacy.
2:49
And finally something that we take for granted, the mRNA is a natural compound in our body.
2:57
Still there are safety challenges if there is changed or is metabolised to something that is more dangerous for us.
3:11
So the product that I wanted to tell you about in that can be used to be evaluated in the formulation delivery is vitamin E TPGS. And vitamin E TPGS is really a derivative of the natural D alpha tocopherol and is the main form of vitamin E.
3:36
But actually in our diet in nature, there are 8 compounds with vitamin E activity four tocopherols and for tocotrienols in our diet to get all 8 of them.
3:49
And actually the mixed tocopherols are a very powerful antioxidant that is used in the food and industry in maintaining stability of lipids and other formulations.
4:03
Also in many vitamins, the natural and the synthetic molecule are identical in the case of alpha tocopherol.
4:13
There are three asymmetric carbons and the synthetic is a mixture of 8 stereoisomers.
4:21
So officially the United States Pharmacopeia and the FDA recognised the natural versus synthetic with different potency and other characteristics.
4:33
So the TPGS is a d-alpha tocopherol with a succinic acid attached to it and polyethylene glycol 1000 on the other side.
4:43
So we have an amphibolic molecule with the lipophilic and one side and hydrophilic other side.
4:56
Because of the interest in exposure to temperature and other characteristics, we wanted to give you some of the properties.
5:05
It's a waxy solid.
5:07
It melts at the low temperature.
5:10
It's soluble in water up to 20% and it forms micellar components, and the particle size is in the one Micron.
5:21
But with special formulations is used in making nanoparticles, the thermal stability is extremely stable compound.
5:34
Actually, the shelf life is 4 years, but even after eight years is pretty stable because the active site of the tocopherol, the hydroxy group is [sterified].
5:48
So it has to be hydrolysed before it's become sensitive to changes.
5:53
So if they're stable and we can be sterilised.
5:57
So this exposure to temperature not a concern for TPGS.
6:05
And as I mentioned before, the primary mechanism that it was considered for the oral application because that has been the most common application until now is the formation of micellar particles which will migrate, microencapsulate the lipid material, and they will form these particles which will be absorbed essentially simulating the function of our body that we use the bile to really for my cells for the absorption of the lipids in our bodies to essentially simulate that mechanism.
6:49
And as you may know, micelles are key component of the absorption of lipids in humans.
6:57
And of course we have the transfer transport mechanism with the cholesterol compounds. Just to appreciate this important function in our body for the absorption of lipids.
7:12
These are diseases that cause significant malabsorption of lipids, cholestasis.
7:20
Essentially the liver doesn't produce sufficient bile which is used for the formation of micelles for absorption.
7:29
These children prior to invention of this water soluble form of vitamin E, they had a lifespan of less than 20 years.
7:38
Now they take this from all for all their lives with pharmacy water soluble absorbable form of vitamin E.
7:46
Cystic fibrosis also, because the function of the liver and the pancreas are not normal, we have a very serious absorption issues when they're pretended for the anaemia, which of course has to do more with the transport of them.
8:04
It was first used as a water soluble form of vitamin E for malabsorption.
8:10
Really that was there in children with Cholestasis.
8:13
It was observed that not only will correct the vitamin E deficiency in these children but will enhance the absorption of other fat soluble vitamin specifically starting with vitamin B. And that prompted the research and development in its use in the formulation of drugs for increasing absorption and bio availability.
8:39
And still studied extensively.
8:42
The number of studies are very large.
8:44
I of course I show the drug with rats and please note that this is a logarithmic scale on this side here and some exciting research that I was involved by accident.
8:59
The zoo animals were observed to have a very dramatic deficiency of vitamin E, although they were fed large amounts in their diet.
9:13
Apparently they changed the diet from the wild to the zoo and the absence of grazing different trees and so on that had natural excipients that created a very simple malabsorption. Actually it is used in feeding providing vitamin E to elephants and of course it has in the absorption of other vitamins and of course it is a very well established pharmaceutical excipient. It was first considered in the pharmaceutical industry some years back in the late 1990s in terms of a HIV protease inhibitor and we can see here that of course the effect on solubility, something that is very important also for mRNA cell permeability and the absorption flux.
10:18
Although we focused, we thought originally that increasing the solubility and formation of micelles was really the critical mechanism for increasing absorption bioavailability.
10:33
The same mechanism did not provide the same efficacy in other formulations.
10:39
So further study it showed that it has an effect on P-glycoprotein, which is the key mechanism of efflux, like how our body pumps back to the gut compounds that are not recognised.
10:57
And also it is very much related to the first and the double is in the multiple drug resistance.
11:07
It's currently used in a large number of drug formulations.
11:15
The latest, one of the late ones was the hepatitis drugs by AbbVie that were introduced few years ago and it's of course evaluated in very large other formulations.
11:28
But we were very surprised to find out that companies that were developing mRNA therapies very interesting this product and we started to find out why this interest. And now we'll be discussing briefly this.
11:47
But it's just an illustration of its safety is used extensively, as I mentioned as a dietary supplement for children with serious malabsorption, for patients with serious malabsorption. And is used also in other formulations ranging from cannabis product to dietary supplements for foods and beverages and the personal care products.
12:13
So the major application of course, is in the pharmaceuticals, but is used also in these other areas.
12:22
Just looking specifically now at the mRNA, I mentioned those things before is the areas that are challenges in the formulation of RNA therapies.
12:37
And first one of course is the route of administration.
12:41
Briefly discuss that, currently vitamin E TPGS has been used in oral applications and topical ocular and there's been very limited number of products for parenteral application.
13:00
Now there is tremendous focus on using TPGS in this, and include from injectable infusion to intravenous, intramuscular, intraperitoneal and so on, nasal and transdermal, nasal inhalation and so on.
13:19
So there is very new strong interest in this route of application.
13:30
As I mentioned before, the key mechanism by which it increases absorption bioavailability of the formation of micelles and the exhibition of efflux mechanism which is related to multi drug resistance and of course increasing bioavailability.
13:48
But as you have heard in many of the presentations today, the chylomicrons is not the main delivery system for mRNA.
13:57
Actually liposomes and nanoliposomes are the main ones and why there are challenges there with schedules.
14:06
And of course you may have about polymeric delivery systems and other ones that are very new and very exciting in this area.
14:18
And we see a lot of interest in evaluating vitamin E TPGS in these delivery systems.
14:30
In general why it’s used in the system is to enhance the encapsulation activity, to support the controlled release and to increase efficacy, and we’ll cover those. And examples include those formulations and then I'm sure we have before that includes polymers and other ones.
14:59
Liposomes are really the ones that are as I mentioned the primary delivery system for mRNA that is vitamin E TPGS has been used in this in the formation of in the delivery systems that include liposomes and of course in primary soft nanoparticles.
15:25
So the stability and the effect of TBGS and stability is one of the primary benefits from a synergistic effect.
15:37
Of course the enzymes, and the mRNA can be broken down by enzymes.
15:45
You have endonucleolytic and the exonucleolytic, but we have also these other mechanisms that are well known. Something that was discussed by previous speakers today, but really needs to be emphasised.
16:04
Liposomes obviously are made from lipids and lipids can get oxidised and the factors that can feed oxidation as you know is light, oxygen, temperature and other components there.
16:19
And in this case, in the case of mRNA, we take up this for that oxidation and oxidation produces free radicals.
16:32
And oxidation is really an autocatalytic reaction.
16:37
Once it starts, it progresses slowly, but then it progresses in geometric fraction.
16:44
So it's very important to prevent it because once it starts, it's very difficult to control oxidation.
16:53
And the formation of free radicals is not only that affect the stability, but free radicals a key component of inflammation and other mechanisms in the body.
17:04
So a lot of the side effects of some of these therapies are related to the production of free radicals and controlling this production is very important.
17:22
Vitamin E TPGS currently specifications of United States pharmacopoeia can have up to 1.5% free tocopherol this powerful antioxidant, but in addition, the metabolism of TBGS produces any additional tocopherol, which will be an antioxidant, and it will control the production of free radicals.
17:49
And the something that is very surprising and I think it needs to be studied further have been reports that the effect on the free radical production is different in normal cells versus cancer cells.
18:07
I'm not sure I understand why, but for cancer cells it's a blessing to have more free radicals because they kill the cancer cells.
18:16
In normal cells, it's a blessing to have less free radicals.
18:21
These are early reports, but it has been more than one report and something that take a lot of further study whether this really happened in the mechanism that involved. And of course, as I mentioned before the three rounding answer key component of the inflammation mechanism.
18:40
So vitamin E has been reported to have anti-inflammatory effects which may be applicable also in the case of mRNA therapies.
18:50
So the effect on stability is the antioxidant effect and that can be in the formulation how long you can stay on the shelf before it's administered.
19:04
But also it has, it's important what happens after administration and during the efficacy process, whether we control the production of free radicals and if it improves the functionality of the mRNA.
19:25
Related to this, but indirectly, it's of course that TPGS have been used in formation of prodrugs and conjugates to reduce toxicity and they improve the stability and efficacy; reduce the adverse effects.
19:49
So that showed before we're surprised to find also that they've been reported synergies in anti-cancer therapies and they reported that it enhances the lymphatic passage, increases the tissue permeation, but more important in facilities reactive oxygen species action.
20:12
And enhances the efficacy and I mentioned observation; this is the studies that reported a different effect on the cancer cells versus the normal cells.
20:25
There have been also reports of having some functionality that may be similar to that of an API and this is the induction of course, the reactive oxygen species within the tumour cell, The down regulation of anti-apoptotic proteins and the caspase dependent and other DNA damage.
20:48
Again, these are the early stages of evaluation and there will be additional reports.
20:55
These are the safety challenges that other speakers have started talked about and I don't need to repeat them, but I want to just to emphasise how safe is vitamin E TPGS in the United States?
21:15
It’s considered, it’s treated like a GRAS generally regarded as same.
21:19
That means that can be used as any other vitamin and food ingredient.
21:24
Canada also considers it form of vitamin E. In the European Community it’s considered a drug but has been an approval for special medical uses including the cystic fibrosis in adults and children. And there is now 1 application to make it into a better supplement. In other countries it’s still considered an API.
21:47
And I will finish with this one to appreciate it that the official not adverse effect level is 1000 milligrammes per kilogramme of body weight.
22:03
That's a very large amount.
22:06
The oral dose, the LD50 dose is very high also, but just to a newborn child with cholestasis.
22:16
The official guideline is 25 international units per day for the rest of their life.
22:24
If 40 kilogramme a child, that's 1000 are used.
22:27
That's equivalent to 2.5 grammes of the TPGS every day for all their life.
22:34
It has been some reports today and others about the inflammatory effect of polyethylene glycol.
22:43
We have not studied it in the mRNA, but we have not heard any report of from the oral use.
22:50
Of course, the parenteral is a different story.
22:54
But with the oral use there is significant absorption of PEGs.
22:57
So I'm not sure if a combination has an effect, but that's an area I'm sure that there would be evaluated further.
23:07
So in summary, we believe that vitamin E TPGS can be evaluated in mRNA therapies because it's a safe and biocompatible product. And I was told that we are out of time.
23:22
So thank you very much for coming this afternoon.