Anavex Life Sciences Corp (AVXL)

[Biostockclub]

Of Mice and Men

The “system” is broken. This fact is apparent from BP’s to FDA, including academic research mindset, and funding from federal government spent on a single hypothesis for 30 years, which has failed more times than we can count, yet is still the touted approach to a cure today - despite nary a single drug being approved using the Amyloid plaque hypothesis and actual clearing of plaque...no approved “mabs”.

Here’s some DD which I investigated in an attempt to learn more about our MS Petri dish in vitro assays in conjunction with BIIB and how that particular process might be carried out from a practical standpoint. What turned up was not what I had intended to find. Still, I followed where the research led and I think what turned up sheds an awful lot of light on the entire process of CNS drug testing. A lot of our Anavex posters have been getting things right, and everyone gets his/her credit, but this one piece of the puzzle may pull together much which has been misunderstood or simply not acknowledged before. Once we have a greater understanding of our fundamental lack of success in this area, the cause, whether it was deliberate or simply ordained by inadequate tools, will remain open for debate. Whether it was BP’s tight control over FDA or academics wishing to secure further grants is open to debate as usual. However, we can see that the our company/approach warrants the concern and/or adulation of all, as many have rightly postulated it is the death knell of more than the Amyloid theory...the entire system. Precision medicine is the fix so good will come of it, even if the change is slow and we are the spearhead, whether we are permitted to hit the target or earmarked for doom by the big players, there will be no denying we changed the system. Note: this is not about precision medicine which is the future and the fix, this is about the past and one of the interesting ingredients which caused the problem, a less discussed flaw in the system but it also bodes well for our drug’s success - in fact, given the chinks in the system’s armor - we shine like never before. But we need to discuss the problem first, and I apologize if it is long, but it is understandable and necessary before we can fully appreciate what we have accomplished. (You’ve indulged me before...up for one more thought provoked read? C’mon. This is a big part of our story and I’ve included narrative, links, quotes, jokes, reasons to believe in us...it’s even interesting in the field of science.)

And so we begin:
Nidan, first to you, buddy. You have maintained with great steadfastness that our drug discovery and approval system is BROKEN. Nobody fails to wake up to reality after 30 years of the same embarrassing mishaps. You are correct, btw. I have some information to add to the “HOW did this happen?” Sorry, I can’t answer the Why? question. Greed? Corruption? Incompetence? Necessity? All of the above? Puzzle piece coming.

Sokol, this research and the puzzle pieces it provides, followed on the heels of our discussion of how our drug is being tested for MS by Biogen. Thank you for getting this ball rolling...even if it ended up turning into another ball but still leaving some room for the answer to our original question.

Xena, you were right about certain points which you continue to drive home much to your credit, at least that’s what I gather from this evidence. I think many will see this too.

Power, and others in this camp, can’t be sure of what our friendly giant BP’s have been up to with our drug for the past 2 years but some dots here to connect if you like.

Aero-man: “There is enough blame to go around between the academics and the for profit corporations”...you could not be more correct.

jimmy- mcyoloswag: Dashed decent contribution as to the different types of neuronal cells and I’m sure this won’t surprise you.

Neither will it surprise falconer.
Nor the many loyal long contributors who consistently raise great points and DD.

GrassyKnoll - yes, the book does parallel the Anavex story - amazing...I want your reading list!

And, to dado, your post containing pro-inflammation and anti-inflammation turns out to be the key to the wisdom of an investment in this stock.

Anavex community, please consider the following. It is interesting information, imo, and meaningful with our best hope for the future already in motion.

Why is our system broken? Why do processes fail? At which point in the process can one determine and declare it is inadequate and there is a drastic need to start over and try a new/different approach?
Don’t get me wrong, in a “break and replace” world, I value proper upkeep and maintenance. I would rather repair than replace almost anything , and invest attention to it from the start to avoid a state of disrepair...everything from relationships, to mechanical devices, to scientific discoveries. But, in a case where there is no baby in the bath water...I’m for throwing it all out.

The transgenic mice model:
(No baby, all bath water)
In matters of scientific discovery, one starts with a situation which is pervasive and problematic, observes common features thought to be significant, develops a hypothesis, sets out to test the hypothesis, then either is able or fails to validate the theory. If able, process over. If fail, try, try, again, hoping to tweak whatever appeared out of place or demonstrated faulty logic within each iteration hoping to get closer and closer to an outcome which passes. Even if that means revising entire theories.*

So, we have Alzheimer’s Disease in human brains. Along the way, we also notice that these brains have certain hallmarks: Beta Amyloid peptides, derived from Amyloid-B precursor proteins(APP), aggregate into plaques, and microtubule- protein tau which forms neurofibrillary tangles, both accumulate in the aberrant brain of the Alzheimer’s patient. Other than these visible pathologies, the patients demonstrate cognitive impairment, memory loss - short term and long term, and gradually progressing failure of neural function impacting mood, sleep, agitation, aggression, which, along with the cognitive impairment - confusion, disorientation, memory loss, render the sufferer eventually unable to carry out daily activities or care for themselves, and they spiral downward to total neurodegeneration until they reach a point of demise.

The observation phase, searching for causal evidence, began with autopsies of the diseased demised: Amyloid plaque build up was the first noted visible hallmark. At a point when testing of this cause remained unsuccessful (9 years) and instrumentation became more sophisticated, additional causes were looked for, although Amyloid plaque was never abandoned as one, it appeared it was not alone and more were needed. It was then that tau tangles were added to the observable.
This led to a hypothesis that the presence of these factors cause the disease. Further testing on the new amyloid/tau theory began. That is not irrational. So far, rational science.

Interesting to note that at some point, Amyloid plaque was found in brains of people without symptoms of Alzheimer’s. That could have meant something akin to “wrong path” but perhaps still was within the realm of “exceptions” so, maintain the hypothesis but allow for exceptions. That’s not entirely crazy either.

In order to test the hypothesis that removing Amyloid plaque from diseased brains would restore normal function, a model was needed, because you can’t test drugs on human brains without adhering to the “do no harm” pledge first.

Here is an interesting thing learned: there are no animals who suffer Alzheimer’s disease naturally in which to test drugs. Xena, this supports your points that we may well be doing this to ourselves with medicines which have as nasty side effects, brain damage...leading to Alzheimer’s. (As well as ‘bad’ lifestyles which contribute.) Animals don’t have ‘bad’ lifestyles, only inadequate abilities which cull them from the pack way before they are candidates for Alzheimer’s and they definitely don’t take maintenance drugs - they rarely have the health coverage to pay for them*). Perhaps, socialized medicine will help...?:)
But, more to the point, they also do not generally live as long as humans (at least the laboratory test models which need quick turnaround time), and they have an entirely different brain structure...they have primitive brains which are completely instinct driven. They have little cognition as we know it. (More to maximize reflexes and reactions - to territorial stimuli - than to dedicate to reflections and reminiscences such as enjoying sunsets, or showing each other photos of their grandchildren.) Primitive brains differ in anatomy and function from more highly developed brains with cognition. Wild animals can be domesticated and trained to learn certain routines - but they are responding to stimuli, not want of knowledge. (Two rabbits escaped from a laboratory and ended up in a lettuce patch. After being out for 2 hours and eating all the lettuce they could, one said, “I can’t believe this! We are the luckiest rabbits in the world...look at this enormous lettuce patch. Have you ever seen anything like it? And we are free! No more cages! The second rabbit shrugged his shoulders turned around and began hopping away. “Why are you leaving and where on earth are you going? What’s better than this?” asked the first rabbit. The second rabbit said, “I’m going back to the lab. I really need a cigarette.”)

Mice do not have Alzheimer’s or Amyloid plaque or tau.
Non-human primates have brains with Amyloid plaque in them. FACT. But they do not develop Alzheimer’s. Even as they age..........
Now, I can see Nidan, virtually, smh (smack his* head)...kind of a dead giveaway. Plaque - but no disease. (As Investor quotes, “Undaunted by knowledge, we moved on.” Applies.) Theory continues.
But wait, nidan, stick with me, it gets worse...

So, no animals to test this on. No problem. We will GIVE the animals (mice) Alzheimer’s in order to test cure it. After all, we give them cancer, and polio, and diabetes, and viruses, and other pathologies which enable us to develop cures to other maladies.
True. But, we know what strains of bacteria and viruses, and cancer cells to inject for mice to develop those pathologies. How do you give a mouse a Neurodegenerative disease which usually appears in the elderly after the brain malfunctions and suffers the continual insult of stress leading to inflammation over years and years? Do we wear out these mice brains in the hope of inducing Alzheimer’s? No. We think Alzheimer’s arises from Amyloid plaque so we just inject Amyloid plaque into their brains. (Even though that’s not how humans sporadically get Alzheimer’s - a diety doesn’t come and dump a bunch of plaque in our brains.) Well, we don’t dump it in the mice either. More specifically, in transgenically altered mice (the kind we use in preclinical research), in order to simulate Alzheimer’s, we create a gene which can be implanted in a mouse which overexpresses production of APP (see link at the end) the precursor protein which causes the mouse to create Amyloid peptide plaque. They refer to these as transgenic mice but they might as well call them plaque mice because they are simply “plaque-ing” them and that is all the further they looked for 30 years as a model to test Alzheimer’s drugs on. (Now, Nidan, cue the tsk tsk tsk!) Researchers (BP and academics alike...Aero-man and jimmy-mcyoloswag) have been stacking the deck this way: let’s put so much APP into the mice brains that they become cognitively impaired, just like Alzheimer’s patients, and then try to find ways to remove it. SMH! And THEY DID! For 30 years and counting (but starting to get better) unlike the mice who were successfully cleared of plaque a number of times but who never got better - cognitively. (Then these drugs went into humans, again and again, who also had clearing of plaque but never got improved cognition - function and memory. Starting to get it?)

That’s really not much different than injecting antifreeze or some other toxin to the point of brain damage, but not death, then removing the antifreeze, and the brain damage is still there. Garbage in - garbage out. That’s garbage science! And that’s what’s been going on.

Based on a faulty notion? Based on a clever way to keep funding the same project? Your call.

So, after we repeated this procedure sans success in humans (cleared the plaque but no joy in the cognition, wonder why?) we began to look for other contributing factors - not actual causes or throw out the model. At this point along the path, it had to be clear that the model did not resemble the problem, imo. And I’m sure Nidan would agree, your system needs some tweaking.)

Enter tau tangles. But mice don’t have tau tangles. Not even the transgenic mice! They never go on to produce tau tangles from the Amyloid which is another hallmark in humans! (Btw, you do know who was funding this research in academic labs from the beginning...the NIH. The good old government - us! And, contrary to some, the FDA could have determined at some point that clearing plaque was not a valid endpoint. They are allowed to look at the science that far! and based on previous failures say there is no connect and not approve a trial to go forward, just as they would not see any point in approving a trial for Alzheimer’s if a designated endpoint were to be able to sit long enough to have your blood drawn. It removes blood from the system and can be demonstrated to perform that but that has zero correlation to cognition. Same with removing other things from your body, like plaque. The endpoint should have been cognitive improvement or nothing and after 2-3 years of no gain with the A-b removal, they should have reviewed those trials very carefully before approving. You are right, nidan, egg is all over the FDA’s face now. But, if they were being paid off...just sayin’)
No tau tangles, you say? No problem. We took the closest thing we could get to work in mice to get their Amyloid to produce tau tangles...which happened to be a gene mutation from another CNS disease frontotemporal dementia, and injected the mutation into the already gene altered mice to get brains which contain plaque and tau. And it’s not even the same structure of tau as human brains diseased with Alzheimer’s. Mutated mice express only a 4-repeat tau variant whereas the human brain has a 4-repeat tau variant and a 3-repeat tau variant expressed in a 50:50 ratio.

It’s quite easy to see, as stated in the article, at this point one begins to question exactly what we are modeling. This sounds a lot like putting a bunch of stuff in which you know doesn’t belong in there just so you can take it out and say you show promise in curing the problem. It’s very bad science, if science at all.
(Now you can say it, nidan: system broken! Or, Situation normal all “cooked” up! Completely cooked up! With an “F”!) So perhaps this helps explain why drugs which looked so promising in the mouse models preclinical - failed at a rate of 99.6%. What in the Hell were we testing? I was truly at a loss when I read about this.

Here’s a summary of what is coming which is better than the past and supports our approach and what Anavex has done preclinically to make our results more meaningful. (with links after...this is becoming long-ish even by Bio’s standards, but it’s a complicated historical topic to delve into and do it justice without some substance).
None of this is a guarantee of success but when you look at the modeling we have, you will know why your money is in this company (I do!), and, you will wonder why the FDA hasn’t approved our trials yet (I will leave that to each of you, as well as why countries who want a cure DID approve our trials. And if and when a BP or 3 will drop the phony mouse model charade and buy the drug that treats the cause.) Those who say we just haven’t demonstrated enough with low double digit humans sample are guilty of looking out for themselves until there is nothing more to see and then looking away...we have demonstrated modeling in mice which is orders of magnitudes closer to human brains than anything else ever before seen. Maybe that’s why we have a person driving a car again? And a Nobel Scientist with the smarts joining our trial?)

Building a better mouse-model:
First, let’s agree that while other drugs claimed promise based upon the fact that they removed plaque and tau from the transgenic mouse brains (I keep trying to decide which is correct - mouse brains or mice brains?) , that was their objective, which, once met, justified using these to treat humans with Alzheimer’s and restore cognition. A jump to improved cognition was inferred by removal of plaque and tangles but never demonstrated.
Anavex A2-73 sigma 1 receptor agonist which was postulated to restore cellular homeostasis and correct mitochondrial dysfunction which our company’s science believed was the genesis of the disease, showed less in the area of plaque/tau clearing alone and initially, but demonstrated improved or regained cognition in the transgenic mice.
Here’s the A PLUS +++ takeaway: Even though these mice’s brains’ neurons and mitochondria and ER were damaged rather quickly and through a process altogether different from Alzheimer’s in humans (akin to just picking them up by their tails and traumatically smacking them down on the lab counters until they saw Tom and Jerry stars circling their heads - the equivalent of introducing mutations such that either way they never knew what him them), our approach STILL RESTORED COGNITION! That’s big! That suggests a lot: our S1r agonist can help with all types of brain problems be they Neurodegenerative or sudden concussion or due to hypoxia (X!), restoration appears to take place. That’s a bit huge, really!

Next, Anavex went further in their attempt to nail down the hypothesis. We took regular mice and removed their Sigma 1 receptors. These are called knock out mice. We knocked the S1r out. Then we crossbred them with the transgenically altered mice who have the Alzheimer’s hallmarks and cognitive impairment. Then we administered A2-73 to mice with no S1r to agonize. Know what? They never got better.
(Are you loving this, Nidan??? Bet if I were a gal, you’d kiss me right now lol!)
(Cite and quote at bottom: Anavex)

Finally,
“Rudy Tanzi, a neuroscientist at Harvard Medical School in Boston, Massachusetts, reported in 2014 that his lab had grown human neural stem cells containing APP and PSEN1 mutations in a 3D culture system supported by a gel matrix (S. H. Choi et al. Nature 515, 274–278; 2014). The results were startling: the tissue that developed contained neurons that deposited amyloid-ß into the gel, and plaques formed after 5 to 6 weeks. A week or two later, the elusive tau tangles also appeared” - the exact tau replica seen in humans with Alzheimer’s in the exact ratio. It’s remarkable. And lower cost, and much quicker...in fact, companies can test tons of compounds on these brain models in a very condensed period of time. Anyone want to speculate about the BIIB MTA Petri dish assays going on, now’s your chance! Someone posted it would require more A2-73 than they could get their hands on...au contraire, friend.
(Cite at bottom: Nature 2014)

Now, to dado’s post and the hope of all long investors reading (and not, for that matter):
“The race is on to add different cell types to organoids, so that they can better represent real brain tissue. Tanzi has already incorporated microglia into his model. Initially held in a reservoir outside the dish in which the organoids grow, the microglia travel down microfluidic channels into the main plate in response to the release of pro-inflammatory compounds by stressed neurons (J. Park et al. Nature Neurosci. 21, 941–951; 2018). The microglia then respond to any plaques by pruning the junctions between neurons — effectively trimming the connections that underpin memory. Tanzi thinks that this is how the immune system responds to plaques and tangles, and inflicts harm on the brain.” EMPHASIS MINE
(Cite at bottom: Nature)

Repeat for added emphasis:
In response to release of proflammatory compounds by stressed neurons, microglia travel down microfluidic channels then respond to any plaques by pruning the junctions between neurons - effectively trimming the connections that underpin memory.
Tanzi (Harvard) thinks this is how the immune system works and inflicts harm on the brain.
(Cite at bottom: Nature)
Nidan - cool (thumbs up)

Brain junctions are also referred to as gaps or synapses. (Google brain cell junctions)

“Neurotransmitter (NT) receptors are located on the surface of neuronal and glial cells. At a synapse, one neuron sends messages to the other neuron via neurotransmitters. Therefore, the postsynaptic neuron, the one receiving the message, clusters NT receptors at this specific place in its membrane.” (Google synapse)

“Microglial cells are responsible for clearing and maintaining the central nervous system (CNS) microenvironment. Upon brain damage, they move toward injuries to clear the area by engulfing dying neurons. However, in the context of many neurological disorders chronic microglial responses are responsible for neurodegeneration. Therefore, it is important to understand how these cells can be "switched-off" and regain their ramified state. Current research suggests that microglial inflammatory responses can be inhibited by sigma (s) receptor activation.”
2015 article.
https://www.ncbi.nlm.nih.gov/m/pubmed/25666889/


Nidan and all - the “How” this happened is, we had a really bad model. The “Why” is known to some as always. But, your gift today is to see that the broken model is being FIXED!!!! (And A2-73 passed with flying colors and even checked our math, using the knockout mice, BEFORE we turned in our test paper. We should get an A+ wink! Hell, we should be teacher’s pet! Lol!)

This should give some sense of how important the preclinical mouse work is and if we choose to laugh at “just another mouse model paper being presented” we can do so in context. These are very important when done correctly!
All this adds up to a great hope for our success and I didn’t even have to rely on painters, pianos, golfers, drivers...strong responders or genetic biomarkers. Imagine how we look once we add in those metrics. Yeah, baby! (Austin Powers)

AVXL Community,
I think we have a live horse, remain mounted! A champion thoroughbred. Maybe even a Triple Crown Winner???
FYI, I’m not betting to place or show...all my money is on the big purse...running for them roses:)


Thanks all,
Bio (the LONG long! sorry)

FIN

Citations:

The article in Nature:
https://www.nature.com/articles/d41586-018-05722-9

Anavex knock out cite:
https://www.anavex.com/newlypublished_data_alzheimer/
“The transgenic mice, bred to express human amyloid precursor protein (APP) carrying the Swedish and Indiana mutations, APPSweInd (also known as the J20 line), were crossbred with S1R knockout (S1RKO) mice (APPSweInd/S1RKO). Compared to APPSweInd or S1RKO mice, APPSweInd/S1RKO exhibited an increased vulnerability to amyloid toxicity and more pronounced deficits in both short and long term memory.”