 Good morning everybody. Hope everybody's doing great this morning. I hope you feel fit in the environment that you're in today I loved it. I love the terminology that she had in that last talk. I think it was great and Just so you understand my background. I'm a drug design chemist by training. I was actually designing oncology drugs in a PhD program at Arizona State and From that I had to do a lot of genetic work and so we're gonna we're gonna be looking at a lot of gene sequencing today and In the context of gene sequences, we're gonna be looking at phylogenetic trees of the endocannabinoid system So I want to kind of get an idea of the background. It's pretty much everybody in here a medical professional MDs NDS. No Do we have any evolutionary biologists in here today? No, thank God because oh So I have to give the you're not gonna believe anything I say I Have to give the disclaimer my perspective on evolutionary biology is when I was doing drug design, we were working on a Enzyme that had not been Crystallized and so we had no structure to work with and so we use gene sequences to look for what are known as conserved regions in the gene sequence because the conserved region of the gene sequence often is a sequence that is important for the active site of an enzyme and That was what our focus was in drug design was enzyme active sites and so I'm not an evolutionary Biologist, so don't expect this to be a good evolutionary biology talk. This is a biochemistry talk from a a Conservation of gene sequences perspective So it's important to understand the the function of endocannabinoids because these molecules over time have evolved in different species and Not all species can produce all of these endocannabinoids for example. We put we produce Arachid on a ethanol amide from arachidonic acid Well, there are some species of insects that don't produce arachidonic acid So they can't produce like Drosophila cannot produce arachidon ethanol amide, but Drosophila can produce To arachidon and glycerol and they don't have cannabinoid receptors So we really don't know why the Drosophila is producing an endocannabinoid when it doesn't have an endocannabinoid receptor and we're going to be looking at some of the Receptors that are non endocannabinoid receptors that bind to endocannabinoids as a result of that so when I look at these I see these as derivatives of arachidonic acid and You probably know that arachidonic acid is an important essential fatty acid I mean we don't necessarily produce enough of it We can produce some arachidonic acid from omega-6's, but one of the problems that I have right now with our understanding of the endocannabinoid system is that most literature research in Endocannabinoid system is focusing on just these two endocannabinoids Hmm, and if they're both derived from arachidonic acid does anybody see the problem with that? Well, yeah Because we get a lot more other essential fatty acids in our diet other than arachidonic acid, right? and it turns out that these these arachidonic acid derivatives are strong cannabinoid receptor agonists and They bind differently to CB1 and CB2 and they have different functions in the human body You'll hear neurobiologists talk about arachidonateethanolamide, which is also known as anandamide they refer to this as a partial agonist and They they talk about it as being tonic in nature. So two arachidonic glycerol is referred to as phasic because What these molecules do is they modulate neurotransmission and so This one is referred to as tonic because it's kind of always there And it's always kind of tuning in neurotransmission and this one Kind of comes in when there are bursts of neurotransmission and it's really important and this is one of the most complex You know the human endocannabinoid systems one of the most complex that is that has been characterized so far so When we look at some of these other molecules This is a derivative DHA and so now we transition from Looking at just omega-3 or omega-6 is now we're looking at some of the essential fatty acids that are derived from omega-3 fatty acids And it turns out that the DHA derivative of ethanolamine is referred to as synaptomide Because when it was originally discovered it was discovered by neurochemists that were looking for things that stimulated synaptogenesis So who wants to go out and get some DHA supplements now? Good lord if I could make myself smarter By changing my omega-3 to omega-6 ratio Sounds promising to me And you'll hear me refer when I talk about the endocannabinoid system you'll hear me refer to what's known as endocannabinoid tone You know and our endocannabinoid tone is largely built upon our intake of essential fatty acids and our ability to convert those fatty acids well convert those fatty acids into These molecules that we refer to as endocannabinoid And so if we are efficient at producing endocannabinoids from our essential fatty acids We would have a positive endocannabinoid tone and if we're inefficient we would say that we have a negative endocannabinoid tone and It's important because the balance of omega-3s and omega-6s is so Important in this one of my favorite researchers Oliver Manzoni has done some groundbreaking work with mice And one of the things that he's shown with mice is that mice that are fed a diet that's deficient in omega-6s or pardon me Wow, there's a slip diet deficient omega-3s There is no diet deficient omega-6s is there no doesn't exist But there it's easy to have a diet that's deficient omega-3s That's a standard American diet because of all of our consumption of the industrial seed oils I won't call them food oils because they're not they're industrial seed oils and they're good for one thing lubrication But what Oliver Manzoni has shown is that mice lose Neuroplasticity in the absence of omega-3s who in here would like to have less neuroplasticity Sign me up Maybe I could become president so We know that in mice they lose neuroplasticity if they don't get omega-3s and we know That the DHA ethanol amide endocannabinoid was discovered by people that are looking for things that stimulate synaptogenesis Paints a compelling picture for me. Does it not for you to quit drinking or eating omega-6s? so The last molecule that's up here is another one that is a serotonin derivative of Arachidonic acid that has endocannabinoid function There's also a dopamine endocannabinoid and so the point is I want you to understand that your diet influences your endocannabinoid system Dramatically, and we're just scratching the surface on these molecules And we need a lot more research that doesn't just focus on The first two that were discovered and actually if if I put this slide presentation into historical order We're going backwards Because the endocannabinoids were actually discovered after the receptors were discovered and the receptors were scubbed were discovered after THC was characterized and so Our whole understanding of the endocannabinoid system is backwards because we discovered we meaning Raphael McCoolam discovered THC in the 60s, and then we thought THC just had some magical property with neural membranes until 1990 when the first cannabinoid receptor was discovered the second cannabinoid receptor was discovered in 1993 and then the endocannabinoids were discovered in the late 90s and so our understanding of this is built backwards and we have a lot of laws associated with cannabis that put a lot of roadblocks in front of the Researchers that were doing this kind of work Okay, so when we look at the the gross Physiology of cannabinoid receptors You'll often hear people saying that CB2 receptors are found in the immune system. Well, they are that's true And they're found in a lot of other places in the body, and they have a lot of diverse functions one of the primary functions of CB2 is in the blood-brain barrier and in the gut barrier CB2 is absolutely essential For normal permeability it maintains normal blood-brain barrier permeability by controlling cytochrome or cytochrome cytochine inflammation and so CB2 you'll often hear that it was that it is in the immune cells But it was it was discovered in the rat spleen, which is why people think it's only in the immune cells It is it is found in a large number of other cells besides immune cells and has some pretty diverse function CB1 I focus primarily on The synaptic function of CB1 because I think that that's probably the most important function of CB1, but CB1 also has influences on some of our very Fundamental physiological homeostatic processes like eating Everybody knows that you know cannabis is is really important in the oncology population And I want to be one of the few people that are out here saying cannabis does not cure cancer It's not a magic bullet It's a really important toolbox in the in the toolbox of the oncologist because Western medicine has nothing for The anti-medic effect that cannabis has Western medicine has nothing to improve appetite in patients with catechia And so like every other thing in the toolbox of the oncologist Cannabis has one thing that needs to be applied with all of the other tools So when you hear people say that cannabis cures cancer turn and walk Please that's not good science. That's bro science and I work in the cannabis industry But I am unpopular in the cannabis industry because of my stance on good science versus bro science so Some of the other things that are really important within CB1 function our motility in the gut This is why CB CB1 antagonists like like THC are really good for patients with Crohn's and IBS those Spastic conditions of the gut can be quieted down with CB1 receptor agonists Okay, a little bit more Closer look I've been doing a lot of talks about meta metabolism nation. I just spoke at low-carb USA and I Tailored this talk to fit the the overall metabolic function of CB1 and CB2 And it's interesting because when you look at things like CB1 stimulates glucose uptake in adipose tissue This is not a sunny side up egg. That's supposed to be an adipose cell and In adipose cells or sunny side up eggs CB1 increases glucose uptake it increases insulin sensitivity and it increases the storage of fats But one of the weird things about the population studies of cannabis users and Animals is that in in these studies cannabis users are typically lower in BMI Than the general population. Well, does that make sense when you think about? CB1 stimulating the uptake of glucose and the storage of fat. Well, no it doesn't I call this a stoner paradox We'll come back and look at this a little bit later in the talk, but I just want to confuse you up front and Bring it back. I mean it's confusing because we have these conflicting things between cell line tests and animal models And this is often true in biochemistry because you know, we probe these type of activity using pharmacological methods like can't cannabinoid receptor antagonists and we probe with with agonists Pharmacologically and we you know, we look at different cellular functions and often they don't add up in organism level studies So we'll come back and take a look at this because there's more to this when you look at the organism Because of all of the things that are previous speak spoke or Speaker spoke about in terms of how different cells within the organism have to cooperate And so whenever you look at cell line or enzyme level Data you always have to recognize it There's a limitation and when we put that data into the context of an organism level study it will likely not necessarily fit Okay, so this is where the rubber beats the road in terms of CB1 function This is what I think is the most important part of CB1 function the synaptic model. So in this model on the left We have a an excitatory Neuron and on the right we have an inhibitory neuron and they come together and they they Intervate with a post-synaptic neuron to do their thing and their thing is neurotransmission What we what we see with endocannabinoids is the endocannabinoids are produced post-synaptically By specific enzymes and we'll talk we won't talk too much about these enzymes in terms of the Genetic evolution, but just know that there are Enzymes that produce these endocannabinoids and not all organisms produce these enzymes And so what we see here is the most complex endocannabinoid system that exists in nature as we know it today And so these endocannabinoids are produced post-synaptically. They're transported. They go to receptors presynaptically and They modulate the release of neurotransmitters They don't stop the release of neurotransmitters. They modulate it. And so like the anti-lock brakes on your car they They tune the release of these neurotransmitters so that your central nervous system does not overload itself with information or or neurotransmission and so there's there are Direct relationships between these receptors and the release of neurotransmitters and and when the The receptor sites have been bound the end of cannabinoid is then broken down into its It's individual synthetic components. And so we have enzymes that do that as well And so there are organisms that have the enzymes that to produce some of these endocannabinoids that don't have cannabinoid receptors and There are organisms that have the enzymes that break down endocannabinoids that don't have You know connect the cannabis the cannabinoid receptors so in general when we look at This it's a really important thing for the overall longevity of the central central Excuse me of the central nervous system and because we have such a complex Good lord. Sorry. We have such a complex central nervous system It's important that our bodies can regulate neurotransmission and especially excitatory neurotransmission because excess glutamate neurotransmission is associated with chronic neurodegenerative diseases like Alzheimer's ALS And so this is where the neuro protective effects of phyto cannabinoids are important Okay, are we ready for For the gene sequences So this is what's known as phylogenetic tree and this is a phylogenetic tree for The CNR1 CNR1 is the name of the gene that encodes for CB1 receptor and so You can see from this that there are some differences in spacing and if you're not an evolutionary biologist What what this tells us is that there is a large number of species that produce the cannabinoid receptor and You know some of these are You know advanced organisms organisms like the primates and some of these are simple organisms and You know some of these organisms the elephant shark is referred to as an evolutionarily resistant Species because it has not evolved in you know hundreds of thousands of years and so when you see Evolution evolutionarily resistant species like the elephant shark that are expressing cannabinoid receptors that have a high degree of homology with the human receptor It means that this receptor has something very fundamental with our basic physiology That's the main thing. I want you to come out of this is That when you see a phylogenetic tree like this that spans from Mammals to invertebrates it means that that gene is really important in terms of the basic Function and it's it's funny because in different organisms You know, I look at a lot of different studies and there are bird studies that have shown when birds are treated with cannabinoid receptor Antagonists it changes their song pattern So by blocking their cannabinoid receptor, it's their song patterns are fundamental to their entire social structure And if you can interrupt that with a cannabinoid receptor antagonist, that's a pretty profound evolutionary Effect right there in mice Cannabinoid receptor antagonists in mice pups inhibits suckling Well, that's a horrible thing to change evolutionarily, you know when we when we look at how Receptors and different genes evolve over time There's a lot of these receptors that are you know genes that evolve faster than others And this is one of the ones that has had evolved very slowly over time and Part of it is because there's a lot of crossover and what I mean by crossover is this so this is the enzyme Monoacylglycerolipase that breaks down to arachnidolglycerol Cox2 is the enzyme that that we are familiar with because of the the class of drugs Nonsteroidal and anti-inflammatories right cox2 inhibitors block the production of inflammatory Prostaglandins and remember that prostaglandins are fatty acid derivatives as well and it turns out that cox2 can metabolize the endocannabinoids as well and this is why When I look at the phylogenic tree, I want to look at things like cox2 I also want to look at some of the other enzymes in The human endocannabinoid system that are bound by the endocannabinoids So the the last one here FAH is the enzyme that breaks down arachnolaptenol amide also known as anandamide and so there's there's some some Authors that believe that by blocking cox2 We actually have a little bit of an antidepressant effect because remember anandamide was derived the name anandamide It was derived from Sanskrit Ananda which is bliss okay, so This is the phylogenetic tree for the cannabinoid receptor 2 and CNR 2 the gene CNR 2 encodes for CB2 receptor and here is where there is a lot of disagreement within evolutionary biology. There's There there was a split at one point where there was one organism in the phylogenetic tree that Produced a mutant CB1 and There are some people that believe that that mutant CB1 is what became CB2 evolutionarily and the reason they believe that is because there is a high degree of Homology and everybody get what I mean when I say homology. There's a lot of you can see The similarity in each of these gene sequences. That's what homology is There's a high degree of homology between CB1 and CB2 Which suggests that there's an evolutionary relation between the two But there's only 44 percent homology which which means that it It probably is within the same species that the split happened and so This is one area where there needs to be a lot more study. I mean I think the first Study that I actually could pull up on the evolutionary biology of the anokinabinoid system was 2000 And then there was really nothing till 2005 and there was 2007 and I think you know You could count on both hands the number of published articles on the evolution of the endocannabinoid system, so This is a representation that I used at a conference on low-dose naltrexone and the reason I wanted to put it up here is because endocannabinoids and phyto cannabinoids are referred to as promiscuous receptor binding because they bind to cannabinoid receptors they bind to Vanilloid receptors they bind to opiate receptors and because of that when we talked about The evolutionary biology we have to think about okay if an organism produces endocannabinoids, but it doesn't have Cannabinoid receptors well, there must be some other function in that organism and likely You know TRPV is a suspect to look at in organisms that don't have cannabinoid receptors And the vanilla receptors are important for the sensation of pain and heat and over their evolution the vanilla receptors have actually split into receptors that can that can sense heat pressure and acidity and This is the evolutionary tree for the TRP receptors and it looks like It's not as diverse as the cannabinoid receptors, but there's a large number of organisms that have these receptors and It's important. There's the elephant shark again It seems like because it's evolutionarily conserved. It's an ancient gene It's probably pretty important for a lot of fundamental processes Um CB2 I wanted to come back. We're not going to look at much more in terms of phylogenetic trees I want to take this back into physiology again and leave you with some of Some of the physiological thing that you as practitioners can be looking at to help your patients that have issues with their endocannabinoid system. So CB2 we talked a little bit earlier about The Neurological importance of CB2 so microglia and astrocytes CB2 has a profound effect on cytokine inflammation As we all know cytokine inflammation is the backbone of most chronic neurodegenerative diseases One last phylogenetic tree FAH this is the enzyme that breaks down the the endocannabinoid Raccoonolethanolamide Just notice that there's a lot of different enzymes that have this but to me when I look at this I see there's less Single cell organisms and there's more mammals on this. So there's something about this enzyme that tends to be more It's found more often in the complex organisms like us Okay when we look at metabolism we talked a little bit earlier about Obesity and BMI This study was looking at mice and what they were saying was that They concluded the inability of omega-3 fatty acids to reduce adipose tissue and serum levels of classic endocannabinoids Might contribute to lack of beneficial effect of these lipids on glucose homeostasis and type 2 diabetes Clients will remember that the classic endocannabinoids are from omega-6s so they kind of missed the boat on this and When we look at CB1 and CB2 receptor function in terms of How they influence? fat mass we see that there are some Influences based on the expression of RNA for insulin and Glucose uptake proteins and so this once again I Don't think that this fully explains The the role of the endocannabinoids system in how our our body Distributes and stores adipose tissue when we look more deeply into this the CB1 receptor actually is down regulating mitochondrial function Well, that doesn't fit into what we would think is positive for overall health But when you look at it It seems like the insulin receptor is Central to the ability of the endocannabinoid system to either have a positive or negative effect on our disposition of adipose tissue and Modiculant and it's directly related to how insulin influences mitochondrial function and there is an interplay between that toll like receptors and the cannabinoid receptors We look at in a cannabinoid system function at adipose tissue and It is actually CB1 receptors actually blocking the carnitine shuttle Well, that doesn't bode well for the ketogenic diet as it would seem right We look at the inter cannabinoid system and brown adipose tissue versus white adipose tissue and we see things like Increase in fatty acid uptake in white adipose tissue, which we would think would probably lead to obesity We look at fasting fasting is influenced and influences the endocannabinoid system and the stoner paradox We we see in mice that Adult male mice that have dietarily induced Obesity and remember when you're inducing dietary Obesity omega-6 is a really really good at that But omega-3s are not We look at the gut gut function There's a strong interplay between cannabinoid receptors and the immune system in terms of how gut permeability is controlled and it turns out that gut permeability has an interplay with With the microbiome and so The endocannabinoid system influences the microbiome and the microbiome influences the endocannabinoid system There's a lot of great mouse studies on how there are specific Gut microbiota that are influenced by phyto cannabinoids and there seems to be a protective effect when we look at the phyto cannabinoids Delta 9 THC And this is for clinicians. I think this is a really important thing to look at. There's a genetic reason why Oral consumption has a 7 to 72 hour range of half-life for the physiological effects of THC Horrible horrible Pharmacological profile and that's oral inhaled really predictable three to four hour half-life and Really predictable response. There's a genetic variation in Cytochrome 2C9 which is the cause of this because oral to consume THC goes through first-class metabolism and if you have Polymorphisms in your 2C9 you are likely going to be either an ultrafast or ultra slow metabolizer of Delta 9 THC CBD which is how to cure everything Is really important. I think that it should be treated like an essential nutrient, but I wouldn't say that it's going to cure your cancer Has an incredibly long half-life. So when you're thinking about dosing these things are really important I love beta-carry-off-lane and the reason I have this in here is because beta-carry-off-lane is a CB2 selective agent And it's also found in Some of my favorite foods. So clove Black pepper shiso anybody know shiso from your Japanese restaurant if you don't next time you get sushi order shiso With one of your sushi rolls. It's one of the most brilliant herbs that exists on the planet Mercy mercy and lining I put these two here to contrast them because most modern quinovars of cannabis have been bred to either produce mercy and are called Indicas or they produce high levels of limonene or piney and they're called sativa's well the indica and sativa are How many f-bombs can I drop when I think about How much of a gross generalization they are the thing is Strain names don't mean anything you need to think about these is quinovars because Mercy has Profound pharmacological activity. We're gonna everybody mind if we go a little bit over into the Q&A We got a couple more slides. We don't have a lot, but I think that this is really important So mercy's activity can actually be blocked by the drug naltrexone Which implies that mercy is an opiate receptor Agnest and so when you think about Pharmacological utility high mercy and strains which have been bastardly called Indicas can be really good for pain management and especially if you're doing an oral preparation that has an extremely long half-life This can be something that has very profound positive effects for cancer patients that have metastatic pain Because that pain is not touched by many things and as you know Opiates are probably not the thing that you want to manage that pain on the the ups the positive side I refer to mercy high mercy and strains as being kind of Heavy and dark They're really good for sleep, and they're really good for pain But you don't necessarily want to go drive a front-end loader when you're consuming a high mercy and strength Especially if you're consuming it orally And I refer to limonene is being more light everybody Everybody knows the smell of citrus and everybody knows that it makes you feel good, right? I mean, this is why citrus is in cleaning products because limonene has a positive uplifting feeling and high limonene strains tend to give you a kind of mindful and you know uplifting high and You'll often hear them refer to a sativa which doesn't really mean anything, but There was a study in Japan in an institution, so it was pretty well controlled They showed that just by smelling limonene essential oil in a cotton ball in a mental institution They reduced the consumption of pharmacological antidepressants. That's pretty powerful. I think so To tie this back into evolution and how we co-evolve with our foods I really would have loved to have been able to do a co-evolution of how phyto cannabinoids produced by plants Influenced our evolution, but there's really no data on it yet, and I would be sitting up here making stuff up But what we do know is that part of our Agriculturalist evolution we've included a lot of things like spices And this is one of the things that I want to caution you about when you're reading mouse studies How many mice are consuming beta karyothelene rich herbs when they're eating a high-fat diet? Well, probably not unless there's their Ayurvedic mice, right? You may know any Ayurvedic mice. I bet they smell good But it's important when you look at these studies that are looking at high-fat diet Number one recognize is that high-fat diet just omega-6 is well for mice. It likely is or is it just saturated fat? Well, probably and there are some issues with that. I mean, I don't want to know what a high-fat diet does I mean we already see that in the general American population, right? It's not a good outcome When we look at high omega-6 fats But the thing that's often overlooked is how do these phytochemicals from our culinary herbs Influence our metabolism and our microbiome and I think that that's the take-home message is we've evolved to develop receptors and enzymes For endocannabinoid system that that influences our basic things like hunger, you know extinction of fear these things are primal In terms of our neurological development and it's important that we can include These kind of things in our diet to modify our endocannabinoid system so that we can have a healthy endocannabinoid system Okay with that we have any questions It was interesting. You mentioned the Neltruxone LDN low-dose Neltruxone five milligrams a day is as maybe in my Antiquated history I've been touted as a anti-aging cure all no downsides But you just mentioned it a block something perhaps useful But is there a daily a normal daily function our body? Do you have any comments on now? LDN in a good or positive or negative way so low-dose Neltruxone one of the positive effects of low-dose Neltruxone that's not observed in high-dose is Upregulation of the opiate receptors and you know There's other there's other things that low-dose Neltruxone does it is it antagonist of toll-like receptors So there's an immune component. That's really important The probably the most important thing that low-dose Neltruxone does is it also is blocking the Opiate receptor the opiate growth factor receptor, and I think that that's probably one of the more important things I don't think that you know when I when I mentioned that that the effects of Mercy were blocked by Neltruxone. That's high dose typically a pharmacological dose of Neltruxone that will block Opiate receptor agonist is high dose Do you have any Do and or don't Thoughts on LDN as a long-term and the aging protocol? My understanding is not from the clinical side So I can't really comment on that nature would be a better person that she has like the clinical experience with low-dose Neltruxone Thank you very much for your talk super interesting Obviously CBD is everywhere and there's tons of hype around it That could be a whole talk, but would in your opinion What do you think that there are benefits for CBD and what would be kind of the top ones in your opinion neuro protective effects? I mean the the US Department of Health and Human Services Has a patent on the neuro protective effects of CBD and they name the NMDA receptor system and the ampere receptor system as the primary targets and remember the NMDA receptor system is Link to almost every chronic neuro degenerative disease and so I would say Number one. I'm not gonna say that CBD cures anything But it's the if I were to take any, you know phyto compound on a daily basis It would be CBD and simply because it's just really it's a really good tonic for so many different things Yeah, well, thank you Steve. It's gonna be a big round of applause for that great talk