Plant Chemistry, Cannabinoids, Cannabis Terpenes & the Entourage Effect | Ryan Vandrey | #152

About the guest: Ryan Vandrey, PhD is a Professor of Psychiatry & Behavioral Sciences at Johns Hopkins University, where his lab studies the behavioral pharmacology of cannabis (marijuana), nicotine/tobacco, and other substances.Episode summary: Nick and Dr. Vandrey discuss: marijuana; cannabinoids like THC; terpenes like limonene, myrcene, and pinene; the entourage effect; anxiety and other side effects of cannabis consumption; and more.*This content is never meant to serve as medical advice.

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* Episode transcript below.

Full AI-generated transcript below. Beware of typos & mistranslations!

Ryan Vandrey 2:59

empirical data buying some of the claims around cannabis, whether it's good, whether it's bad, or somewhere in between, and we try to be caustic to all of that and, and let our data do the speaking for us.

Nick Jikomes 3:14

And do you guys work with humans animal models? What's your modality?

Ryan Vandrey 3:18

So we in the lab here, there's a translational approach here. So there are some scientists part of the cannabis science lab that do preclinical work. And I personally do work with humans almost exclusively.

Nick Jikomes 3:35

And so a lot of your work informs policy how, you know, before we get into the science of it, how direct is that connection? Do you work directly with policymakers? Or do you just sort of work in an area that's of interest to policymakers who you hope then read it? It's

Ryan Vandrey 3:50

a little of both. And you know, when I started, I started out doing research with cannabis, it was only us doing things that we thought would be helpful. But you know, that's 20 years ago before widespread legalization or decriminalization, and changes in in retail marketplace really coming about. So it was pretty tangential. But now, you know, we're very intentional in monitoring and seeing what's happening and seeing what kinds of changes in the laws are being put forth and seeing changes in product types. And then we basically use that as a foundation for determining what kinds of research projects we think are the most important that what we go after. So, I guess to go back to your original question, you know, we do research that we believe is very helpful to policy and decision making and things like that, but we also have worked directly with legislators on Language in draft bills and legislation. I was part of a task force work group that helped design the medical cannabis law in the state of Maryland. And we've been called off by the FDA and other regulatory agencies to kind of come in and speak to what we've seen in the laboratory and how that might help them inform their policies and regulations at the state and federal level.

Nick Jikomes 5:25

So you study cannabis, also known as marijuana, and it goes by many names. Obviously, you know, it's a plant people use it for recreational and medicinal purposes. As a scientist as a biology guy, how do you, what is cannabis to you? A lot of people think of it as being synonymous with THC, that from a chemical or pharmacological perspective, what is it

Ryan Vandrey 5:48

as I try to not even use the term cannabis anymore, because it's an umbrella term that 20 years ago was meaningful, in that if you said cannabis 97% of the time, we were talking about dried flowers of a plant that were rolled up and smoked, or stuffed into a pipe or a bong and smoked. And that was it. And it was always THC dominant in terms of chemical composition. But the reality is, once we opened up a retail marketplace, there are hundreds of different kinds of cannabis based products that exist now. And so I try as much as I can to talk about cannabis as a generality. But when we talk about specific things, and whether it's good, whether it's bad, how do you use it once the decision making, I try to be much more precise, because when we talk about those kinds of things, health impacts, we need to know what the primary chemical constituent is what the intended route of the stration is. And there's further nuance beyond that. But those are the two key things were I try as hard as I can to say, THC dominant cannabis CBD dominant cannabis CPG, dominant cannabis, and then talk about whether it's intended to be inhaled, orally ingested, topically applied, or some other manner of assumption.

Nick Jikomes 7:24

And, you know, one of the things that's always been interesting, but also tricky from a research perspective, about cannabis, of any kind is, you know, is that it's, it's not one thing, people consume different types. And we have to be specific about what exactly the differences are. They consume it in different ways. The research historically has not always but often been. We often see researchers using preparations where subjects are consuming something like pure THC or something that is different from what people are using out in naturalistic settings out in the real world when they go buy cannabis legally or illegally, and then consume it. Can you talk a little bit about the pros and cons of doing that of studying sort of the isolated molecules versus the actual products that are out there in the real world?

Ryan Vandrey 8:17

Yeah, so and this will be a recurring theme throughout our conversation is there's a lot of nuance to cannabis science and to retail cannabis goods that have to be factored in and are important and how you talk about it, how you think through these things. So we have studied both whole plant products as well as isolated individual chemical constituents, I think it's very important to do both. Because we want to have science that is reflective of whole plant botanical products. And there's a whole theory behind, you know, is the whole greater than the sum of its parts kind of thing. And there's an entourage theory about how, you know, the natural plant product is somehow superior to the individual chemical constituents. But you need the science to back that up. But at the same time, if all of the science is done with this whole plant, botanical product, which is made up of hundreds of individual chemical entities, that vary from one plant to the next to the next to the next, can we say anything very confidently about what the individual constituents do without studying them. And so part of what we are doing here is to understand the effects of individual chemical entities and how they interact with each other. And you can kind of take some of that data to then go back to whole plant modalities of say, okay, A well in, in this scenario according to Ana entourage hypotheses, these two things balance each other out, whereas this one exacerbates something else. Can we take those specific hypotheses and pull them in to the lab and say, Is this true or is it not? And I think that that's important and becoming more important, because what we're seeing are cannabis industry claims about these things in the absence of empirical data.

Nick Jikomes 10:32

And, I mean, you know, one of the one of the remarkable things about cannabis is the variety or at least the apparent variety that's out there, if you go into a retail store, so I live in Washington, there's there's cannabis shops all over the place, you go, and you've got hundreds of different products, many different product types. For now, let's just let's just focus on the most common type, which is flower THC dominant flower, then the most most common method of consumption, which is inhalation either by smoking or vaporization. So but even within that, there's a lot of variety. There's a lot of different strains or cultivars or chemo VARs, depending on the nomenclature you want to use, that are out there or at least, marketed as such. And, you know, there's there's purportedly hundreds, hundreds, hundreds of different varieties of Canna THC dominant cannabis flower. And there are phenotypic, like morphological differences. Some of them, they vary by color, they vary by you know, the shape and the density of the buds and so forth. They vary by aroma, you can smell differences often oftentimes is best done side by side, you can smell one smell the other. And they often have a common core to them that that marijuana smell, but there's a lot of variety there in the aroma. And people say that there are a lot of different effects that different cultivars will be better or worse for giving you a certain type of psychoactive effect, or might work for one medical application where another strain doesn't work at all. Some might have side effects and others don't, et cetera, et cetera. So there's all of this apparent or purported variety out there. And a key question is, how much of that is real? And how much of that is marketing? to sort of get into this question? If we're thinking about THC dominant flower, the most common type of cannabis product consumed? What is it chemically? So it's obviously got THC in it. But what else is in there that accounts for some of that variation in aroma, and potentially some of those differences in effects that people claim? Yeah.

Ryan Vandrey 12:35

So when you think of the cannabis plant, and again, I am not a botanist, at all on the macro Well, right. So there are three kind of Common Core types of chemical entities in the cannabis plant. There are phyto cannabinoids, and these are the things that are unique to the cannabis plant. Many of them but not all of them directly engaged with the body's endocannabinoid system, albeit in different ways. Then you have terpenes or terpenoids. And these are kind of the flavor, the aroma, the scent type, chemicals. And again, some are unique to cannabis and some are not you know, a lot of them are common to other types of plants, citrus plants, things like that. And then you have so you got your cabinet leaves your terpenes, and then your flavonoids, right. And so a lot of it is the structure and integrity of the plant itself and waxes and things like that, that really, there's no reason to believe that they're pharmacologically active. But they're important parts of the skeleton of the plant so to speak, right? And so, it when, when a behavior pharmacologist like myself comes in, you know, I'm drilled in and focused on the phyto cannabinoids and the terpenes. And you know, the terpenes, or there are certain ones that, again, in the cannabis industry, you will see these claims about this variety is better for anxiety. And this variety is better for insomnia because of these terpenes. Right. And there there's a basis for that largely preclinical work, but a little bit then a growing literature of clinical work that kind of backs some of that up. But most of that science is research on those particular entities in isolation. And very little research has gone kind of gone the way of if you go into the dispensary and you say okay, I'm gonna get OG cush for whatever health condition and that's going to be better than The Purple Haze or blue dream, or were any of the other strains of cannabis available at the time. And going back to your point, yes, these different strains can look different, they smell different. But by and large, there's an abundance of chemical overlap in the presence of certain chemical entities and the ratio and actual concentration of these things. So when you go into a dispensary, I estimate probably about 85% of those things. The dominant phyto cannabinoid is going to be delta nine THC. The next one is going to be CBD or CBN, or CBG. But those are going to be much, much, much smaller concentrations, compared to Delta nine THC, we're talking 18 to 24% versus one, maybe 2%, most likely less than 1% on all of this other stuff, right? terpenes, half, one, maybe 2%. So we're talking incredible amounts of overlap in the chemical composition of all of these different THC dominant flowers. And the other thing that many people don't recognize is that even if you were to take one particular strain, right, let's call it blue dream. Blue Dream purchased from a dispensary in Washington, versus blue dream purchase from a dispensary in Maryland where I live might be very chemically different. And it could be because again, the growing conditions, how they were handled these things from a, from a I guess the trying to find what the right word is, but you know, the generational growing and cultivating of these things, these plants change over time. And the chemical expression of the plants based on their genetics can change based on the environment in which they're grown, when they're harvested, and all of that stuff. And then individual bugs very chemically from even the same plant, THC concentration, and above, at the top of the plant closest to the lights or the sun or wherever it's grown, is going to be different from the bugs, the bottom of the plant, the left side of the right, so even if we were to say okay, Blue Dream is X percent THC X percent CBD X percent pinene, limonene, linalool, whatever that's happens to be the chemical signature of the one Bud was tested from that crop, and is not necessarily exactly the same for all of the other bugs from that crop much less than next crop, much less the crop grown across the country 18 generations later. So the naming convention for strains is interesting from a marketing perspective, I think that there's likely some consistency. But it's not precise. And I think there's more overlap the difference, and there's inadequate research to show how much of that is true chemical composition driving pharmacology versus I'm buying a cannabis plant that's, I'm buying it for a reason I intend to have some certain effect. The guy that sold it to me said it's great for this. So I have this whole bias. Yeah, go into it of this is going to do this for me, this other guy say yet definitely going to do that. Yeah, it's more likely to do that, or do something completely different.

Nick Jikomes 18:50

Right. So so there's this expectation effect, mispricing come in based on what you're what you're expecting the product to do, it's reinforced by what people tell you to do, often people with an interest in selling you that effect. Before we get into entourage effects and things like that, you know, another another key variable here is dose dependent effects. So I want to talk about that a little bit. And let's just focus on just THC to start with. So you know, there's, there's a common belief out there that some strains will reliably make you sleepy, and some strains will reliably make you energized, sort of, you know, the upper and the downer version of cannabis strains. How much of that, in principle can be explained by just you know, people self titrating the dose that they're consuming? So for example, if I, if I have, I could take the exact same strain, the exact same THC concentration, the exact same everything I could smoke, or inhale or take in some other ways, a little bit of that. I could also take a larger dose of the exact same product with the exact same chemical composition. Could you drive both types of effects using the same thing just at two different doses?

Ryan Vandrey 19:57

Absolutely. So You know th C's effects are very dose dependent. You know, the, your example in terms of arousal is one key thing. Another one is anxiety, right? low doses of THC can be anxiolytic high doses can be angiogenic, right. And so where you are on the dose response curve is going to drive a very different user experience. The other thing is, is where you are on the the dose curve in terms of the drug composition. So similar to alcohol, you can have a different experience on the ascending limb. So as the drug is increasing in concentration, your body versus when it's decreasing alcohol is a stimulant as you're increasing blood alcohol, and then it's a massive sedative on the descending limb. And so you can get different effects in different depending on how long it's been since you took your dose, in addition to how big a dose you took,

Nick Jikomes 21:03

yeah, I mean, you know, as someone who worked in the cannabis industry for a number of years and studied the chemistry and and the responses people have to this stuff, I would say that a fair and conservative statement is if you know, and we're talking about THC, dominant cannabis here, a significant I'm not gonna say all but a significant proportion of the variation in the effects people report from THC dominant cannabis can be explained by the amount of cannabis you consume on any given occasion. And that's going to be a function of the dose, how much you take, and your history of taking it. In other words, your sensitivity to the THC. But you know, a lot of the effects out there that people map on to different products that are supposedly the sleeping product versus the energizing product. A lot of that, not necessarily all of it, but a lot of it can be explained simply by how much THC they've consumed, and how sensitive they happen to be to it at the time they take it.

Ryan Vandrey 22:00

Yeah, other thing that's, that's often overlooked is the environment that you're in, and the intention of the use experience, right. And so if you take a small dose and then go outside on a bright sunny day, you might feel more invigorated and or energized, and hey, let's go for a run, let's be active. But, you know, if you take your dose and you're curled up in a dark basement on a cow, are you going to want to go for a run, now you're gonna sit down and play Xbox or watch a movie or just be lazy, right. And so a lot of it can be dependent on that, you know, the psychedelic world is notorious for talking about the importance of set and setting in a drug experience. But I think the same can be applied to cannabis as well, I think it very much is important. And we've seen this across drug classes, not just cannabis and psychedelics, but, you know, the the environment, the intent, whether you're in a social situation versus using on your own, all of those things can impact the overall user experience, you know, are you at a party? Or are you hanging out with your grandparents, right, you know, your response and how you behaviorally, function and feel is going to be impacted by all of those things.

Nick Jikomes 23:25

So even though we can explain a lot of the variance in effects to THC, dominant cannabis products, with the dose of the THC, the sensitivity and the use history of that person with THC, and the set and setting the contextual factors in which people consume it, there are still a lot of very interesting molecules in these products, many of them have pharmacological properties that would suggest the possibility that they're contributing to the psychoactive and or medicinal effects. And many people certainly make those claims there's a lot of there's so many anecdotes out there about this, that you can't really ignore them. The idea here is often referred to as the entourage effect. It's the idea that all of or many of the cannabinoids terpenes and or other things in the plant, interact with each other. Many of them are pharmacologically active at different sites, different receptors, things like this. And it's really the constellation of these things that you consume, together with the other factors like dose and context that are going to dictate what the experience is like or what the medical effects might be. Can you summarize what the entourage effect is conceptually, in your own words, and sort of just start to give us a perspective on how much evidence has existed for this historically, before we go into some of your more recent work? Yeah,

Ryan Vandrey 24:46

so the entourage effect has been around for a long time and this goes back to research done in the 70s. I think he but then maybe even before late 60s 70s, definitely by the early 80s where, you know, people would do research studies with a pure THC, and then like a cannabis extract, or a hashish or something like that. And they would see, you know, unexpected or uniquely different effects in one study versus another. And there have been some studies that have directly compared these things, but very, very few. And so again, the idea behind the entourage effect, and the importance of it to the cannabis industry really emerged in the late 90s and early 2000s, when people started talking about making the push for cannabis legalization for medicinal purposes at the state level, there had to be an argument that cannabis as a whole plant entity was uniquely different, was perhaps in some cases better and safer, and more applicable for medicinal use or therapeutic benefit, compared with just THC, which is FDA approved in the form of dramatical. Right, so there is a pharmaceutical formulation of THC that is approved by the FDA that's legal and can be prescribed. And so there was a big push to really emphasize that it's

Nick Jikomes 26:26

essential. There's there's not so conceptually, there's not just sort of the scientific idea about the interaction of chemicals. There was also a specific policy motivation for searing like this being true. Yeah,

Ryan Vandrey 26:38

very big policy motivation. And again, when you look at the science, there's evidence to support the entourage effect or certain hypotheses there. And there's also evidence to refute it. And I, as we talked about the beginning, Nuance becomes important, right? So a lot of the interactions and the differences between strains of cannabis or cannabis combinations are whole plant cannabis versus just THC, by itself are going to be driven by you know, what relative concentrations are we talking about what else is in the plant and how it's taken. So one really important one, a good example of this is CBD, right? And so right now, the common theory is that high CBD strains or high CBD containing plants or cannabis products are safer and less potent than high a THC with very little or no CBD products. And a lot of that was driven by flour that came out where CBD was overexpressed, you know, just over the last 1015 or so years, right? And while I think the data show that that's the case, when it's inhaled, in most cases, not all cases, but where we have to kind of look at the the nuance is, you know, is CBD actually attenuating THC as effects in that situation? Or is, you know, when you overexpress CBD in a plant, are you over or under expressing something else? You know, the THC concentration in a one to one flower is much lower. It's usually around eight, nine 10% of each.

Nick Jikomes 28:39

So there's basically there's a confound, if you've got higher CBD, you also have lower THC. So it's lower THC

Ryan Vandrey 28:46

and you might have higher or lower other things that are very important pharmacologically that we just don't understand yet. Though, flip of that, is that I Well, before I go on the flip, the important thing there too, is that there have been a number of studies that have specifically gone after that does CBD attenuate the effects of THC? And for every study that says yes, there's a study that says no, and there's probably two that say, it doesn't really matter. So it's, it's in between, right. So it's a very mixed bag. And when you so why is there a mixed bag, let's drill down on the research methods. So in many cases, the approach was to pre dose individuals with oral CBD, and then have them inhale THC.

Nick Jikomes 29:38

And that's usually with a very high dose of CBD, right, different

Ryan Vandrey 29:40

routes of administration here, the timing of the onset of the drugs is going to be different. doesn't fully represent how it's used in the real world, right. So there's there's methodological differences across these studies that likely explain why some of them say yes, it attenuates some some It doesn't. And then you get some that say not only does it not attenuate THC effects, some say it exacerbates it. And that route of administration comes in. So the flip of this is that when it's inhaled, we see more evidence of entourage type effects where CBD reduces or you see less strong THC effects. But on the oral dose side, there's more research and a couple of studies from my lab in particular that have come out recently where we say simultaneous oral dosing of a high CBD product, if you match the THC dose to get a high THC and a high CBD and THC dose is exactly the same same route, same everything we see an exacerbation of the THC effects, and it's through a metabolic interaction between CBD and THC. Yeah,

Nick Jikomes 30:54

that's what I was gonna ask you about. So like, in principle, you know, in theory, I could explain this, you know, a couple different ways. One, you know, if you think about concurrent administration through through the inhaled route of THC and CBD, and we know pharmacologically THC is activating CB one receptors. CBD is not doing that it's probably actually interfering with the ability of something like THC to activate the CB one receptor. Now you've got apply a lot plausible biological means by which CBD could interfere with THC and potentially mitigate it side effects. On the other hand, I think there's there's something interesting metabolically going on with respect to how some of these molecules affect liver metabolism. Can you talk about that a little bit? Yeah, so

Ryan Vandrey 31:36

CBD has been shown to be a pretty potent inhibitor of drug metabolism, and specifically the two substrates that are used to metabolize THC. And so in one of our recent studies, again, identical doses of THC given the same way to the same people in the same study, just on two different occasions, when we had high doses of CBD given with a pretty high dose of THC, 20 milligrams of THC. When we looked at blood levels of THC, and these individuals over crossed the course of the study, when the same dose of THC was given with a high dose of CBD, we saw double the THC blood levels, and again, times the 11 hydroxy metabolite THC blood level. So THC is metabolized two times the first time the parent molecule is metabolized to another psychoactive substance, 11 hydroxy THC. And again, so that was increased tenfold, at peak levels, about a substantially different drug effect based on just this metabolic interaction.

Nick Jikomes 32:49

I see. So so the idea there is if you take these things orally, CBD is affecting enzymes in the liver responsible for the metabolism of THC. And so you're actually getting higher concentrate,

Ryan Vandrey 33:02

you're reducing your body's ability to eliminate THC. So therefore your drug effects are stronger and longer lasting than if you take it without CBD. Again, coming back to one of our other things that we talked about earlier, this interaction is entirely dependent on dose and concentration in your brain and your blood and all of that stuff too. Right. And so, you know, we saw this really strong interaction. But this was in a study with pretty high doses of both drugs. And we gave people some other drugs on top of all of that to probe which specific enzymes C. CBD was in or interrupting. But we've seen it in other studies as well as have other scientists. So we know CBD inhibits THC metabolism through oral route to the administration. Now some colleagues of ours that at the University of Washington in Washington state have done kind of preclinical modeling of this interaction. And their science suggests that this is unique to oral dose, and that you wouldn't see the same effects within elation. So, again, we need to come back we need more research to find out, okay. across different routes of administration, what's the level of pharmacological interaction and metabolic interaction between these drugs? And at what dose concentration does this happen? And at what ratio of CBD to THC does the ratio of factor into the total doses factor in and if so, in what way? And once we do a few more studies like that, I think we can use statistics and modeling to really kind of fill in the blanks on exactly how this is going to happen to predict the outcome of a particular product being used by a particular individual.

Nick Jikomes 34:58

So why exactly what do we see a different such a different effect from oral versus inhaled routes of administration, is it because when you consume these things orally, they go through the portal vein and a higher proportion is metabolized by the liver before entering systemic circulation,

Ryan Vandrey 35:16

it has to do with the rate of absorption and the rate of elimination. So when you inhale a drug, you're getting your lungs are very efficient drug delivery mechanisms, you basically get almost all of the drug in to your blood, within minutes, you know, it gets absorbed very, very fast. And so you're getting this peak drug effects right away, your peak concentrations happen almost immediately, and then your body works very quickly to eliminate. And so that's why with inhalation, the peak drug effects happen really fast. And then you get a pretty linear decline and drug effects. If it lasts, you know, an hour, two hours, maybe three hours. When you take THC orally, you don't feel anything for usually the first 3045 hour, right, and then the peak drug effects are sustained for a longer period of time. And there's a slower elimination is because there's a slower absorption of the drug by your gut into your blood. There's metabolism that happens in your gut, and as well as after it's absorbed. And so it's a very different level, your body's continually absorbing the drug, in different states of metabolism and breakdown, and for a longer period of time. And so when you have that inhibition happening on something that's happening slower, you can get these bigger, larger differences in terms of impact of liver and design functioning.

Nick Jikomes 36:53

So, you know, when we start to think about these things in combination, you know, we've already seen from, from what you've told us, like it gets very complicated very quickly, the dose matters. Probably the relative ratio of compounds matters, the way you consume, it matters. There's a whole other class of compounds that we're going to talk a little bit about, that we haven't even started to talk about yet. And that's the terpenes. So can you say a little bit more about, you know, what are terpenes? And what are some of the more prominent cannabis terpenes. And in the literature historically, you know, do any of them have pharmacological properties that would make us think or hypothesize that they could be contributing to the psycho activity of cannabis?

Ryan Vandrey 37:31

Sure. And before I go there, just one more thing to add on that last piece is that the difference in the time force and the rate of the impact on the metabolic stuff, the other factor is that the viscosity of cannabinoids are lipophilic. And when you orally ingested, it gets it sticks to tissue, including your brain. So we find, interestingly enough, when you orally adjust cannabis, even at the peak level highest drug effect, they're very low concentrations of the drug in your blood. And so it's not really circulating, it's a much slower elimination because of that. So these things get in and get into your brain and just stay there they camp. And so it's a little bit different there as well. Yeah,

Nick Jikomes 38:21

I mean, what just one more thing, before we move on, that might be worth adding there is you I very often hear people say things like, well, I took 10 milligrams in the form of a cookie. And then I took 10 milligrams in the form of a gummy or a brownie. And it was a very different effect. And presumably, that has to do with the fat composition of the substrate that these fat soluble molecules are in, which would affect the rate of absorption.

Ryan Vandrey 38:44

Yeah, yeah, for certain. And, you know, we've seen that we've heard that now, to add another layer of complexity. On top of that you have the matrix the drug is in is going to drive, how quickly it gets absorbed through the gut, how much is going to get absorbed versus gets metabolized right then and there. What the person ate and how long ago they ate, it also impacts it considerably. So if you take a cannabinoid on an empty stomach, you're gonna get far less drug, see the opposite of alcohol and many other medications, you get far less strict abortion than if you take it with a meal, and in particular, a high fat meal. So high fat meals actually protect the drug and get it through your body faster than if you were to take it on an empty stomach or with a low fat meal. And then there's other impacts there too, where there you know, there are there's nanoemulsion technology to try to speed up and protect the drug. So there's a lot of innovation in the cannabis industry right now in this space, again, without much published science. So my colleague Tory spindle here at Hopkins, just got a really nice grant from the At the National Institutes of Health, to study this very thing. So we're going to be he's going to be doing a research study starting up the summer, where we're going to be giving people you know, the same high THC distillate extract, in a gummy in a beverage and in a brownie, little look at fat content or macro nutrient content as well as the use of nanoemulsion technology versus not. So it'll be at nanomoles beverage versus just a straight distillate in a gummy and the distillate in a high fat brownie formulation. So I'll be fascinating. Yeah,

Nick Jikomes 40:39

that sounds Yeah, no, no one that I'm aware of has done that type of study in a nice head to head way so far, it's it's pretty clear, there's going to be differences, but what those differences are, and knowing that with precision, I don't think we have that information yet.

Ryan Vandrey 40:53

Yeah. So again, important, because when you go back and you talk about regulatory decision making, if you if if a state decides their unit dose, or max unit doses, 10 milligrams or five milligrams, is it appropriate to have that be the same across formulations, or edible subtypes? Or does there need to be some kind of labeling difference, you know, fast acting, will hit you harder than this other thing, right. So, you know, we need that, what and whether or not it impacts regulation at all, it's important from a public education perspective. So the user knows, going into this. And there are folks that are savvy to this already, they've experienced that they, they know it, and they adjust their doses and what they buy based on that. But as legalization, especially for non medical purposes, is rolling out widely, many people don't. And so that's the kind of thing that we need to worry about, we need to get the message out there, we need good data behind it. And then we need some kind of, you know, packaging, labeling, the warnings, so to speak, or at least something that conveys that kind of nuance to the to the user. So you don't end up with the people going in and saying, Well, five milligrams was fine in this shouldn't be fine with five at that. And then you go past the point of comfort, and instead of a happy, relaxed time, you're curled up in a ball on a couch scared and paranoid and wondering what did I do the sky is falling, right? So in, THC can go very quickly from one of those to the other, we're trying to minimize

Nick Jikomes 42:47

switching gears to terpenes. So again, remind people what these things are, give us maybe two or three or four examples of the more prominent ones that typically are found in cannabis. And historically, you know, are any of these things known to affect brain receptors and other aspects of physiology, that would make us think that they could be relevant to the psychoactivity?

Ryan Vandrey 43:08

Yeah, so you know, there again, there's probably more than 100 Different terpenes in the cannabis plant. And they're going to be at different levels, depending on the the genetics of the plant, how it was grown, and when it was harvested, and all of that stuff. But, you know, when you look at the science behind these particular terpenes, you look at the relative concentrations that they exist naturally in the plant, or at least exist in current retail flour products. The heavy hitters are limonene, pinene, linalyl, beta caryophyllene, those kinds of things, there's probably a list of about 20 or so that people think are important in some way, shape, or form. The ones that we've been interested in and studying in the lab here are limonene, pinene, and mercy. And we got interested in that based on published papers by Ethan Russo and a couple other people that kind of pulled together and triangulated preclinical data and some clinical data of these things and other formats, not necessarily in cannabis and some cannabis industry stuff and anecdote. You know, these are the most among the more abundant terpenes that are found in cannabis plants and they're the ones where we see when we look at, you know, a cannabis dispensary profile or a marketing page or whatever. These are the ones that people are calling out saying this one or this particular strain has high amounts of this therefore it does that this person ticular terpene is associated with this kind of effect. And so for limonene, a lot of that is it helps with anxiety, it helps increase arousal with does this. And I will say this, I've seen probably 10 different claims about what limonene does in different areas of different media here. So it's still one of those cases. And this is, I think, the case across the board with cannabis broadly, there are lots of claims, but there's not a lot of data. It's hard to refute claims in the absence data. So, you know, limonene the big thing is anxiety and some arousal. So it's a citrus terpene found in you know, lemons, limes, oranges, grapefruits, all of that kind of stuff. It's if you walk down the cleaning aisle at your grocery store, lemon pledge, that's limonene you know, so it's, it's there, it's abundant. It's in other food products, it's in other household products. So it's everywhere. It's lemon scent. Right and it's an essential oils that people already are using homeopathic Lee in their homes, you know, you get a diffuser, you put the essential oil in there. A lot of them have lime and laminate have piney have little lavender scent, right? So these things are, there's a whole history of human use of these substances, various forms for health and wellness promotion. They also happen to be in the cannabis plant. So there's a fusion of these claims and feelings and beliefs around them. What we don't know is okay, say one cannabis plant has point 7% limonene in it. Another has barely detectable levels, and one has 2% limonene. Is there a functional difference between those three plant cultivars simply based on the concentration of limonene? Who knows? Right? And and even if limonene of those varying concentrations, if the dose that you take is the same? What about the variation in all of the other chemicals? Can those trump the impact of climate? We have no idea. Right? And so part of the research we do here is to try to understand that how how important are these individual chemical entities? Are they strong enough to actually impact the effects of THC? And we're doing that because THC is largely the by far dominant chemical entity in cannabis products. And it is the chemical entity that is driving most of the effects that people associate and seat when they use cannabis products. What is normal or non medicinal purposes? Right? And so, you know, our research is really saying, okay, are these interactions meaningful? If so, at what doses I want relative concentrations, and what is actually going to happen?

Nick Jikomes 48:23

So that starts to set us up for this recent study that you did with vaporized THC and limonene. Can Can you set that up for us a little bit more? What were you guys setting out to do? And what was the basic setup of this study?

Ryan Vandrey 48:34

Sure. So again, this comes back to largely theoretical work that Dr. Russo had done, and others have done but and written about and that the industry has latched on to and made claims about. But again, you know, this, this all started, where Ethan called me up and said, Brian, you know, I wrote this paper was published in the British Journal of pharmacology and, gosh, I don't even remember how 2013 It's been six years, and no one has done the science to support it or refute it to back it up. And so I said, that's crazy. Why don't we do it that way. And so it set us out on on this line of he lays out very many very specific hypotheses about the entourage effect and about how certain terpenes interact with THC to produce the entourage effect. And he came in as the believer, the one who is, you know, promoting this idea. And I came in at the time of being the scientific skeptic. I was like, There's no way right, but I liked that challenge. I liked that interaction. And I think that Ethan the mind complemented each other very well in approaching this. And you know, the advantage of of me joining forces with him is that we have a lab set up that we could actually go in and test these things in a very controlled manner. So we felt confident in in our approach to it methodologically, and we had the capabilities to do it. So we said about finding funding, we looked a lot of different places, it took us a couple of years to get it going. But like I said, we landed with a grant from the National Institute on Drug Abuse. And the idea was to take three different terpenes, limonene, pinene, and Mersing. And to test individual hypothesized effects of how these things would interact with each other. Going into it, based on preclinical data, as well as some clinical studies with humans of limonene. Not as part of cannabis, but as an individual entity, that limonene would attenuate the anxiety inducing effects of THC. And that's based on data from again, citrus oils being used homeopathic Lee to lift mood and there was a hospital study that showed it reduced anxiety and mood disorders wing of the hospital when it was sent out into the air. And it follows the claims of a lot of the cannabis products and marketing. So can limonene reduce the anxiety inducing effects of THC pinene was selected because the hypothesis there is that pining could reduce the impairment of working memory induced by THC. And myrcene was believed to not attenuate THC, but to exacerbate the sedating effects of THC. And so when you look biochemically at all of the claims about indica and sativa 's and things like that, the the hypothesis right now in in terms of Dr. Russo's beliefs in science and research, and in the cannabis industry claims is that myrcene is sedating. And we'll be associated with that couch lock phenomena of Lincoln indica type of cannabis product. So we have three individual terpenes that are available, they're generally recognized as safe, although that doesn't really apply to inflation. So on the regulatory side, we had hurdles to jump into clear to get things going. But you know, we had three very clear distinct hypotheses that could be tested in the laboratory by so we got we went limonene pinene myrcene. In order a limonene study has done and was recently published, the pinene study finished about two weeks ago. And we're just getting it analyzing the data. And myrcene study is just getting going. So we're kind of in the middle of all of that at the moment. So I'm happy to talk about the results of the limonene study. And we can talk about general generalities for pining and immersing, but I have no data to share just yet on that. Yeah,

Nick Jikomes 53:29

let's just let's focus on the one that's done with limonene. So I've got it right here, I've looked at it. So the hypothesis is that for escalating doses of THC, you're gonna have especially the higher doses, a side effect of anxiety. So anxiety will be more common at a higher dose of THC. And the hypothesis with respect to eliminate is that if you add limiting into the mix, at some level, potentially, it will mitigate the anxiety provoking effects of THC. So you should see less anxiety and people who inhale THC plus limonene than you do in people who inhale THC only, or THC with less limonene. So what was the setup of the study? How many people what type of people and how did you actually deliver this stuff to them?

Ryan Vandrey 54:16

Sure. So to kind of go back and talk about our methods. So one of the things that I think is really important that that I think we do well is we control dosing. And again, we were very intentional along not just going and getting a high limonene whole plant cannabis product versus a low limonene cannabis product comparing the two because again, as we talked before, there could be a multitude of other chemical differences between those two plants. It could be driving any difference we see. So

Nick Jikomes 54:49

you really you really want me to isolate THC and alignment and you want

Ryan Vandrey 54:53

to be very specific does limonene actually do anything to THC so we purchase Synthetic THC we purchased, plant derived but purified, highly purified limonene. So both were greater than 99% purity. We then modeled a dose delivery method that we validated that if we heated up a THC at a certain temperature in a certain device, we would deliver all of that THC. And the same thing with the limonene. And we made sure that when we heated it at this temperature, this device, we didn't convert these things into something else. So we were very careful to get our dosing methods secure, and to work with this. So we opted for inhalation and vaporization in particular. So we know that our doses are being delivered fully and completely. And we're not converting things to other other chemical entities through this process. And then we had to figure out what doses do we see now we've done a dozen dozens of studies with an ITC cannabis here in the lab, we know pretty much how many milligrams of THC it takes to get people pretty high and very high. And so we kind of did that we picked two doses of THC, 15 milligrams and 30 milligrams, which to the occasional cannabis user will get people moderately high at pretty dang high. And in we wanted to do that because we weren't sure we didn't want to Sledgehammer everybody and give limonene and conclude that, ah, well, it doesn't do anything because we completely destroyed with THC. So we wanted variation in there, you know, can limonene overcome a little bit of anxiety? Or can it overcome quite a bit of anxiety and some of those, see, we're

Nick Jikomes 57:02

here to help orient people here. So you're gonna use 15, or 30 milligrams of THC, approximately how many milligrams would be in a typical joint that you would buy at a recreational marijuana shop.

Ryan Vandrey 57:14

So again, you got to do some math there. If you're taking something that's like, say 20% THC, and you buy a pre roll joint, pre rolled joints are usually about a gram. So that whole joint is going to have about 200 milligrams of THC in it. Now, few people will go in and buy a one gram pre roll joint that smoked the whole thing by themselves. Or if

Nick Jikomes 57:40

they do that they'll probably have frequent consumer who has decreased their sensitivity,

Ryan Vandrey 57:44

not our population. So we want to eliminate high levels of tolerance in this experiment, because people with a high level of tolerance, it can give them a high dose of THC, they tend not to get too anxious. So we recruited people into this study who were healthy adults, had prior experience inhaling cannabis, high THC cannabis, and at least at some point in their past history, had inhaled THC and gotten anxious, apparently, right, we wanted to maximize the likelihood that we would see this effect, so that we can really tell if limonene could reverse it or attenuate it. And so, again, that's just good science, I think. And so, so we had our target THC doses based on prior studies that we had run. And then we had to figure out what doses of limonene we could use. So I mentioned before, there had been some research on limonene exposure in ambient air. But we couldn't find anything published anywhere in any way, shape, or form of anybody directly inhaling limonene. So we were kind of pushing the envelope there and had to justify ecologically relevant doses of limonene that we've believed would be safe to these individuals. We never want to hurt people in our experiments, even though we're talking about inducing a little bit of anxiety. So what we did is we