Groundbreaking COVID-19 Study Shows CBD May Help Inhibit Infection

Groundbreaking COVID-19 Study Shows CBD May Help Inhibit Infection March 30, 2021 Madeline Colli Recently, researchers performed a study which discovered that a cannabis plant compound inhibited COVID-19 infection in human lung cells. Researchers in the US recently conducted a study which disclosed that a cannabis plant compound inhibited infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in human lung cells. SARS-CoV-2 is the virus that causes COVID-19, threatening global health and world economies (1,2). Marsha Rosner, PhD (3), and other colleagues from the University of Chicago (Chicago, Illinois) discovered that cannabidiol (CBD) and its metabolite 7-OH-CBD potently blocked SARS-CoV-2 replication in lung epithelial cells. Through the combination of CBD and 7-OH-CBD, the expression of certain genes within the viral cells were inhibited and changes in gene expression within the lung cells that resulted from the presence of COVID-19 were reversed (4). Thus, CBD and 7-OH-CBD serve in both protective and therapeutic roles. CBD was also found to have the ability to block viral ribonucleic acid (RNA), including the coding for the spike protein, which is the tool the virus uses to enter our cells. ―This study highlights CBD, and its active metabolite, 7-OH-CBD, as potential preventative agents and therapeutic treatments for SARS- CoV-2 at early stages of infection,‖ said Rosner and the team (1). Even with recently approved vaccines being rolled out in many countries, the virus continues to spread rapidly, heightened by more-

transmissible variants such as the B.1.1.7 variant. Rosner and her colleagues say that this highlights the need for alternative approaches, especially among populations with limited access to vaccines. Few therapies to date have been identified as being able to block SARS- CoV-2 replication and viral reproduction. Primarily, the SARS-CoV-2 virus enters host cells through the binding of a surface viral protein—called spike protein— to the human host cell receptor angiotensin-converting enzyme 2 (ACE2). From there, the viral genome is then translated into two large polypeptides that are severed by the viral proteases main protease (MPro) and papain-like protease (PLPro) to produce the proteins necessary for viral replication, assembly, and budding (1). To analyze the effect of CBD on SARS-CoV-2 replication, the researchers pretreated A549 human lung carcinoma cells expressing ACE-2 (A549-ACE2) with 0-10 μM CBD for 2 hours before infecting them with SARS-CoV-2. Evaluation of the cells 48 hours later expressed CBD had potently inhibited viral replication in the cells. When CBD was assessed to possibly prevent proteolytic cleavage by MPro and PLPro, it was observed that CBD had no effect on the activity of either protease. This revelation led the team to hypothesize that CBD targets host cell processes. Consistent with their hypothesis, RNA sequencing of infected A549-ACE2 cells treated with CBD for 24 hours showed significant suppression of SARS-CoV-2-induced changes in gene expression. CBD was shown to have effectively eliminated viral RNA expression, which included RNA coding for the spike protein. It was also found that both SARS-CoV-2 and CBD triggered significant changes in cellular gene expression, such as the expression of several transcription factors. Further analysis of host cell RNA expressed that the virus-induced changes were almost completely reversed, though rather than the cells returning to a

normal cell state, the CBD plus virus-infected cells resembled those treated with CBD alone. Another critical finding researchers discovered was that CBD had ―effectively reversed‖ the triggering of a hyperinflammatory response, also known as a ―cytokine storm‖ which is brought on by the presence of the virus, restoring cells not to a previral level of inflation, but a state as if the cells had been treated with only CBD. A study completed near the end of 2020 by the Dental College of Georgia (DCG) (Augusta, Georgia) and the Medical College of Georgia (Augusta, Georgia) also had similar findings to the team from the University of Chicago (5). Cytokine storms have been one of the principal causes of death resulting from a COVID-19 infection. CBD was identified in reducing damage in the lungs spurred by cytokine storms caused from COVID-19 by normalizing levels of apelin, a peptide known to reduce inflammation, decreasing physical lung damage associated with adult respiratory distress syndrome (ARDS), and improving oxygen levels. During a COVID-19 infection, these apelin levels were seen to be at very low levels. Rosner and her team stated that ―CBD has the potential not only to act as an antiviral agent at early stages of infection but also to protect the host against an overactive immune system at later stages,‖ (1). During the University of Chicago’s study, a range if other cannabinoids were also tested, but by the end of the trial, CBD was the only cannabinoid found to have any effect at all on COVID-19-infected cells. Limited-to-no antiviral activity was exhibited by the other cannabinoids investigated. CBD is often consumed as part of a Cannabis Sativa extract, which is also what led the team to consider investigating other cannabinoids with closely related structures to see if they would reveal similar or stronger results. The team hopes that CBD will continue to be investigated as a potential COVID-19 treatment following the research from their article about the study,

which is currently under peer-review. ―We advocate carefully designed placebo-controlled clinical trials with known concentrations and highly-characterized formulations in order to define CBD’s role in preventing and treating early SARS-CoV-2 infection,‖ the authors stated (4). Even with these impressive scientific breakthroughs, more is still needed to be researched on the medicinal properties of the cannabis plant and how it can be used to fight the global coronavirus pandemic. References 1. https://www.news-medical.net/news/20210314/Cannabis- compound-inhibits-SARS-CoV-2-replication-in-human-lung- cells.aspx 2. https://www.cancer.gov/publications/dictionaries/cancer- terms/def/sars-cov-2 3. https://benmay.uchicago.edu/faculty/marsha-rosner-phd 4. https://www.goodnewsnetwork.org/cbd-was-found-to-prevent- viral-rna-expression-and-reverse-gene-changes-in-infected- lung-cells/ 5. https://www.cannabissciencetech.com/view/covid-19-lung- inflammation-reduced-by-cbd Cannabis Science and Technology® and Nutritional Outlook® Launch Expert-Driven Webcast Series on CBD Quality Control, Testing, and More March 10, 2021 Madeline Colli This unique series will take place during 2021, leveraging the expertise of these industry-leading publications and delivering insight and best

practices on critical issues such as cultivation, extraction, processing, testing, third-party certifications, and much more. Quality control is a crucial problem in the hemp cannabidiol (CBD) market. The quality gap is widening as more products enter the space. Government regulators, industry critics, and responsible market players understand many products in the market are failing to meet label claims. As competition grows in the CBD market, and as industry scrutiny increases, responsible companies must stand out by ensuring they employ industry best practices for putting credible, high-quality products on the shelf. The first webcast, ―Controlling THC Content,‖ kicks off on Tuesday, March 16, 2021, at 3 p.m. EDT/2 p.m. CDT/12 p.m. PDT. During this session, hemp cultivation experts will explore the best practices and methods for controlling tetrahydrocannabinol (THC) content. Experts will also provide an update on the legal landscape and challenges facing the industry. CBD: The Road to Quality webcast series programs and dates are as follows:  Chapter 2: “Extraction and Processing Excellence” (May 2021).  Chapter 3: “Testing: Methods, Requirements, and Best Practices” (July 2021).  Chapter 4: “How to Choose the Right Testing Lab” (September 2021).  Chapter 5: “Meeting Label Claims and the Role of Certification” (November 2021). Details on upcoming sessions will be announced in the next few months. ―Delivering direct access to trusted scientific news that is relevant and practical is key to our mission at MJH,‖ said Mike Hennessy Jr.,

president and CEO of MJH Life Sciences™, the parent company of Cannabis Science and Technology® and Nutritional Outlook®. ―Together, with the expertise of Cannabis Science and Technology® and Nutritional Outlook®, we can provide those making and selling CBD products with the tools they need to ensure the quality of their products. Join us as we tap into the expertise of the CBD industry’s leading players and advisers.‖ For more information and to register, click here. About Cannabis Science and Technology® Cannabis Science and Technology® is a multimedia platform that focuses on educating the legal cannabis industry about the science and technology of analytical testing, quality control, extraction and cultivation. The magazine is accompanied by an online component that provides relevant information and tutorials for all members of the cannabis industry. About Nutritional Outlook® Nutritional Outlook® is the industry’s No. 1 resource for news and insights on dietary supplements, healthy foods and beverages. An award-winning business-to-business resource, Nutritional Outlook® provides manufacturers with the nutrition and healthy ingredient news they need to get their business done right. Contacts Cannabis Science and Technology® Media Alexandra Ventura, 609-716-7777 [email protected]

A Market Overview: CBD Topicals December 3, 2020 Alissa Marrapodi Cannabis Science and Technology, January/February 2021, Pages: 52-54 CBD is a growing industry. In this article, topicals and their growing popularity are discussed. The market for cannabidiol (CBD) products is ever growing. And within this market are topicals—one of the hottest product formats. The Brightfield Group reported, in 2019, U.S. sales of CBD topicals exceeded $703 million, second only to CBD tinctures. By 2025, topicals are projected to reach $4.5 billion. During the summer of 2020, CWI Consulting Services—a consumer packaged goods (CPG) planning and development consultancy that focuses on CBD products for the mainstream market—conducted research and gathered data on the topicals CBD market. It surveyed 360 hemp-based topical CBD products from 112 brands. The top 53 brands were identified as market leaders by the Brightfield Group— this group included every company that sold more than $350,000 in topical CBD products in 2019. What’s interesting is sales from the top 53 brands comprised almost 60% of all sales in 2019 in the CBD sector. Furthermore, the top 10 brands—American Shaman, CBD Clinic, CBD FX, CBDMD, Charlotte’s Web, Green Roads, Medterra, PlusCBD, Reliva, and Sagely Naturals—comprised only 36% of sales in 2019. What’s driving the topicals market? According to CWI, the top-two reasons consumers purchase CBD topical products are pain relief and skin health.

Let’s take a look at some of the key highlights from CWI’s report, CBD Topicals in the USA, including various formulations, ingredients, concentrations, a little on pricing, and a few notes on claims and other regulatory matters. (1) How Do Users Want CBD Topicals? When it comes to topicals, the sky is the limit—CBD has made its way into almost every personal care topical format—from lotions to face masks. Lotions and creams largely outweighed all other topical formats, accounting for 43% of all formats. Next was balms, making up 26% of the SKUs. Together, these two categories made up 69% of the topical products reviewed. Other major formats were sticks (7.5%), roll-ons (6%), oils (4%), gels (3.6%), and patches (3%), making up a total of 24.1%. Curious about size? The most common size for CBD topicals is 2 oz. Next in line are 1 oz offerings. But sizes do vary widely. ―The relatively small package sizes suggest that, because CBD products are more expensive than similar non-CBD topicals, and many consumers have yet to try them, brands are balancing quantity with price in order to encourage customers to try these products,‖ according to CWI. What’s in CBD Topicals? The survey results revealed three main types of CBD ingredients used in CPG products: full-spectrum CBD oil, broad-spectrum CBD oil, and isolate, which is 99% or more pure CBD crystals. The survey defined each as the following:  Full-spectrum CBD oil is a sticky oil that has been extracted from hemp plants, usually using supercritical CO2. It typically contains 70% or more CBD, along with other cannabinoids such

as tetrahydrocannabinol (THC) and cannabigerol (CBG), as well as terpenes and other natural constituents of the hemp plant.  Broad-spectrum CBD oil is full-spectrum oil that has been further processed to have the THC removed to below detectable levels. Though THC can be an important component of cannabis medicines, helping CBD do its job, many retailers and consumers are wary of THC and prefer CBD products without it.  Isolate is 99% or more pure CBD crystals, created by further purification of CBD oils. Isolate is easiest to formulate with, as its CBD content is consistent, and it does not impart a scent or flavor. However, since it does not include other hemp constituents, consumers seeking a more natural, complete “entourage effect” consider isolate less appealing. It appears as a variety of names such as pure CBD, ISO99 CBD, and pharmaceutical-grade CBD. Of the three ingredients, full spectrum was the most commonly used CBD ingredient, featured in 35% of all SKUs. Broad spectrum came in second at 21% and isolate at 17%. The fourth most common declared CBD ingredient was hemp extract at 12%. However, CWI said this ingredient suffers from nebulousness and isn’t very helpful to consumers. Finally, ―other‖ made up the remaining 15% of SKUs with unspecified CBD sources. These percentages changed, however, when CWI only looked at the top 10 CBD topical brands. Broad spectrum was the most common, found in 49% of all the products reviewed, followed by full-spectrum at 26%, and isolate at 11%. ―This observation, in which the top-selling brands prefer broad- spectrum CBD as an input, points to a wariness of the pharmaceutical- like isolate, as well as the THC-containing full spectrum,‖ the CWI report stated. ―This suggests that these brands have found a comfortable middle ground in broad spectrum between the other two major alternatives, each of which has its own marketplace drawbacks.‖

Of course, CBD topical products are formulated with other ingredients. And, overall, topical formulations lean heavily on natural ingredients. The top non-CBD ingredients used are coconut oil, water, and menthol. The top essential oils used are peppermint, eucalyptus, and lavender. And the top base oils are coconut, shea, and jojoba. Only 25% of the SKUs used the word organic but none were certified organic. How Much Do CBD Topicals Cost? CWI found even though the wholesale cost of CBD has gone down, due to changes in regulations and increased production, the prices of CBD topicals have not. Why? According to CWI, because formulations have gotten stronger. However, 80% of CBD topical products contain less than 500 mg of CBD, with 26% below 100 mg—the lowest was 5 mg; the highest was 3000 mg. Note: 10% of the SKUs surveyed did not specify CBD content and were excluded from this analysis. In terms of concentration, the most common amount was 5 mg/mL (11%). Most (60%) concentrations were between 1 mg/mL and 7 mg/mL. The mean concentration of CBD in topicals was 9.6 mg/mL and 6.42 mg/mL in the top 10 brands. Two-thirds of the products surveyed cost between $10-50. Among the top 10 brands, prices tend to be lower. According to CWI’s data, 28% were priced between $10-20, while 25% were between $20-30. A total of 77% were priced between $10-50. What’s interesting about CWI’s findings is that products made with CBD isolate tended to be more expensive than full- or broad-spectrum oils, which is odd given that isolate is the lowest priced of the three ingredients and also the easiest to formulate with. Say What?

As is well-known, product claims, and more specifically structure- function claims, are highly regulated by the U.S. Food and Drug Administration (FDA). Products that claim to diagnose, cure, mitigate, treat, or prevent any condition, ailment, or disease are considered a drug. Naturally, these regulations also apply to CBD topical products. Regardless of regulations, CWI’s research found brands use four main ways to communicate CBD’s ability to reduce pain and inflammation: 1. Ignore the rules and take a chance—make an ―inflammation‖ or ―arthritis‖ claim 2. Make careful, nonspecific claims about products being ―soothing‖ or ―revitalizing‖ 3. Formulate with ingredients that are permitted to make claims. For example, lidocaine (1%) can use the claim ―pain relief‖ or ―fights pain,‖ so formulating with it allows for a pain claim to be made, as long as it’s within the guidelines of over-the-counter (OTC) drug rules. 4. Use claims that are not formally defined such as ―recovery‖ and ―anti-aging‖ ―The only way for brands to mitigate regulatory risk related to claims is to have all packaging and communications reviewed by qualified compliance professionals who are well-versed in FDA regulations,‖ according to CWI. ―It is clear that far too many CBD topicals brands are skipping this crucial step in product development.‖ Conclusion It’s clear the CBD market overall has a lot going on and more oversight and regulations are needed. Until those rules and governing bodies come in to play, watchdog reports such as CWI’s are here to

help the public learn more about the products they are taking. To view the full report, click here. Reference: 1. CWI Consulting Services. ―CBD Topicals in the USA: Results and Analysis from a Survey of 100+ Brands‖ 2020. About the Author ALISSA MARRAPODI is a custom content writer for Cannabis Science and Technology. Direct correspondence to: [email protected] How to Cite this Article A. Marrapodi, Cannabis Science and Technology 4(1), 52-54 (2021). COVID-19 Lung Inflammation Reduced by CBD October 30, 2020 Madeline Colli The Dental College of Georgia (DCG) and the Medical College of Georgia, recently performed a study which produced results showing that cannabidiol (CBD), a non-psychoactive cannabinoid found in cannabis, reduces damage in the lungs spurred by cytokine storms caused by coronavirus (COVID-19). A recent study from the Dental College of Georgia (DCG) (Augusta, Georgia) and the Medical College of Georgia (Augusta, Georgia) produced results showing that cannabidiol (CBD), a non-psychoactive cannabinoid found in cannabis, reduces damage in the lungs spurred by cytokine storms caused by coronavirus (COVID-19) (1). In an earlier

study (2), the researchers illustrated that CBD can reduce inflammation and physical lung damage associated with adult respiratory distress syndrome (ARDS), along with improved oxygen levels. The new study revealed the mechanisms behind these earlier results, exhibiting that CBD normalizes levels of apelin, a peptide known to reduce inflammation. During a COVID-19 infection, levels of the apelin peptide are low. Results of this study have been published in the Journal of Cellular and Molecular Medicine (3). In the authors ARDS model, blood levels of apelin (a crucial piece in regulation, bringing both blood pressure and inflammation down) had dropped close to zero and increased 20 times with CBD. In the instance that blood pressure gets too high, apelin levels rise up to help alleviate the pressure. It is assumed that apelin should also help normalize inflammation in the lungs and other related breathing difficulties linked to ARDS. ―Ideally with ARDS it would increase in areas of the lungs where it’s needed to improve blood and oxygen flow to compensate and to protect,‖ said Dr. Babak Baban, DCG immunologist (1). In their ARDS model, apelin did neither of these. When CBD was administered, apelin increased. Although with their current study, the authors reiterated that much more is still being studied and there is some animosity in regard to whether CBD or COVID-19 have a direct effect on apelin but they are continuing on their mission to find out. ―It is an association; we don’t know yet about causative, but it is a very good indicator of the disease,‖ Baban said (1). What they have found so far is that the reductions in apelin could potentially be an early biomarker for ARDS and response to treatment efforts.

Based on their earlier study, the authors mentioned that the data supported the idea that the anti-inflammatory function of CBD may assist in reducing cytokine storm and mitigate the effects of exaggerated inflammation (2). Further adding that, ―considering all potential regulatory effects of CBD as well as the vast distribution of endocannabinoid system in the body, it is plausible that CBD may be used as a therapeutic candidate in the treatment of various inflammatory conditions including COVID-19 and other virus-induced ARDS.‖ Expanding from the researchers’ newest study, the next steps for their research would be to comprise a stronger understanding of the interactions between apelin, CBD, and COVID-19 as well as studying why apelin decreases during COVID-19 infections and how CBD increases those numbers. They will explore how eliminating apelin affects ARDS and if CBD is able to produce the same lung benefit without the peptide. It is assumed that COVID-19 suppresses some sort of reaction that then suppresses apelin and CBD interferes with this process. Nonetheless, the authors doubt the apelin-CBD interaction is the only means by which the compound works in this or other scenarios. Moving forward with their studies, the authors have refined a safe, relatively inexpensive model of ARDS by giving a synthetic analogue of double-stranded ribonucleic acid (RNA) called POLY (I:C). COVID-19 has double-stranded RNA, while human deoxyribonucleic acid (DNA) is single-stranded. The researchers’ model propagated a response similar to the virus, along with the extreme lung damage and cytokine storm, reflecting an overactive immune response in the lungs. Gathering data for these studies, a control group received intranasal saline consecutively for three days while the COVID-19 model received POLY (I:C) intranasally for three days. Then there was a third group,

known as the ―treatment group,‖ who received POLY (I:C) and CBD over the same timeframe. The results produced significantly reduced apelin levels in the mice who had developed COVID-19-like symptoms compared to controls, and treatment with CBD normalized the immune response, apelin levels, oxygen levels, swelling, and scarring in the lungs. CBD has shown itself to be a natural apelin agonist. Apelin levels are consistently measurable in the lungs, which is one of the reasons it proves to be a good biomarker. Aside from CBD, other cannabinoids, such as tetrahydrocannabinol (THC), are proving themselves beneficial in the fight against COVID-19 by decreasing the risk of cytokine storms, ARDS, and mortality (4,5). With future research, cannabis may prove itself useful in strengthening the side of those battling the virus and others fighting to rid the world of COVID-19. References: 1. https://www.healtheuropa.eu/cbd-reduces-covid-19-lung- damage-with-protective- peptides/103374/#:~:text=In%20a%20new%20study%2C%20scie ntists,levels%20of%20a%20natural%20peptide 2. https://www.cannabissciencetech.com/view/evidence-grows-for- cbd-as-potential-covid-19-treatment 3. https://onlinelibrary.wiley.com/journal/15824934 4. https://www.cannabissciencetech.com/view/a-storm-of-research- activity-into-cannabis-and-coronavirus-a-closer-look-at- expanding-research-of-cannabinoids-and-terpenes-in-covid-19 5. https://www.cannabissciencetech.com/view/treating- immunocompromised-patients-during-the-covid-19-pandemic- a-conversation-with-dr-bonni-goldstein

Medical Cannabis and Better Patient Care: Breaking the Stigma with Science and Education October 30, 2020 Madeline Colli Dr. Junella Chin, the Founder of MedLeaf RX and the Chief Medical Advisor for Yesterday Wellness, discusses with us the beneficial aspects CBD and how the medical field is beginning to incorporate the concept of medical cannabis into patient treatments. The cannabis plant was used as a medicinal herb for thousands of years before its prohibition in the 20th century. As legalization for cannabis and hemp has spread across the US and globe since 1996, medical professionals are left to fill in the gaps of knowledge with their patients on its use. For instance, cannabidiol (CBD) is hailed as a treatment option for many conditions but many doctors are not taught about it or the endocannabinoid system in medical school. Dr. Junella Chin, the Founder of MedLeaf RX and the Chief Medical Advisor for Yesterday Wellness (1), recently spoke to us about the beneficial aspects CBD and how the medical field is beginning to incorporate the concept of medical cannabis into patient treatments. What inspired you to pursue CBD as a treatment option for your patients? Dr. Chin: As a teenager, I was diagnosed with ankylosing spondylitis (AS), a progressive type of arthritis that affects the spine, pelvis, hips, and back and causes extreme stiffness and nerve pain.

I spent my younger years trying conventional treatments— epidurals, narcotics, muscle relaxants, acupuncture, physical therapy—but the pain was unrelenting. By the time I got to medical school in San Francisco, I was having difficulty standing for long periods in the operating room. One of the attending physicians saw this and asked me about it. I told him I had AS but that I couldn't take the meds I needed for relief while doing rounds or while attending a four-hour hip-replacement surgery because they made me drowsy and foggy. Here, I was in a hospital surrounded by great medical minds, but I was disheartened to find nothing could help my condition. The attending physician and my mentor Dr. Levine pulled me aside and handed me a bottle containing a tincture. \"This is marijuana,\" he told me, \"but it won't make you high.\" He didn't call it CBD oil. He just said it was a different type of cannabis plant. I was mortified but desperate. As a medical student, training to be a doctor, my first thought was, \"You are offering me pot? You want to make me a drug addict?\" I grew up in the Bronx, where there was a huge social stigma around marijuana. Kids that smoked weed were either the dropouts or involved in gangs. I also grew up in a very traditional Chinese household. My parents believed in ―Reefer Madness‖—that weed would led to psychosis and schizophrenia. The little brown dropper bottle Dr. Levine handed me smelled like a combination of alcohol, wet dog, and grass, and I didn't know what to think. To my amazement, the tincture worked very well. The pain and inflammation of my arthritis decreased dramatically, my AS stopped progressing, and my health improved. Even though California legalized medical cannabis in 1996, I didn't dare tell anyone I was using it. I was a young physician and didn't want to jeopardize my career. But once I got my health back, I decided to

learn more about cannabis and how it helps manage pain and improve people's overall health and wellness. Having suffered in pain for so long, I know what it feels like to say to your doctor, \"I've tried everything, and nothing has helped.\" I have been helping patients integrate medical cannabis into their health and wellness for more than a decade. Cannabis changed the trajectory of my life. I would not have been able to finish medical school and become a doctor had it not been for cannabis treatment. I had an educational advantage in doing my medical school training in California. California legalized medical cannabis in 1996. I was in the middle of a switch box and was able to engineer my circumstances to learn holistic and integrative cannabis medicine. Do you face any scrutiny or stigma from other medical professionals for using medical cannabis or CBD as a treatment option? Dr. Chin: In medical school, there is a \"this is how we've always done it\" syndrome. That is the conventional, allopathic medicine model. How did we come to believe that prescription medication is the only or most effective way to treat disease? We are taking a reductive approach. In medical school, we are trained in identifying what's bad in your body and then to get rid of it. This is only part of the puzzle. We often get locked into this one-size-fits-all thinking. There is no one-size-fits-all when it comes to working with plant medicine like cannabis. Botanical remedies have been mainstays of folk medicine in many cultures throughout history. Phytocannabinoid medicine is on that list. We also have to remember there are synergistic effects of the complex compounds found in plants. When we try to isolate one of those molecular compounds and take its’ pure form, not only may you

not get all the benefits, but you increase risk by taking them out of context. What resources do doctors and other medical professionals have to learn more about cannabis as a treatment option? Dr. Chin: Unfortunately, there are very little resources since it’s still not taught in medical schools. What benefits have you seen with CBD in your patients? Dr. Chin: Over the last 15 years, my medical cannabis career has been based on anecdotal and clinically applied evidence. I help patients integrate medical cannabis in the context of a full-scope general and holistic medical practice. CBD is an empowering medicine. The cannabis plant is unique. There is no precedent, no other drug in the world that we are using recreationally and medicinally for therapeutic uses. The historical record of safe use is unparalleled. The endocannabinoid system (ECS) modulates and interfaces with all of the other systems throughout your body. It regulates physical functions, such as movement, pain sensation, immune responses, and cognitive or mental capacities, such as perception, mood, and memory. The ECS naturally produces cannabinoid-like molecules that stimulate the body's endocannabinoid receptors. These receptors are in the brain, muscles, and fat (adipose tissue). There is a galaxy of cannabinoid receptors in the digestive system. How does CBD affect the way the brain processes emotional memories (for example, in patients suffering from PTSD and other psychiatric disorders)? Dr. Chin: Anxiety, stress, and chronic sleep deprivation all inhibit Gamma-Aminobutyric acid (GABA), a naturally occurring brain

chemical that directs neurons to slow down or stop firing. This neurotransmitter also helps to induce sleep, relax muscles, and calm us down. In essence, GABA directs the body to chill out. Cannabis modulates GABA, helping return the body to its more normal functions. Careful dosing may help stem the racing thoughts that cause disrupted sleep and panicked awakenings during the night. It can also be used to treat what are called ―parasomnias,‖ sleep disorders such as jaw grinding, sleepwalking, or nightmares. The ―cannabis molecules‖ produced by your own body (endogenous cannabinoids) make you resistant to stress, similar to the way endorphins provide natural relief from pain. Integrating optimally- dosed cannabis products can help bring the body back into balance. Cannabinoids also work to mitigate stress (2). CBD directs the brain to activate the parasympathetic nervous system, which induces a state of calm. It works well for social anxiety and everyday stress, plus more extreme forms of anxiety such as panic attacks. One small study (3) showed it even ―reduced anxiety in public speakers‖ so if you’re gearing up for that online TED Talk, take note! How do the different state regulations the US impact your work? For instance, is it more difficult to treat patients in New York compared to California? Dr. Chin: Every state has its own medical cannabis laws that we have to adhere to as MDs. I have not found it more challenging to practice in New York versus California. Do you feel that CBD may become more sought out and the future for treating not only neurological disorders but other ailments as well? Have you studied any other cannabinoids?

Dr. Chin: At last count, the cannabis plant contains more than 500 chemical compounds, many of them only in trace amounts. We need more research, but the research needs to focus on the fundamental understanding of what is going on with the whole plant and not selecting out individual molecules like we know what we are doing. Again, we are looking at this field from a \"disease-target-kill model.\" Allopathic medicine takes the reductive approach. We are separating disease into its parts, element by element, component, and subpart, and then using drugs to attack these separate targets. CBD is part of a whole plant. There's a whole universe of regenerative medicine, nutritional medicine, preventative, and integrative medicine. Cannabinoid science and the push for cannabis legalization is teaching us a vital lesson. It is reframing our thinking about medicine and healthcare. It also raises some of the most critical questions about how we think about medicine in the future. What would you like to see change in medical cannabis regulations in the United States? Dr. Chin: I think we needed a guide to cannabis and CBD a while ago—it almost feels like we are racing to keep up with the explosion in popularity of CBD in particular and nobody understands what it is, how to use it, or even what they are actually getting. As states legalize and as the federal government is trying to figure out what a \"legal CBD market\" looks like, the general public is caught in the middle with very little quality information about cannabis. For example, most people don't realize that CBD comes from the cannabis plant, it is a chemical compound extracted from theCannabis sativa plant. In the US, the CBD we can \"legally\" purchase is defined as coming from \"industrial hemp,\" however, the hemp plant is Cannabis sativa, it is just a cultivated variation of the plant that is being farmed and used in legal medical and recreational states to produce the

cannabis containing THC, another chemical compound in the cannabis plant. The main difference is that the hemp plant is a low resin plant containing very low amounts of THC. Of course, there is, and will continue to be, stigma around cannabis. First, it is still federally illegal and most people don't realize that it isn't illegal because it is dangerous but it was made illegal for economic reasons. There were decades of deliberate propaganda that resulted in movies like \"Reefer Madness\" in the 1930s and the \"Just Say No\" campaign in the 1980s. Fear, bigotry, and greed led to the vilification of the medicinal cannabis plant and the versatile hemp plant. It is hard to abolish decades of misinformation, not to mention the decades of criminalization of cannabis and the imprisonment of mostly people of color for possessing cannabis. As long as the plant is federally illegal and people believe the misinformation, there will still be a stigma. Patients are looking for reliable information but there are few, trusted healthcare-provided resources. Healthcare practitioners receive little or no education regarding medical cannabis. As doctors, we need to be capable of engaging in meaningful discussions with patients regarding the potential harms and the benefits of cannabis and safely help them manage their diseases and improve their quality of life. I always say I'd like to see \"cannabis in every medicine chest\"—that it is accessible to all people who need it. But we still have a long way to go. We do have to remember that there are counter indications and cross reactions of CBD and cannabis with other prescription medications and certain health conditions. Cannabis is not a cure-all, and it works differently for each person so just because it helped their

friend or neighbor with a particular ailment or condition doesn't mean it will work the same way for them. Rescheduling of cannabis to Schedule II, III, IV, or V presupposes that there is an FDA-approved pharmaceutical that can be prescribed. That is not the case. Even with Epidiolex (CBD extract) approval for severe epilepsy in 2018, it does not carry over to other forms of CBD or cannabis in any way. In the University of London’s study (4), what are your opinions and hopes of their findings? Dr. Chin: I think this UL study is promising and more research is needed. When vessels that supply blood to the brain are too clogged or damaged, this can result in a decline in cognitive function, memory loss, and difficulty making executive decisions. In addition, mobility and balance get impaired, too. Increasing blood flow to areas in the brain help to maintain cognitive faculties as we age and perhaps increasing memory in Alzheimer’s disease patients. How does CBD increase the cerebral blood flow throughout the brain? Dr. Chin: The mechanism of action with respect to increase cerebral blood flow on the brain is unclear. References: 1. https://yesterdaywellness.com/ 2. https://pubmed.ncbi.nlm.nih.gov/28249817/ 3. https://pubmed.ncbi.nlm.nih.gov/21307846/ 4. https://medicalxpress.com/news/2020-08-cannabidiol-blood- brain-hippocampus.html

New Study Links Cannabidiol’s Multitudinous Effects to Disruption of Cholesterol Homeostasis October 14, 2020 Cindy Orser Cannabis Science and Technology, October 2020, Pages: 50-53 Op-ed on a new study related to how cannabidiol (CBD) exerts its clinical efficacy. HQUALITY/adobestock.com A significant contribution to our understanding of how cannabidiol (CBD) exerts its clinical efficacy has recently been published online by researchers at the University of Colorado (1). The evidence for how CBD affects more than 20 membrane targets had potentially implicated that the influence of CBD on membrane fluidity was responsible for its pharmacological effects, but the actual mechanism had not been demonstrated until now. The key finding from the multi-

omic systems level analysis in human cell lines is that CBD partitions into cellular membranes, which disrupts cholesterol homeostasis along with membrane spanning ion channels and receptors, triggering a myriad of downstream consequences. CBD affected cytosolic calcium, cholesterol transport and storage, cellular energy sensors, and oxidative stress responses in a dose-dependent manner (1). In addition, CBD-exposed cells were sensitive to extracellular cholesterol, which induced cell death, or apoptosis, in a dose dependent manner. Cholesterol interacts with the fatty acid tails of phospholipids to moderate the biophysical properties of the plasma membrane, reducing fluidity and making the membrane less permeable to small water-soluble molecules that would otherwise freely cross. While cholesterol’s bad press has led to the widespread prescription of statins that block synthesis of cholesterol, it is essential for both membrane permeability and also in metabolism as the starting point for the synthesis of bile salts and steroid hormones including cortisol, estrogen, and testosterone. Cholesterol makes up around 25–30% of the lipid content of the cell membrane and it is the dominant lipid in the brain. Cells either synthesize cholesterol from acetyl coenzyme A (acetyl-CoA) or take it up from the food we eat. While cholesterol deficiency results in several diseases, we are just beginning to investigate and appreciate the regulatory role that cholesterol plays in its relationship to cellular homeostasis, inflammation, and pain much like how we think of CBD today. Of the proposed list of CBD targets, 22 are membrane-localized channels and receptors. Mutations in one voltage-dependent sodium channel, NaV1.1, are responsible for epilepsy syndromes including Dravet syndrome for which Epidiolex is prescribed (2). Many other proposed targets are calcium channels or receptors, in fact postsynaptic calcium mobilization is the probable mechanism for the anticonvulsant activity of CBD (3). The recent study brings strong

evidence to a previously proposed theory that this interaction of CBD with membrane-localized channels is indirect through biophysical alteration of the lipid bilayer with cholesterol as the effector molecule (4,5). Bioactive Lipids Growing evidence suggests that pain sensation caused by inflammation is in part regulated by the pro- or anti-nociceptive actions of lipid mediators which include endogenous cannabinoids as well as agonists of peroxisome proliferator-activated receptor-α (PPAR-α), platelet activating factor (PAF), and various products of oxidative polyunsaturated fatty acids (PUFAs) metabolism (6). An increasing number of lipid molecules have been shown to suppress the inflammatory process, restore homeostasis in damaged tissues, and attenuate pain sensitivity by regulating neural pathways that transmit nociceptive signals from the periphery of the body to the central nervous system (7). The endocannabinoids, anandamide and 2- AG, are synthesized locally on-demand to mitigate the effects of exogenous and endogenous pro-analgesic agents by attenuating nociceptor excitability and contrasting local pro-inflammatory signals from another group of bioactive lipids, including membrane cholesterol (7). Loss of Cholesterol Homeostasis Let’s take a closer look at how loss of cholesterol homeostasis mitigates a wide range of physiological change. Normally, cholesterol levels are tightly regulated between the plasma membrane (PM) sensors and the endoplasmic reticulum (ER). When cholesterol is depleted, low density lipoprotein (LDL) molecules transporting cholesterol bind to receptors and enter the cell through endocytosis to reach lysosomes where the cholesterol is released from the LDL and delivered to the PM and ER membranes. If there is excess cholesterol, it is removed from the PM and delivered to the ER where it is esterified

for storage in lipid rafts and not available for normal cellular processes (8). When this normal feedback inhibition of cholesterol levels is disrupted there is subsequent activation or deactivation of raft- associated proteins, such as death receptor proteins, protein kinases, and calcium channels. Furthermore, increased production of cholesteryl esters, has been reported across human tumors compared with normal tissue. Cholesterol, Cancer, and CBD Changes in cholesterol metabolism play a role in carcinogenesis. It is well established that cancer cells have a much higher concentration of cholesterol and the levels of cholesterol determine their resistance to chemotherapeutic drugs (9). This increased capacity to esterify and accumulate cholesterol in tumor cells has been associated with a higher growth rate, suggesting a link between cholesteryl ester production and cell proliferation (10,11). There does not appear to be a correlation with the levels of cholesterol consumed, but rather de novo synthesis from dysregulation of the cholesterol pathway. Several anti-cancer drugs are able to suppress growth and induce apoptosis of tumor cells through alteration of cholesterol containing lipid raft contents via disrupting lipid raft integrity and thereby sensitizing cancer cells to therapeutics and also influencing cancer drug resistance (12). Similar findings have been reported for cancer cells treated with CBD, essentially robbing cancerous cells of their energy source, cholesterol. In earlier reporting, primary microglial cells treated with CBD resulted in a concentration dependent induction of apoptosis that was not counteracted by CB1 or CB2 receptor antagonists but was attenuated by a lipid raft disruptor. This finding suggests that CBD induces a pro- apoptotic effect in primary microglia through lipid raft coalescence and elevated expression of GM1 ganglioside, involved in neuron repair, and caveolin-1, a structural component of lipid rafts (13). In the

case of unregulated cholesterol accumulation in cancer cells, CBD is eliciting the same apoptotic outcome as anti-cancer drugs without the toxic side effects. Likewise, statins have proven therapeutic in some cancers—including gastric, hepatic, prostatic, and esophageal— through their inhibition of cholesterol synthesis (14). Cellular Calcium, Cancer, and CBD CBD treatment was associated with high levels of Ca2+ in the cell (1). An influx of Ca2+ ions can damage cells through disruption of neural circuitry through excitotoxicity which can occur in neurodegenerative diseases or brain trauma such as traumatic brain injury (TBI) or stroke even causing cell death. Cellular calcium homeostasis is necessary for stress responses in metabolic organs including the heart, liver, and adipose tissue. Multiple lines of evidence demonstrate that inappropriate consumption of nutrients and chronic metabolic diseases leads to dysregulation of cytosolic and organelle Ca2+ fluxes and disrupted equilibrium in metabolic tissues and immune cells, which in turn alters organelle homeostasis, signaling pathways and autophagy (15). Cellular Ca2+ homeostasis fails to maintain its adaptive integrity in the face of structural and signaling defects introduced by chronic metabolic challenges whether through over consumption of nutrients or through treatment with CBD. The effects rendered by CBD treatment cannot escape the interrelatedness of all processes in the human body. In a recent study by Olivas-Aguierre and colleagues (2019) the treatment of acute lymphoblastic leukemia of T-cells (T-ALL) with CBD resulted in mitochondrial dysfunction and cell death by altering their calcium homeostasis, which was directly demonstrated as the inhibitor of mitochondrial Ca2+ uptake protected the T-ALL cells from CBD-induced cell death (16). Importantly, and of even further interest open to speculation, is that healthy, resting T-cells were resistant to the effects

of CBD and avoided cell death! These data provide provocative evidence for CBD as a candidate for chemotherapy trials. CBD-Induced Mitochondrial Dysfunction We now understand that not only does CBD intercalate into the plasma membrane, disrupting ion channels, but it also activates internalization of PM cholesterol which in turn destabilizes the ability of mitochondria to manage its own calcium homeostasis affecting multiple cellular processes including Ca2+ signaling the downstream cell death. CBD also causes the translocation of other membrane proteins leading to decreased cellular energy (ATP) production. At play here is the translocation of a key mitochondrial membrane protein, hexokinase I (HK1). Loss of functioning HK1 results in the decoupling of glycolysis from mitochondrial respiration through diminished transformation of glucose to glucose 6-P (17). Through multi-omic profiling in combination with fluorescence resonance energy transfer (FRET)-based biosensor screening, the Colorado group’s results confirm an earlier report by Alharris and colleagues (2019) that CBD- induces mitochondrial dysfunction (18). Mitochondrial dysfunction leading to apoptosis or cell death, reported in the Colorado study, has also previously been shown in pancreatic b- cells with associated insulin resistance prior to cell death a hallmark in type 2 diabetes (19). Findings from another relevant study this time involving neurons and platelet-activating factor (PAF), which is also influenced by calcium levels and PM cholesterol is worth citing here (20). PAF binds to its receptor (PAFR) located in the PM and nuclear membrane which mobilizes Ca2+ and activates numerous signaling pathways including inflammation, which can result in cell death. If cells are treated with statins or CBD to reduce the level of cholesterol, then PAFR is displaced, PAF doesn’t bind to its receptor and neurons become resistant to apoptosis. This is a significant finding in light of another study where deletion of PAFR in a mouse knockout

experiment abolished the inflammatory response and neuronal apoptosis caused by TBI (21). As a side note, Etizolam, a benzodiazepine analog, is a PAF-receptor antagonist used to treat anxiety. Far Reaching Implications The widespread self-medication with CBD, now accessible in a dizzying array of consumables readily available to the public, has preceded any clear understanding of the mechanisms behind CBD’s apparent benefit across a wide range of ailments. The far-reaching cellular disruptions observed in the Colorado study were corroborated by transcriptomics, metabolomics, and proteomics showing the downstream impact from disruption in cholesterol biosynthetic and regulatory pathways including decoupling of glycolysis from mitochondrial respiration, upregulation in oxidative stress response, increase in Ca2+ cytosolic levels, and the increased production and storage of cholesterol sensitizing cells to apoptosis making cholesterol the effector molecule of CBD across many intracellular pathways rather than any specific target. The Colorado study has shown that not all CBD effects on cells are beneficial. While there had been earlier reports from human clinical trials of the now Food and Drug Administration (FDA)-approved CBD product, Epidiolex, indicating that CBD treatment could lead to risk of liver injury as indicated by increased levels of aminotransferase activity and inhibition of P-450 (CYP) enzymes (22), the molecular mechanisms of that potential toxicity were not fully understood (23). It should be mentioned that the Colorado study was carried out on two immortalized human cell lines: SK-N-BE(2) neuroblastoma cells and HaCaT human keratinocyte cells with CBD treatment in the micromolar range which would be less than the prescription dosing of Epidiolex, but probably more than self-administered CBD dosing. Their

analyses based on transcriptomics showed that the enriched transcription factors shared oxidative stress as a stimulus, resulting in upregulation of oxidative stress response including fatty acid and cholesterol biosynthetic gene transcripts. It was known that CBD upregulates mRNA transcripts in microglial cells involved in fatty acid metabolism and cholesterol biosynthesis (24). This alteration in fatty acid metabolism was also shown in a mouse chronic alcohol feeding study where CBD reduced the infiltration of liver cells with fat, a distinct benefit (25). If anything, this study has brought attention to the need for basic research on cannabinoids and the biological systems they effect to clarify why endocannabinoids are synthesized locally on an as needed basis in nanomolar amounts and then are quickly degraded. Fundamentally, the endocannabinoid system presents a unique opportunity to look at how a lack of essential nutrients, specifically lipid-based foods, or an overabundance of low-quality nutrient intake impacts synaptic plasticity and resulting outcomes in physical and mental health. The Role of Cannabinoids in the Global Fight Against Antibiotic Resistance August 19, 2020 Alexia Blake, Gilad Landan, Jeremy Friedberg Cannabis Science and Technology, July/August 2020 , Pages: 40-43

A closer look at some of the current in vitro evidence suggesting that cannabinoids possess potent activity against a range of gram-positive bacteria. Chudakov / adobestock.com Antimicrobial resistance is a critical issue among the global healthcare community. Based on reports from global health authorities and hospitals, the World Health Organization (WHO) has identified 12 pathogens with increased antibiotic resistance, including Methicillin- resistant Staphylococcus aureus (MRSA) (1). In the United States, MRSA infections topped 300,000 every year since 2005, resulting in more than 10,000 deaths and $1.7 billion in healthcare costs in 2017 (2). Similarly, the Canadian Antimicrobial Resistance Surveillance System reported that the rate of MRSA infections within sentinel hospitals increased since 2015 from 2.8 to 3.17 cases per 10,000 patient-days in 2017 (3). The prospect of losing lives to avoidable infections is real, particularly during spikes in healthcare use that may overburden healthcare systems, compromise sanitation practices, and further exacerbate the emergence of antimicrobial resistance. COVID-

19 has created such a spike in healthcare use on a global scale, and in preparation for future healthcare crises, novel antibiotic tools need to be developed now to support strained healthcare systems. Current in vitro evidence suggests that cannabinoids such as tetrahydrocannabinol (THC), cannabidiol (CBD), cannabigerol (CBG), cannabinol (CBN), and cannabichromene (CBC) possess potent activity against a range of gram-positive bacteria including MRSA, as well as gram-negative bacteria when supplemented without membrane penetrating agents (4–6). In particular, CBG presents superior activity in the real-world applications of biofilm inhibition, biofilm eradication, and antibacterial activity at concentrations well below toxic levels that are achievable in topical formulations with utility in over-the-counter and acute care clinical settings (5). Cannabinoids present an opportunity to develop a novel and highly effective antibiotic pipeline. Further in vivo work is required to validate the antibacterial efficacy of cannabinoids when administered orally, intravenously, or through other routes for systemic treatment. Cellular agriculture offers a way to produce pure and consistent cannabinoids for further research and integration into end products for this specific utility (7). Early Studies Show Effectiveness of Cannabinoids Against Gram-Positive Bacteria In response to these healthcare challenges, researchers have focused their efforts on evaluating cannabinoids as potential antibacterial agents against both gram-positive and gram-negative bacteria, which have different cell wall structures (Figure 1, click to enlarge). Gram- positive bacteria have a cytoplasmic membrane surrounded by a thick peptidoglycan layer, while the more numerous and varied gram- negative bacteria have three layers, including a thin peptidoglycan layer surrounded by an elaborate outer membrane studded with complex sugars.

One of the first studies in this domain evaluated the antibacterial properties of delta-9-THC and CBD against seven different pathogens that included both gram-negative and gram-positive bacteria (4) (Table I, click to enlarge). This early work suggested that 1–5 µg/mL THC and CBD possess both bacteriostatic (prevent further growth) and bactericidal (kill existing cells) properties against gram-positive bacteria but have no effect on gram-negative bacteria because they are unable to cross the thick bacterial outer membrane unassisted. These findings were in agreement with more recent research, which utilized in vitro susceptibility models to assess the antibacterial activity of five cannabinoids (THC, CBD, CBG, CBC, and CBN) against gram- positive MRSA (5). In this work, the minimum inhibitory concentration (MIC) required to prevent bacterial growth of THC, CBD, CBG, CBC, and CBN was reported as < 2 µg/mL (Table II). This activity was found to be comparable to several commonly used antibiotics, suggesting that cannabinoids are potent antibacterial compounds when applied directly to the pathogen site, as is the case with topical administration. A separate study reported similar results for the same cannabinoids against a different strain of MRSA (USA300), with the exception of CBC (Table II) (6). The agreement between these two studies serves as promising evidence of the potential that cannabinoids offer as topical antibacterial agents for gram-positive bacteria. Cannabinoids May Also Inhibit and Eradicate Biofilm Formation While bacterial growth is a concern for bacterial spread, biofilm formation is regarded as a critical factor contributing to infection duration because of its heartiness in the face of traditional hygiene and subsequent accumulation on surfaces and in reservoirs, particularly in hospitals and other healthcare settings. In this context,

one study explored the ability of THC, CBC, CBG, CBN, and CBC to inhibit MRSA biofilm formation, and to eradicate preformed biofilms of MRSA (6). The results indicated that these cannabinoids were able to inhibit biofilm formation, and that this inhibitory activity was correlated with their antibacterial activity as measured by MIC against gram-positive pathogens. In particular, CBG was identified as the most effective cannabinoid capable of inhibiting 50% of biofilm formation at 0.5 µg/mL. CBG was also able to eradicate preformed biofilms (minimum biofilm eradication concentration of 4 µg/mL) and kill MRSA persister cells (MIC of 5 µg/mL). Furthermore, MRSA did not develop resistance to CBG, even when treated with lethal concentrations of CBG ranging from 2 to 16x MIC and cultured for 15 days, an important finding in the context of antibacterial resistance. Collectively, these findings suggest that the antibacterial activity of CBG may extend beyond topical antibiotic applications and be suited for antibacterial coating applications for medical equipment, such as stents. With Some Assistance, Cannabinoids Show Promise Against Gram-Negative Pathogens Gram-negative bacteria are frequently to blame for infections and are often difficult to treat in part because of the protective polysaccharide shell that surrounds their outer lipid membrane. Indeed, eight of the 12 WHO Priority Pathogens previously mentioned are gram-negative. While previous findings suggested that the antibacterial activity of cannabinoids was limited to gram-positive pathogens, a more recent study showed how the coadministration of a commonly used gram- negative antibiotic can improve the antibacterial activity of cannabinoids against gram-negative pathogens (6).

The study determined the effectiveness of CBG, CBN, CBD, CBC, and THC against a laboratory strain of gram-negative E. coli following coadministration of Polymyxin B, a topical antibiotic that disturbs the outer membrane characteristic to gram-negative bacteria and is used in the commercially available topical ointment Polysporin (Johnson & Johnson). In all cases, coadministration of Polymyxin B enabled the cannabinoids to act as effective antibacterial agents against the laboratory strain with MIC values below 1 µg/mL (Table III, click to enlarge). However, the dose of Polymyxin B required to enable this activity varied with each cannabinoid. In particular, CBC and THC required the lowest amount of Polymyxin B to prevent bacterial growth, while all other cannabinoids required twice the amount of Polymyxin B to achieve the same synergistic effect. In the same study, CBG was selected for further testing against additional gram-negative pathogens given its superior effects against biofilms and persister cells (Table III). Coadministration of Polymyxin B proved to enable CBG’s antibacterial activity against a clinical strain of E. coli, K. pneumonia, P. aeruginosa, and A. baumannii at varying concentrations not exceeding 1 µg/mL. Cellular Agriculture Offers a Way to Further Understand Antibiotic Cannabinoids Crucially, the existing pool of antibiotic cannabinoids—together with the barely explored minor cannabinoids—form an extensive and coherent pipeline that awaits development. The current research suggests that cannabinoids are well-suited antibiotic agents when applied topically, particularly against gram-positive pathogens, while action against gram-negative bacteria is enabled by coadministration of a secondary antibiotic. Ongoing in vitro and in vivo work will undoubtedly further our understanding of the topical and systemic antibiotic properties of cannabinoids; however, it can be very difficult to source high-quality,

pure cannabinoid ingredients at exact dosages to meet requirements for this research. Cellular agriculture offers a means to produce consistent, uncontaminated cannabinoid ingredients at scale to keep pace with the anticipated increase in research around antimicrobial use cases and demand by the health and pharmaceutical markets. Further research will undoubtedly cement their utility in the global fight against antibiotic resistance. About the Authors Alexia Blake is the director of product development at LAVVAN in Markham, Ontario, Canada. Gilad Landan is a senior scientist at LAVVAN in Santa Clara, California. Jeremy Friedberg is the chief science officer at LAVVAN in Guelph, Ontario, Canada. Direct correspondence to Jeremy Friedberg at [email protected]. How to Cite this Article A. Blake, G. Landan, and J. Friedberg, Cannabis Science and Technology 3(6), 40-43 (2020). FDA Releases Cannabis Research Guidelines August 6, 2020 Madeline Colli The U.S. Food and Drug Administration (FDA) recently released a draft guidance on developing cannabis-based drugs. These guidelines come weeks after the White House announced it had completed reviewing the draft guidance. In December 2018, Congress passed the 2018 Farm Bill (1), which was a groundbreaking piece of legislation that legalized the sale and cultivation of industrial hemp and its derivatives after decades of

prohibition. For quite some time, lawmakers, advocates, and stakeholders have pressed the U.S. Food and Drug Administration (FDA) to create a regulation structure to assist the industry. Recently, the FDA did just that by releasing a draft guidance on developing cannabis-based drugs (2). These FDA guidelines come weeks after the White House announced it had completed reviewing the draft guidance (3). The new document (2) mentions that the guidance is ―limited to the development of human drugs and does not cover other FDA regulated products.‖ The draft documentation is meant to provide an outline for drug manufacturing. The agency is currently still working on developing guidelines that will allow cannabidiol (CBD) to be marketed and sold as a food item or supplement. CBD has increased in popularity in recent years because of its potential for treating medical conditions. ―A range of stakeholders have expressed interest in the development of drugs that contain cannabis and other compounds found in cannabis. Recent legislative changes have also opened new opportunities for cannabis clinical research. As that body of research progresses and grows, the FDA is working to support drug development in this area,‖ said FDA Principal Deputy Commissioner Amy Abernethy (4). Some of the legislative changes Abernathy is referring to is the 2018 Farm Bill, legalizing industrial hemp with less than 0.3% tetrahydrocannabinol (THC). Researchers are now able to use industrial hemp from any source as long as it doesn’t exceed the allowed THC limit and not limiting them to use from the University of Mississippi, which is the country’s only federally authorized cannabis manufacturer.

In the new draft guidance, there is an emphasis on THC testing, mentioning that investigators and sponsors may find it beneficial to calculate the level of delta-9 THC early in the development process of their proposed investigational drug product so that they can gain better insight into the potential control status. The document also provides step-by-step guidance on how to test for THC on a dry weight basis according to the FDA’s standards (4). The FDA is receptive to the public and has opened a 60-day comment period for stakeholders to display any opinions on the proposed research guidelines. A Tribute to the Life and Legacy of Charlotte Figi June 23, 2020 Josh Crossney Cannabis Science and Technology, June 2020 , Pages: 23-24 A tribute piece to Charlotte Figi, a medical cannabis and CBD pioneer who inspired the name of the medical cannabis strain Charlotte’s Web. A tribute piece to Charlotte Figi, an American girl with Dravet syndrome who took cannabidiol (CBD) oil to prevent seizures and inspired the name of the medical cannabis strain called Charlotte’s Web. Cannabidiol (CBD), is now available everywhere-from grocery stores to gas stations. Millions of people around the globe use CBD to combat a

variety of illnesses, disorders, and diseases. Yet many CBD users do not know that the awareness of the medical benefits of CBD, as well as their access to it, was due in large part to the heroic battles of a young, courageous girl named Charlotte Figi. The cannabis industry has had many pioneers and trailblazers. We recently lost one of our greatest: Charlotte Figi, the young woman who became a hero in the CBD movement. On April 7, 2020 Charlotte passed away at the age of 13. Nichole Montanez, a close family friend of the Figi’s, posted a message on Paige Figi’s (Charlotte’s mother) Facebook page that read (1): ―Charlotte is no longer suffering. She is seizure-free forever. Thank you so much for all of your love.‖ We herein honor Charlotte and not only share her amazing story, but also explain the incredible inspiration that Charlotte has been to so many pediatric cannabis families. Charlotte’s Legacy Charlotte and her twin sister, Chase, were born on October 18, 2006 to parents Paige and Matt and big brother Max. Charlotte suffered from Dravet syndrome. According to the Dravet Syndrome Foundation (2), Dravet syndrome is a rare catastrophic, lifelong form of epilepsy that begins in the first year of life with frequent or prolonged seizures. There are currently very limited options to treat Dravet syndrome. At the tender age of 5, Charlotte suffered up to 300 grand mal seizures a week and had extreme difficulty speaking. Paige first searched for CBD oil in 2012 and Charlotte began taking high-CBD/low-tetrahydrocannabinol (THC) oil from the cannabis strain called ―Hippie’s Disappointment.‖ This oil immediately reduced her epileptic seizures and the oil was later named ―Charlotte’s Web‖ in her namesake. Her story was the focus of the CNN documentary Weed hosted by Sanjay Gupta in 2013 (3), bringing

worldwide attention to the medicinal benefits of CBD, and making Charlotte a key figure in the medical cannabis movement. Sanjay Gupta, in a statement shortly after Charlotte’s passing said (4), ―Charlotte changed the world. She certainly changed my world and my mind. She opened my eyes to the possibility of cannabis being a legitimate medicine. She showed me that it worked to stop her crippling seizures, and that it was the only thing that worked.‖ The Stanley brothers, who bred and commercialized Charlotte’s Web, the high-CBD/low-THC cannabis strain named after Charlotte, paid tribute to her on their website (5), stating: ―Charlotte was ten feet tall and carried the world on her shoulders. Inspiring is a lacking word, as are courageous and vivacious and strong and beautiful. She was divine. She grew, cultivated by a community, protected by love, demanding that the world witness her suffering so that they might find a solution. [...] Her story built communities, her need built hope, and her legacy will continue to build harmony.‖ Charlotte inspired the medical cannabis revolution by championing the life-changing effects of CBD for patients with seizures and other debilitating illnesses and influenced legislation reform across the United States and worldwide. Congressman Matt Gaetz (R-FL) one of the leading GOP champions for broad marijuana reform on Capitol Hill, provided this statement on Twitter (6): ―Charlotte lived a life of tremendous significance. Her story inspired me to completely change my views on medical cannabis and successfully pass legislation so that patients could get help in Florida.‖ In addition to championing that CBD bill in Florida, a bipartisan congressional bill, ―The Charlotte’s Web Medical Access Act‖ was named after her and introduced in 2015 (7).

Charlotte’s Influence For this special edition of ―Cannabis Crossroads,‖ I want to honor Charlotte not by reciting the legislation that she impacted, but by taking a closer look at the amazing influence and inspiration that she provided to so many pediatric cannabis families. These families became pediatric cannabis trailblazers themselves, forging ahead from inspiration drawn from Charlotte. (See upper right for Figure 1, click to enlarge.) Gail Rand, whose son Logan was diagnosed with epilepsy at age one and ADHD at age two, spoke about how she first heard about Charlotte in June 2012 and her life was forever changed. ―Her family’s willingness to share their story in a time where the stigma around medical cannabis was huge and the Federal government was still raiding many facilities was so incredibly brave,‖ said Rand. ―Her mom, Paige, shared Charlotte’s story with Maryland legislators and regulators in a way that was incredibly powerful. They got to ask questions and learn that this is real medicine that could have a massive impact on Marylanders. For me, it is personal-my son is seizure free and his life was saved because of Charlotte. I know she is a daughter, sister, and friend, but to millions of people she is an icon of change and I’m forever grateful.‖ I also spoke with Tracy Ryan, founder of CannaKids and Saving Sophie. Tracy’s daughter Sophie was diagnosed with an optic pathway glioma brain tumor at eight months old. Ryan explained that it was the documentary, Weed 1, that not only changed the course of medical cannabis forever, but also gave family’s like hers legitimacy in their choice to use this plant on their small children. ―It quelled judgement and paved a way that only a story like Charlotte’s could do. When Sophie was honored with being one of the first 20 children in California to receive Charlotte’s Web oil over 6 years ago, we forever

felt connected to this family that we owe so much to,‖ said Ryan. ―Our entire industry, worldwide, has this little girl’s struggles to thank for advancements in the understanding of this plant that could otherwise have taken a decade. God gave Charlotte a very important mission to fulfill, and she not only did so, but saved countless lives globally and forever changed the world leaving it a healthier and happier place than when she entered it.‖ I also spoke with mother and daughter powerhouses, Janie and Rylie Maedler, about how Charlotte influenced them. Janie’s daughter Rylie was diagnosed with an extremely rare, aggressive giant cell granuloma in 2013. ―Charlotte Figi was and still is an inspiration to me,‖ said Rylie Maedler. ―Charlotte was living proof that cannabis can help treat those with severe untreatable epilepsy. She helped introduce cannabis as a treatment for pediatrics. Although Charlotte may not physically be with us now; what she accomplished will always be with us.‖ ―I know speaking as a mom that in your gut, you know what has to be done to save your child and this does away with fear,‖ said Janie Maedler. ―Paige and Charlotte have given parents the strength they have needed to follow that gut. I’m forever thankful to them for this.‖ Finally, I spoke to Moriah Barnhart whose daughter, Dahlia, was diagnosed with aggressive brain cancer at the age of two. ―Sharing your child’s story in order to change public perception can come with stifling political, social, and psychological repercussions,‖ said Barnhart. ―To embrace those repercussions while dealing with the daily struggles of caring for a severely special needs child takes unimaginable bravery and resilience. The life and death of Charlotte Figi embody the idea that suffering should not be in vain.‖

Conclusion In closing, I would like to share my deepest appreciation and gratitude to Charlotte and her family for advocating for their daughter’s healthcare and access to medicinal cannabis as a treatment, not only for their family, but for families all around the world. Charlotte has had a massive impact on the global cannabis community that will never be forgotten. The Figi’s are true pioneers that have opened so many minds about cannabis treatments with their advocacy and story sharing. Colorado Governor Jared Polis proclaimed April 7th, ―Charlotte Figi Day‖ in Colorado. The loss of Charlotte Figi was felt around the world and she will be greatly missed by our entire community, but her legacy will live on forever and will continue to inspire many more individuals for years to come. References 1. https://www.cnn.com/2013/08/08/health/gupta-changed-mind- CBD and Horses: Paving the Path for Future Research May 1, 2020 Madeline Colli Cannabis Science and Technology, May 2020 , Currently, little information exists on the calming and painkilling effects on animals using cannabidiol (CBD). Dr. Kimberly Guay, an Associate Professor at Tarleton State University in Stephenville, Texas, is hoping to change that by conducting research on the effects of CBD in horses and other livestock.

Currently, little information exists on the calming and painkilling effects on animals using cannabidiol (CBD). Dr. Kimberly Guay, an Associate Professor at Tarleton State University in Stephenville, Texas, is hoping to change that by conducting research on the effects of CBD on horses and other livestock. Dr. Guay and her group hope that their future findings can benefit animals during transport and other times of stress. Here, Dr. Guay discusses her interest in this area, some of the current research, and her plans for the future. What enticed you to research the effects of cannabidiol (CBD) on horses? Dr. Kimberly Guay: My area of research details evaluating methods of monitoring and improving well-being in both companion animals, as well as livestock. Many of my students were already using CBD to help ease anxious behavior in their horses. CBD products for both horses and dogs have been available online for quite some time, so it seemed like a logical place to start. Can you tell us more about the science behind your study (how were the horses selected, how frequently are they given treatment, what dose, how are the monitored, and so forth)? Guay: We have multiple ongoing projects: An acute dose study-which is more controlled in that we use university horses, dose them once and monitor how quickly the CBD is detected in their system, as well as how long it took to metabolize it. Another single dose study we are performing measures a change of behavior (such as anxious or stereotypic behavior) after a single dose. We are researching several daily dose studies that will document any changes in behavior, immune response, and stress indices over time. Lastly, we will be doing several transport studies that will evaluate how CBD (both single and daily doses) may help mediate the stress of transport.

What form of CBD have you found to produce the best results? Guay: Currently, it is too early to tell what methods deliver the best results. How were the CBD products chosen? Guay: Most CBD companies are very confident in their products, we have many who are eager to let us analyze their products. Have you done quality control testing on the products? Guay: Although it is evident that there are differences to products from different origins, we require the companies to send certificates of analysis for each batch of product that we use. Also, we only use one product in one project at a time so as not to confound the results. Has the stigma around cannabis affected your work at all? Guay: You mean aside from the jokes? Yes. Primarily, people are very interested and open to the potential of what CBD can offer, however some of my student workers have received hang-ups when we call looking for potential subject horses for studies. What results has your research revealed so far? Guay: Unfortunately, it is still too early to tell. We do anticipate releasing our initial findings in early 2021. Do you think that CBD has the same effect on animals as it does on humans? Does that imply that animals also have an endocannabinoid system? Guay: Most animals have an endocannabinoid system, where all of the receptors are still unknown. I expect much research to be seen in the near future that will verify and document similarities and unique differences of cannabinoid receptors in various species. How will your research affect future CBD studies with animals? Guay: Firstly, I hope to gain insight as to if, and how, cannabinoids

and industrial hemp may be utilized to increase human and animal well-being. Then with my work, provide other researchers direction to focus on further studies. What are the next steps in your CBD research? Do you plan to study any other cannabinoids as treatment options for animals? Guay: Yes, we have multiple projects in the pipeline, but we are also planning to work with multiple species-not just horses. There is potential for work in transport and weaning stress in livestock species such as pigs and cattle. I am also very interested in the potential benefits of cannabinoids in wild-captive animals. How to Cite this Article M. Colli, Cannabis Science and Technology3(4), 30-31 (2020).