Cbga And CBD Oil

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What is CBD? What is CBDA? Learn CBDA benefits, CBD benefits and how to look for the best CBDA oil before you buy. Learn the difference between CBD vs CBDA. Cannabigerolic Acid CBGA, in turn, is oxidocyclized by flavin adenine dinucleotide-dependent oxidases, namely, cannabichromenic acid (CBCA) synthase, cannabidiolic acid (CBDA) synthase, and Positive effect of some cannabinoids in the treatment and prophylaxis of a wide variety of oxidation-associated diseases and growing popularity of supplements containing cannabinoids, mainly cannabinoid oils (e.g. CBD oil, CBG oil), in the self-medication of humans cause a growing interest in the an …

Everything You Need to Know about CBDA.

CBD vs CBDA: Surely you’ve heard of CBD. But now we have CBDA? What is CBDA? CBDA benefits, are there any? Also, what is the best CBDA oil out there? You’ve likely seen CBD products on shelves in stores near you. CBD is everywhere. Now, CBDA and other cannabinoids are emerging on the health supplement scene.

No, CBDA is not a typo. Also, no: CBDA is not interchangeable with CBD. They’re different compounds, although they do have a close relationship.

In this post, we’re going to take a deep dive into CBDA and tell you everything you need to know about CBDA and even CBD before you buy a bottle. We will go over CBDA benefits as well as CBD benefits and more.

But before we talk about what CBDA is, let’s give a quick overview of the more popular cannabinoids, THC, and CBD.

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A Quick Overview Of Cannabinoids

To get to CBDA, we must first discuss the most well-known cannabinoids on the health and wellness scene:

THC and CBD.

Actually, let’s back it up even a step further —

What is a cannabinoid?

A cannabinoid is a naturally occurring compound found in the cannabis plant. There are currently over 110 known cannabinoids, with more likely to be discovered as studies continue. Cannabis really is a remarkable plant!

These cannabinoids are what causes the effects of marijuana (THC), CBD, CBDA, and other cannabinoid products users have come to enjoy.

But how?

Cannabinoids interact with the human body through receptors in the endocannabinoid system or ECS. The ECS is actually a fairly recently-discovered system that regulates our homeostasis and health functions like mood, anxiety, stress, sleep, inflammation, pain, and more.

Within this ECS, there are two primary receptors:

CBDA vs CBD: Raw CBDA hemp growing on MONTKUSH Farms.

CB1 and CB2.

While CB1 primarily binds to the brain and nervous system, the CB2 receptor mainly interacts with our immune system. Our body naturally produces its own endocannabinoids, but when they are out of balance, we feel things like anxiety, stress, other mood disorders and may even have trouble sleeping.

Cannabinoids from the cannabis plant like THC, CBD, CBDA, and others can mimic our body’s natural endocannabinoids. They bind to our CB1 and CB2 receptors to produce their many potential health benefits.

The various cannabinoids produce different benefits because they each have unique characteristics and bind to our receptors differently. Often, cannabinoids work even better in tandem with other cannabinoids and compounds, a phenomenon is known as the entourage effect.

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Why Are THC & CBD More Popular?

Mainly because more is known about them, research on CBDA is still in its very early stages, and it is more difficult to extract CBDA than THC or CBD (more on that later).

THC has been used medicinally and recreationally for centuries, while CBD has recently burst onto the scene.

Despite its recent boom and popularity, a stigma still exists about CBD. This is due to its relationship with THC and marijuana. But despite sharing an origin, CBD and THC perform different functions.

THC, or tetrahydrocannabinol, is the psychoactive compound found in Cannabis Sativa. In short, THC is the compound that gets you high. THC is most often consumed by smoking strains of marijuana that are rich in THC, but you can also find it in oils, tinctures, capsules or softgels, and edibles like a CBD gummy.

On to CBD.

Meanwhile, CBD, or cannabidiol, is found in the same plant but is actually non-psychoactive, meaning CBD will not get you high while producing its therapeutic benefits. For this reason, it has become popular. CBD is often extracted from CBD-rich strains of cannabis, like hemp. It is typically seen in oils, tinctures, supplements, extracts, food and beverage products, gummies, and seemingly everything.

CBD products that have high THC content come from marijuana plants, while the majority of CBD products you see on the market (and the only ones technically allowed by the FDA) are those that have a THC content of 0.3% or less.

So what about CBDA? Where does it fit into the crowded cannabinoid market?

What Is CBDA?

All cannabinoids in cannabis and hemp come from cannabigerolic acid (CBGa), the mother of all cannabinoids.

Plant enzymes then convert the CBGa into a combination of the three major cannabinoid precursor compounds: tetrahydrocannabinolic acid (THCA), cannabichromenic acid (CBCA), and cannabidiolic acid (CBDA). The combination will depend on the unique cannabis strain they are derived from.

CBDA is a non-psychoactive compound that serves as a precursor to CBD. More specifically, CBDA is decarboxylated to create CBD, meaning it is heated. This can happen instantly if smoked or vaped, or slowly if the plant material is left to dry in the sun or even at room temperature.

Cannabidiolic acid can, therefore, be thought of as raw CBD. CBDA is most often found in the live or raw hemp plants bred for high CBD levels.

Cannabidiolic acid is often consumed as CBD, but it can also be beneficial in its raw form. CBDA oil can be consumed or absorbed via tinctures, raw cannabis juice, topical creams, and raw CBDA oil.

To date, CBDA has attracted much less public attention and is seen less on the market. However, the raw juicing cannabis trend is bringing CBDA into the spotlight. People are wondering about the differences between CBD and CBDA and if CBDA is better than CBD.

So is it?

Let’s take a look.

The Differences Between CBDA & CBD

The major difference between CBDA and CBD is actually the amount of heat applied to the substance.

As we mentioned, the main difference is that CBDA is a precursor to CBD. You create CBD by heating CBDA or raw CBD.

While research in CBDA is in much earlier stages than even CBD, we know that they share some similarities.

First,

They are both non-psychoactive, meaning users won’t get high or stoned. This is because CBD and CBDA both do not directly interact with our endocannabinoid receptors.

Second,

CBD and CBDA are thought to cause their signature effects by activating our 5-HT1A serotonin receptors. You’ve likely heard of serotonin, as it is a vital neurotransmitter in our brains that is closely involved in regulating our mood, sleep, anxiety, and even nausea.

CBD is noted to help with things like anxiety, depression, inflammation, and even a rare form of childhood epilepsy. You see CBD in such a wide variety of health supplements because of its versatility and because it has been studied much more than CBDA.

CBDA Research

Meanwhile, CBDA is in the very early stages of research but shows just as much, if not more, promise than CBD. For example, one study found that CBDA produces antidepressant effects on rats at doses 10 to 100 times lower than CBD.

This is due to CBDA’s relationship with serotonin receptors. For this same reason, CBDA is further researched as an anti-nausea drug, with one study finding CBDA more effective at reducing nausea than CBD.

UPDATE 1/12/2022: An article was written on Forbes.com “Study Finds Cannabis Compounds Prevent Infection By Covid-19 Virus” shares some new, interesting findings of CBDA and mentions – “…researchers found that two cannabinoid acids commonly found in hemp varietals of cannabis, cannabigerolic acid, or CBGA, and cannabidiolic acid, also known as researchers found that two cannabinoid acids commonly found in hemp varietals of cannabis, cannabigerolic acid, or CBGA, and cannabidiolic acid, also known as CBDA, can bind to the spike protein of SARS-CoV-2, the virus that causes Covid-19. By binding to the spike protein, the compounds can prevent the virus from entering cells and causing infection, potentially offering new avenues to prevent and treat the disease.” The article goes on to say – “Orally bioavailable and with a long history of safe human use, these cannabinoids, isolated or in hemp extracts, have the potential to prevent as well as treat infection by SARS-CoV-2,” the researchers wrote in an abstract of the study.”

Raw CBDA Oil now available.

What Are The Potential Benefits?

The reason that cannabinoids like THC and CBD have been studied so much more than CBDA is because CBDA and other acidic forms of cannabinoids are not considered to be pharmacologically active. This means that they don’t affect the ECS in the same way that their decarboxylated forms do. Therefore, most research has focused on CBD instead of CBDA.

Recent CBDA Research

But recent research shows that raw CBDA oil has its own unique potential. One study showed that CBDA could act as an effective anti-inflammatory agent.

In this study, CBDA was found to be more proficient than THC at blocking COX-2, an enzyme produced when inflammation is present. The same study found that the acidic component of CBDA plays a vital role in its ability to inhibit COX-2.

Another study found that CBDA was a thousand times more powerful than CBD for anti-nausea and anti-anxiety effects. In this study on animal models, CBDA displayed “significantly greater potency at inhibiting vomiting in shrews and nausea in rats” when compared to CBD.

Part of the issue with CBDA when it comes to potential medical use is that it is an unstable compound. This is evident when you consider that it gradually decarboxylates, even just at room temperature. However, Dr. Raphael Mechoulam, the cannabis scientist who first synthesized THC and CBD, said at the 2019 CannMed conference that his research team discovered a way to transform unstable CBDA into a more stable compound.

Additionally, CBDA appears to share most of the benefits CBD users seek, such as anti-anxiety properties and more.

Is CBDA Better Than CBD?

Well, until we have concrete evidence from human testing, it’s too early to say! However, early research suggests that raw CBD oil can be just as, if not more, effective at treating things like depression, nausea, and inflammation. It may also have special properties that CBD does not, making it an option for different treatments.

CBDA shows promise as an anticonvulsant and may even have antibacterial, antioxidant, and cancer-preventing potential (specifically breast cancer). So while “better” is a hard judgment to come by, there is enough early evidence to suggest that CBDA has its own distinct qualities that may set it apart from CBD in certain areas.

Summary

As you can see, CBDA does indeed have its own unique potential in the growing cannabinoid industry. While research into much of CBDA’s potential is still in the early stages, enough evidence gives scientists and avid supplement users great optimism.

For those of you who are just dipping your toes into the CBD/health supplement waters, CBDA may indeed offer another good option to add to your health and wellness routine. CBDA is thought to be risk-free and comes without any high or potential risks related to THC (such as if your company drug tests).

Raw CBDA might be what you need to optimize your wellness routine!

Raw CBDA Oil now available.

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CBDA vs CBD: Major Differences You Should Know

When you first think about cannabis derivatives, you probably think of two different components: THC and CBD. Most people are familiar with how both of these substances work. THC produces the “high” associated with cannabis use, although it also has some therapeutic applications. CBD, on the other hand, produces no “high” and has its own separate benefits.

You might not be aware that there are more than a hundred other compounds in the cannabis plant. One of these is Cannabidiolic acid, also known as CBDA. So, what’s the difference between CBDA vs. CBD? Like CBD, CBDA is found naturally in the cannabis plant, both in its hemp and marijuana variations. In fact, CBDA is the precursor chemical that produces CBD.

Even though these are two very similar compounds, important differences between them need to be discussed. Here, we’ll explain why you might want to use CBDA vs. CBD, as well as what benefits you can gain from both compounds. Let’s take a closer look!

What is Cannabidiolic acid?

Cannabidiolic acid, or CBDA for short, is a natural compound that the cannabis plant produces. CBDA is a relatively recent discovery and was not even isolated until 1996. It’s primarily found in raw, unprocessed cannabis plants.

The main difference between CBDA and CBD is that CBDA is a precursor chemical to Cannabidiol (CBD), similar but not acidic. What that means is that it’s a natural compound found in the raw plant. As with many other cannabinoids, CBDA undergoes a transformation when cannabis is processed. When the plant is heated, cured, or dried, acidic compounds break down into new chemicals. This is the process that produces large amounts of CBD from CBDA.

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In raw cannabis, by contrast, you’ll find a large quantity of CBDA. You can also find it in most raw hemp, unprocessed cannabis products. For instance, raw hemp oil is rich in CBDA. This is also true for regular CBD oils that haven’t been filtered or otherwise processed.

Is it the Same as CBD?

CBD and CBDA are both cannabinoids. This is a class of chemicals that are found in the cannabis plant. In fact, they’re also found in the human body and are used by the endocannabinoid system to regulate normal bodily functions. This is why cannabinoids are often considered healthy dietary supplements. In total, the cannabis plant contains at least 113 cannabinoids. However, the exact concentration will depend on the type of plant and whether or not the plant has been decarboxylated.

What is Decarboxylation?

Decarboxylation sounds like a complex process, so let’s break it down as simply as possible. As we mentioned, a major difference between CBDA vs. CBD is that CBDA is an acid. This is because there’s a chemical chain called “carboxyl” attached to the molecule. When the cannabis is left to cure, the carboxyl chain will slowly break down. When this happens, CBDA and other acidic cannabinoids lose their acidity. They also become more active. Decarboxylation can be accelerated by other processes, like heating in particular.

In live, growing cannabis plants, CBD is only found in relatively small quantities. By drying or heating the plant, producers can convert the CBDA into CBD. However, one thing is the same between these two compounds: both of them are non-intoxicating. Neither one will get you “high,” which means they’re safe to take before work or even before driving.

The Endocannabinoid System

Cannabinoids all affect your body through your natural endocannabinoid system. This system activates neurotransmitters, which can help moderate your brain function. It also helps regulate many other bodily systems. However, the effects of CBDA vs. CBD are different since CBD is active and CBDA is not.

What Benefits Does CBDA Offer?

Okay, we’ve talked a bit about the science of how CBDA works. But what does it actually do when you ingest it? The short answer is that we know a lot less about the effects of CBDA compared to the effects of CBD. The reason is that CBD has been studied for many years, while scientists are only just beginning to study the effects of CBDA.

Why the lack of study? The simple answer is that scientists didn’t think that cannabinoids had any effect in their acidic, non-active form until very recently. As a result, they didn’t bother to run many studies on CBDA and other inactive cannabinoids.

Back to 2008

All of this started to change in 2008. That year, some researchers noticed that CBDA had a very similar structure to commonly-used non-steroidal anti-inflammatory drugs (NSAIDs). This class of drugs includes everyday painkillers such as acetaminophen and ibuprofen. These researchers followed up on their discoveries by investigating whether CBDA actually worked the same way as common NSAIDs. Amazingly, they found that CBDA inhibits COX-2, the same body chemical that’s inhibited by NSAIDs.

Can Help with Inflammation and More

However, CBDA isn’t just used to reduce inflammation. You can also use it to treat nausea and anxiety. In this case, there’s a strong similarity between CBDA vs. CBD. In fact, CBDA is actually more effective. It’s more than a thousand times more effective than CBD in activating a serotonin receptor that reduces nausea and anxiety. This effect is even more pronounced in chemotherapy patients using ondansetron (OND) to treat nausea.

Furthermore, CBDA doesn’t act directly on the endocannabinoid system. Instead, by inhibiting COX-2 activity, it has an indirect effect. It also acts on 5-HT receptors, which influence serotonin production. Serotonin production is essential to overall health, including reduced anxiety and general well-being.

Cannabigerolic Acid

CBGA, in turn, is oxidocyclized by flavin adenine dinucleotide-dependent oxidases, namely, cannabichromenic acid (CBCA) synthase, cannabidiolic acid (CBDA) synthase, and Δ9-THCA synthase, producing CBCA, CBDA, and Δ9-THCA, respectively.

Related terms:

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Analysis of Cannabis

Cannabigerol (CBG)

Cannabigerol comes from cannabigerolic acid (CBGA) after decarboxylation via heat or light. The protonated molecule [M + H] + is 317.2475. The fragmentation MS-MS spectra of cannabigerol is probably the simplest and most straightforward of all the cannabinoids studied here. As seen in Fig. 13 , only two major fragments are observed: the ion at m/z 193.1215 (F1), which is common to all the other major cannabinoids and corresponds to the olivetol moiety with the ortho methyl group, and the ion at m/z 123.0438 (F2), which again corresponds to the resorcinol moiety with the ortho methyl group. A smaller ion is seen at m/z 207.1369, which probably corresponds to the breakage of the isoprenoid moiety between atoms C1-C2.

Fig. 13 . MS-MS spectrum for cannabigerol (CBG) at 30 V.

Cannabidiol in Refractory Epilepsy

Phytocannabinoids: Biosynthetic Pathway

Around 113 phytocannabinoids are identified in C. sativa plant [27] . These cannabinoids are biosynthesized as carboxylic acid, with the precursor being cannabigerolic acid 1 abbreviated as CBGA ( Fig. 14.1 ). The synthesis of CBGA is an enzymatic reaction that involves an alkylation of olivetolic acid (alkylresorcinolic acid responsible for the polyketide nucleus of the cannabinoids) with the help of geranyl pyrophosphate [28] . The CBGA formed is sequentially converted into tetrahydrocannabinolic acid 2 (THCA), cannabidioloic acid (CBDA) 3, and cannabichromenic acid (CBCA) 4 with the help of synthase enzymes (THCA synthase, CBDA synthase, and CBCA synthase) [27] . These cannabinoids, especially THCA, secrete in the storage cavity of glandular trichomes [28] and are responsible for the self-defense mechanism of the cannabis plant with its insect repellant, absorption of ultraviolet radiation, and cell death properties [28] . These acids are decarboxylated into their neutral homologous (THC, cannabidiol, and cannabichromene) by a nonenzymatic reaction involving heat or light, such as during storage or smoking ( Fig. 14.1 ). The two important neutral cannabinoids are THC 5 and cannabidiol 6. Cannabidiol has both neuroprotective and antiseizure properties. A THCA synthase polymorphic gene determines the fate of cannabidiol/THC ratio in plants. A single locus (B) that possesses two codominant alleles B(T) and B(D) controls these genes. Plants expressing the homozygous condition of B(T)/B(T) have predominant THC phenotype, while homozygous of B(D)/B(D) has predominant cannabidiol phenotype. The heterozygous condition of B(T)/B(D) has the intermediate phenotype situation [29] . Besides cannabinoids, the plant also has other ingredients such as terpenes and phenolic compounds. Both mono- and sesquiterpenes present in the plant are synthesized in the same glandular trichomes (where cannabinoids are generated) and participate equally toward identifying cannabis species. Moreover, around 20 different flavonoids are identified in cannabis plants. Both terpenes and polyphenolic compounds (such as flavonoids) may contribute toward the medicinal properties of this plant [30] .

Fig. 14.1 . Biosynthesis of cannabinoids.

Cannabis preparation containing the higher concentration of cannabidiol and lower levels of THC is desirable in clinics for treating seizures. With this background, the plant named “avidekel” has been cultivated [31] . This plant has a high amount of cannabidiol (about 16.3%) with a very low amount of THC (about 0.8%). Horticultural practices and differences in the breeding condition may result in plant varieties with varying contents of THC and cannabidiol [32] . Plants cultivated with less light and more tightly grown plantations have more cannabidiol contents, while plants grown with more light and at a greater distance between samplings have more of THC content [32] . Also, cannabidiol hemp oil available in the market has a high quantity of cannabidiol and lower levels of THC contents and could be useful therapeutically.

Biosynthesis and Pharmacology of Phytocannabinoids and Related Chemical Constituents

Brian F. Thomas , Mahmoud A. ElSohly , in The Analytical Chemistry of Cannabis , 2016

Phytocannabinoid Constituents in Cannabis

Phytocannabinoids are a structurally diverse class of naturally occurring chemical constituents in the genus Cannabis (Cannabaceae). This chemical classification is broadly based on their derivation from a common C21 precursor ( cannabigerolic acid , 4 CBGA), or its C19 analog (cannabigerovaric acid, 5 CBGVA), the predominate phytocannabinoid precursors formed through the reaction of geranyl pyrophosphate with olivetolic and divarinic acid, respectively ( Fig. 2.1 ).

Figure 2.1 . Biosynthesis of phytocannabinoids.

Enzymatic conversion of cannabigerolic and cannabidivaric acid produces a wide variety of C21 terpenophenolics, 6 including (−)-trans-Δ 9 -tetrahydrocannabinol (Δ 9 -THC), cannabigerol (CBG), cannabichromene (CBC), cannabicyclol (CBL), cannabidiol (CBD), cannabinodiol (CBND), and cannabinol (CBN), and their C19 homologs Δ 9 -tetrahydrocannabivarin (Δ 9 -THCV), cannabivarin (CBV), and cannabidivarin (CBDV). More than 100 phytocannabinoids across 11 chemical classes have been isolated and identified to date. 7 In the growing Cannabis sativa plant, most of these cannabinoids are initially formed as carboxylic acids (eg, Δ 9 -THCA, CBDA, CBCA, and Δ 9 -THCVA) that are decarboxylated to their corresponding neutral forms as a consequence of drying, heating, combustion, or aging ( Fig. 2.2 ). There are also different isomers of phytocannabinoids resulting from variations or isomerization in the position of the double bond in the alicyclic carbon ring (eg, (−)-trans-Δ 8 -THC). It is important to note that CBN is not formed biosynthetically, but is an oxidative degradant of Δ 9 -THC. 2

Figure 2.2 . Primary phytocannabinoid constituents in cannabis.

Regulation of cannabinoid content in each plant phenotype (chemotype) has been proposed to involve genetic control of the expression of a variety of synthetic enzymes by four independent loci. 8 Qualitatively, the cannabinoid chemotype is controlled by the variation in expression of these phytocannabinoid synthetic enzymes, resulting in progenies and populations that have discrete distributions of chemical composition (ie, chemical ratios of phytocannabinoids, such as the Δ 9 -THC/CBD ratio). Quantitatively, the phytocannabinoid content is controlled by polygenic mechanisms, and is strongly influenced by environmental factors, such that a Gaussian distribution of total cannabinoid content is typically observed. In addition, the cannabinoid content and profile changes over time as the plant grows, matures, and ages. 9 Wild-type chemotypes can therefore differ between Δ 9 -THCA predominance and CBDA predominance in discrete populations, but vary dramatically in total cannabinoid content, with clones of both types reaching total cannabinoid content levels of up to 25–30% (w/w) of the dry and trimmed inflorescences. Spontaneous mutations and selective breeding have produced unique chemotypes that show CBGA-, 10 CBCA-, 11 or Δ 9 -THCVA predominance, 8 as well as cannabinoid-free chemotypes. 12 Selective breeding has produced hundreds of strains that differ in appearance and chemical composition, and patients and recreational users often prefer specific strains for their purported ability to produce specific pharmacological effects. De Meijer speculates that future breeding might produce novel terpenophenolic compounds such as those with branched alkyl or aromatic side chains, or chemotypes with increased ratios of currently minor constituents such as methyl, butyl, or farnesyl cannabinoids. 8

The current variation in phytocannabinoid content across and within chemotypes has important implications in medicinal cannabis and cannabis-based formulations and dosing. This has become increasingly apparent and can be recognized by the plethora of varieties of cannabis being cultivated, manufactured, and marketed as dosing formulations in the medicinal and recreational market. Similarly, the nonphytocannabinoid composition of cannabis is receiving increasing pharmacological attention, particularly terpenoids and flavonoids. 13

Analysis of Cannabis

2 Biosynthesis of phytocannabinoids

Through two polyketide pathways, fatty acids and coenzymes synthesize olivetolic acid and divarinolic acid. These acids react with the substrate geranyl pyrophosphate and geranyl-diphosphate:olivetolate geranyltransferase synthesize cannabigerovarinic acid (CBGVA) and cannabigerolic acid (CBGA), respectively. The Cannabis spp. genome further encodes for THCA synthase, cannabidiolic acid (CBDA) synthase, and cannabichromenic acid CBCA synthase which react with CBGVA and CBGA to synthesize six acid phytocannabinoids as shown in Fig. 1 [5] .

Fig. 1 . Phytocannabinoid acid synthesis pathways from CBGVA and CBGA.

The acid phytocannabinoids decarboxylate to the neutral compounds after harvest and upon exposure to heat and light. An example of this is shown in Fig. 2 for THCA to THC + CO2 decarboxylation. Therefore, in the living plant, THC, CBD, cannabigerol (CBG), or the other decarboxylated neutrals are not present in large concentrations. Other processes such as photo-irradiation converts CBCA and cannabichromene (CBC) to cannabicyclolic acid (CBLA) and cannabicyclol (CBL), respectively, and like other acid phytocannabinoids, CBCA decarboxylates to create a secondary pathway to CBL. Isomerization of THC to △ 8 -tetrahyrocannabinol (△ 8 -THC) can also occur, and oxidative aromatization transforms THC into cannabinol (CBN) which can be photochemically rearranged into cannabinodiol (CBND). Photo-oxidation and exposure to heat of CBDA transform it into cannabielsolic acid A (CBEA-A) which decarboxylates into cannabielsoin (CBE). Photo-oxidation of CBD also transforms into CBE with exposure to heat [6] . This is just a fraction of the potential number of cannabinoids that can be found in cannabis plants.

Fig. 2 . THCA decarboxylation to neutral THC.

Development & Modification of Bioactivity

Arno Hazekamp , . Renee L. Ruhaak , in Comprehensive Natural Products II , 2010

3.24.2.2 Biosynthesis of the Cannabinoids

The cannabinoids most commonly detected in herbal Cannabis materials are shown in Figure 4 . For the chemical numbering of cannabinoids, five different nomenclature systems have been used so far, 43 but the most commonly used system nowadays is the dibenzopyran numbering, which is also adopted by Chemical Abstracts. In Europe, the monoterpenoid system based on p-cymene has also been widely used. As a result, the major cannabinoid delta-9-THC is sometimes described as delta-1-THC in older manuscripts. In this chapter, the dibenzopyran numbering is consistently used, therefore THC is fully described as (−)-trans-Δ 9 -tetrahydrocannabinol ( Figure 5 ).

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Figure 4 . Structures of the cannabinoids most commonly found in Cannabis plant materials. All cannabinoids have the (6aR,10aR)-orientation, according to the chemical numbering shown in Figure 5 .

Figure 5 . Two most commonly used numbering systems for the cannabinoids. The dibenzopyran system is used in this chapter.

It is commonly thought that cannabinoids are unique compounds only found in Cannabis. However, some exceptions exist in the plant kingdom: In Helichrysum umbraculigerum Less., a species from the family Compositae, the presence of CBGA, cannabigerol (CBG), and analogues to CBG was reported. 44 Moreover, in liverworts from Radula species the isolation of geranylated bibenzyls analogous to CBG was reported, 45 suggesting the homology of genes from the cannabinoid pathway in at least some other species.

3.24.2.2.1 The acidic cannabinoids

In all biosynthetic pathways for cannabinoids that were postulated until 1964, CBD or cannabidiolic acid (CBDA) was regarded as the key intermediate, which was supposedly built from a monoterpene and olivetol or olivetolic acid (OA), respectively. However, Gaoni and Mechoulam 46 showed that CBG is the common precursor of cannabinoids, biosynthesized through the condensation of geranyldiphosphate and olivetol or OA. Subsequently, they concluded that CBD, THC, and CBN all derive from CBG and differ mainly in the way this precursor is cyclized. 47–50 A further improvement of our understanding of cannabinoid biosynthesis came when Shoyama et al. 51,52 concluded that neither the free phenolic (noncarboxylic acid) forms of the cannabinoids nor cannabinolic acid (CBNA) were produced by the living plant. Instead, they postulated a biosynthetic pathway based on geraniol and a polyketoacid, resulting in the production of the acidic cannabinoids. The same conclusion was reached by Turner and Hadley 53 after the study of African Cannabis types.

It is now known that cannabinoids are produced by the metabolism of the plant in the form of carboxylic acids, where the substituent at position 2 is a carboxyl moiety (–COOH). 52 Incorporation studies with 13 C-labeled glucose have confirmed that geranyl diphosphate (GPP) and OA are specific intermediates in the biosynthesis of cannabinoids. 54,55 The first specific biosynthetic step is the condensation of GPP with OA into CBGA, catalyzed by the prenylase enzyme geranyldiphosphate:olivetolate-geranyltransferase (GOT). 54 Furthermore, biosynthetic pathways finally became clear by identification and subsequent cloning of the genes responsible for the conversion of CBGA to THCA, CBDA, and cannabichromenic acid (CBCA), respectively. 56–58 Further oxidation of THCA leads to the formation of CBNA, which is still formed after the plant material is harvested and high levels could be due to poor storage conditions ( Figure 6 ).

Figure 6 . General overview of the biosynthesis of cannabinoids and putative routes. Reproduced with permission from I. J. Flores-Sanchez; R. Verpoorte, Phytochem. Rev. 2008, 7, 615–639.

The terpenoid GPP is derived from the deoxyxylulose phosphate/methyl-erythritol phosphate (DOXP/MEP) pathway. 55,59 Not much is known about the biosynthesis of OA yet, but it has been proposed that a polyketide synthase (PKS) could be involved. 59 However, a PKS specifically yielding OA has not been found to date. Interestingly, OA itself has never been isolated from the plant material, possibly indicating it to be a very short-lived intermediate.

Mahlberg and Kim 60 reported that glandular trichomes are exclusively specialized to synthesize high amounts of cannabinoids and that other tissues contain only very low amounts. These authors distinguished three types of glandular trichomes in Cannabis, with different localization, morphology, and cannabinoid content. Cannabinoids are deposited in the noncellular, secretory cavity of glandular trichomes. However, after confirming the presence of the central precursor CBGA, as well as THC synthase activity in the secretory cavity, it was suggested that this is not only the site of cannabinoid accumulation, but also the site of cannabinoid biosynthesis. 35

3.24.2.2.2 Occurrence of short-chain cannabinoids and other homologues

Most commonly, the acidic cannabinoids produced by plant metabolism contain a pentyl side chain, derived from the OA moiety. Cannabinoids with propyl side chains result if GPP condenses with divarinic acid instead of OA, into cannabigerovarinic acid (CBGVA). The three known cannabinoid synthase enzymes are not selective for the length of the alkyl side chain, and will convert CBGVA into the propyl homologues of CBDA, THCA, and CBCA. 61 All chain lengths from –methyl to –pentyl have been found in naturally occurring cannabinoids, probably all arising from the incorporation of shorter chain homologues of OA. The side chain is important for the affinity, selectivity, and pharmacological potency for the cannabinoids receptors.

Many other minor acidic cannabinoids have been identified over the years, including monomethyl and other types of esters. 38,62 The biosynthetic pathways explaining this variation have been studied. 59

Analysis of Cannabis

James W. Favell , . Matthew Noestheden , in Comprehensive Analytical Chemistry , 2020

3.4 Method performance

For method validation, extracts were fortified at 0.008%, 0.4%, and 4% (w/w; cannabinoid/chocolate) after extraction (vide supra). While this approach is not an ideal way to perform method validation, the cost of infusing chocolates with the phytocannabinoids of interest at levels representative of the concentration ranges in cannabis edibles was prohibitively high. Notwithstanding this, all analytes were quantitated over a simulated range of 0.008–4% (w/w) with quadratic fit and inverse concentration weighting. The same calibration functions and dynamic ranges were evaluated for each day of validation (n = 3), with all correlation coefficients (R 2 ) being > 0.99. Carry-over was determined to be ≤ 0.1% by area (data not shown) for all analytes evaluated after a high calibration sample at 4% (w/w).

3.4.1 Accuracy

For the purposes of this study, recoveries were determined accurate if they varied by less than or equal to ± 20% at the low and mid concentrations and by less than or equal to ± 10% at the high concentration ( Tables 3–5 ). In the dark chocolate matrix ( Table 3 ) all compounds fall within the prescribed region except for CBG and CBN. In milk chocolate ( Table 4 ) it is apparent that the phytocannabinoids were biassed high at the low fortification level. Similarly, recoveries are biassed low at the low fortification level in white chocolate ( Table 5 ) with five cannabinoids (CBDV, CBDVA, CBDA, CBGA, and THCA) outside of the target recoveries.

Table 3 . Method validation summary for cannabinoids in a dark chocolate matrix.

Recovery Conducted at midlevel Method limits (% w/w)
ID Analyte 0.008% (w/w) 0.4% (w/w) 4% (w/w) % RSDr % RSDR MDL MRL
1 CBDV 93.5 96.1 94.0 6.52 15.9 0.005 0.008
2 CBDVA 90.0 92.4 97.8 6.05 15.4 0.003 0.008
3 THCV 102 97.6 97.3 5.69 13.2 0.004 0.008
4 CBD 101 97.4 103 6.29 11.4 0.004 0.008
5 CBG 101 92.8 87.3 6.40 14.9 0.005 0.008
6 CBDA 99.3 91.3 95.8 6.79 12.2 0.003 0.008
7 CBGA 102 92.5 93.9 3.64 9.80 0.003 0.008
8 CBN 113 93.7 89.3 7.63 12.1 0.004 0.008
9 THCVA 104 93.3 91.6 7.66 10.4 0.004 0.008
10 Δ9-THC 101 94.8 91.3 5.96 9.14 0.005 0.008
11 Δ8-THC 96.8 97.6 92.6 6.51 17.6 0.004 0.008
12 CBC 103 94.8 90.4 5.94 10.3 0.004 0.008
13 THCA 92.3 88.7 92.1 6.15 8.64 0.003 0.008
14 CBCA 107 85.4 98.1 6.57 7.63 0.003 0.008

Table 4 . Method validation summary for cannabinoids in a milk chocolate matrix.

Recovery Conducted at mid level Method limits (% w/w)
ID Analyte 0.008% (w/w) 0.4% (w/w) 4% (w/w) % RSDr % RSDR MDL MRL
1 CBDV 112 95.5 98.8 6.60 7.26 0.003 0.008
2 CBDVA 117 92.5 101 6.03 7.05 0.003 0.008
3 THCV 116 95.4 99.0 6.03 7.59 0.003 0.008
4 CBD 116 97.9 99.6 7.05 7.75 0.003 0.008
5 CBG 114 97.8 99.2 6.00 7.44 0.003 0.008
6 CBDA 115 93.8 99.6 5.51 7.27 0.003 0.008
7 CBGA 119 97.9 101 6.79 6.89 0.003 0.008
8 CBN 114 97.6 96.3 6.39 7.76 0.003 0.008
9 THCVA 123 97.3 98.4 5.90 7.44 0.004 0.008
10 Δ9-THC 114 102 97.6 5.97 7.07 0.003 0.008
11 Δ8-THC 115 101 95.0 5.96 7.82 0.003 0.008
12 CBC 109 97.6 97.0 5.63 8.64 0.003 0.008
13 THCA 118 98.0 97.7 9.50 11.5 0.003 0.008
14 CBCA 109 100 103 9.26 10.3 0.002 0.008

Table 5 . Method validation summary for cannabinoids in a white chocolate matrix.

Recovery Conducted at midlevel Method limits (% w/w)
ID Analyte 0.008% (w/w) 0.4% (w/w) 4% (w/w) % RSDr % RSDR MDL MRL
1 CBDV 77.6 94.7 91.2 4.13 12.0 0.006 0.008
2 CBDVA 76.1 92.5 95.8 4.12 10.2 0.004 0.008
3 THCV 86.1 96.6 97.1 3.42 9.60 0.005 0.008
4 CBD 86.1 96.0 104 3.74 8.56 0.005 0.008
5 CBG 84.8 91.9 82.8 3.79 12.9 0.006 0.008
6 CBDA 77.4 91.5 95.3 4.17 10.9 0.005 0.008
7 CBGA 76.0 90.8 88.9 3.18 8.66 0.006 0.008
8 CBN 98.0 92.0 84.7 3.16 10.2 0.005 0.008
9 THCVA 85.0 89.8 83.4 3.49 14.4 0.006 0.008
10 Δ9-THC 86.9 94.9 90.5 4.44 7.08 0.006 0.008
11 Δ8-THC 84.0 97.4 92.1 3.80 16.1 0.006 0.008
12 CBC 86.5 93.8 92.2 5.09 7.59 0.005 0.008
13 THCA 78.0 87.5 89.3 3.55 4.73 0.005 0.008
14 CBCA 93.5 85.7 91.6 4.06 5.24 0.004 0.008
3.4.2 Repeatability and intermediate precision

Repeatability, expressed here as relative standard deviation (RSDr), was determined across five replicate samples prepared at a fortified concentration of 0.4% (w/w). In all three matrices, repeatability was acceptable, with an average RSDr across all compounds of 6.3% in dark chocolate, 6.6% in milk chocolate, and 3.9% in white chocolate ( Tables 3–5 ). Similarly, intermediate precision was assessed as the relative standard deviation (RSDR) of 15 replicates over the course of 3 days (five replicates per matrix per day). As might be expected, RSDR was greater than RSDr. Across all 14 analytes in a given matrix, the average RSDR was found to be 12.1% in dark chocolate, 8.0% in milk chocolate and 9.5% in milk chocolate. For all compounds in all three matrices, both RSDr and RSDR yielded Horwitz ratios (HorRat) [27] between 0.3 and 1.3, indicating suitable method repeatability and intermediate precision.

3.4.3 Method detection limit and method reporting limit

The MDL and MRL were determined in a similar fashion, the primary difference being that the verification of the MDL was conducted across three separate days of analysis while the MRL verification was conducted on a single day. For this method, the successfully verified MRL was 0.008% (w/w) for all analytes in all matrices, whereas MDLs ranged from 0.002% to 0.006% (w/w; Tables 3–5 ).

LC–MS/MS quantitation of phytocannabinoids and their metabolites in biological matrices

Wessam H. Abd-Elsalam , . Raimar Löbenberg , in Talanta , 2019

2.10 Cannabigerol (CBG)

CBG is the decarboxylated form of cannabigerolic acid (CBGA), which is the first biogenetic phytocannabinoid [ 89 ]. It is usually present in the least amount within most analyses, but can be detected in the urine of cannabis users [ 90 ]. CBG has sedative, antibacterial, antiproliferative, and bone-stimulating properties, but clinical studies of CBG in humans are lacking [ 83 ]. CBG can be detected and quantified below the 1.0 ppb level in blood [ 91 ], plasma and urine [ 83 , 88 ], where the lowest LLOD and LLOQ were reported in blood samples (0.1 and 0.2 ppb, respectively).

Cannabis sativa bioactive compounds and their extraction, separation, purification, and identification technologies: An updated review

3.3.4 Supercritical fluid chromatography coupled with mass spectrometry (SFC-MS)

In recent years, SFC-MS has been used as a novel technology to identify cannabinoids in C. sativa plants, but it still requires further improvement. In a study, 11 cannabinoids in C. sativa extracts, including CBL, CBD, Δ 8 -THC, THCV, Δ 9 -THC, CBC, CBN, CBG, THCA-A, CBDA, and CBGA, were well identified by SFC-ESI-MS [ 91 ]. In another study, some cannabinoids, like CBD, Δ 8 -THC, THCV, Δ 9 -THC, CBN, CBG, THCA-A, CBDA, and CBGA, were quantitatively determined from C. sativa plant extracts using ultra-high-performance supercritical fluid chromatography/photodiode array detection-mass spectrometry (UHPSFC/PDA-MS) [ 92 ]. In addition, a study developed a generic SFC-MS method suitable for the analysis of a large panel of substances using seventeen synthetic cannabinoids from multiple classes as model compounds [ 93 ]. Furthermore, SFC-MS seems to be a useful alternative to GC-MS and LC-MS for illegal drug detection, such as cannabinoids [ 94 ].

Synthetic Biology ● Synthetic Biomolecules

Sean Romanowski , Alessandra S. Eustáquio , in Current Opinion in Chemical Biology , 2020

Cannabinoid production and structure diversification in yeast

A recent example of expanding on the artemisinin success is provided by the production of natural and engineered cannabinoids in yeast by Luo et al. [ 19 ]. Luo et al. [ 19 ] started by establishing olivetolic acid production from hexanoyl-CoA and malonyl-CoA by using two previously characterized Cannabis enzymes ( Figure 1 a).

To improve on the endogenous hexanoyl-CoA supply, the authors then introduced a previously reported pathway employing genes from different bacteria. A strain that overproduces geranyl pyrophosphate was then engineered by upregulating the mevalonate pathway to isoprenoids. A suitable prenyltransferase to generate cannabigerolic acid from geranyl pyrophosphate and olivetolic acid was identified by screening 10 candidate enzymes (seven from Cannabis, two from hops, and one from Streptomyces bacteria). Finally, Δ 9 -tetrahydrocannabinolic acid (THCA) was produced by using a Cannabis THCA synthase previously identified. After increasing the copy number of THCA synthase and the two Cannabis olivetolic acid pathway genes ( Figure 1 a), final THCA titers observed were 8 mg/L. For reference, the THCA content of Cannabis sativa plants of chemotype I (drug-type) has been shown to reach up to approximately 80 mg/g in leaves and 200 mg/g in flowers [ 20 ]. Moreover, the authors explored the promiscuity of the pathway toward hexanoyl-CoA analogs to produce derivatives with different substitutions at C-3 of THCA ( Figure 1 a). From the 19 fatty acids fed to yeast strains, five were incorporated (pentanoic, heptanoic, 4-methylhexanoic, 5-hexenoic, and 6-heptynoic acid), including terminal alkene and alkyne derivatives that can be further modified through semisynthesis. As cannabinoids have increasing therapeutic interest [ 21 ], the established synthetic biology route to natural and engineered derivatives may facilitate future investigations of cannabinoids as therapeutics, although yield improvement will be required to reach the gram-per-liter scale required for commercialization.

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Biocatalysis and Biotransformation ● Bioinorganic Chemistry

Anna Fryszkowska , Paul N. Devine , in Current Opinion in Chemical Biology , 2020

Building complexity: C-C, C-N, O-P bond-forming biotransformations

Enzymatic coupling of small molecular fragments can build complexity in a convergent and selective manner. Such reactions can set multiple stereogenic centers while obviating the need for the protection groups or oxidation–state readjustments. Consequently, in recent years, industrial chemists have adopted and developed a number of new enzymatic classes adept at forming C–C, C–N, and O–P linkages ( Schemes 2 and 3 ). Ultimately, access to these diverse biotransformations enables the construction of multienzyme, multistep pathways [ 7 , 8 ], further expanding the structural complexity that can be achieved through biocatalysis ( Scheme 3 ).

Scheme 2 . Building complexity: selected examples of diverse C–C and C–N bond-forming biotransformations used for the synthesis of active pharmaceutical ingredients or their intermediates: (a) Tandem enzymatic process to (m)ethylate coumarin scaffolds with known biological activity catalyzed by an evolved SAM-dependent transferase coupled with a SAM-forming enzyme SalL [ 36 ]. The use of 5′-chloro-5′-deoxyadenosine (CIDA), an analog SAM-precursor, significantly improved the performance and the scope of the C-alkylation; (b) Cyclopropanation catalyzed by a hemoprotein catalyst derived from thermophilic bacterial globins that react with diazoacetone and an unactivated olefin substrate to furnish a cyclopropyl ketone, an intermediate in the synthesis of grazoprevir [ 38 ]. This proof-of-concept study demonstrates a previously unreported non-natural reaction for enzyme catalysts enabled by enzyme evolution. (c) Large-scale reductive amination to access GSK2879552 compound, catalyzed by an evolved imine reductase (IRED). The process employs glucose dehydrogenase (GDH) to recycle nicotinamide redox cofactors NAD(P)H [ 46 ]; (d) Traceless peptide macrocyclization catalyzed by engineered peptidase, omniligase-1 [ 51 ]. Efficient ligation was achieved for ring sizes 12–16 amino acids (>90% conversion) and a number of peptides containing nonpeptidic moieties. Hydrophobic or slightly polar amino acids are preferred at position P4 and in positions P1, P1’, and P2’ proline should be avoided. (e) n vivo production of a key amide intermediate in the synthesis of losmapimod [ 53 ]. The formation of the amide bond between 6-chloronicotinic acid and neopentyl amine via tandem coenzyme A (CoA)-dependent ligase CBL/transferase 66CaAT proceeded in >80% conversion and 74% isolated yield.

Scheme 3 . In vivoI biosynthetic pathways and in vitro enzymatic cascades creating architectures of non-natural molecules. (a) Engineered in vivo pathway for synthesis of cannabinoids and their analogs in yeast [ 37 ]. The natural hexanoyl-CoA (R = C5H11) was produced via a heterologous biosynthetic pathway, using genes from Ralstonia eutropha, Cupriavidus necator, Clostridium acetobutylicum, and Treponema denticola. Geranyl pyrophosphate (GPP) was produced by introducing the Enterococcus faecalis pathway. Expression of the Cannabis sativa genes encoding acyl activating enzyme (CsAAE1), synthase, and olivetolic acid (OA) cyclase (TKS-OAC) produced olivetolic acid analogs, which conjugated with GPP by prenyltransferase CsPT4. The resulting cannabigerolic acid (CBGA) was transformed into the Δ 9 – tetrahydrocannabinolic acid (THCA) using the cannabinoid synthase THCAS. After exposure to heat (ΔT), THCA decarboxylated to Δ 9 -tetrahydrocannabinol (THC). To probe the analog production capability of the engineered strain, an array of fatty acids of various chain lengths (R = C4–C7), branching and degrees of saturation were fed into the culture, providing six novel derivatives of THCA. (b) Fully assembled in vitro biocatalytic pathway to islatravir [ 17 ]. Evolved enzymes are in colored boxes and wild-type auxiliary enzymes are in white boxes; BB denotes the backbone of the respective evolution. Desymmetrizing oxidation catalyzed by galactose oxidase (GOase) from Fusarium graminearum in the presence of bovine catalase and horseradish peroxidase (HRP) from Amoracia rusticana provided chiral aldehyde. Its phosphorylation by pantothenate kinase (PanK) from Escherichia coli in the presence of acetyl kinase (AcK) from Thermotoga maritima to recycle adenosine triphosphate (ATP) produced 2ethynylglyceraldehyde 3-phosphate. Deoxyribose 5-phosphate aldolase (DERA) from Shewanella halifaxensis; phosphopentomutase (PPM) from E. coli; purine nucleoside phosphorylase (PNP) from E. coli enabled the one-pot assembly of islatravir, where the reaction equilibrium was driven by phosphate removal with sucrose phosphorylase (SP) from Alloscardovia omnicolens. (c) Sequential reactions combining chemo- and biocatalysis enabled by surfactant TPGS-750-M. A rhodium complex-catalyzed 1,4-addition reaction followed by a nitro group reduction with carbonyl iron powder (CIP), and ketone reduction by alcohol dehydrogenase ADH101. The reactions were performed as a tandem, one-pot process.

C-C bond formation

The construction of carbon skeletons can be achieved with mechanistically diverse enzyme families catalyzing aliphatic C–C bond formation or aromatic substitution. Many of these biocatalysts have been successfully employed in the synthesis of drug intermediates in the discovery and development fields ( Schemes 2 and 3 ). An evolved S-adenosyl methionine (SAM)-dependent methyltransferase NovO coupled with a SAM-forming enzyme SalL enabled late-stage functionalization of coumarin analogs to modulate their biological function ( Scheme 2a ) [ 36 ]. Deoxyribose 5-phosphate aldolases have been engineered for the stereoselective synthesis of complex carbohydrates [ 17 ] ( Scheme 3b ). Native prenyl transferase CsPT4 and two flavin-dependent synthases THCAS and CBDAS were used to produce cannabinoid analogs Δ 9 – tetrahydrocannabinolic acid (THCA) and cannabigerolic acid (CBDA), respectively [ 37 ] ( Scheme 3b ). A chiral cyclopropyl intermediate of grazoprevir was obtained at preparative scale using a hemoprotein-catalyzed carbene addition to an unsaturated carbon–carbon bond [ 38 ] ( Scheme 2b ). This novel approach is one of the first examples of industrial chemists using engineered biocatalysts to perform non-native chemistry which was pioneered by Arnold et al. [ 39 ]. Such biotransformations further expand the chemical matter accessible through biocatalysis to structural motifs rarely seen in nature, yet highly desired in pharmaceutical development.

C–-N bond formation: amination and amide formation

Reductive amination, amide bond formation, and C–N bond formation in nucleoside synthesis are fundamental reactions in biology and chemistry ( Schemes 2 and 3 ). There is a need to improve our capabilities in their synthesis to access therapeutic modalities [ 4 ]. Imine reductases and reductive aminases have been rapidly adopted by the pharmaceutical industry [ 40–45 ]. A recent process example uses an engineered imine reductase to access drug candidate GSK2879552 in excellent stereoselectivity and yield on a kilogram scale ( Scheme 2c ) [ 46 ]. The enzymatic transformation led to a greener manufacturing process, reducing the number of synthetic steps and waste.

The renewed interest in peptide-like therapeutics [ 1 , 3 , 4 ] has been driving the demand for green and efficient routes to form amide bonds and access noncanonical amino acids. Traditional methods for peptide syntheses either require significant protecting group manipulation or the use of a solid phase peptide synthesizer which suffer from sustainability issues [ 47 ]. Hydrolases, amidases, and peptidases have been heavily used in biocatalytic amide bond–forming reactions because of the efficient and selective manner in which they react [ 14 ]. The selectivity of these enzymes has been exploited for more complex transformations such as bioconjugation and macrocyclization [ 48 , 49 ]. Engineered transglutaminases, sortases, butelases, and peptiligases have been shown to mediate ligations of diverse peptides and proteins by acting selectively under the mild conditions required for the functionalization of biomolecules. Particularly noteworthy are versatile and broadly applicable engineered ligases such as thymoligase used in the synthesis of a 28-mer peptide [ 50 ] and omniligase-1, which quantitatively forms macrocyclic peptides ( Scheme 2d ) [ 51 ].

Novel ATP-dependent amide ligations have gained attention [ 52 , 53 ] and are becoming practical with the development of industrially relevant methods for ATP recycling [ 17 , 54 , 55 ]. Recently N-acyltransferases coupled with CoA ligases were shown to display activity on diverse carboxylic acid and amine partners ( Scheme 2e ). These biocatalysts optimized for activity, stability, and selectivity on non-natural substrates hold great potential to become a more general amidation platform. In parallel, diverse biocatalytic strategies have enabled access to a growing array of amino acid analogs [ 56 , 57 ]. These novel enzyme classes will play an important role in discovery and development of therapeutic peptides [ 3 , 4 ].

C-N and O-P bond formation in nucleoside chemistry

Nucleosides, oligonucleotides, and their analogs possess a wide variety of biological activity and have been extensively used in the pharmaceutical industry [ 2 , 58 ]. Their syntheses typically involve iterative protecting group manipulations and multiple oxidation state adjustments which contribute to high step count resulting in low overall yield. Biocatalysis is uniquely suited for the syntheses of these compounds as enzymes can be used to mimic the metabolic pathways used to make the natural compounds [ 59 ]. Two main enzyme classes, nucleoside transferases and nucleoside phosphorylases, have been effectively used to prepare modified nucleosides by catalyzing the exchange of the unnatural base and/or sugar moieties [ 59 ]. The recently published manufacturing route to islatravir features stereospecific glycosylation catalyzed by an evolved phosphorylase, which directly couples non-natural 2-fluoroadenine with a biocatalytically prepared deoxyribose 1-phosphate derivative ( Scheme 3b ) [ 17 ]. This example demonstrates that enzymatic syntheses can address the lack of efficient methods to access C1-4’-substituted sugar anaolgs, as well as thio-sugars, carbo-sugars, aza-sugars, and their respective nucleosides.

Phosphorous is a key element in many small and large molecules possessing biological activity. Syntheses of phosphorylated sugar and nucleotide and oligonucleotide derivatives present a challenge for traditional synthetic methodologies, particularly where regioselective and stereoselective installation of the phosphorus is necessary [ 59–61 ]. Biocatalytic methods are beginning to play an important role in solving these complex problems faced by medicinal and process chemists alike ( Scheme 3b ) [ 4 , 17 , 62 , 63 ]. Progress in scalable ATP formation and recycling [ 54 , 55 ] has advanced kinase-catalyzed phosphorus chemistry providing access to phosphorylated sugars [ 17 ] and nucleotides [ 55 ]. Novel enzymatic strategies are emerging which allow for convergent assembly of non-natural nucleic acids and have been used in the synthesis of oligonucleotides [ 61 , 62 ]. Polymerases, ligases, and synthases have been successfully engineered to efficiently couple nucleotide analogs [ 59 , 61–63 ]. We anticipate that biocatalytic approaches for X–P bond formation will lead to accelerated development of nucleoside and (oligo)nucleotide-based therapeutics.

CBG, CBD, Δ9-THC, CBN, CBGA, CBDA and Δ9-THCA as antioxidant agents and their intervention abilities in antioxidant action

Positive effect of some cannabinoids in the treatment and prophylaxis of a wide variety of oxidation-associated diseases and growing popularity of supplements containing cannabinoids, mainly cannabinoid oils (e.g. CBD oil, CBG oil), in the self-medication of humans cause a growing interest in the antioxidant properties of these compounds, especially those not showing psychotropic effects. Herein, we report the antioxidant activity of cannabigerol (CBG), cannabidiol (CBD), Δ9-tetrahydrocannabinol (Δ9-THC), cannabinol (CBN), cannabigerolic acid (CBGA), cannabinolic acid (CBDA) and Δ9-tetrahydrocannabinolic acid (Δ9-THCA) estimated by spectrophotometric methods: ABTS, DPPH, ORAC, beta-carotene CUPRAC and FRAP. The presented data prove that all the examined cannabinoids exhibit antioxidant activity manifested in their ability to scavenge free radicals, to prevent the oxidation process and to reduce metal ions. Although the intensity of these activities is not the same for the individual cannabinoids it is comparable for all of them with that of E vitamin. As results from the research, the significance of the two types of electron sources presenting in examined cannabinoids, phenolic groups and double bonds transferring electrons, depends on the type of electron-accepting species – radicals/metal ions.

Keywords: Acidic cannabinoids, antioxidant activity; Cannabinoids; Cannabis; Hemp; Reduction potential.

Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.

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