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WO2019070885A1 - Procédés de préparation de compositions à base de plantes encapsulées, solubilisables, produits à base de celles-ci - Google Patents

Procédés de préparation de compositions à base de plantes encapsulées, solubilisables, produits à base de celles-ci Download PDF

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Publication number
WO2019070885A1
WO2019070885A1 PCT/US2018/054216 US2018054216W WO2019070885A1 WO 2019070885 A1 WO2019070885 A1 WO 2019070885A1 US 2018054216 W US2018054216 W US 2018054216W WO 2019070885 A1 WO2019070885 A1 WO 2019070885A1
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Prior art keywords
mixture
aqueous solution
spirulina
encapsulated
finished product
Prior art date
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English (en)
Inventor
Sarah RODRIGUEZ
Ali JAMALIAN
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Spirulinex LLC
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Spirulinex LLC
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/5005—Wall or coating material
    • A61K9/5021—Organic macromolecular compounds
    • A61K9/5052—Proteins, e.g. albumin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/748—Cyanobacteria, i.e. blue-green bacteria or blue-green algae, e.g. spirulina
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/348—Cannabaceae
    • A61K36/3482—Cannabis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts

Definitions

  • the present disclosure relates to Cannabis-based products and formulations, and methods of preparing said Ccmnabis-based products and formulations.
  • Cannabis is a genus of flowering plants that includes at least three species, Cannabis sativa, Cannabis indica, and Cannabis ruder alis. Cannabis plants include secondary metabolites called cannabinoids. Cannabinoids are hydrophobic, and can act on cannabinoid receptors in tissues and cells of the human body.
  • formulations e.g., liquid solutions and/or suspensions
  • cannabinoids and/or other hydrophobic compounds suitable for medicinal, pharmaceutical, nutraceutical, edible materia], cosmetic, topical, and food applications.
  • the production of such formulations can include the preparation of a plant-based and microbial-based suspension, and a microencapsulation of the plant material with the microbial material.
  • production of a formulation includes the preparation of a suspension including a synthetic material (e.g., including one or more synthetic cannabinoids) encapsulated with a microbial material.
  • a method includes providing a Cannabis extract material including a shatter, a crumble, a budder, an oil (e.g., butane hash oil, propane hash oil), nug run concentrate, C02 concentrate, rosin, trim run, live resin, sap, dry sift, ice water hash, full melt, wax, pull and snap, fraction, THC, CBD, or any other product resulting from an extraction of hydraulic press operation applied to plant, or a synthetic material that includes a Cannabis derivative.
  • the method also includes providing distilled water, and raising a pH of the distilled water to a value of about 9 to about 10, to produce a raised-pH water.
  • An aqueous solution including the raised-pH water and Spirulina (Arthrospira platensis, and/or Arthrospira maxima) or components thereof, is prepared, the aqueous solution having about 3% weight Spirulina per volume. The aqueous solution is blended.
  • the blended aqueous solution is cavitated (e.g., using one or more of an ultrasonicator or one or more other instruments (e.g., sequentially) to accomplish one or more of cavitation, the application of impact force(s), or the application of shear force(s)) to at least partially liberate proteins of the Spirulina, thereby producing a membrane-disrupted Spirulina solution (i.e., where at least a portion of the Spirulina cell membrane has been disrupted, e.g., resulting in liberation of cellular component/contents of the Spirulina).
  • the Cannabis extract material and the disrupted Spirulina solution are combined, in a ratio of about 1:1, to form an intermediate mixture.
  • the intermediate mixture is cavitated to form a first mixture including the Cannabis extract material encapsulated by the Spirulina or a derivative thereof.
  • the first mixture e.g., having a pH value of about 9 to about 10
  • the layer is then dehydrated form a finished product.
  • citric acid can be added to the first mixture to produce a reduced-pH mixture having a pH of about 3.
  • the reduced-pH mixture is then dispensed (e.g., poured) onto a substrate to form a layer including about 2.5 mL of the reduced-pH mixture per square inch.
  • the layer is dehydrated form a finished product.
  • a method for the micro-encapsulation of terpenes, lipids, sterols, antioxidants, cannabinoids and/or other hydrophobic compounds includes providing an extract material (e.g., a shatter, a crumble, or abutter) and preparing an aqueous solution including a cellular material (e.g., Spirulina). Preparing the aqueous solution can include increasing a pH of distilled water to produce a raised-pH water, and combining the raised-pH water with the cellular material.
  • an extract material e.g., a shatter, a crumble, or abutter
  • Preparing the aqueous solution can include increasing a pH of distilled water to produce a raised-pH water, and combining the raised-pH water with the cellular material.
  • a disruption of the aqueous solution is performed (e.g., via ultrasonication, or one or more of cavitation, the application of impact force(s), or the application of shear force(s)), to at least partially liberate proteins of the cellular material.
  • the extract material and the disrupted aqueous solution are combined to form an intennediate mixture.
  • the intermediate mixture is disrupted to form an encapsulated mixture, for example via cold cavitation (i.e., at or below room temperature, e.g., for a duration of about 2 hours, or about 3 hours, or about 4 hours, or about S hours, or from about 3 hours to about 4 hours, or from about 3.S hours to about 4.S hours), or via heated cavitation (e.g., at a temperature of about 40°C, about 50°C, about 60°C, about 62°C, about 64°C, about 66°C, about 68°C, about 69°C, about 71°C, about 70°C, about 73°C, about 75°C, about 79°C, about 81°C, about 83°C, or about 8S°C, or between about 60°C and about 7S°C, or between about 60°C and about 75°C) (e.g., for a duration of not more than about 10 minutes, not more than about 15 minutes, not more than about 20 minutes, not more
  • the method also includes decarboxylating the extract material before combining the extract material and the disrupted aqueous solution.
  • drying the encapsulated mixture includes pouring the encapsulated mixture onto a substrate (e.g., silicone), and dehydrating the poured encapsulated mixture.
  • the dehydrating can be performed at a temperature of about 165°F and for a duration of at least about 15 hours.
  • drying the encapsulated mixture includes lyophilization of the encapsulated mixture.
  • drying the encapsulated mixture is performed under vacuum.
  • drying the encapsulated mixture is performed at a temperature of from about 10°C to about 100°C.
  • drying the encapsulated mixture includes nano-spray drying.
  • the method also includes blending the aqueous solution before performing the disruption of the aqueous solution.
  • a method for me nano-encapsulation and/or microencapsulation of terpenes, lipids, sterols, antioxidants, cannabinoids and/or other hydrophobic compounds includes receiving distilled water and one or more Cannabis extract materials, such as a shatter, a wax, a crumble, or a butter. A pH of the distilled water is raised to a value of about 9 to about 10, to produce a raised-pH water.
  • aqueous solution including the raised-pH water and Spirulina, is prepared such that the aqueous solution has about 3% weight Spirulina per volume.
  • the Cannabis extract material includes the wax, and the wax has about 3% THC.
  • Preparing the aqueous solution can also include at least one of mixing and cavitating (e.g., using an ultrasonicator and/or one or more other instruments, such as a high pressure homogenizer, to accomplish one or more of cavitation, the application of impact force(s), or the application of shear force(s)).
  • the method can also include diluting the aqueous solution to a predeteimined/desired dilution level (e.g., about 1:1,000, about 1:10,000, about 1:20,000, about 1:100,000, about 1:200,000, etc.).
  • a predeteimined/desired dilution level e.g., about 1:1,000, about 1:10,000, about 1:20,000, about 1:100,000, about 1:200,000, etc.
  • a method for the nano-encapsulation and/or microencapsulation of terpenes, lipids, sterols, antioxidants, cannabinoids and/or other hydrophobic compounds includes receiving distilled water and a Cannabis extract material including a shatter, a crumble, or a butter.
  • a pH of the distilled water is raised to a value of about 9 to about 10, to produce a raised-pH water.
  • An aqueous solution, including the raised-pH water and Spirulina, is prepared such that the aqueous solution has about 3% weight Spirulina per volume.
  • the aqueous solution is blended and cavitated to at least partially liberate proteins of the Spirulina, thereby producing a disrupted Spirulina solution.
  • the Cannabis extract material and the disrupted Spirulina solution are combined, in a ratio of about 1:1, to form an intermediate mixture.
  • the intennediate mixture is sonicated to form a first mixture including encapsulated Cannabis extract material.
  • the mixture is dispensed onto a substrate to form a layer including about 2.5 mL of the mixture per square inch.
  • the layer is men dehydrated to form an encapsulated material.
  • lecithin e.g., soy lecithin, sunflower lecithin, or any other plant-based lecithin
  • ⁇ l% may be added to the first mixture before dispensing onto the substrate and forming the layer (e.g., to impact "mouthfeel" of a finished product).
  • citric acid may optionally be added to me first mixture to produce a reduced-pH mixture having apHof about 3 before dispensing onto the substrate and forming the layer.
  • the encapsulated material can subsequently be combined with a solvent to form a liquid finished product, which, for example, can be a solution or a suspension.
  • a centrifuging step is performed after cavitation, to remove broken cell membrane from the disrupted Spirulina solution, leaving only or substantially only protein behind for the subsequent solubilization/encapsulation of the wax (e.g., to modify a texture of the finished product).
  • a method for the nano-encapsulation and/or microencapsulation of terpenes, lipids, sterols, antioxidants, caimabinoids and/or other hydrophobic compounds includes providing an extract material, and preparing an aqueous solution including a cellular material. A disruption of the aqueous solution is performed, to at least partially liberate proteins of the cellular material. The extract material and the disrupted aqueous solution are combined to form an intermediate mixture, and a disruption of the intermediate mixture is performed, so as to form an encapsulated mixture.
  • the encapsulated mixture is a liquid finished product (e.g., a solution or a suspension).
  • a pH of the encapsulated mixture is reduced to form a liquid finished product, which, for example, can be a solution or a suspension.
  • FIG. 1 A is a process flow diagram illustrating a method for encapsulation of an extract material, according to some embodiments.
  • FIG. IB is a tree diagram illustrating methods for encapsulation of an extract material, according to some embodiments.
  • FIG. 2 is a process flow diagram Ulustrating a method for preparing an extract material, according to some embodiments.
  • FIG.3 is a block diagram showing components of a system for encapsulation of an extract material, according to some embodiments.
  • FIGS.4A-4E are images showing example finished products, according to some embodiments.
  • FIG.4F is a photographic image of a powder, according to an embodiment.
  • FIG. 5 is an illustration of an example microparticle, according to some embodiments.
  • FIG. 6 is a plot of absorbance as a function of pH, for an encapsulant-based solution according to an embodiment.
  • FIGS. 7A-7D are scatterplots showing the distribution of particle sizes within the encapsulant-based solution of FIG. 6.
  • FIGS. 8A-8B are microscopic images of a second encapsulant-based solution according to an embodiment.
  • FIGS. 9A-9B are plots snowing the distribution of particle sizes within the encapsulant-based solution of FIGS. 8A-8B.
  • the present disclosure is directed to methods and systems for the microencapsulation of hydrophobic compounds and/or lipophilic compounds, and/or materials, such as extracts or synthetics, that include one or more hydrophobic and/or lipophilic compounds as well as one or more neutral and/or hydrophilic compounds, including but not limited to terpenes, terpenoids, lipids, sterols, antioxidants, cannabinoids, and/or the like, and/or various combinations of various neutral, hydrophilic, and lipophilic/hydrophobic compounds.
  • Embodiments includes methods for microencapsulation and/or otherwise stabilizing such compounds using microbial (including algae, for example a cyanobacteria, eukaryote, prokaryote, etc.), bacteria, protozoa, fungi (unicellular or multicellular), plant cells, cellular material, and/or components derived therefrom, and/or one or more naturally or synthetically derived compounds related to one or more microbe and/or plant cellular compounds (e.g., proteins), as well as to the resulting outputs of such methods, and products including/lncorpoiating outputs of the method(s) in finished end products thereof.
  • Applications of the methods, compounds, microencapsulated compounds, and finished end products as set forth herein include, by way of non-limiting example, health and medicinal, pharmaceutical, supplements, nutraceuticals, cosmetics, edible materials, and food applications.
  • Cannabis and cannabinoids may relieve pain, lower inflammation and decrease anxiety. Cannabis and cannabinoids have been implicated in the relief of anxiety, nausea, pain, and the improvement of appetite. Due to these properties, Cannabis and cannabinoids may have use in treating symptoms of cancer and the side effects of cancer therapies.
  • the U.S. Food and Drug Administration (FDA) has approved two cannabinoids, dronabinol and nabilone, for the treatment and prevention of nausea and vomiting, which are possible side effects of chemotherapy. Furthermore, laboratory studies have shown the capacity of Cannabis to selectively kill cancer cells.
  • cannabinoids can provide a well-tolerated, promising therapeutic for me treatment of seizures in those with epilepsy.
  • cannabinoids have been implicated for the treatments of Alzheimer's Disease, ALS, Chronic Pain, Diabetes Mellitus Dystonia, Epilepsy, Fibromyalgia, GI Disorders, Gliomas/Cancer, Hepatitis C, HIV, Huntington's Disease, Hypertension, Incontinence, MRS A, Multiple Sclerosis, Osteoporosis, Pruritus, Rheumatoid Arthritis, Sleep Apnea and Tourette's Syndrome.
  • Water-based solutions having compounds dissolved therein, are easier to deliver orally and via injection, when compared to non-aqueous solutions.
  • the incorporation of compounds that are hydrophobic and/or lipophilic into water-based solutions has historically been difficult; as discussed below.
  • tipophilic/hydrophobic compounds such as most cannabinoids, as well as many other hydrophobic medicinal and nutraceutical compounds
  • incorporation of a hpophihc/hydrophobic compound (or compounds) in aqueous solution formulations can be difficult, as well as time and resource intensive.
  • such solutions can be difficult to reproduce consistently, distribute evenly/uniformly, etc.
  • it can even lead to poor adsorption within an aqueous environment, and this poor solubility in aqueous or polar solutions can, in turn, lead to low or inconsistent dosing and/or bioavailability (e.g., oral bioavailability).
  • lipophilic/hydrophobic compounds can be difficult to dispense and accurately measure, often sticking to containers (e.g., glass or plastic containers), including when in aqueous solution, and thus further increasing the difficulty in delivering and/or integrating such compounds as part of a formulation.
  • Embodiments of the present disclosure address the problems outlined above, and facilitate the solubilization, suspension, and/or micro-encapsulation of lipid, terpene, sterol, cannabinoid, and/or other generally non-polar, lipophilic and/or hydrophobic compounds, as well as mixtures/compounds containing such compounds as well as one or more neutral and/or hydrophilic compounds therein, in some embodiments, without the use of added or synthetic detergents and/or surfactants, to produce finished products, including in some instances, finished products that are suitable for consumption.
  • micro-encapsulation/micTo-encapsulation/ stabilization methods set forth herein utilize whole cells and extractions of whole cells ("cellular material”), rather man a single substance or molecule, thereby utilizing a mixture of one or more of lipids, proteins, and/or nucleic acid for encapsulation of hydrophobic material.
  • a finished product (also referred to as a "final substance,” a “consumable,” a “composition,” or a “concentrate”) produced by methods set forth herein can include an extract, oil, or wax (e.g ., one or more lipid, fatty, sterol, non-polar, lipophilic, and/or hydrophobic compounds) that is micro-encapsulated by cellular material and/or components of cellular material, in some embodiments, mat is micro-encapsulated by cellular material and/or components of cellular material in one or more polar solvents.
  • an extract, oil, or wax e.g ., one or more lipid, fatty, sterol, non-polar, lipophilic, and/or hydrophobic compounds
  • finished products produced and/or facilitated by methods set forth herein exhibit high uniformity (e.g., in texture, in low occurrence of lipid droplets, etc.), and/or can be configured to be partially, substantially, or fully water-soluble, and/or can be used as a "concentrate" (including one or more components of extract material), e.g., for incorporation into one or more edible, topical, or otherwise consumable products.
  • hpophilic/hydrophobic compounds of the disclosure can include terpenes, sterols, cannabinoids, and other compounds of medicinal and nutraceutical value/application.
  • terpenes include but are not limited to nerolidol, limonene, pinene, alpha-pinene, d-linalooL, myrcene, ocimene, terpinolene, terpineol, valencene, caryophyllene, BCP, alpha humulene, phellandrene, carene, fenchol, terpinene, borneol, bisabolol, phytol, camphene, sabinene, camphor, isobomeol, cedrene, guaiol, pulegone, isopulegol, cymene, terpineol, and/or the like.
  • Examples of classes of plant sterols types include but are not limited to sterol elsters, stands and pro-sterols.
  • Examples of lipids include but are not limited a-linolenic acid, arachidonic acid, eicosapentaenoic acid, and docosahexaenoic acid.
  • Examples of antioxidants include zeaxanthin, lutein, a-carotene, fucoxanthin astaxanthin, canthaxanthin, and b-carotene.
  • Cannabinoids according to the disclosure can include, while not being limited to those obtained or obtainable from cannabis plants, which can include, by way of non- limiting example: ⁇ 9-Tetrahydrocannabinol ( ⁇ 9-THC), ⁇ 8-Tetrahydrocannabinol ( ⁇ 8- THC), Cannabichromene (CBC), Caimabicyclol (CBL), Cannabidiol (CBD), Cannabielsoin (CBE), Cannabigerol (CBG), Cannabinidiol (CBND), Cannabinol (CBN), Cannabitriol (CB1), and their propyl homologs, including, by way of non- limiting example cannabidivarin (CBDV), ⁇ 9-Tetrahydrocannabivarin (THCV), cannabichromevarin (CBCV), and cannabigerovarin (CBGV).
  • Cannabinoids according to the disclosure can also include those produced by plants (also known as phytocannabinoids, natural cannabinoids, herbal cannabinoids, or classical cannabinoids).
  • Cannabinoids of the disclosure can include synthetic cannabinoids, and/or cannabinoids isolated from cannabis plants, by way of non-limiting example, Tetrahydrocannabinol (THC), Cannabidiol (CBD) (e.g., derived from Cannabis indica, Cannabis ruderalis, or Cannabis saliva (“hemp")), CBG (Cannabigerol), CBC (Cannabichromene), CBL (Cannabicyclol), CBV (Cannabivarin), THCV (Tetrahydrocannabivarin), CBDV (Cannabidivarin), CBCV (Cannabichromevarin), CBGV (Cannabigerovarin), and CBGM (Cannabigerol Monomethyl Ether).
  • THC Tetrahydrocannabinol
  • cannabinoids are synthesized and accumulated as cannabinoid acids (e.g., cannabidiolic acid (CBDA)), and drying and/or heating can cause such acids to decarboxylize (e.g., CBDA to CBD).
  • CBDA cannabidiolic acid
  • THC is the principal psychoactive constituent (or cannabinoid) of the cannabis plant.
  • the initially synthesized and accumulated form in plant is THC acid (THC A).
  • THC has mild to moderate analgesic effects, and cannabis can be used to treat pain by altering transmitter release.
  • Other effects include relaxation, alteration of visual, auditory, gustatory, and olfactory senses, fatigue, and appetite stimulation, as well as anti-nausea effects.
  • THC occurs mainly as tetrahydrocannabinols acid (THCA, 2-COOH-THC).
  • THCA tetrahydrocannabinols acid
  • 2-COOH-THC tetrahydrocannabinols acid
  • Geranyl pyrophosphate and olivetolic acid react, catalyzed by an enzyme to produce cannabigerolic acid, which is cyclized by the enzyme THC acid synthase to give THCA.
  • THCA is decarboxylated producing THC.
  • the pathway for THCA biosynthesis is similar to that which produces the bitter acid humulone in hops.
  • THC variants include:
  • CBD is another cannabinoid found in cannabis, and has been shown to act as an indirect antagonist of cannabinoid agonists, and has also been shown to act as a 5-HT1A receptor agonist, an action which is potentially involved in antidepressant, anxiolytic, and neuroprotective effects.
  • CBD is also an allosteric modulator at the Mu and Delta opioid receptor sites.
  • Cannabis produces CBD-carboxylic acid through the same metabolic pathway as THC, until the last step, where CBDA synthase performs catalysis instead of THCA synthase.
  • Non-limiting examples of CBD variants include:
  • CBG is another cannabinoid found in cannabis, and is generally considered to be low or non-psychoactive .
  • CBG can be found in higher concentrations in hemp rather than in varieties of Cannabis cultivated for high THC content and their corresponding psychoactive properties.
  • CBG has been found to act as a high affinity ⁇ 2-adrenergic receptor agonist, moderate affinity 5-HT1A receptor antagonist, and low affinity CB1 receptor antagonist.
  • CBG also binds to the CB2 receptor.
  • CBG has been shown to relieve intraocular pressure, useful in the treatment of glaucoma.
  • Non-limiting examples of CBG variants include:
  • CBN is a psychoactive cannabinoid found in cannabis, particularly Cannabis sativa and Cannabis indica/afghanica. It is also a metabolite derived from tetrahydrocannabinol (THC). CBN acts as a weak agonist of the CB1 and CB2 receptors, with lower affinity in comparison to THC.
  • CBN variants include
  • CBC has structural similarity to the other natural cannabinoids, including tetrahydrocannabinol, tetrahydrocannabivarin, cannabidiol, and cannabinol, among others.
  • CBC may play a role in the anti-inflammatory and anti-viral effects of cannabis, and may contribute to the overall analgesic effects of cannabis.
  • Non-limiting examples of CBC variants include:
  • CBV is a cannabinoid found in minor amounts in cannabis, and is generally considered to have low or no psychoactivity. It is an analog of cannabinol (CBN) with the side chain shortened by two methylene bridges (-CH2-). CBV is an oxidation product of tetrahydrocamiabivarin (THCV, 1HV).
  • CBDV is a cannabinoid found in Cannabis, and is generally considered to have low or no psychoactivity. It is a homolog of cannabidiol (CBD), with the side-chain shortened by two methylene bridges (CH2 units).
  • CBDV cannabidiol
  • THCV or THV is a bomologue of tetrahydrocannabinol (THC) having a propyl (3-carbon) side chain. This terpeno-phenolic compound can be round naturally in Cannabis, sometimes in significant amounts. Plants with elevated levels of propyl cannabinoids (mcluding THCV) have been found. THCV has been shown to be a CB1 receptor antagonist
  • Cannabicyclol is a cannabinoid found in the Cannabis species.
  • CBL is a degradative product like cannabinol. Light converts cannabichromene to CBL.
  • Non- limiting examples of CBL variants include: [0049]
  • Non-limiting examples of CBE variants include:
  • Encapsulation can occur in cells and viruses.
  • encapsulation is generally performed by lipid-based organelles and vesicles, while prokaiyotes use large proteinaceous shells (e.g., nucrocompartments and other protein-based compartments) to encapsulate proteins, enzymes, and other molecules.
  • proteinaceous shells e.g., nucrocompartments and other protein-based compartments
  • mis compartmentalization keeps components of various cell mechanisms together to increase efficiency.
  • cells and these compartments may also be useful as a delivery mechanism for exogenous molecules and compounds.
  • Spirulina (Arthrospira) and/or its protein-based compartments may be especially useful for oral delivery of exogenous molecules and compounds.
  • Spirulina is a free-floating, filamentous bacteria that offers several advantages as an encapsulation system. First, it is safe for human consumption, and can be used as a nutritional supplement. Second, the cell wall/protein compartment can protect the encapsulated exogenous molecule or compound until the low-pH environment of the stomach degrades the encapsulating proteins and releases the exogenous material.
  • polypeptides are needed for encapsulation of the exogenous material.
  • these polypeptides are parts of the cell wall or a protein-based compartment.
  • these polypeptides tether or otherwise attach the exogenous material to the cell wall or a protein-based compartment.
  • these polypeptides insert the exogenous material into protein-based compartments.
  • the protein-based compartments are microcompartments .
  • varieties of microbial, plant, and/or algal species having a cell wall, or components thereof can be utilized as suspension and/or encapsulation material(s) (i.e., cellular materials), or as additives, and can include: Algal species such as Dunaliella salina, Haematococcus pluvialis, Coelastrella striolata, Diatoms such as Phaeodactylum iricomutum, and Isochrysis galbana, such as Cyanobacteria such as Arthrospira platensis and Arthrospira maxima; Chlorella species such as Chlorella autotrophica, Chlorella colonials, Chlorella lewinii, Chlorella minutissima, Chlorella pituha, Chlorella pulchelloides, Chlorella pyrenoidosa, Chlorella rotunda, Chlorella singularis, Chlorella sorokiniana, Chlorella variabilis, Ch
  • radula G. acicularis, G. pistillata, Eucheuma spinosum, Polyides rotundus, Osmundea pinnatifida, Pterocladiella capillacea, Sphaerococcus coronopifolius, and Gelidium microdon, Rhodophyta; Cystoseira abies- marina and Fucus spiralis, Phaeophyta; Ulva compressa, Chlorophyta; Kelp species such as Ascophyllum nodosum, Laminara Digitata; Other plants species such as aloe, alfalfa, turmeric root (Curcuma longa), ginger root, wheatgrass, and Cocos nucifera: Nutritional yeasts such as S.
  • Probiotic strains such as Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus gasseri, Lactobacillus paracasei, Lactobacillus bulgaricus, Lactobacillus brevis, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus salivarius, Bifidobacterium lactis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium longum; and Species often used in Kombucha production such as species of the genus Zygosacchaiomyces, Acetobacter and Gluconacetobacter.
  • the cellular material used for encapsulation is Spirulina (e.g., Arthrospira platensis).
  • Spirulina is an ecologically sound, nutrient-rich dietary supplement. Dried spirulina often includes about 5% water, about 24% carbohydrates, about 8% fats, and about 51% to about 71% (e.g., about 60%) protein (amino acids), along with vitamins and minerals. More specifically, in some embodiments 100 grams (g.) of dried Spirulina has an energy of about 290 kcal, about 23.9 g. of carbohydrates (including about 3.1 g. of sugars and about 3.6 g. dietary fiber), about 7.72 g.
  • fat including about 2.65 g. of saturated fat, about 0.675 g. of monounsaturated fat, and about 2.08 g. of polyunsaturated fat
  • 57.47 g. of protein including about 0.929 g. of tryptophan, about 2.97 g. of threonine, about 3.209 g. of isoleucine, about 4.947 g. of leucine, about 3.025 g. of lysine, about 1.149 g. of methionine, about 0.662 g. of cystine, about 2.777 g. of phenylalanine, about 2.584 g. of tyrosine, about 3.512 g. of valine, about 4.147 g.
  • aiginine about 1.085 g. of histidine, about 4.515 g. of alanine, about 5.793 g. of aspartic acid, about 8.386 g. of glutamic acid, about 3.099 g. of glycine, about 2.382 g. of proline, and about 2.998 g. of serine), as well as various vitamins and minerals.
  • Spirulina used in methods of the present disclosure may be commercially available and/or artificially cultivated. Fresh Spirulina and/or dry Spirulina may be used, with fresh Spirulina being preferable in some embodiments since fresh Spirulina is enriched in nutrients of Spirulina.
  • Spirulina may be cultivated by any known method used in the cultivation of blue-green algae and, for example, viable Spirulina can be inoculated in a culture medium in a concentration of 20 to 500 mg/L based on the weight of the algal dry biomass, and then cultivated.
  • a light source used in the cultivation may be sunlight or artificial light, for example a light source of 1,000 to 100,000 ⁇ x.
  • the cultivation temperature can be from 20°C to 65°C, for example from 30°C to 40°C.
  • the cultivation period can be from about 5 days to about 10 days.
  • an amount of carbonate ions in the solution can be increased by appropriately blowing a carbon dioxide gas or an air.
  • Spirulina can be synonymous with "Arthrospira.”
  • the genus Arthrospria includes at least 57 species of which 22 are currently taxonomically accepted.
  • reference to “Spirulina” or “Arthrospira” without further designation can include reference to any of the following species: A. amethystine, A. ardissonei, A. argentine/, A. balkrishnanii, A. baryana, A. boryana, A. braunii, A. breviarticulata, A. brevis, A. curia, A. desikacharyiensis, A. fimiformis, A. fusiformis, A. ghannae, A.
  • a product or finished product according to the disclosure can take on any of a wide variety of forms, including but not limited to: flake, granule, shred, pellet, tablet, powder, stick, wafer, or sheet.
  • Flake material for example, can be configured to be high in Tetrahydrocannabinol-H (“THC-H”) and/or have particular cannabinoid profiles.
  • a finished product can be further modified or incorporated into a modified "end" product which can also take on any of a wide variety of forms, including (but not limited to): a baked good (e.g., a cookie, brownie, cake, etc.), a spice rub, a pastille, lozenge, gum, spread, oil, etc., and/or be configured as a product for buccal delivery, such as a lozenge or pastille, a snus-like product or pouch (e.g., woven or non-woven, and/or having a dissolvable pouch), etc.
  • a baked good e.g., a cookie, brownie, cake, etc.
  • a spice rub e.g., a pastille, lozenge, gum, spread, oil, etc.
  • a snus-like product or pouch e.g., woven or non-woven, and/or having a dissolvable pouch
  • a dissolvable pouch such that the inner contents (i.e., the finished product) is eventually directly exposed to a mouth of a user, such mat one or more components is delivered via buccal, sublingual, and/or oral administration.
  • products and/or finished products of the present disclosure are one or more of alcohol-free, substantially alcohol-free, essentially alcohol- free sugar-free, substantially sugar-free, essentially sugar-free (i.e., free from added sugar), gluten-free, substantially gluten-free, fat-free, or substantially fat-free.
  • the finished product is alcohol-free, sugar-free, gluten-free, and fat-free.
  • an extract material e.g., a Cannabis extract
  • the extract material can include one or more of: Shatter, a transparent Cannabis extract.
  • Shatter is a highly stable concentrate that can be easily broken/shattered. Most shatters are transparent in color and can be handled without considerable residue. However, exposing shatter to heat, even room temperature, can cause shatter to become oil. If a concentrate is exposed to enough heat, it can decarboxylate permanently/irreversibly.
  • Crumble a dry extract with a cheese-like structure that can be crumbled and is not transparent like shatter. Crumble is formed by agitating an oil until it reaches a point of crystallization, which removes its translucent color. Crumble is often stored using parchment paper.
  • Budder has a consistency that is between wax and crumble. Budder is versatile for use bom indoors and outdoors, and is often manipulated using a tool.
  • BHO butane hash oil
  • the BHO process extracts the cannabinoids, terpenes, waxes, concrete oils, and occasionally, chlorophyll, from the plant itself. After the BHO process, the butane is removed from the final product. The BHO process is often used for mass production.
  • PHO propane hash oil
  • propane hash oil a kind of hash oil that is extracted from Cannabis through the use of propane.
  • PHO can have a consistency similar to that of budder, because propane extraction processes use elevated pressures as compared with BHO, and thus can remove different ratios of oils and wax from the plants compared to the butane process.
  • PHO can preserve more terpenes and remove residuals, depending on the Cannabis strain mat you use.
  • Nug Run Concentrate This form of extract can have more trichomes and terpenes than the Cannabis plant itself, making it rich in flavor and less harsh to smoke.
  • CO 2 Concentrate This form of extract has a liquid amber or golden appearance, and is often used in cartridges used for vape pens. A benefit of using the CO 2 method is that it is safer than butane or propane extraction methods. In its final form, CO 2 oils do not include any harmful residual components, since the process itself is effective in killing any bacteria or mold that might have been present in the plant material. A disadvantage of CO 2 concentrate preparation methods is that its flavor profile is not as potent as BHO and PHO.
  • Rosin extraction is not suitable for mass production, but it is safer man many other extraction methods. Rosin techniques often do not include the use of solvents.
  • Trim Run This form of extract is made using Cannabis plant trimmings, and thus may be used to supplement other extraction techniques to reduce or eliminate waste of the Cannabis plant. However, trim run is often less potent than other concentrates/extracts, and can contain more chlorophyll than other extracts.
  • Live Resin This form of extract is produced using frozen Cannabis instead of cured plant material. Dabbing live resin can preserve more terpenes and THC that the curing process usually removes. Live resin can have a more potent flavor and aroma, as compared with BHO produced using dried cannabis. Several methods can effectively extract frozen buds, but the rosin technique is what gives you "Live Rosin," which has a better and more powerful flavor profile compared to other kinds of concentrates.
  • Sap This form of extract is a sticky concentrate that is more suitable for use indoors or in lower-humidity conditions. Sap can be easier to handle than oils, but is often handled using tools, since manual handling easily cause melting of the sap.
  • Dry Sift This form of extract is one of the finer forms of concentrates. Dry sift is made using silk screens in varying sizes, meshes and/or microns to isolate the trichome heads from the bud. The silk screens are used to ensure that only the finest form of material ends up in the final product Dry sift can be a lower-yielding process than some other extraction techniques. Dry sift can have a desirable flavor profile since the dry sift production process is highly effective at preserving terpenes. Dry sift has a soft, sand-like consistency that is pourable.
  • Ice Water Hash This form of extract is prepared using ice water extraction, and has a texture/consistency similar to that of dry sift.
  • Ice water hash can be made by hand or with the use of a washing machine, after which a filter can be used to separate the material and water using screens with different microns or meshes. The screens are used, for example, to prevent any unwanted particles from ending up in the final product because they isolate the glandular trichome heads.
  • a metal strainer can be used to break down the hash so that it can be cured once all moisture has been removed. The ice water bash can then be stored in an air-tight glass container for curing.
  • Full Melt This form of extremely high-quality/purity extract can be made using either the dry sift or ice water hash method. In full melt concentrate, no plant material at all is included in the end product. Full melt is often made using only, or substantially only, isolated trichome heads (e.g., with no residual components). Full melt is susceptible to melting at slight/low exposure to heat
  • Wax This form of extract has a form that is between a solid and liquid, can have an earwax appearance, and is relatively easily mampulable (often with tools).
  • Pull and Snap This form of extract is a moderately stable, solid form of Cannabis concentrate. Pull and snap is flexible, and can be bent or rolled (e.g., into ball form). Pull and snap is suitable for outdoor use, and is often stored in a non-stick container.
  • the extract material can include a predetermined amount of cannabinoids (e.g., about 40% to about 99% cannabinoids, or about 55% to about 90% cannabinoids, or about 70% to about 90% cannabinoids).
  • FIG. 1 A shows obtaining an already-prepared extract material, optionally, prior to beginning the method 100, Cannabis material (e.g., Cannabis plant trim and/or flower) can be obtained and an extraction thereof is prepared using one or more extraction methods, including (but not limited to): solvent extraction (e.g., butane extraction, ethanol extraction, or low-grade alcohol extraction, supercritical carbon dioxide (CO 2 ) extraction, olive oil extraction, etc.), a rosin or resin extraction, a "press” extraction, etc.
  • the extract material can optionally be decarboxylated. Once decabroxylated, the extract material can have a honey-like, adherent or adhesive-like (i.e., "sticky”) consistency.
  • a polar liquid such as an aqueous solution, a water solution, water (for purposes of illustration and not limitation, distilled water) is obtained, and a pH of the distilled water is increased ("tip-adjusted), at 102D, for example to a pH of about 9 to about 10, resulting in a "high pH water.”
  • the pH adjustment can be accomplished, for example, by adding sodium hydroxide (NaOH), a bicarbonate, and/or other food-safe pH- adjusting additive.
  • Spirulina is obtained.
  • the Spirulina and the high pH water are combined, at 102F, into an aqueous Spirulina solution (e.g., to about 3% weight of Spirulina per volume, or about 5% weight per volume, or about 7% weight per volume, or from about 0.5% to about 30% weight per volume, or from about 1% to about 9% weight per volume, or from about 1% to about 10% weight per volume, or from about 2% to about 4% weight per volume, or from about 3% to about 5% weight per volume, or from about 3% to about 7% weight per volume).
  • an aqueous Spirulina solution e.g., to about 3% weight of Spirulina per volume, or about 5% weight per volume, or about 7% weight per volume, or from about 0.5% to about 30% weight per volume, or from about 1% to about 9% weight per volume, or from about 1% to about 10% weight per volume, or from about 2% to about 4% weight per volume, or from
  • Spirulina can be found in nature, particularly in alkaline lakes. Spirulina as utilized herein can, according to some embodiments, be cultured/maintained at alkaline pH levels.
  • the pH of the aqueous Spirulina solution is alkaline (pH greater than 7).
  • the pH of the aqueous Spirulina solution is between about 8.S and about 11.
  • the pH of the aqueous Spirulina solution is between about 9 and about 10.
  • the aqueous Spirulina solution is processed to liberate (e.g., by disrupting the membrane) the cell contents of the Spirulina (e.g., to liberate the contents, including protein contents, of the Spirulina cells) while maintaining a relatively low temperature (e.g., at a temperature of about 69°C or lower, or at room temperature or below, where "room temperature” refers to a temperature of between about 15°C (59°F) to about 25°C (77°F)).
  • a relatively low temperature e.g., at a temperature of about 69°C or lower, or at room temperature or below, where "room temperature” refers to a temperature of between about 15°C (59°F) to about 25°C (77°F)
  • the processing of the Spirulina can be performed in an ice bath.
  • the temperature is regulated to prevent denaturing of one or more cell contents, such as one or more proteins.
  • the Spirulina solution is blended or mixed.
  • Such blending can be performed, for example, using an immersion blender (e.g., a hand immersion blender), mixer, or the like, either continuously or periodically (e.g., for about 30 seconds or for a series of 30-second durations with waiting periods in between to avoid excessive temperature increase), via agitation (e.g., using a stir rod or stir bead), sonication (e.g., low- frequency sonication at about 500 Hz), ultrasonication, etc., in some embodiments, while the temperature is monitored and/or regulated.
  • an immersion blender e.g., a hand immersion blender
  • mixer e.g., a series of 30-second durations with waiting periods in between to avoid excessive temperature increase
  • agitation e.g., using a stir rod or stir bead
  • sonication e.g., low- frequency
  • Ultrasonication can cause cavitation, the application of impact forces, and the application of shear forces.
  • any other instrument (apart from a sonicator) or series of instruments (e.g., employed serially) designed to impose such forces can instead be employed.
  • the spirulina solution or any other intermediate spirulina-based product set forth herein can be subjected to cavitation using a first instrument, followed by the application of impact force(s) via a second instrument, followed by the application of shear force(s) via a third instrument
  • the spirulina solution or any other intermediate spirulina-based product set forth herein can be subjected to the application of impact fbrce(s) using a first instrument, followed by cavitation via a second instrument, followed by the application of shear force(s) via a third instrument.
  • the spirulina solution or any other intermediate spirulina-based product set forth herein can be subjected to the application of shear force(s) using a first instrument, followed by cavitation via a second instrument, followed by the application of impact fbrce(s) via a third instrument, or any other sequential combination thereof.
  • the aqueous Spirulina solution may be disrupted using any appropriate process, including mechanical or/or non-mechanical methods or processes.
  • Methods of disrupting the cellular material include, but are not limited to lysis, acid hydrolysis, desiccation, enzymatic treatment, use of detergents, solvents, or antibiotics, physical means (e.g., osmotic shock, pressure) to solid shear, use of a bead mill, cell press (e.g., French press), liquid shear, uhrasonication, sonication, high-pressure homogenizer(s), ultrafine shearing, alkaline pretreatment, acidic pretreatment, homogenization, high-pressure homogeni ration, thermal, hydrothermal, pulsed-electric field, microwave-assisted extraction, or a combination thereof.
  • mechanical disruption of the cells may be preferable to chemical disruption as it can reduce or prevent chemical contamination while preserving (or substantially preserving) the functionality of the cell contents.
  • the aqueous Spirulina solution is optionally ultrasonicated or otherwise cavitated (e.g., at greater than about 20kHz, for example about 40kHz) for a predetermined duration (e.g., about 30 minutes).
  • This cavitation step can be referred to as a "first disruption," in that at least a portion of the Spirulina proteins are liberated.
  • a "disruption" can refer to an at least partial liberation of proteins of the Spirulina or other cellular encapsulant material.
  • the aqueous Spirulina solution can be cooled (e.g., by adding ice, refrigerating, etc.) during one or both of 102G and 102H.
  • the cavitation step 102H can be performed cold or hot.
  • high temperatures may be avoided, for example so as to niinimize protein denaturing.
  • high temperatures may be desirable, for example, for antimicrobial purposes.
  • Hie disruptions of method 100 may be performed for any of a variety of durations, depending upon the implementation. For example, in some embodiments, the disruptions are performed for durations sufficient (either individually or collectively) to liberate the components, such as proteins, desired or needed for encapsulation.
  • Liberation of proteins and/or other components f om the Spirulina cell can be measured by any of a variety of mechanisms or methods appropriate for a particular application according to the disclosure.
  • the amount of proteins in solution can be measured by determining the absorbance of the solution at 280nm; as whole cells are disrupted, this absorbance measurement should increase.
  • protein concentration can be measured by methods including but not limited to: Bradford assay, colorimetric assays, biuret test, Lowry protein assay, and bicinchoninic acid assay (BCA).
  • the extract material (optionally decarboxylated) and the aqueous Spirulina solution are combined in a predetermined ratio to form an "intermediate mix."
  • the extract material and the Spirulina solution are combined shortly or immediately after the extract material is removed from a decarboxylation oven, while the extract material is still warm and flowable, e.g., for improved incorporation of the extract material into the aqueous Spirulina solution.
  • the aqueous Spirulina solution can be heated while being combined with the extract material, however in such cases the heating can be monitored and/or regulated, and/or controlled such mat it is applied for limited duration (e.g., at a temperature of about 69°C or lower and/or for a duration of not more man 3 hours) to avoid unwanted microbial growth. It should be noted mat the temperatures and times discussed herein can be varied based on the particular implementation, and any number of applicable factors, such as ambient humidity, controlled environment, pressure, etc.
  • the extract material is added to the aqueous Spirulina solution, while in other embodiments, the aqueous Spirulina solution is added to the extract material.
  • the predetermined ratio of extract material to Spirulina solution can be, for example, about 10: 1, about 9: 1, about 8: 1, about 7: 1, about 6: 1, about 5: 1, about 4: 1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:6, about 1:7, about 1:8, about 1:9, or about 1:10 (weight: eight).
  • the intermediate mix undergoes a "second disruption," during which encapsulation / stabilisation occurs, and which is performed via one or both of cold cavitation (e.g., ultrasonication) (102K) or heated cavitation (e.g., ultrasonication) (102L). As shown in FIG.
  • cold ultrasonication 102K (or any other method of cavitation) can optionally be performed for a predetermined duration (e.g., about 2 hours) and/or at a predetermined temperature (e.g., room temperature or below).
  • a predetermined duration e.g., about 2 hours
  • heated ultrasonication can be performed for a predetermined duration (e.g., about 45 minutes) and/or at a predetermined temperature (e.g., about 69°C).
  • heated ultrasonication is performed for a shorter duration man cold ultrasonication, for example to mitigate microbial growth.
  • the intermediate mix only undergoes one of cold ultrasonication 102K or heated ultrasonication 102L as part of me encapsulation step 102J. In other implementations, the intermediate mix only undergoes both at least one cold ultrasonication 102K and at least one heated ultrasonication 102L, in any order, as part of the encapsulation step 102J.
  • a "fully disrupted, encapsulated mixture" is produced, in which all or substantially all of the Spirulina proteins have been liberated, and in which all or substantially all of the extract material is encapsulated by the Spirulina solution.
  • a pH of the fully disrupted, encapsulated / stabilized mixture is optionally reduced (e.g., to a pH of about 7, about 6.S, about 6, about 5.5, about S, about 4.5, about 4, about 3.3, about 3, or about 2.S), for example by adding an acid, such as citric acid, ascorbic acid, phosphoric acid, tartaric acid, lactic acid, a combination of citric acid and ascorbic acid, any other food grade acid or combinations thereof, or one or more salts thereto.
  • an acid such as citric acid, ascorbic acid, phosphoric acid, tartaric acid, lactic acid, a combination of citric acid and ascorbic acid, any other food grade acid or combinations thereof, or one or more salts thereto.
  • Addition of a pH-adj listing additive can have one or more benefits, including: mitigation of microbial growth in the finished product, enhancement to salivary response, improved taste, action as a masking agent, increase buccal and/or sublingual absorbance/uptake, etc. In other embodiments, however, no pH reduction step 102M is performed, and the encapsulated mixture has a high pH of about 9 to 10.
  • the encapsulated mixture (e.g., in the form of a slurry) is poured onto a substrate to form a layer or film, for example such mat the poured layer has a predetermined volume per unit area (e.g., about 2.5mL per square inch of substrate area, or from about lmL to about SmL, or about 7mL, or about 8mL, or about 10mL, or about 12mL, or about 14mL, or about 16mL, or about 18mL, or about 18mL-22mL, or about 20mL, or about 22mL per square inch of substrate area).
  • a predetermined volume per unit area e.g., about 2.5mL per square inch of substrate area, or from about lmL to about SmL, or about 7mL, or about 8mL, or about 10mL, or about 12mL, or about 14mL, or about 16mL, or about 18mL, or about 18mL-22mL, or about 20mL, or about 22m
  • the substrate can be a glass, a coated surface (e.g., TEFLON-coated, etc.), silicone, or other substrate/non-stick substrate.
  • a predetermined temperature e.g., about 165°F
  • a predetermined duration e.g., about IS hours or longer, from about 1 minute to about 10 days, about 1 minute to about 60 minutes, about 1 hour to about 10 hours, about 2 hours to about 5 hours, about 5 hours to about 10 hours, about 10 hours to about 20 hours, about IS hours to about one day, about 1 day to about S days, or about 5 days to about 10 days
  • a predetermined environment e.g., ambient conditions, low- humidity conditions, nitrogen-filled drybox conditions, vacuum conditions, etc.
  • one or more other drying techniques 102Q can be performed on the fully disrupted, encapsulated mixture.
  • the finished product when prepared using drying steps 102N and 102P, and once removed from the silicone substrate, can have a flat, flake-like texture similar to flake-type fish food.
  • the flake-like finished product can be configured to not fully dissolve in water, and instead may dissociate.
  • the flakes or other product when exposed to an aqueous environment (e.g., sublingual, buccal, or oral environment), can be configured to disperse into particles having an average size from between about 0.5mm to about lnm.
  • the flakes can be graded (e.g., as small, medium, or large size) and sorted for use in a variety of applications.
  • the flakes can be ground, milled or otherwise further mechanically processed, for example to generate a powder for subsequent use in a variety of applications.
  • no step is performed at or above a temperature that promotes microbial growth, and/or such that mean temperature (and/or other environmental parameters) are maintained to avoid or substantially avoid conditions that allow or promote undesired or harmful microbial growth.
  • a sterilization step can be performed, e.g., to the finished product and/or to the Spirulina (e.g., prior to step 102F).
  • Sterilization can be performed, for example, using ultraviolet (UV) sterilization, proton sterilization, microwave sterilization, etc., for example to remove bacterial contamination, fungal contamination, viral contamination, and/or the like.
  • UV ultraviolet
  • sterilization can be performed by drying (102P or 102Q) at an elevated temperature (e.g., about 150°F, about 155 °F to about 165°F, about 165°F, about 150°F or higher, about 150°F to about 170°F, about 100°F to about 200°F, etc.) and/or at pressures and/or in conditions that reduce or neutralize one or more dangerous contaminants.
  • an elevated temperature e.g., about 150°F, about 155 °F to about 165°F, about 165°F, about 150°F or higher, about 150°F to about 170°F, about 100°F to about 200°F, etc.
  • dehydration at 102P when performed on the disrupted, encapsulated mixture without a pH reduction at 102M, can result in a modified consistency (e.g., a non-flake form) of the finished product.
  • a flake-like consistency can still be obtained, for example, by adding a salt, sodium bicarbonate, or other metabolite (e.g., prior to drying), and/or by increasing the ratio of spirulina to extract material (e.g., by increasing the amount of spirulina used and/or by decreasing the mount of extract material used).
  • the finished product is partially dissolvable in water or another solvent
  • the finished product when placed in water, may be about 20% insoluble, about 19% insoluble, about 18% insoluble, about 17% insoluble, about 16% insoluble, about 15% insoluble, about 14% insoluble, about 13% insoluble, about 12% insoluble, about 11% insoluble, about 10% insoluble, about 9% insoluble, about 8% insoluble, about 7% insoluble, about 6% insoluble, about 5% insoluble, about 4% insoluble, about 3% insoluble, about 2% insoluble, or about 1% insoluble.
  • one or more parameters can be modified such that less than all of the extract material is encapsulated by the Spirulina solution.
  • the finished product can be more fully dissolvable in water than the unmodified method 100.
  • a 'Spirulina solution can refer to a solution that includes one or more components (e.g., proteins, waxes, etc.) derived from Spirulina, and does not necessarily require that all parts of the original whole cell Spirulina are retained in the solution.
  • components e.g., proteins, waxes, etc.
  • one or more additives can be added, at one or more steps of the method 100 and/or can be added to the finished product
  • a powder or oil additive may be introduced prior to encapsulation.
  • an oil can be added before pouring, following an agitation step (e.g., to avoid encapsulation of the flavorant).
  • Additives can include, but are not limited to: vitamins, minerals, supplements, flavorants (e.g., peppermint oil, peppermint powder, peppermint extract, spearmint, menthol, cinnamon, ginger, orange, tangerine, peach, lime and lemon extract, pomegranate extract, chili oil etc.), relaxants (e.g., magnesium, zinc, melatonin, chamomile, etc.), and/or colorants (e.g., red, green, yellow, orange, blue, purple, etc.).
  • flavorants e.g., peppermint oil, peppermint powder, peppermint extract, spearmint, menthol, cinnamon, ginger, orange, tangerine, peach, lime and lemon extract, pomegranate extract, chili oil etc.
  • relaxants e.g., magnesium, zinc, melatonin, chamomile, etc.
  • colorants e.g., red, green, yellow, orange, blue, purple, etc.
  • Finished products produced by the methods set forth herein can be used standalone (e.g., as an additive for cooking, as a raw consumable, such as an edible, inhalable, or smokable, etc.) or incorporated into (e.g., used as an ingredient in) another product (e.g., oil-based infusions, inhalers/vaporizers, baked goods, beverages, etc.).
  • Finished products can be configured for admimstration via one or more pathways, including but not limited to mucosal membranes, via insufflation, inhalation, buccal administration, sublingual admimstration, etc.
  • example finished product prepared according to the method 100 of FIG. 1A were "water tested" in a cold, dark environment for 30 days by adding the finished product to water and a xanthan gum stabilizer, and no degradation was observed.
  • the finished product has a shelf life (i.e., where the active does not degrade more than 10%) of at least about 2 weeks, about 3 weeks, about 4 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 1 year.
  • shelf life i.e., where the active does not degrade more than 10%
  • a granulation can be performed after the decarboxylation 102B and before the combination 1021.
  • "granulation” refers to breaking down into smaller pieces, particles, particulates or components, for example by cutting, grinding, agitating, blending, chopping, pulverizing, etc.
  • Any strain of Cannabis e.g.. Black Jack, Gelato, Gorilla Glue #4 ("GG4"), Lemon Garlic OG, Sour Diesel, Sunset Sherbet, etc.
  • Black Jack Cannabis is grown in organic soil, has a 9-week flower cycle, and produces dense buds with a grape-like structure, often covered in THC -filled trichomes.
  • the Black Jack flavor profile begins with an earthy citrus flavor and finishes with a sweet and peppery flavor.
  • the GG4 flavor profile includes a sweet pine finish, earthy aromas and a distinct pungent note.
  • Sour Diesel is a Sativa-dominant strain with a citrus and diesel fuel aroma profile, and can be used to relieve anxiety, depression, and/or chronic fatigue.
  • Lemon Garlic OG is a 20% Sativa/80% lndica hybrid grown from feminized seeds. Lemon Garlic OG often includes low levels of CBD and from 8-20% THC, and is an aromatic strain with citrus, pine and garlic flavors.
  • the genetic profile of Cannabis when used to prepare the extract material for use in a process of the present disclosure, can influence the potency, flavor and/or other attributes of the finished product.
  • adjustments to the ratio of extract material to Spirulina solution to form the 'intermediate mix" can impact the potency (e.g., of the THC, CBD, and/or other active/psychoactive constituents of the extract material encapsulated therein), as well as the physical properties (e.g., texture, color, surface area, surface roughness, solubility, tensile strength, hardness, volume per weight, optical transparency or translucency, pore size, etc.) of the finished product.
  • the proportion of Spirulina solution can result in a less flaky finished product, resembling dried seaweed.
  • reducing the proportion of Spirulina solution can result in a finished product having a yellowish, highly brittle film in which not all of the extract material is encapsulated (i.e., the extract material is less than fully encapsulated).
  • the thickness of the poured encapsulated mixture at 102N afreets the consistency of the finished product.
  • a larger thickness or deeper pour
  • a relatively more flexible, ductile, brittle, resilient, flaky, firm, and/or elastic (i.e., stretchable) finished product depending on the other materials and processes utilized.
  • a relatively smaller thickness can lead to a more brittle finished product.
  • An excessively thick pour may result in a cracker- like product.
  • FIG. 1 A Since the method of FIG. 1 A uses a whole cell microorganism (rather than a single molecule constituent or other subdivision component of the whole cell microorganism) to encapsulate a lipid, there is no need to isolate such components, and the method can be faster and more efficient than known encapsulation methods.
  • a whole cell microorganism rather than a single molecule constituent or other subdivision component of the whole cell microorganism
  • FIG. IB is atree diagram 150 illustrating methods for encapsulation of an extract material, according to some embodiments.
  • the method includes the following steps: disruption of an aqueous solution (e.g., an aqueous solution containing cellular material, such as Spirulina, in a solvent) 204, extraction of constituent materials from the cellular materials 206, drying steps 208 and 214, constituent resolubilization/resuspension 210, combined mechanical disruption 212, use of a microencapsulated solid 216, and use of a microencapsulated liquid 218.
  • an aqueous solution e.g., an aqueous solution containing cellular material, such as Spirulina, in a solvent
  • constituent resolubilization/resuspension 210 e.g., constituent resolubilization/resuspension
  • combined mechanical disruption 212 e.g., use of a microencapsulated solid 216, and use of a microencapsul
  • Each of disruption 204, extraction 206, drying steps 208 and 214, constituent resuspension 210, and combined mechanical disruption 212, as indicated by asterisks in FIG. IB, can be subjected to temperature control (e.g., heating or cooling).
  • temperature control e.g., heating or cooling
  • Each of the microencapsulated liquid and the microencapsulated solid can be referred to as a finished product.
  • FIG. IB there are multiple possible paths through the diagram (i.e., multiple different methods are represented), as indicated by arrows.
  • a first disruption 204 is immediately followed by a combined mechanical disruption 212, for suspension of extraneous material and the formation of a microencapsulated liquid.
  • the microencapsulated liquid is either used, at 218, or dried at 214 to form a microencapsulated solid, which can then be used at 216.
  • a first disruption 204 is immediately followed by an extraction 206 before the combined mechanical disruption 212 for suspension of extraneous material and the formation of a microencapsulated liquid.
  • the microencapsulated liquid is either used, at 218, or dried at 214 to form a microencapsulated solid, which can then be used at 216.
  • a first disruption 204 is immediately followed by an extraction 206, a first drying step 208, and a constituent resuspension 210 before the combined mechanical disruption 212 for suspension of extraneous material and the formation of a microencapsulated liquid.
  • the microencapsulated liquid is either used, at 218, or dried at 214 to form a microencapsulated solid, which can then be used at 216.
  • the water when water or another polar solvent is used as the solvent, the water may be pH-adjusted, e.g., according to a predetermined timing.
  • the pH of the water is adjusted prior to cellular disruption (i.e., before 204), after cellular disruption (after 204), during the extraction process (206), during me constituent resuspension (210), during the combined disruption (212), or prior to the drying at 214.
  • the pH may be adjusted to a value of from about 7 to about 10, at concentrations ranging from about 0.5mM to about SOmM, using a pH-adjusting agent or compound, such as ammonium aluminum sulphate, ammonium bicarbonate, ammonium carbonate, ammonium hydroxide, ammonium phosphate di basic and ammonium phosphate mono basic, magnesium carbonate, magnesium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, sodium hydroxide, and/or calcium hydroxide.
  • a pH-adjusting agent or compound such as ammonium aluminum sulphate, ammonium bicarbonate, ammonium carbonate, ammonium hydroxide, ammonium phosphate di basic and ammonium phosphate mono basic, magnesium carbonate, magnesium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, sodium hydroxide, and/or calcium hydroxide.
  • pH-modification of the aqueous solution may be carried out by pH modifying compounds citric acid, acetic acid, amino acids such as glutamate, glycine, ascorbic acid, sodium hydroxide, tartaric acid, malic acid, fumaric acid, and lactic acid,
  • osmolality may be adjusted by the use of one or more salts and/or buffers, for example sodium chloride (NaCl) or magnesium sulfate (MgSO 4 ), at a concentration ranging from 0mM to 500mM.
  • salts and/or buffers for example sodium chloride (NaCl) or magnesium sulfate (MgSO 4 ), at a concentration ranging from 0mM to 500mM.
  • the amount of polar solvent in the final microencapsulated liquid can range from about 15% to about 99.9%, or about 50% to about 99.9%, or about 55% to about 95%, or about 60% to about 90%, or about 65% to about 85%, or about 70% to about 80%, or about 75% to about 99.9%, or about 75% to about 90%, or about 70% to about 90%, or about 60% to about 80%, or about 50% to about 80%, or about 90% to about 99.9%, or about 60% to about 80%, or about 95% to about 99.9%, or about 50% to about 70%, or about 50% to about 80%.
  • variations of hydrophobic compounds in the form of an oil or a wax, can include one or more terpenes, sterols, lipids, antioxidants, cannabinoids, and/or other compounds of medicinal and/or nutraceutical value.
  • terpenes included in microencapsulated material include bomeol, carophyllene, cineole, delta-3- Carene, limonene, linalool, myrcene, pinene, pulegone, and/or terpineol.
  • classes of plant sterols that may be included in the encapsulated material are type sterol esters, sterols, and pro-sterols.
  • Examples of lipids mat may be included in the encapsulated material include a-linolenic acid, arachidonic acid, eicosapentaenoic acid, and docosahexaenoic acid.
  • Examples of antioxidants that may be included in the encapsulated material include zeaxanthin, lutein, a-carotene, fucoxanthin astaxanthin, canthaxanthin, and ⁇ -carotene.
  • cannabinoids examples include Cannabigerol (CBG), Cannabichromene (CBC), Cannabicyclol (CBL), C ⁇ nnabivarin (CBV), Tetiahydrocaniiabivarin (THCV), Qmnabidivarin (CBDV), Caiinabichromevarin (CBCV), Cannabigerovarin (CBGV), Cannabigerol Monomethyl Ether (CBGM), Tetrahydrocannabinol (THC), Tetrahydrocannbinolic acid (THCA), Cannabidiol (CBD), Cannabidiolic Acid (CBDA).
  • CBD Cannabigerol
  • CBC Cannabichromene
  • CBL Cannabicyclol
  • CBV C ⁇ nnabivarin
  • THCV Tetiahydrocaniiabivarin
  • THCV Qmnabidivarin
  • CBDV Caiinabichromevarin
  • CBDV Cannabi
  • Example non-polar, hydrophobic additives to the oil/wax include fatty acids such as omega-3 fatty acids, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), alpha-Linolenic acid (a-Linolenic acid; ALA), conjugated linoledc acid and oils containing fatty acids such as fish oil, algae oil, krill oil, flaxseed oil, soybean oil, and walnut oil.
  • fatty acids such as omega-3 fatty acids, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), alpha-Linolenic acid (a-Linolenic acid; ALA), conjugated linoledc acid and oils containing fatty acids such as fish oil, algae oil, krill oil, flaxseed oil, soybean oil, and walnut oil.
  • Example additives to the oil/wax include compounds containing and contained in fatty acids such for example, triglycerides, including, polar lipids, for example, phosphoric acid, choline, fatty acid chains, glycerol, glycolipids, triglycerides, fatty acid esters, and phospholipids (e.g., phosphatidylcholine, phosphatidylethanolarnine, and phosphatidylinositol).
  • fatty acids such for example, triglycerides, including, polar lipids, for example, phosphoric acid, choline, fatty acid chains, glycerol, glycolipids, triglycerides, fatty acid esters, and phospholipids (e.g., phosphatidylcholine, phosphatidylethanolarnine, and phosphatidylinositol).
  • polar lipids for example, phosphoric acid, choline, fatty acid chains
  • example additives to the oil/wax are compounds containing fat-soluble vitamins, for example, Vitamins D, E, K, and A, corresponding provitamins and vitamin derivatives such as esters. Additionally, water-soluble vitamins, made lipid soluble may also be included. For example, ascorbyl palmitate, a fat-soluble version of vitamin C.
  • Example non-polar additives to the oil/wax include compounds containing carotenoids such as beta carotene, mixed carotenoids (mixtures of alpha and beta), zeaxanthin, capsanthin, cantfaaxanthin, bixin lycopene, violerythrin, gamma carotene, astaxanthin, and lutein.
  • carotenoids such as beta carotene, mixed carotenoids (mixtures of alpha and beta), zeaxanthin, capsanthin, cantfaaxanthin, bixin lycopene, violerythrin, gamma carotene, astaxanthin, and lutein.
  • herbs and/or spices may be added to wax/oil, such mat they may be microencapsulated with the concentrate, or added to the aqueous solution.
  • Example additives to the "concentrate” include compounds containing micronutrients, such as vitamins, minerals, co-factors, and extracts such as coenzymeQ10, curcurninoids (turmeric extract), ginger extract, niagen, trimethylglycine, inositol, choline, ciucoline N-acetyl-cysteine, astaxanthan, saffron, marigold, magnesium, zinc, melatonin, ginseng extract, glutamate, niacin, b vitamins, folic acid, vitamin bl2, taurine, glucaronic acid, malic acid, and n-acetyl-tyrosine.
  • micronutrients such as vitamins, minerals, co-factors, and extracts such as coenzymeQ10, curcurninoids (turmeric extract), ginger extract, niagen, trimethylglycine, inositol, choline, ciucoline N-acetyl-cysteine,
  • stabilizers mat may be added to the concentrate (i.e., the finished products), for example to improve "mouthfeel” and/or to modify a property thereof (e.g., to increase the density and/or viscosity of the concentrate) include guar gum, xanthan gum, gum acacia, sodium alginate, carrageenan, Saladizer®, and marine red-algae.
  • one or more preservatives such as EDTA, citric acid, acetic acid, sodium benzoate, sodium propionate, potassium sorbate, or sulphur dioxide, may be added to the concentrate or to one or both of the oil/wax material and the aqueous solution.
  • cellular materials may be broken, or "disrupted" (e.g., at 204 and/or 212) through use of mechanical disruption instrumentation such as a high pressure homogenizer, sonication (e.g., using a bath sonicator or a horn sonicator), Dounce homogenization, French press, and vigorous shaking with glass beads, paddle blenders, and industrial blending.
  • mechanical disruption instrumentation such as a high pressure homogenizer, sonication (e.g., using a bath sonicator or a horn sonicator), Dounce homogenization, French press, and vigorous shaking with glass beads, paddle blenders, and industrial blending.
  • disruption of cellular materials is performed via ultrasonication (or any other method of cavitation) at a frequency ranging from about 15kHz to about 75kHz, and at a power of about 120W or about 2S0W. Filtration or centrifugation may be used after cellular disruption, for example to remove unwanted cellular debris.
  • Microencapsulation of extraneous hydrophobic material may be performed before, during, and/or after the breaking/disruption of cellular wall/membrane material (204).
  • extraction of constituent materials from the cellular materials after mechanical disruption via solvent extraction (e.g., using ethanol, methanol, hexanes, ethy-lacetate, chloroform, other solvents, and/or mixtures of any of the foregoing), followed by drying (at 208) of the extracted material, and re solubilization (at 210) in water or another polar solvent
  • solvent extraction e.g., using ethanol, methanol, hexanes, ethy-lacetate, chloroform, other solvents, and/or mixtures of any of the foregoing
  • extraction of constituent materials from the cellular materials (206) after mechanical disruption (204), is followed by removal of unwanted cellular debris via a reduction in pH levels ranging from 3-5, removal of the supernatant, re suspension of the precipitate in a solvent having a pH ranging from 5-10, and drying of the precipitate (208).
  • the material may be resolubilized (at 210) into an aqueous solution having apHof from 3-9. Precipitation of the material is optional.
  • Resolubilization (at 210) of extracted cellular constituents may include one or more mixing, shaking, sonication, vortexing, and use of stabilizers such as glycerol, guar gum, xanthan gum, locust bean gum, acacia gum, karaya gum, and tara gum.
  • stabilizers such as glycerol, guar gum, xanthan gum, locust bean gum, acacia gum, karaya gum, and tara gum.
  • the cellular material includes whole cells, and one or more constituents thereof are left unextracted.
  • natural flocculation or aggregation of cellular material and encapsulation of a dense oil/wax material may lead to settling.
  • Reduction of coalescence can be abated by the addition of sulfuric acid, phosphoric acid, EDTA, and/or another chelating agent
  • cellular aggregation can be reduced by increasing the density of the solution using one or more salts, sugars, gums, or stabilizers.
  • no extraction of the constituent cellular material is performed, and the cellular material is directly used as a microencapsulant for a suspension of extraneous hydrophobic material (i.e., extract material) though use of mechanical cellular disruption at concentrations of cellular material ranging from about 0.1g/100ml (0.1%(w/v)) to about 10g/100ml (10% (w/v)) for maximal disruption of the cellular material.
  • extraneous hydrophobic material i.e., extract material
  • a selected ratio e.g., at 212 of microencapsulated extraneous hydrophobic material (in grams) to dry cell weight (in grams) of cellular material (whether extracted or not extracted) can range from about 0.001 to about 1 (w/w) for optimal microencapsulation of the extraneous material.
  • a selected ratio of compound to cellular material or extracted cellular material can depend on the type and characteristics of the extraneous hydrophobic material (extract material) to be encapsulated.
  • pre-heating, simultaneous heating and/or post-heating of cellular material is performed via a method such as gravimetric convection heating or water bath heating, before, during and/or after one or more cellular disruption mechanisms are implemented.
  • This heating may range from about 30°C to about 100°C (e.g., about 50 °C).
  • a selected heating temperature or temperature range can vary according to the type of cellular material, its recalcitrance, lipid, protein, cell wall type, content, and/or concentrations .
  • cooling of the cellular material before, during, and after the mechanical disruption process is performed, for example to a temperature ranging from about -20°C to about 15°C, to preserve protein structures that stabilize cellular membrane and lipid structures.
  • the polar solvent is altered before, during and/or after mechanical disruption of the material via addition of compounds, e.g., citric acid, acetic acid, trehalose, various polymeric gums, amino acids such a glutamate, glycerol, ethylenediarninetetraacetic acid (“EDTA”), phosphoric acid, ascorbic acid, sodium hydroxide, tartaric acid, malic acid, fumaric acid, and lactic acid, which alter pH, oxidation state, osmolality, activity, or viscosity.
  • compounds e.g., citric acid, acetic acid, trehalose, various polymeric gums, amino acids such a glutamate, glycerol, ethylenediarninetetraacetic acid (“EDTA”), phosphoric acid, ascorbic acid, sodium hydroxide, tartaric acid, malic acid, fumaric acid, and lactic acid, which alter pH, oxidation state, osmolality, activity, or viscosity.
  • mechanical cellular disruption is performed using a method that can depend on the type of cellular material being disrupted. Variations in lipid, protein, and cell wall content and type, as well as general recalcitrance, can influence the type and time of mechanical disruption.
  • the time for mechanical disruption can range from about 5 minutes to about 180 minutes, or about S minutes to about 180 minutes, or about 1 hour to about 3 hours, or about 5 minutes to about 15 minutes, or about 10 minutes to about 25 minutes, or about 20 minutes to about 30 minutes, or about 30 minutes to about 45 minutes, or about 60 minutes to about 90 minutes, or about 60 minutes to about 120 minutes, or about 90 minutes to about 120 minutes, or about 2 hours to about 3 hours, or about ISO minutes to about 180 minutes.
  • Reducing disruption time can help to maintain the integrity of the cellular material, whereas increasing disruption time can break/disrupt the cellular material while still allowing for micro-encapsulation of the extraneous, hydrophobic (extract) material.
  • preparation of the extraneous, hydrophobic (extract) material to be microencapsulated includes heating of the extraneous, hydrophobic (extract) material at a predetermined temperature and for a predetermined duration.
  • the extraneous compounds may undergo changes in physical form, e.g., from solid to liquid, or from liquid to gas, to facilitate complete or substantially complete microencapsulation.
  • Extraneous compounds may undergo structural changes catalyzed by heat, e.g., decarboxylation to improve encapsulation efficiency.
  • the temperatures for altering the extraneous compounds can range from about 50°C to about 270°C, and the heating duration for altering extraneous compounds can range from about 0 min. to about 120 min.
  • alteration of the microencapsulation is performed through the addition of one or more emulsifiers, polymers, and/or sugars, for example to form gels, jelly-like compounds, or gummy materials.
  • Microencapsulated extraneous material i.e., finished product
  • the suspension can be dried to produce a solid form of micro-encapsulated material.
  • the drying process may be carried out via one or methods such as, but not limited to: dehydration using vacuum, e.g., at a temperature ranging from about 10°C to about 100°C, laminar air flow, e.g., at a temperature ranging from about 10°C to about 100°C, one or more fans operated at ambient room temperature, lyophilization, and nano-spray drying.
  • microencapulated extraneous compounds in both liquid and solid from
  • dry goods such as cereals, grains, cakes, confections, spices, salts, seasonings for edible and medicinal consumption
  • microencapulated extraneous compounds in either liquid or solid form
  • Example uses of the concentrate/finished product include: liquid form "as-is,” diluted, or concentrated; solid form “as-is,” diluted, or concentrated; powder form as-is, diluted, or concentrated; incorporation into film(s); incorporation into flake(s); incorporation into sublingual dissolvable pill(s); incorporation into pressed pill(s); incorporation into sublingual dissolvable film(s); incorporation into gum(s), and gummies; incorporation into dried (dehydrated and freeze dried) fruits and flour(s); incorporation into tea(s), coffee(s), and juice(s); production of salt crystal(s) for edible consumption; production of salt crystal(s) for topical use; production of sugar crystal(s) for edible consumption; production of sugar crystal(s) for topical use; and incorporation into gel(s), ointment(s), salve(s), cream(s), and/or lubricants) for topical and/or mucosal absorption.
  • Example percentages of final components ("substances") in a dried/solid form of the concentrate are provided for powder, flake, and salt forms (Examples 1, 2, and 3, respectively) are as follows:
  • Example percentages of final components in a liquid form of the concentrate are as follows:
  • Hydrophobic or lipophilic material in aqueous solution will often separate into a bilayer solution, or seen as droplets (as a liquid) or will often stick to walls of a glass or plastic container (as a solid). With increased concentration of disrupted cellular material in aqueous solution, microencapsulation of hydrophobic material increases (up to a specific maximal point), as seen by reduced bilayer separation of hydrophobic material.
  • the hydrophobic material and disrupted cellular material are characterized by particular wavelength (nm) absorbance signatures. With increased concentration of disrupted cellular material (up to a specific maximal point), rnicroencapsulation of hydrophobic material increases (up to a specific maximal point), as determined by shifts in absorbance signatures at A280nm (e.g., general protein absorbance). Although the A280 signature can be used to determine the degree to which the Spirulina proteins have been liberated (associated with the degree of cell membrane disruption), other wavelengths are also of interest for characterization purposes and/or for comparing materials before and after encapsulation and/or before and after other steps of the preparation process (e.g., any wavelength between 240nm and 700nm).
  • Increased microencapsulation can be determined by microscopic evidence of structural and aggregation state changes of cellular material before and after implementation of the methods set forth herein.
  • Disruption of cellular material can be assessed by increased protein absorption at 280 nm, decrease in turbidity as determined by absorption at 600 nm, and/or direct observation of breaking cellular material via microscopy (e.g., brightfield or phase contrast microscopy).
  • microscopy e.g., brightfield or phase contrast microscopy
  • Desirable ranges for cellular disruption range from 40% to 100% lysis, as assessed by methods described in (d), depending on the type of cellular material disrupted.
  • Desirable concentration of cellular materials to increase percentage disruption ranges from about 0.1% to about 10% (w v). However, desirable encapsulation is often achieved using a greater percent of total cellular material, e.g., ranging from about 0.1% to about 40% (w/v).
  • encapsulant materials of the present disclosure are protein-based, and are fully or substantially de-fatted.
  • the encapsulant materials do not include a water phase carrier.
  • methods for encapsulation of an extract material, as described herein can readily emulsify at least 3% of a selected hydrophobic compound, using about 3% of whole cell mass or about 5% de-fatted, protein- rich whole cell mass.
  • a method for encapsulating of an extract material does not include heating to a temperature greater man 75 °C, since doing so can destabilize the encapsulation process.
  • a method for encapsulating of an extract material includes mixing and/or emulsification at a rate of less man about 5,000 rpm, since higher mixing rates can result in the unwanted introduction of excess air, and can lead to foaming of the solution.
  • a method for encapsulating of an extract material includes ultrasonicating (or any other method of cavitating) at 10kHz or greater.
  • a method for encapsulating of an extract material does not include a filtration step.
  • FIG. 2 is a process flow diagram iUustrating a method 200 for preparing an extract material, according to some embodiments.
  • a plant e.g., a Cannabis plant
  • One or more of the stems 220B, full leaves 220C, and trim 220D can be dried, at 220E, and, optionally, granulated (220F).
  • the dried plant material undergoes an extraction according to one of solvent extraction (220G), CO 2 extraction (220H), oil extraction (220J), or solventless extraction (e.g., rosin, resin, or live resin process) (220K).
  • solvent extraction 220G
  • CO 2 extraction 220H
  • oil extraction 220J
  • solventless extraction e.g., rosin, resin, or live resin process
  • two or more of solvent extraction (220G), CO 2 extraction (220H), oil extraction (220J), a solventless extraction (220K) can be performed on the dried plant material as part of an overall extraction process flow.
  • distillation can refer to any process of separating one or more components or substances from a liquid mixture by selective boiling and condensation, to produce a "purified" product.
  • the distillation is performed by molecular distillation, to minimize degradation of one or more of the extract component "Winterization” can refer to any process by which cannabinoids and/or terpenes or other fats are separated from Cannabis plant material. Winterization can include an alcohol "wash.”
  • FIG. 3 is a block diagram showing components of a system 300 for encapsulation of an extract material, according to some embodiments.
  • the system 300 can include an oven 322B (e.g., for performing drying as shown at 208 and/or 214 in FIG. IB), a dehydrator 322C (e.g., for dehydrating as shown at 102P in FIG. 1A), a blender 322D (e.g., for blending of Spirulina solution as shown at 102G in FIG. 1A), a decarboxylator 322E (e.g., for performing decarboxylation of extract material as shown at 102B in FIG.
  • an oven 322B e.g., for performing drying as shown at 208 and/or 214 in FIG. IB
  • a dehydrator 322C e.g., for dehydrating as shown at 102P in FIG. 1A
  • a blender 322D e.g., for blending of Spirulina solution
  • an ultrasonicator 322F e.g., for optional uhrasonication (or cavitation via any other means set forth herein), as shown at 102H, 102K and/or 102L in FIG. 1A
  • a sonicator 322G e.g., for blending of Spirulina solution as shown at 102G in FIG. 1A
  • an extraction unit 322A e.g., for blending of Spirulina solution as shown at 102G in FIG. 1A
  • FIGS. 4A-4D are images showing example finished products, according to some embodiments.
  • the finished product shown in FIG. 4A was prepared using a ratio of 1:1 (w:w) Spirulina:Wax (e.g., at step 1021 of FIG. 1A), a pour thickness (e.g., at step 102N of FIG. 1A) of about 2.5mL/in 2 , a drying duration of about IS hours.
  • the finished product shown in FIG.4B was prepared using a ratio of 3:2 (w:w) Spirulina: Wax (e.g., at step 1021 of FIG. 1A), a pour thickness (e.g., at step 102N of FIG.
  • the finished product shown in FIG. 4C was prepared using a ratio of 1 : 1 (w:w) Spirulina: Wax (e.g., at step 1021 of FIG. 1A), a very thin pour thickness (e.g., at step 102N of FIG. 1A) (i.e., a pour thickness significantly less than 2.SmL/in 2 ), and a drying duration of about 39 hours.
  • the finished product can have a flake-type appearance and texture, and can be highly brittle. Flakes can be irregularly shaped and, unless sorted or otherwise further processed (e.g., broken up, ground, cleaved, filtered, etc.), can vary dramatically in size.
  • a fast-dissolve powder can be generated following a process set forth herein (such as the process illustrated in FIG. 1 A), but using a spray dryer instead of a dehydrator or other drying technique.
  • shaped products can be produced by adding one or more thickening agents, such as agar-agar, gelatin, acacia gum, lerithin, or guar gum, and/or one or more edible plasticizers, such as glycerol, to the encapsulated mixture (e.g., at step 102J of process 100 in FIG. 1A.
  • such additives can result in an encapsulate mixture that is mechanically shapeable (e.g., via cutting, molding, extruding, bending, folding, etc.).
  • An example, hexagonally-shaped product is shown in FIG.4E.
  • An example powder was prepared as follows: 1.2 grams of whole-cell Spirulina powder were mixed by band with 300 mg of distillate oil, men macerated using a mortar and pestle (although other methods of maceration, mixing, or grinding, including using industrial equipment, are also contemplated), to yield a powdery substance, having a final appearance as shown in FIG.4F. Mixtures of the spirulina powder with one or more flours, such as rice flour, tapioca flour, potato starch, corn starch, and/or other polymer sugars, such as mahodextrin, also allow for a powder-like consistency to be formed. In some embodiments, a protein extraction can be performed prior to the preparation of the powder, and the resulting extracted protein extract spirulina powder can be used in lieu of the whole- cell Spirulina powder.
  • flours such as rice flour, tapioca flour, potato starch, corn starch, and/or other polymer sugars, such as mahodextrin
  • FIG.5 is an illustration of an example microencapsulated extract material (e.g., of a finished product), also referred to herein as a "microparticle'' or "microcapsule,” according to an embodiment.
  • a microparticle can have a size (e.g., a diameter) of about 1 micrometer ( ⁇ m) to about 1 millimeter (mm), or about 1 ⁇ m to about 5 ⁇ m, or about 5 ⁇ m to about 10 ⁇ m, or about l ⁇ m to about 10 ⁇ m, or about 1 ⁇ m to about 100 ⁇ m, or about 10 ⁇ m to about 100 ⁇ m, or about 100 ⁇ m to about 500 ⁇ m, or about 500 ⁇ m to about lmm, or about lmm to about 5mm, or about lmm to about 2mm, or about 50 ⁇ m to about 75 um, or about 100 ⁇ m to about 250 ⁇ m, or about 10 ⁇ m to about 500 ⁇ m, or about 100um to about 5mm.
  • a microparticle can have a core/shell structure (a microcapsule structure) including an extract material core 524 surrounded by an encapsulant 526 (e.g., including Spirulina).
  • a microparticle can have any other shape, such as irregular, capsule-shaped, rod-shaped, etc.
  • each microparticle in a finished product can have a different composition. Characterization of microparticles can be performed using one or more of scanning electron microscopy (SEM), optical microscope, laser diffraction, fluorescence microscopy, etc.
  • the microcapsule and/or finished product containing microcapsules of the disclosure are configured to not include, not include any added, be free from added, be substantially free from added, or be essentially free from added one or more surfactants, one or more artificial surfactants, one or more detergents, one or more emulsifiers, one or more lecithins, one or more lysolethesins, one or more glycolipids, one or more saponins, one or more monoglycerides, one or more sorbitan esters, one or more sucrose esters, one or more saturated fatty acids, one or more unsaturated fatty acids, and/or one or more hydrocolloid polymers.
  • the nucrocapsule/microparticle and/or finished product does not include, or is substantially free from, polyethylene glycol ("PEG"). In some embodiments, the nucrocapsule/microparticle and/or finished product does not include, or is substantially free from, a polysorbate (e.g., polysorbate 20, 40, 60, 65 and/or 80). In some embodiments, the nucrocapsule/microparticle and/or finished product does not include, or is substantially free from, any Tween compound (e.g., Tween 20, Tween 40, Tween 60, or Tween 80).
  • PEG polyethylene glycol
  • a polysorbate e.g., polysorbate 20, 40, 60, 65 and/or 80.
  • any Tween compound e.g., Tween 20, Tween 40, Tween 60, or Tween 80.
  • the macocapsule/microparticle and/or finished product does not include, or is substantially free from, Polyoxyethene (8) stearate. In some embodiments, the microcapsule/microparticle and/or finished product does not include, or is substantially free from, brorninated vegetable oil. In some embodiments, the microcapsule/microparticle and/or finished product does not include, or is substantially free from, succistearin (stearoyl propylene glycol hydrogen succinate). In some embodiments, the microcapsule/microparticle and/or finished product does not include, or is substantially free from, Propanediol 1-2 esters of fatty acids. In some embodiments, the macocapsule/microparticle and/or finished product does not include, or is substantially free from, propylene glycol esters of fatty acids.
  • a finished product is in the form of a dissolvable or semi- dissolvable strip or patch for absorption through me gums, cheek, and/or oral mucosal region of a user, for example to reduce pain associated with dental work (e.g., removal of wisdom teeth, dry socket conditions, etc.).
  • a finished product is configured for use in one or more of a topical salve, a cream, a gel, a solution, a bath salt, a liquid and/or a solid.
  • contemplated additives can include (but are not limited to) one or more of the following: arnica, calendula, California poppy, olive oil, sunflower oil, avocado oil, beeswax, sage oil, lavender oil, cypress oil, red cedar oil, Chamomile, Lavender oil, Aloe, Mugwort, Himalayan salt, Epsom salt, eucalyptus, horsetail, eyebright, shea butter, sweet almond oil, rosehip seed oil, pomegranate seed oil, and cypress essential oil.
  • the finished product is a liquid, including an aqueous liquid, that includes microencapsulated materials, nanoencapsulated materials, microcapsules, and/or nanocapsules, according to the disclosure.
  • Such embodiments include but are not limited to suspensions, solutions, and/or emulsions.
  • the microencapsulation/nanoencapsulation provides enhanced stability over time, such as less than a 5%, 10%, 15%, 20% or 25% loss of potency of the active microencapsulated and/or nanoencapsulated active ingredient(s) over time, such as 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, or more.
  • the microencapsu mecanic/nanoencapsulation provides enhanced stability in environment factors, such as exposure to air, light, heat (and/or temperature changes).
  • the nucroencapsulanon/nanoencapsulation provides enhanced bioavailability.
  • the finished product (including one or more cannabinoids or cannabis extracts/components) can be used in treatment regimens for and/or treating one or more of chronic pain, paralysis, neuropathy, Crohn's Disease, inflammatory bowel disorders (i.e., UBS and IBD), glaucoma, post-traumatic stress disorder (PTSD), anxiety, seizures, epilepsy, autoimmune disorders, autism, tumors, and/or one or more types of cancer.
  • the finished product can additionally or alternatively be configured for treating nausea and vomiting mat are unresponsive to other medications, particular given the improved absoiption/bioavailability provided by the disclosed products.
  • the finished product (including one or more cannabinoids, cannabis extracts/components, including THC and/or CBD) can be used for treatment regimens for dependency on opioids.
  • the disclosure includes methods of alleviating a symptom associated with anxiety, post-traumatic stress disorder, chronic pain, or opiate dependency, paralysis, neuropathy, Crohn's disease, inflammatory bowel disorders, glaucoma, seizures, epilepsy, autism, or cancer comprising administering to a subject one or more formulations/final products according to the disclosure.
  • a formulation/final product is administered once a day, twice a day, three times a day, four times a day, five times a day, six times a day, or more.
  • a fbrmulation/final product is administered in the morning, afternoon, evening and at bedtime.
  • active components of the formulation(s)/final produces) can be configured for a particular time (e.g., a morning formulation could have relatively more or less active, such as THC, either by quantity or relative proportion to another active, such as CBD, when compared to an evening formulation).
  • liquid finished product includes microencapsulated material dissolved within, substantially dissolved within, and/or suspended within a solvent or supernatant.
  • the final product can be a solution, a suspension, or a mixture.
  • An example solution-type formulation was prepared, including 3% Spirulina and 3% THC wax, by premixing with a Silverson high speed mixer for 30 seconds, followed by 45 minutes of uhrasonication (or any other method of cavitation) at 10°C. The formulation was then diluted with water (having a pH of ⁇ 9), to form a final solution having a ratio of 1 : 20,000 (foimulation:water). The final solution can also be referred to as a THC- infused liquid, or a Spiiulinex solution. A volume of 1 mL of the final solution was men analyzed, as described below. pH Range Testing of Example Solution
  • Microscopy images of the final solution were obtained using instrumentation developed by Fluid Imaging Technologies, model FlowCam VS1.
  • a 300uM capillary flow cell was used, with a 4X objective lens and an imaging resolution cutoff of 2 ⁇ m. Details pertaining to the instrumentation and experimental settings are provided in Table 1 ("Liquid Imaging Parameters”) and Table S ("Additional Experimental Details”), below. Particles less man 2 ⁇ m were not imaged during microimaging.
  • FIGs. 7A- 7D Graphical representations of the particle population based on log frequency, volume %, circle fit, and intensity, each as a function of diameter, are shown in FIGs. 7A- 7D (respectively). Furthermore, two distinct populations of particles were identified using edge gradient (a high edge gradient population and a low edge gradient population) .
  • Nanoscale measurements of a second final solution prepared using the formulation described above with reference to the microscale imaging, but further diluted in water (the water having a pH of ⁇ 9) to 1:200,000 (instead of 1:20,000 as was done for microimaging), were recorded using a Nanosight instrument designed for Nanoparticle Tracking Analysis, version 2.3. All measurements were performed in triplicate. Sample information for each of three samples is provided in Table 6.
  • a "D value” is defined as the diameter of a particle at which a defined percentage of the sample's mass is comprised of a diameter less than that value.
  • the D50 value is the diameter at which 50% of the particles of the sample have a diameter less than the value.
  • Particle count analysis also reveals total concentration of particles/mL of solution.
  • Table 9 shows the concentration-weighted nanoimaging size data for the second final solution.
  • Key features of the analyzed second final solution particle population include: a mean diameter of 174nm +/- 4. lnm, a mode of 79nm +/- 0.3nm, and a DS0 value of 13 lnm +/- 3.2nm.
  • Plots of the particle concentration per particle diameter are presented in FIGS.9A and 9B, with FIG 9B showing plots for each of the three sample measurements.
  • the raw data used to generate the plots of FIGs.9A-9B is presented in Appendix B.
  • nanoscale particle/droplet sizes in final products prepared according to the present disclosure can impact the bioavailability and/or absorption rate of one or more active ingredients (e.g., THC, CBD) when the final product (e.g., solution) is ingested.
  • active ingredients e.g., THC, CBD
  • smaller droplet size may facilitate faster absorption or increased oral bioavailability, such that the related onset of the therapeutic or medicinal effects) occurs sooner, whether the final product being consumed is in solution form or in a dry (e.g., flake) form.
  • absorption mechanisms relating to nanoscale materials can be found in: Bapi Gorain, Hira Choudhury, Amit Kundu, Lipi Sarkar, Sanmoy Karmakar, P.
  • a range of masses (0.15 grams, 0.20 grams, 0.25 grams, 0.30 grams, 0.40 grams, 0.50 grams, 0.60 grams, and 0.70 grams) of protein powder were mixed with 150mg distillate oil and water (to 5 mL, and readjusted to pH 8.5), and were homogenized using high shear mixing and heated ultrasonic cavitation, followed by filtration using a 1uM glass syringe filter. Solutions were men stored at 4°C overnight and examined for visible oil precipitate (as evidenced by sticking to plastic) and general precipitate (slurry at bottom of tube), as summarized in Table 11 below. As shown in Table 11, there is a general trend me increased protein correlates with decreased oil precipitation.
  • Table 12 below shows protein sequences for the top 10 proteins and their associated Grand Average of Hydropathy ("GRAVY”)- GRAVY index scores (averaging hydrophobicity and hydrophilicity) for the proteins were determined using the Kyte- Doolittle and Hopp Woods formula. Hydrophobicity scores (arbitrary units) below 0 are more likely to be globular (hydrophilic protein), and scores above 0 are more likely to be membrane-interacting (hydrophobic). In some instances, proteins encapsulating hydrophobic materials in aqueous solution can be amphipathic, thus having regions of both hydrophobicity and hydrophilicity.
  • GRAVY Grand Average of Hydropathy
  • encapsulation occurring during the preparation of the formulations described herein is accomplished, at least in part, by virtue of one or more key proteins (e.g., apcE Phycobilisome core- membrane linker polypeptide (Anchor polypeptide LCM), cpcA C-phycocyanin alpha subunit, apcB Allophycocyanin beta subunit, cpcB C-phycocyanin beta subunit, ribH 6,7- dimethyl-8-ribityllumazine synthase protease, ATP-dependent zinc-metallo (fragment), etc.) of Aithrospira platensis or edible versions of the genus Arthrospira, which are liberated during the preparation process.
  • apcE Phycobilisome core- membrane linker polypeptide chor polypeptide LCM
  • cpcA C-phycocyanin alpha subunit e.g., apcB Allophycocyanin beta subunit, cp
  • the term ''automatically is used herein to modify actions mat occur without direct input or prompting by an external source such as a user. Automatically occurring actions can occur periodically, sporadically, in response to a detected event (e.g., a user logging in), or according to a predetermined schedule.
  • determining encompasses a wide variety of actions and, therefore, “determining” can include calculating, computing, processing, deriving, mvestigating, looking up (e.g., looking up in a table, a database or another data structure), ascertaining and the like. Also, “deterrnining” can include receiving (e.g., receiving information), accessing (e.g., accessing data in a memory) and the like. Also, “determining” can include resolving, selecting, choosing, establishing and the like.
  • Hardware modules may include, for example, a general -purpose processor, a field programmable gate array (FPGA), and/or an application specific integrated circuit (ASIC).
  • Software modules (executed on hardware) can be expressed in a variety of software languages (e.g., computer code), including C, C++, JavaTM, Ruby, Visual BasicTM, and/or other object-oriented, procedural, or other programming language and development tools.
  • Examples of computer code include, but are not limited to, micro-code or micro-instnictions, machine instructions, such as produced by a compiler, code used to produce a web service, and files containing higher-level instructions that are executed by a computer using an interpreter.
  • embodiments may be implemented using imperative programming languages (e.g., C, Fortran, etc.), functional programming languages (Haskell, Erlang, etc.), logical programming languages (e.g., Prolog), object-oriented programming languages (e.g., Java, C++, etc.) or other suitable programming languages and/or development tools.
  • Additional examples of computer code include, but are not limited to, control signals, encrypted code, and compressed code.
  • Various concepts may be embodied as one or more methods, of which at least one example has been provided. Hie acts performed as part of the method may be ordered in any suitable way. Accordingly, embodiments may be constructed in which acts are performed in an order different than illustrated, which may include performing some acts simultaneously, even though shown as sequential acts in illustrative embodiments. Put differently, it is to be understood that such features may not necessarily be limited to a particular order of execution, but rather, any number of threads, processes, services, servers, and/or the like that may execute serially, asynchronously, concurrently, in parallel, simultaneously, synchronously, and/or the like in a manner consistent with the disclosure. As such, some of these features may be mutually contradictory, in that they cannot be simultaneously present in a single embodiment. Similarly, some features are applicable to one aspect of the innovations, and inapplicable to others.
  • the disclosure may include other innovations not presently described. Applicant reserves all rights in such innovations, including the right to embodiment such innovations, file additional applications, continuations, continuations-in- part, divisionals, and/or the like thereof.
  • advantages, embodiments, examples, functional, features, logical, operational, organizational, structural, topological, and/or other aspects of the disclosure are not to be considered limitations on the disclosure as defined by the embodiments or limitations on equivalents to the embodiments.
  • the terms "about” or “approximately,” when preceding a numerical value, indicate the specified value plus or minus a range of 10%.
  • a duration of “about 30 minutes” refers to a duration of from 27 minutes to 33 minutes.
  • a reference to "A and/or B", when used in conjunction with open-ended language such as “comprising” can refer, in one embodiment, to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); etc.
  • the phrase "at least one,” in reference to a list of one or more elements, should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. Ibis definition also allows mat elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase "at least one" refers, whether related or unrelated to those elements specifically identified.
  • At least one of A and B can refer, in one embodiment, to at least one, optionally including more than one, A, with no B present (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements); etc.

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Abstract

L'invention concerne un procédé pour la microencapsulation de terpènes, de lipides, de stérols, d'antioxydants, de cannabinoïdes et/ou d'autres composés hydrophobes, lequel procédé consiste à fournir un extrait de matériau et à préparer une solution aqueuse comprenant un matériau cellulaire. Une perturbation de la solution aqueuse est effectuée, pour libérer au moins partiellement des protéines du matériau cellulaire. L'extrait de matériau et la solution aqueuse perturbée sont combinés pour former un mélange intermédiaire. Le mélange intermédiaire est perturbé pour former un mélange encapsulé. Le mélange encapsulé est ensuite séché, pour former un produit fini.
PCT/US2018/054216 2017-10-03 2018-10-03 Procédés de préparation de compositions à base de plantes encapsulées, solubilisables, produits à base de celles-ci Ceased WO2019070885A1 (fr)

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