Albumin Protein Used as an Emulsifier and Drug Carrier

Abstract

The present disclosure is directed to a method of using albumin as an emulsifier to aid in blending otherwise immiscible ingredients. The method uses albumin as an emulsifier and carrier. The present disclosure also teaches the use of albumin as a microencapsulated liquid, gel or powder, and as a drug or dietary supplement carrier to enhance bioavailability.

Description

[Technical field]

[0001] This disclosure relates to the use of albumin as an emulsifier, foaming agent or drug carrier. [Background technology]

[0002]

[0002] Albumin is a protein found in the seeds of several plants, including cannabis seeds (Cannabis sativa), kidney beans (Phaseolus vulgaris), and carob beans (Parkia biglobosa). Albumin is also a bioactive protein produced in the human body, making up 55% of the proteins in blood plasma. Albumin has a variety of functions, including maintaining blood pressure and transporting nutrients and other bioactive molecules, such as cannabinoids.

[0003]

[0003] The bioavailability of drugs to reach their intended biological end-point is a known problem in the medical field. In general, the dose of a drug is indirectly proportional to its bioavailability. The use of carrier proteins such as albumin has been shown to help more efficiently deliver drugs and other compounds with low bioavailability, and to increase the solubility of drugs and other compounds that are poorly soluble in water.

[0004]

[0004] These bioactive molecules are important in regulating blood pressure and are injected into patients in cases where emergency blood pressure regulation is required, such as in burns, shock, and liver disease.

[0005] Albumin as a drug carrier (e.g. for cannabinoids) can be injected into the human body or taken orally to increase the blood concentration of the drug. Many compounds, including hormones, drugs, fatty acids, and steroids, are carried in human blood by the albumin protein. Albumin has a short half-life of 20 days in the human body, which can lead to albumin deficiency in the body. This deficiency can be life-threatening.

[0005]

[0006] Proteins from plant seeds are a rich alternative to animal-derived protein sources. In microencapsulation, these proteins are used as wall-forming materials for various active compounds. In most cases, two types of techniques of microencapsulation, spray drying and droplet formation, are used for the preparation of microparticles from plant proteins. Proteins extracted from soybean, pea and wheat have already been investigated as carrier materials for microparticles.

[0006]

[0007] Microencapsulation and nanoencapsulation are techniques used to isolate and deliver liquids or other ingredients to patients in need. The encapsulated material is called the core or inner phase, while the outer material is considered the outer or envelope phase.

[0007]

[0008] Cannabis seeds are used worldwide as a food source and as a dietary supplement. Cannabis inflorescences are rich in non-psychotropic, biologically active cannabinoids. Cannabis seeds have a pleasant nutty taste and are a valuable source of essential fatty acids, minerals, vitamins, fiber, and essential amino acids.

[0008]

[0009] Refined hemp protein has traditionally been a low-quality, undesirable product due to the type of manufacturing methods used to produce it. Some negative attributes include a coarse, grainy texture, insolubility in water, a dark, "earthy" appearance, and an "earthy" flavor.

[0009]

[0010] In the food industry, there is a wide variation in the emulsifying properties reported for different proteins. Meat proteins tend to be good emulsifiers, but using vegetable proteins as emulsifiers may have marketing advantages. In addition to being a drug carrier, albumin can act as both an emulsifier and a foaming agent.

[0010]

[0011] As outlined above, albumin is a versatile, highly soluble, stable, non-toxic, non-toxic, biocompatible, and biodegradable protein. Due to its versatile nature, it can be used to deliver drugs, hormones, metals, and fatty acids by conjugating these molecules to specific binding sites on albumin. The structure, location, size, charge, and hydrophobicity of these drug binding sites are of great importance to optimize the interaction of albumin with the drug.

[0011]

[0012] Until recently, the only source of albumin was donated blood, but since albumin has an inherent risk of contamination and its protein decontamination is difficult, efforts are currently being made to produce albumin from genetically modified rice by isoelectric focusing.

[0012]

[0013] Non-genetically modified, certified organic cannabis is rich in albumin (35% of total protein). Potential uses of cannabis-derived albumin include emergency medical procedures to stabilize blood pressure and to increase the bioavailability of drugs and dietary supplements that are not normally absorbed by the body. These facts are not well known in the cannabis and dietary supplement industries. A source of non-genetically modified albumin also offers marketing advantages.

[0013]

[0014] Recent studies demonstrate the ability of albumin to enhance the water solubility of other molecules. Albumin plays the role of a "biosolubilizer" and allows the solubilization of a wide range of biomolecules and drugs in hydrophilic media, i.e., plasma. The solubilization-enhancing properties of albumin are mainly due to its ability to form reversible binding complexes with ligands. This allows the bound molecule to flow through the blood in a higher concentration than its initial solubility. Albumin has two main sites that bind ligands mainly by hydrophobic and electrostatic interactions. At physiological pH, the overall charge of albumin is negative, but the two main binding sites are positively charged, facilitating the binding of anionic molecules. In addition, albumin has several secondary binding sites, which increase the number of molecules that it binds, for example, up to seven fatty acid molecules. Among the substances that show the highest affinity for albumin are anionic molecules (weak acids) and hydrophobic molecules of medium size (100-600 Da); poorly soluble drugs. In addition, the albumin molecule has numerous accessible free amino and carboxyl groups suitable for forming highly soluble salts with acidic or basic drugs, respectively. Summary of the Invention

[0014]

[0015] Embodiments of the present disclosure also provide for the use of albumin from any source, including but not limited to any plant-derived, human-derived, or animal-derived source, and synthetic sources such as yeast or bacterial fermentation, for use as an emulsifier to aid in the blending of otherwise immiscible ingredients.

[0015]

[0016] Some embodiments provide for the use of albumin from any source, including but not limited to any plant-derived, human-derived, or animal-derived source, and synthetic sources such as yeast or bacterial fermentation, for use as a drug carrier.

[0016]

[0017] Additional embodiments provide for the use of albumin from any source, including but not limited to any plant, human, or animal derived source, and synthetic sources such as yeast or bacterial fermentation, for use in enhancing the bioavailability of poorly water soluble drugs or food compounds.

[0017]

[0018] In some embodiments, a method of blending immiscible components using an emulsifier is provided, comprising using a protein as the emulsifier. The source of protein may be albumin from any source, and blending results in an emulsion. The protein fraction may be used in dry, gelled or aqueous form and may be stabilized by the addition of a flow agent. The protein fraction may be wet milled, resulting in proteins greater than 5 kDa and with solubility in the pH range of 8.0-12.0, at concentrations ranging from 40-99%. Furthermore, the emulsion may have a loading capacity of up to 60%. The stability of the emulsion may be extended by the addition of nanocellulose, with a particle size of about 50 nm.

[0018]

[0019] In some embodiments, a method for producing microencapsulates and nanoencapsulates is provided, comprising dissolving albumin in water, adding lipid-based components, and mixing by sonication or high pressure homogenization. The encapsulated product may be soluble in oil or water and may contain a surfactant composed of a water-soluble protein. The encapsulated product may be coated with water-soluble plant-derived proteins and used as a drug or dietary supplement carrier. The encapsulated product may be used to enhance the bioavailability of drugs. In some embodiments, the albumin may be obtained by using algae-derived polymers. The albumin may be concentrated by isoelectric focusing, water-salt dialysis, or ultrafiltration. In some embodiments, the albumin may be enzymatically or chemically hydrolyzed. The albumin may be treated with absorbents or chemicals to remove color and flavor. Additional embodiments include a method in which the lipid-based component is selected from the group consisting of cannabinoid oil, edible oil, pharmaceutical lipids, and combinations thereof, wherein the lipid-based component remains liquid at ambient temperature.

[0019]

[0020] In an additional embodiment, a method of using albumin as a drug carrier is provided, comprising binding the albumin to a drug and orally delivering the drug carrier to a mammal. The drug may be ingestible or injectable. The drug may be a cannabinoid.

[0020]

[0021] In some embodiments, a method is provided for improving the water solubility of a compound by adding emulsified albumin to the compound, the compound being a food compound. The food compound may have a lipid-based component. [Brief description of the drawings]

[0021] [Figure 1]

[0022] Albumin proteins (shown as light spheres) are shown carrying fatty acid molecules (shown as dark spheres and also indicated by arrows). [Diagram 2]

[0023] FIG. 1 is a flow chart showing the process of isolating and extracting cannabis protein from cannabis seeds. The raw material (cannabis seeds) can be replaced with a myriad of plant and / or animal raw materials. The isolated protein has a molecular weight above 5 kDa. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022]

[0024] Reference will now be made in detail to exemplary embodiments of the present disclosure. While the present disclosure describes several embodiments, it will be understood that it is not intended to be limited to these embodiments. On the contrary, the present disclosure is intended to cover all alternatives, modifications, and equivalents that may be included as defined by the claims.

[0023]

[0025] One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of this disclosure and are within the scope of the present disclosure, but are in no way limited to the methods and materials described.

[0024]

[0026] All publications, published patent documents, and patent applications cited in this application are indicative of the level of skill in the art to which this application pertains. All publications, published patent documents, and patent applications cited in this specification are herein incorporated by reference to the same extent as if each individual publication, published patent document, or patent application was specifically and individually indicated to be incorporated by reference.

[0025]

[0027] In some embodiments, methods are provided for the microencapsulation and nanoencapsulation of products, where the emulsified product contains an oil soluble active ingredient and a surfactant composed of water soluble proteins from polymeric separations (animal, fungal, whole plant, spent / processed materials, leaves / seeds).

[0026]

[0028] In some embodiments, sources with higher concentrations of protein, such as hemp seeds, almonds, or chia seeds, are more preferred.

[0029] In some embodiments, the water-soluble protein fraction must maintain solubility over a broad pH range of 8-12. In some embodiments, the water-soluble protein fraction must maintain solubility over a broad pH range of 9-12. In some embodiments, the water-soluble protein fraction must maintain solubility over a broad pH range of 10-12. In some embodiments, the isolated protein will be greater than 5 kDa. In some embodiments, the isolated protein will be greater than 4 kDa. In some embodiments, the isolated protein will be greater than 4.5 kDa. In some embodiments, the isolated protein will be greater than 5.5 kDa. In some embodiments, the isolated protein will be greater than 6 kDa. Algae-derived polymers (described in US2020 / 0231928A1) can be used to separate the desired protein from the feedstock. Concentration of the isolated protein can be achieved by isoelectric focusing, water-salt dialysis, or ultrafiltration. An example of the method is shown in FIG. 2, but the unhulled hemp seeds in FIG. 2 can be replaced with any of the substances listed above as surfactant starting materials.

[0027]

[0030] In some embodiments, the isolated protein fraction may be used as an aqueous solution, a gel-like suspension, or a dry product. The effective protein concentration may range from 40 to 99% (w / w); this range includes a mixture of proteins including the protein of interest (albumin). In one embodiment, the effective protein concentration may range from 50 to 99% (w / w); this range includes a mixture of proteins including the protein of interest (albumin). In one embodiment, the effective protein concentration may range from 60 to 99% (w / w); this range includes a mixture of proteins including the protein of interest (albumin). In one embodiment, the effective protein concentration may range from 70 to 99% (w / w); this range includes a mixture of proteins including the protein of interest (albumin). In one embodiment, the effective protein concentration may range from 40 to 50% (w / w); this range includes a mixture of proteins including the protein of interest (albumin).

[0028]

[0031] The isolated protein fraction can also be treated with absorbents or chemicals to remove color and flavor. The structure of the protein can be altered by hydrolysis. Hydrolysis can be performed using enzymes such as pancreatin, pepsin, papain, ficin, bromelain, alcalase, and / or chemicals (such as pH adjustment).

[0029]

[0032] The protein fraction demonstrates its applicability for the encapsulation of lipid-based ingredients (nano and micro). Lipid ingredients include cannabinoid oils, pharmaceuticals, and edible oils (omega). The encapsulated oils may be crude, filtered, distilled, or refined, as long as the material remains liquid at ambient temperature.

[0030]

[0033] In one embodiment, the stable emulsion has a lipid load of greater than 20%. In one embodiment, the stable emulsion has a lipid load of greater than 10%. In one embodiment, the stable emulsion has a lipid load of greater than 30%. In one embodiment, the stable emulsion has a lipid load of greater than 40%.

[0031]

[0034] The encapsulated product is created by dissolving the concentrated protein fraction in water in a first vessel and combining the lipid-based component in a second vessel. The contents of the first vessel are slowly added to the second vessel with constant agitation. In one embodiment, effective agitation is achieved by sonication (20 kHz) or (10-30 kHz), or high pressure homogenization (25,000-40,000 psi), or high pressure homogenization (30,000-40,000 psi), or high pressure homogenization (20,000-40,000 psi).

[0032]

[0035] In some embodiments, as shown in FIG. 2, proteins involved in albumin protein production are isolated by stirring in the presence of algal polymers and peroxide.

[0033]

[0036] In some embodiments, liquid emulsions can be stabilized by adding additional oils or thickeners to match the hydrophobic lipophilic balance (HLB) and viscosity of lipid and aqueous phases.Powder encapsulates can be stabilized by adding flow agents such as dextrin or silicon dioxide.

[0034]

[0037] In some embodiments, the separation process is followed by the formation of a stable emulsion of the protein and lipid-based active ingredients.

[0038] In some embodiments, a stable emulsion of microencapsulated lipids is dried by spray drying or evaporative plate drying.

[0035]

[0039] In some embodiments, a stable emulsion of microencapsulated lipids is incorporated into a gelatinous matrix. definition

[0040] Unless otherwise defined, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods, devices, and materials similar or equivalent to those described herein can be used in the practice or testing of the present methods, the preferred methods, devices, and materials are described below.

[0036]

[0041] As used in this application, including the appended claims, the singular forms "a," "an," and "the" include plural references unless the content clearly dictates otherwise and are used interchangeably with "at least one" and "one or more." Thus, reference to a "protein" includes a plurality of proteins, and so forth.

[0037]

[0042] As used herein, the term "about" refers to an insignificant modification or variation of a numerical value that does not change the basic function of the item to which the numerical value is related.

[0043] As used herein, the terms "comprises," "comprising," "includes," "including," "contains," "containing" and all variations thereof are intended to extend to a non-exclusive inclusion, such that a process, method, product-by-process, or composition of matter that comprises, encompasses, or contains an element or list of elements does not include only those elements, but may include other elements that are not expressly listed or that are inherent to such process, method, product-by-process, or composition of matter.

[0038]

[0044] As used herein, the term "homogenizer" refers to an agent or device that aids in the process of converting immiscible liquids into an emulsion.

[0045] As used herein, the term "emulsifier" refers to an agent that helps immiscible liquids blend into a stable solution. In general, emulsions with smaller particles tend to be more stable.

[0039]

[0046] As used herein, the term "bioavailability" refers to the extent and rate at which a substance is absorbed into a biological system or becomes available at a site of physiological activity.

[0047] As used herein, the term "absorption" refers to the transfer of a substance from the blood into a cell, tissue, or organ and then to the rest of the body.

[0040]

[0048] As used herein, the term "carrier" refers to a base material used to deliver a substance that serves to improve the selectivity, efficacy, and / or safety of administration of the substance to a patient in need thereof.

[0041]

[0049] As used herein, the term "loading capacity" refers to the amount of oil that can be emulsified when mixed with albumin as a weight / weight percentage.

[0050] As used herein, the term "flow agent" refers to a substance used to improve the physical properties of a product, such as dextrin or silicon dioxide.

[0042]

[0051] As used herein, the term "sorbent" refers to materials such as carbon, activated charcoal, and the like.

[0052] As used herein, the term "nutraceutical" refers to any product derived from food sources that contains health-improving additives, such as ubiquinone, s-adenosylmethionine, glucosamine, and the like, and has medicinal properties.

[0043]

[0053] As used herein, the term "drug" refers to any chemical that induces a change in the physiology or psychology of an organism. The term "drug" includes, but is not limited to, THC, cannabinoids, NSAIDs, nicotine, antipsychotics, antiemetics, statins, and the like.

[0044]

[0054] As used herein, the phrase "foaming agent" refers to a surfactant which, when present in small amounts, promotes foam formation or increases its stability by inhibiting foam coalescence. Foaming agents can be inorganic chemicals such as sodium bicarbonate, ammonium carbonate, ammonium bicarbonate, and calcium azide, as well as organic foaming agents such as azodicarbonamide, benzenesulfonylhydrazide, and dinitrosopentamethylenetetramine.

[0045]

[0055] As used herein, the phrase "isoelectric focusing" refers to the isolation of proteins by precipitation at a pH that reduces the net charge of the protein to zero.

[0056] As used herein, the phrase "emulsion stability" refers to the ability of an emulsion to prevent coagulation, flocculation, sedimentation, or phenomena similar to Ostwald ripening. Stability can be affected by pH or temperature adjustment.

[0046] Processing Steps 1. Purified cannabis albumin protein was wet-pulverized using a high-pressure homogenizer, and the average particle size was measured to be 50 nm (nanometers). At a concentration of 2.1%, the resulting protein product was tested as an emulsifier and found to be capable of emulsifying vegetable oils with a very high loading capacity: with 1% cannabis albumin, up to 60% of the oil could be kept in an emulsified state. 3. This protein product was tested as an emulsifier for cannabinoids and was found to produce very stable emulsions with small particle sizes of less than 100 nm. By adding a small amount (e.g., 0.5%) of crystalline nanocellulose, the stability of the emulsion was further extended to more than 24 months without emulsion separation. 4. Albumin cannabinoid emulsions were further tested for bioavailability studies. Emulsification with albumin protein was found to enhance the absorption of cannabinoids. Specifically, bioavailability was increased by 20-fold, with nearly 100% of ingested cannabinoids being absorbed within 6 hours. 5. Safety studies have shown that cannabis albumin has negligible side effects when injected into the human blood circulation, making pure cannabis albumin suitable for use as a pharmaceutical drug carrier for both ingestible and injectable drugs. 6. When cannabis albumin is dispersed in water at a concentration ranging from 5 to 20%, more specifically 10%, and heated at a temperature between 60 and 90°C, more specifically 85°C, for 15 minutes, and then homogenized with the addition of a small amount of salt, usually 2% of the total mixture, a transparent gel can be formed. This gel can be applied in food, medicine, and cosmetics. EXAMPLES

[0047] 1. Use of cannabis-derived albumin as a plant-based emulsifier: Traditional saponin-type emulsifiers such as Quillaja saponaria have limitations in emulsion particle size (250 nm) for rational processing. Substituting albumin (cannabis) allows end users to reach particle sizes of about 50 nm using high-pressure homogenizers. This technology and ingredients can be combined with bioactive ingredients, resulting in enhanced biological uptake and improved efficacy. EXAMPLES

[0048] 1. Use of hemp-derived albumin as a plant-based amino acid for agriculture: a. Amino acids chelate minerals (make minerals bioavailable), strengthen the plant immune system, stimulate plant growth, and improve the quality of fruits and vegetables. Currently, amino acids used in agriculture are derived from fish, which is not sustainable. EXAMPLES

[0049] 1. Use of cannabis-derived albumin for emergency medical procedures: a. Albumin is the most abundant protein in circulating plasma. Albumin accounts for half of the total protein content in plasma in healthy individuals, making up about 5% of plasma. There are 140 g of albumin in an adult weighing 70 kg (154 lbs). Albumin exerts osmotic pressure to keep the blood hydrated and maintain blood pressure. In the event of blood loss, albumin is administered to maintain blood pressure and keep the patient alive. Uncontaminated sources of albumin are needed for emergency medical care. It is very difficult to keep donated blood free from contamination. Currently, genetically modified rice is used as a source of albumin. Cannabis could easily become a much better source of this essential protein. EXAMPLES

[0050] 1. Use of cannabis-derived albumin as a drug carrier: a.When drugs and other bioactive molecules enter the bloodstream, they bind to albumin molecules to remain water-soluble and be transported throughout the body. Examples include hormones, fatty acids, and cannabinoids. Albumin can be used to create nanoemulsions that can be introduced into the body intravenously or orally, making it an ideal carrier for administering drugs and other therapeutic agents into the body. EXAMPLES

[0051] 1. Use of cannabis-derived albumin as a source of bioactive peptides: a.Enzymatic and microbial hydrolysis of albumin produces protein fragments called peptides. These peptides are used in medical applications such as recovery from high blood pressure and dementia.

Claims

[Claim 1] The invention as described in the specification.

Images