Lipid polymer composition and method of use

Abstract

A composition comprising a plurality of lipid-polymer composite particles encapsulating a bioactive agent, the lipid-polymer composite particles comprising a block copolymer, a lipid selected from the group consisting of neutral lipids, cationic lipids, and anionic lipids, and a sterol, the plurality of lipid-polymer composite particles having an average particle size between 10 and 1000 nanometers. A method for preparing a plurality of lipid-polymer composite particles encapsulating a bioactive agent, the lipid-polymer composite particles comprising a block copolymer, a lipid selected from the group consisting of neutral lipids, cationic lipids, and anionic lipids, and a sterol, the plurality of lipid-polymer composite particles having an average particle size between 10 and 1000 nanometers, the method comprising homogenizing the bioactive agent with a polymer to form a homogenized solution and injecting the lipid and sterol into the homogenized solution.

Description

[Background technology]

[0001] The delivery of bioactive agents with low solubility in aqueous media poses a challenge to drug delivery. Approaches for targeted delivery of insoluble bioactive agents have included the use of liposomes and / or micelle nanoparticles as carrier systems. However, challenges related to stability, degradation, and bioavailability have hindered the widespread implementation of such delivery systems. Therefore, novel formulations of nanoparticle delivery systems are needed for the effective delivery of bioactive agents. [Overview of the Initiative]

[0002] In one embodiment, the present invention provides a composition comprising a plurality of lipid-polymer composite particles that encapsulate a bioactive agent. The lipid-polymer composite particles may comprise a block copolymer, lipids (e.g., neutral lipids, cationic lipids, or anionic lipids), and sterols. The plurality of lipid-polymer composite particles may have an average particle size of about 10 nm to about 1000 nm (e.g., about 10 nm to about 500 nm, about 10 nm to about 100 nm, and about 500 nm to about 1000 nm).

[0003] In some embodiments, the bioactive agent is a therapeutic agent, a nutritional supplement, or a pleasure agent. In some embodiments, the bioactive agent is cannabinoid or cannabinoid derivative, terpenes, flavonoids, antibiotics, preservatives, antifungal agents, antimicrobial agents, analgesics, anti-inflammatory agents, antiprotozoal agents, steroids, antiviral agents, lipophilic drugs, anti-VEGF agents, anti-glaucoma agents, essential oils, immunogens (e.g., vaccine components), nicotine or nicotine analogs, cyclosporine A, tacrolimus, isotretinoin, propofol, griseofulvin, or any combination thereof. In some embodiments, the essential oils include tea tree oil, myrrh oil, eucalyptus oil, clove oil, lavender oil, peppermint oil, Roman chamomile oil, German chamomile oil, frankincense oil, helichrysum oil, cypress oil, angelica oil, labdanum oil, petitgrain bigarade oil, orange bigarade oil, bergamot oil, sweet orange oil, palmarosa oil, lemon iron bark oil, may chang oil, basil oil, sweet marjoram oil, geranium oil, patchouli oil, valerian oil, sandalwood oil, neroli bigarade oil, grapefruit oil, coriander oil, and citronella oil. Includes Nera oil, black peppermint oil, garly gum oil, juniper yig oil, spearmint oil, scotch pine oil, rosemary oil, clary oil, ginger oil, lemon oil, mandarin oil, cumin oil, juniper berry oil, lemon balm oil, myrtle oil, ravensara oil, sweet thyme oil, everlasting oil, manuka oil, dwarf pine oil, oregano oil, vetiver oil, melissa oil, white fir oil, cassia oil, lemongrass oil, lime oil, wintergreen oil, fennel oil, ylang-ylang oil, or a combination thereof. In some embodiments, the concentration of essential oils is 0.01% to 95% by weight of the composition (e.g., 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%). In some embodiments, the composition comprises multiple essential oils. In some embodiments, the multiple essential oils comprise 2 to 10 essential oils.In some embodiments, the concentration of the multiple essential oils is between 0.01% by weight and 95% by weight of the composition (e.g., 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%). In some embodiments, the composition is formulated as an eye drop formulation.

[0004] In some embodiments, the block copolymer is a poloxamer (e.g., poloxamer 407). In some embodiments, the weight ratio of poloxamer to bioactive agent is approximately 2 to approximately 15.

[0005] In some embodiments, the lipid comprises a carbon chain of 4 to 22 lengths and a head group having a neutral, cationic, or anionic head group. In some embodiments, the lipid is phosphatidylcholine, phosphatidylserine, phosphatidylglycerol, phosphatidylethanolamine, or phosphatidylinositol.

[0006] In some embodiments, the lipid concentration is about 0.1 mol% to about 10 mol%. In some embodiments, the sterol is a phytosterol, synthetic sterol, cholesterol, or cholesterol analog.

[0007] In some embodiments, the sterol concentration is about 5 mol% to about 50 mol% of the total lipid composition. In some embodiments, the weight ratio of sterols to lipids is about 0.01 to about 0.50.

[0008] In another embodiment, the present invention is characterized by an immunogenic composition comprising the composition described herein. In another aspect, the present invention is characterized by a method of providing a bioactive agent to a target by administering any of the compositions of the above embodiments to the target.

[0009] In some embodiments, the bioactive agent includes cannabinoids, with a dose ranging from approximately 0.01 mg / kg to approximately 30 mg / kg. In some embodiments, the mode of administration is local, oral, by injection, sublingual, oral, rectal, vaginal, ocular, ear, nasal, inhalation, spray, or percutaneous.

[0010] In another embodiment, the present invention features a method for preparing the compositions of any of the above embodiments. In some embodiments, the preparation of the compositions of any of the above embodiments comprises a multi-step process. In some embodiments, the multi-step process comprises a first step of homogenizing a bioactive agent with the polymer of any of the above embodiments to produce a homogenized solution, and a second step of injecting the lipids and sterols of any of the above embodiments into the homogenized solution of the first step (e.g., immersion injection).

[0011] In another embodiment, the present invention provides a method for treating dry eye disease, inflammation, eye pain, contagious conjunctivitis, dark circles under the eyes, red eyes, bacterial eye infection, fungal eye infection, viral eye infection, nutritional deficiency, macular degeneration, glaucoma, or elevated intraocular pressure, comprising administering to the eye of a subject a composition having a plurality of lipid-polymer composite particles encapsulating a bioactive agent, the lipid-polymer composite particles comprising a block copolymer, lipids selected from the group consisting of neutral lipids, cationic lipids, and anionic lipids, and sterols, the plurality of lipid-polymer composite particles having an average particle size between 10 and 1000 nanometers.

[0012] In another aspect, the present invention features a method for treating a disease or condition selected from inflammation, pain, bacterial infection, fungal infection, protozoal infection, anxiety, agitation, stress, fatigue, insomnia, mental fatigue, memory loss, organ rejection, eczema, acne, and skin infections, the method comprising administering to a subject a composition having a plurality of lipid-polymer composite particles encapsulating a bioactive agent, the lipid-polymer composite particles comprising a block copolymer, a lipid selected from the group consisting of a neutral lipid, a cationic lipid, and an anionic lipid, and a sterol, the plurality of lipid-polymer composite particles having an average particle size between 10 and 1000 nanometers.

[0013] In some embodiments of any of the above aspects, the bioactive agent is a cannabinoid or cannabinoid derivative, terpene, flavonoid, antibiotic, preservative, antifungal agent, antibacterial agent, analgesic, anti-inflammatory agent, antiprotozoal agent, steroid, antiviral agent, lipophilic drug, anti-VEGF agent, antiglaucoma agent, essential oil, immunogen (e.g., a vaccine component), nicotine or a nicotine analog, cyclosporin A, tacrolimus, isotretinoin, propofol, griseofulvin or any combination thereof.

[0014] In some embodiments, the block copolymer is a poloxamer. In some embodiments, the weight ratio of the poloxamer to the bioactive agent is between 2 and 15. In some embodiments, the lipid comprises a carbon chain of length 4 to 22 and a neutral, cationic, or anionic head group. In some embodiments, the lipid is phosphatidylcholine, phosphatidylserine, phosphatidylglycerol, phosphatidylethanolamine, or phosphatidylinositol. In some embodiments, the concentration of the lipid is from about 0.1 mol% to about 10 mol%. In some embodiments, the sterol is phytosterol, or synthetic sterol, or cholesterol, or a cholesterol analog. In some embodiments, the concentration of the sterol is from about 5 mol% to about 50 mol% of the total lipid composition. In some embodiments, the weight ratio of sterol to lipid is from about 0.01 to about 0.50.

[0015] In another aspect, the present invention features a method of use for pleasure, comprising administering to a subject a composition comprising a plurality of lipid-polymer composite particles encapsulating a bioactive agent, wherein the lipid-polymer composite particles comprise a block copolymer, a lipid selected from the group consisting of a neutral lipid, a cationic lipid, and an anionic lipid, and a sterol, and the plurality of lipid-polymer composite particles have an average particle size between 10 and 1000 nanometers.

[0016] In some embodiments of the above aspects, the bioactive agent is a pleasure agent. In some embodiments, the bioactive agent is a cannabinoid or cannabinoid derivative, terpene, flavonoid, antibiotic, preservative, antifungal agent, antibacterial agent, analgesic, anti-inflammatory agent, antiprotozoal agent, steroid, antiviral agent, lipophilic drug, anti-VEGF agent, anti-glaucoma agent, essential oil, immunogen (e.g., vaccine component), nicotine or nicotine analog, cyclosporin A, tacrolimus, isotretinoin, propofol, gliocladin or any combination thereof. In some embodiments, the block copolymer is a poloxamer.

[0017] In some embodiments of any of the above-described features, the essential oils include tea tree oil, myrrh oil, eucalyptus oil, clove oil, lavender oil, peppermint oil, Roman chamomile oil, German chamomile oil, frankincense oil, helichrysum oil, cypress oil, angelica oil, labdanum oil, petitgrain bigarade oil, orange bigarade oil, bergamot oil, sweet orange oil, palmarosa oil, lemon iron bark oil, may chang oil, basil oil, sweet marjoram oil, geranium oil, patchouli oil, valerian oil, sandalwood oil, neroli bigarade oil, grapefruit oil, and coriander oil. Oils including citronella oil, black peppermint oil, garly gum oil, juniper yig oil, spearmint oil, scotch pine oil, rosemary oil, clary oil, ginger oil, lemon oil, mandarin oil, cumin oil, juniper berry oil, lemon balm oil, myrtle oil, ravensara oil, sweet thyme oil, everlasting oil, manuka oil, dwarf pine oil, oregano oil, vetiver oil, melissa oil, white fir oil, cassia oil, lemongrass oil, lime oil, wintergreen oil, fennel oil, ylang-ylang oil, or combinations thereof. In some embodiments, the concentration of the essential oil is 0.01% to 95% by weight of the composition (e.g., 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%). In some embodiments, the composition comprises a plurality of essential oils. In some embodiments, this plurality of essential oils comprises 2 to 10 essential oils. In some embodiments, this plurality of essential oils is 0.01% to 95% by weight (e.g., 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%). In some embodiments, this method includes a composition that is formulated as an eye drop formulation.

[0018] In some embodiments, the weight ratio of poloxamer to bioactive agent is between 2 and 15. In some embodiments, the lipid comprises a carbon chain of 4 to 22 lengths and a neutral, cationic, or anionic head group. In some embodiments, the lipid is phosphatidylcholine, phosphatidylserine, phosphatidylglycerol, phosphatidylethanolamine, or phosphatidylinositol. In some embodiments, the lipid concentration is about 0.1 mol% to about 10 mol%. In some embodiments, the sterol is phytosterol, or synthetic sterol, or cholesterol, or cholesterol analogue. In some embodiments, the sterol concentration is about 5 mol% to about 50 mol% of the total lipid composition. In some embodiments, the weight ratio of sterol to lipid is about 0.01 to about 0.50.

[0019] definition To facilitate understanding of the present invention, several terms are defined below. Terms as defined herein have meanings that are generally understood by those skilled in the art relating to the present invention. Terms such as "a," "an," and "the" are not intended to refer only to singular entities, but include a general set of which specific examples may be used for illustrative purposes. Terms used herein are used to describe specific embodiments of the present invention, but their use does not limit the present invention except as outlined in the claims.

[0020] As used herein, any value provided within a range includes both an upper and lower limit, and any value that falls within the upper and lower limits. Where used herein, the term "approximately" refers to a range of ±10% of the stated value.

[0021] The term "administer" means introducing the formulation of the present invention into the body of a patient who needs it in order to treat a disease or condition. As used herein, the term “bioactive agent” refers to any synthetic or naturally occurring compound (in free form, salt form, or solvated or hydrated form) having a desired biological or physiological effect, such as proteins, drugs, antigens, nutrients, cosmetics, fragrances, flavors, diagnostic agents, pharmaceuticals, vitamins, or nutritional supplements, and is formulated to a level sufficient to provide an in vivo concentration at a functional level (including topical concentrations of topical compositions). Under certain circumstances, one or more components of the lipid matrix i) (e.g., components (a), (b), (c), and / or (d)) may also be activators, but it is preferable that any bioactive agent (iii) is not one of these components (e.g., must not be a component of the lipid matrix). The most preferred activators are pharmaceuticals (e.g., APIs), including drugs, vaccines, and diagnostic agents.

[0022] As used herein, the term “block copolymer” refers to a linear polymer having regions or blocks along its main chain characterized by similar hydrophilic, hydrophobic, or chemical properties.

[0023] The term "diblock copolymer" refers to a block copolymer that contains two blocks. The term "triblock copolymer" refers to a block copolymer that contains three blocks.

[0024] The term "multiblock copolymer" refers to a block copolymer that contains multiple blocks. As used herein, the terms “encapsulate,” “encapsulated,” or “to encapsulate” refer to the encapsulation of a portion within an enclosed polymer assembly structure, such as a micelle (e.g., a bioactive agent as defined herein). An encapsulated bioactive agent (e.g., an encapsulated cannabinoid) is enclosed by a polymer aggregate structure, such as the encapsulated portion being located within the hydrophobic interior of the polymer aggregate structure (e.g., the lumen of a micelle).

[0025] As used herein, the term “anionic head group” refers to a lipid head group that has a net negative charge at physiological pH. As used herein, the term “cationic head group” refers to a lipid head group that has a net positive charge at physiological pH.

[0026] As used herein, the term “neutral head group” refers to a lipid head group that exists in an uncharged form at physiological pH. As used herein, “lipid nanoparticles” or “LNPs” are vesicles comprising a lipid layer that encapsulates a substantially solid lipid core. The lipid core may contain pharmaceutically active molecules. LNPs typically include cationic lipids, non-cationic lipids, and lipids that prevent particle aggregation (e.g., PEG-lipid conjugates).

[0027] As used herein, the term “lipid-polymer composite particles” refers to a complex of molecules held together by non-covalent bonds such as hydrogen bonds, van der Waals forces, electrostatic interactions, hydrophobic effects, and Pi-Pi interactions. Lipid-polymer composite particles may also include, for example, large complexes of molecules that form spherical structures. Examples of lipid-polymer composite particles include lipid nanoparticles and micelles.

[0028] As used herein, the terms “micelle,” “of a micelle,” or variations thereof refer to a polymeric assembly comprising a hydrophilic shell (or corona) and a hydrophobic and / or ionic interior. Furthermore, the term micelle may refer to any polyionic composite assembly comprising a multiblock copolymer having a net positive charge and polynucleotides having a suitable negative charge.

[0029] As used herein, the term "nanoparticles" refers to polymer-based particles having a diameter in the nanometer range (e.g., 1 nm to 1000 nm). The term "nutritional supplement" refers to any substance that is a food or part of a food, and / or provides additional health benefits beyond the basic nutritional value of the food. For example, nutritional supplements may contain ingredients derived from food sources. Exemplary nutritional supplements include, but are not limited to, antioxidants, dietary supplements, fortified dairy products, plant extracts, vitamins, minerals, and herbs.

[0030] As used herein, the term “plural” means two or more, such as at least 2, 20, 50, 100, 1000, 10000, 100000, 1000000, 10000000, or more.

[0031] As used herein, the terms “pleasure agent” or “pleasure substance” or variations thereof refer to compounds that are useful in providing relaxation, enjoyment, and recreational activities to a subject. As used herein, the term “subject” may mean humans, non-human primates, or other mammals, for example, but not limited to dogs, cats, horses, cattle, pigs, turkeys, goats, fish, monkeys, chickens, rats, mice, and sheep.

[0032] As used herein, the term “sterol” includes, without limitation, all sterols, e.g., sitosterol, campesterol, stigmasterol, brassicasterol (including dihydrobrascasterol), desmosterol, carinosterol, polyferasterol, clionasterol, ergosterol, coprosterol, codisterol, isofucosterol, fucosterol, clerosterol, nervisterol, rassterol, sterasterol, spinasterol, chondristerol, peposterol, avenasterol, isoabenasterol, fecostrol, polinastasterol, cholesterol, and all their natural or synthetic forms and derivatives, including isomers. It should be understood that modifications to sterols, i.e., including side chains, also fall within the scope of the present invention.

[0033] The term "therapeutic agent" refers to any substance that possesses therapeutic properties that produce a desired, usually beneficial, effect. For example, a therapeutic agent may treat, alleviate, and / or prevent a disease. Such agents may be synthetic or naturally occurring, non-peptides, proteins or peptides, oligonucleotides or nucleotides, polysaccharides or sugars.

[0034] In this specification, a "vesicle" is defined as a type of lipid-polymer composite particle in which amphiphilic molecules (e.g., lipids) collectively define a volume, for example, a substantially spherical volume. The amphiphilic molecules (e.g., lipids) typically constitute at least one shell of the vesicle. In this shell, the amphiphilic molecules are arranged within a bilayer, with the hydrophilic portion of the amphiphilic molecules oriented outward relative to the plane of this bilayer, and the hydrophobic portion of the amphiphilic molecules primarily located within this bilayer. The opposite arrangement exists when the surrounding medium is hydrophobic. [Brief explanation of the drawing]

[0035] [Figure 1] Preliminary optical microscope images of liposome formulations Fa1, Fa2, and Fa3 are shown. The scale bar is 50 μm. [Figure 2] The images show optical microscope images of preliminary liposome formulations Fb1, Fb2, and Fb3, prepared to evaluate the effect of temperature on particulate formulations. The scale bar is 10 μm. [Figure 3] This table shows the effective diameter and polydispersity of formulations Fc1 to Fc7. Formulations Fc1, Fc2, and Fc9 were loaded with 0.02% (w / w) CBD. Formulations Fc3 to Fc8 were loaded with 0.25% (w / w) CBD. The mean and standard deviation values ​​are reported. [Figure 4] This table shows the mean and standard deviation values ​​of the measured zeta potentials of particles in formulations Fc1 to Fc9. [Figure 5]These are photographic images showing the light transmittance of formulations Fd1 to Fd6. Transparency increased with increasing concentration of poloxamer PLURONIC® F127. The concentration of CBD remained constant at 0.5% (w / w) in all suspensions, while the concentrations of PLURONIC® F127 were 1%, 2%, 3%, 4%, 5%, and 10% for formulations Fd1, Fd2, Fd3, Fd4, Fd5, and Fd6, respectively. [Figure 6] These are photographic images showing the effect of poloxamer concentration on the amount of observable precipitate in formulations Fd6, Fd5, Fd4, Fd2, and Fd1. The concentration of CBD was constant at 0.5% (w / w) in all suspensions, while the concentrations of PLURONIC® F127 were 1%, 2%, 4%, 5%, and 10% for formulations Fd1, Fd2, Fd4, Fd5, and Fd6, respectively. [Figure 7] This graph shows the effective diameter and polydispersity of formulations Fd1 to Fd6. Similar micelle sizes of less than 50 μm were obtained for all formulations. [Figure 8] This graph shows the pH of formulations Fd1 to Fd6 compared to the pH of commercially available eye drops THEALOZ™ DUO and HYABAK™. [Figure 9] This graph shows the viscosity of formulations Fd1 to Fd6 compared to the viscosity of water and two commercially available eye drops, THEALOZ™ DUO and HYABAK™. [Figure 10] This graph shows the size stability of formulations Fd1 to Fd6. Effective diameter and polydispersity were calculated for all formulations on the day of preparation (left bar) and 30 days after preparation when stored at room temperature (20°C, middle bar) or 4°C (right bar). [Figure 11]These are photographic images showing the light transmittance of formulations Fe1 to Fe9. Formulations Fe1, Fe4, and Fe7 were prepared by homogenizing poloxamer and subsequently adding 0.5% (w / w) CBD, phospholipids, and cholesterol by ethanol injection. Formulations Fe2, Fe5, and Fe8 were prepared by homogenizing poloxamer with 0.5% (w / w) CBD and subsequently adding phospholipids and cholesterol by ethanol injection. Formulations Fe3, Fe6, and Fe9 were prepared by homogenizing poloxamer with 0.5% (w / w) CBD and subsequently adding phospholipids and cholesterol in powder form. [Figure 12] This graph shows the particle size, polydispersity, and span value of lipid-polymer composite particles in formulations Fe1 to Fe9. [Figure 13] This graph shows the pH of formulations Fe1 to Fe9. All formulations had a pH close to neutral. [Figure 14] This table shows the tonicity of formulations Fe1-Fe9 compared to physiological saline, control poloxamer and CBD suspension, and three commercially available eye drops: HYABAK® 0.15%, THEALOZ® DUO New, and THEALOZ® DUO. [Figure 15] Figure 15 is a graph showing the size stability of formulations Fe1 to Fe9. Effective diameter and polydispersity were calculated for all formulations on the day of preparation (left bar) and 19 days after preparation when stored at room temperature of either 20°C (middle bar) or 4°C (right bar). [Figure 16] This graph shows the particle size, polydispersity, and span values ​​of optimized formulations Fd5, Fe5, Fe8, and Fe2. [Figure 17] The image shows photographic evidence of excess CBD agitated into formulations containing PLURONIC® F127 at concentrations of 1, 3, 5, 7, 9, 11, 13, and 15% (w / v). The leftmost vial did not contain PLURONIC® F127 and served as a control. The viscosity of the formulations increased with increasing concentration of PLURONIC® F127. [Figure 18] This graph shows the saturated solubility of CBD in water (0%), 1%, 3%, 5%, 7%, 9%, 11%, 13%, and 15% PLURONIC® F127, lipids only (0% + lipids), and 5% PLURONIC® F127 with lipids (5% + lipids). The lipid concentrations used were 0.21 mM DSPC and 0.19 mM cholesterol. CBD solubility increased with increasing PLURONIC® F127 concentration, peaking at approximately 7-11% PLURONIC® F127. There was no substantial difference in CBD solubility due to lipid uptake at the lipid concentrations used. [Figure 19] These are a series of transmission electron microscope (TEM) images of three different formulations containing CBD. In the 5% PLURONIC® F127 formulation (top row), densely packed spherical structures ranging in diameter from 20 to 30 μm were observed. These were likely micelles. In the lipid-only formulation (middle row) (DSPC:Chol; (0.21:0.19 mM)), two distinctly different features were observed: small diffused spherical structures and large aggregated structures. In the formulation containing both lipid and PLURONIC® F127 (bottom row) (5% PLURONIC® F127 + DSPC:Chol (0.21:0.19 mM)), two distinctly different features were also observed: densely packed micelle structures of 20 to 30 μm, and larger monolayer and multilayer structures in which smaller micelles were encapsulated within them. [Figure 20] This graph shows the particle size and polydispersity of formulations containing high concentrations of DSPC and cholesterol. Generally, compared to a combination of PLURONIC® F127 and DSPC:Chol at concentrations of 1.64:1.51 mM, particle size was observed to be larger in the absence of PLURONIC® F127. It was also noteworthy that particle size increased with increasing lipid concentration in the PLURONIC® F127-containing formulation. [Figure 21]This graph shows that there was no substantial difference in pH between formulations with and without PLURONIC® F127, and that the pH did not substantially change even when the concentration of lipids (DSPC and cholesterol) increased. [Figure 22] This graph shows CBD dissolved in formulations without PLURONIC® F127 (left) and formulations containing PLURONIC® F127 (right), while increasing the lipid concentration. [Figure 23] This graph shows the particle size and polydispersity of formulations containing higher concentrations of lipids (DSPC and cholesterol). Here again, the particle size was large without PLURONIC® F127, and the particle size increased as the lipid concentration in the PLURONIC® F127 formulation increased. [Figure 24] This graph shows that there was no substantial difference in pH between formulations with and without PLURONIC(registered trademark)F127, and that the pH did not substantially change even when the lipid concentration increased. [Figure 25] This graph shows CBD dissolved in formulations without PLURONIC® F127 (left) and formulations containing PLURONIC® F127 (right), while increasing the concentration of lipids (DSPC and cholesterol). [Modes for carrying out the invention]

[0036] The present invention features novel lipid-polymer composite particles useful for formulations of bioactive agents for administration to a target, such as a human target. The lipid-polymer composite particles comprise a plurality of nanoparticles (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 100, 1,000, 10,000, 100,000, 1,000,000, 10,000,000, or more) that encapsulate the bioactive agent. These nanoparticles comprise block copolymers, lipids, such as phospholipids, and sterols. Formulations of bioactive agents (e.g., therapeutic agents, nutritional supplements, or stimulants) described herein provide higher drug loading capacity, improved formulation stability, and easier loading of lipid-polymer composite particles with lower water surface tension, enabling lipid coating and encapsulation. This improved process for preparing lipid-polymer composite particles having a lipid coating, such as micelles, enables aqueous loading and controlled drug release of hydrophobic bioactivators. Furthermore, the compositions and methods described herein solubilize hydrophobic bioactivators without the use of organic solvents, such as ethanol. The components of this formulation are described in further detail below.

[0037] Lipid-polymer composite particles Lipid-polymer composite particles may be used to formulate bioactive agents for delivery (e.g., therapeutic agents, nutritional supplements, or stimulants, e.g., cannabinoids). Lipid-polymer composite particles include complexes of defined molecules (such as lipids and polymers) held together by non-covalent bonds such as hydrogen bonds, van der Waals forces, electrostatic interactions, hydrophobic effects, and Pi-Pi interactions. Lipid-polymer composite particles may also include large complexes of molecules that form spherical, rod-shaped, or sheet-like structures. Lipid-polymer composite particles include, for example, micelles and LNPs. Lipid-polymer composite particles may have a predetermined size. The size of the structure may vary based on the size or number of components packed within the structure (e.g., the size of the bioactive agent molecules).

[0038] The size of lipid-polymer composite particles can vary, for example, from about 10 nm to about 1,000 nm. Non-limiting examples of the average particle size with a Z mean include, for example, about 10 nm to about 500 nm, about 10 nm to about 100 nm, and about 500 nm to about 1,000 nm. For example, lipid-polymer composite particles can be found in sizes such as approximately 10nm, 15nm, 20nm, 25nm, 30nm, 35nm, 40nm, 45nm, 50nm, 55nm, 60nm, 65nm, 70nm, 75nm, 80nm, 85nm, 90nm, 95nm, 100nm, 105nm, 110nm, 115nm, 120nm, 125nm, 130nm, 135nm, 140nm, 145nm, 150nm, 155nm, 160nm, and so on. 165nm, approx. 170nm, approx. 175nm, approx. 180nm, approx. 185nm, approx. 190nm, approx. 195nm, approx. , approx. 245nm, approx. 250nm, approx. 255nm, approx. 260nm, approx. 265nm, approx. 270nm, approx. 275nm, approx. 280nm, approx. 285nm, approx. m, approx. 325nm, approx. 330nm, approx. 335nm, approx. 340nm, approx. 345nm, approx. 350nm, approx. 355nm, approx. 360nm, approx. 0nm, approx. 405nm, approx. 410nm, approx. 415nm, approx. 420nm, approx. 425nm, approx. 430nm, approx. 435nm, approx. 440nm, approx. 80nm, approx. 485nm, approx. 490nm, approx. 495nm, approx. 500nm, approx. 505nm, approx. 510nm, approx. 515nm, approx. 520nm, approx. 560nm, approx. 565nm, approx. 570nm, approx. 575nm, approx. 580nm, approx. 585nm, approx. 590nm, approx.Approximately 640nm, approximately 645nm, approximately 650nm, approximately 655nm, approximately 660nm, approximately 665nm, approximately 670nm, approximately 675nm, approximately 680nm, approximately 68 5nm, about 690nm, about 695nm, about 700nm, about 705nm, about 710nm, about 715nm, about 720nm, about 725nm, about 730nm , about 735nm, about 740nm, about 745nm, about 750nm, about 755nm, about 760nm, about 765nm, about 770nm, about 775nm, about 7 80nm, about 785nm, about 790nm, about 795nm, about 800nm, about 805nm, about 810nm, about 815nm, about 820nm, about 825n The average particle size may have a Z-average of approximately 830nm, 835nm, 840nm, 845nm, 850nm, 855nm, 860nm, 865nm, 870nm, 875nm, 880nm, 885nm, 890nm, 895nm, 900nm, 905nm, 910nm, 915nm, 920nm, 925nm, 930nm, 935nm, 940nm, 945nm, 950nm, 955nm, 960nm, 965nm, 970nm, 975nm, 980nm, 985nm, 990nm, 995nm, or approximately 1000nm.

[0039] The average particle size can be measured by zeta potential, dynamic light scattering (DLS), electrophoretic light scattering (ELS), static light scattering (SLS), molecular weight, electrophoretic mobility, size exclusion chromatography (SEC), field flow fractionation, or other methods known in the art. In certain embodiments, lipid-polymer composite particles have an average Z-mean particle size of about 10 nm to about 100 nm. Those skilled in the art will understand that a population of lipid-polymer composite particles (e.g., LNPs or micelles) may have a range of average Z-mean particle sizes within the population. Thus, the population may be polydispersible. The population may have a polydispersity index of 0.3 or less (e.g., 0.05 to 0.3). The polydispersity index can be determined using DLS (see, for example, ISO 22412:2017).

[0040] Lipid nanoparticles The bioactive agents of the present invention can be completely encapsulated in lipid formulations, such as LNPs, or other lipid-polymer composite particles. LNPs are highly useful for systemic application because they exhibit extended circulatory life after intravenous (iv) injection and accumulate at distal sites (e.g., sites physically distant from the administration site). The LNPs include "pSPLPs" containing encapsulated condensant-nucleic acid complexes, as shown in PCT publication number WO2000 / 003683. LNPs may have an average diameter of about 50 nm to about 150 nm, more typically about 60 nm to about 130 nm, more typically about 70 nm to about 110 nm, and most typically about 70 nm to about 90 nm, and are substantially non-toxic. Furthermore, when the bioactive agents are present in the LNPs of the present invention, they are resistant to degradation by nucleases in aqueous solution. Nucleic acid-lipid particles and methods for preparing them are disclosed, for example, in U.S. Patents 5,976,567; 5,981,501; 6,534,484; 6,586,410; 6,815,432; U.S. Publication 2010 / 0324120; and PCT Publication WO96 / 40964.

[0041] In one embodiment, the lipid-to-drug ratio (mass / mass ratio) (e.g., lipid-to-biogenic agent ratio) is in the range of approximately 1:1 to approximately 50:1, approximately 1:1 to approximately 25:1, approximately 3:1 to approximately 15:1, approximately 4:1 to approximately 10:1, approximately 5:1 to approximately 9:1, or approximately 6:1 to approximately 9:1. Intermediate ranges beyond those listed above are also considered to be part of the present invention.

[0042] Non-limiting examples of cationic lipids include N,N-dioleyl-N,N-dimethylammonium chloride (DODAC), N,N-distearyl-N,N-dimethylammonium bromide (DDAB), N-(I-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP), N-(I-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA), N,N-dimethyl-2,3-dioleyloxy)propylamine (DODMA), 1,2-dilinoleyloxy-N,N-dimethylaminopropane (DLinDMA), 1,2-dilinolelenyloxy-N,N-dimethylaminopropane (DLenDMA), and 1,2-dilinole Ilcarbamoyloxy-3-dimethylaminopropane (DLin-C-DAP), 1,2-dilinoleyoxy-3-(dimethylamino)acetoxypropane (DLin-DAC), 1,2-dilinoleyoxy-3-morpholinopropane (DLin-MA), 1,2-dilinoleoyl-3-dimethylaminopropane (DLinDAP), 1,2-dilinoleylthio-3-dimethylaminopropane (DLin-S-DMA), 1-linoleoyl-2-linoleyloxy-3-dimethylaminopropane (DLin-2-DMAP), 1,2-dilinoleyloxy-3-trimethylaminopropane chloride (DLin-TMA.Cl), 1,2-dilinoleoyl-3-trimethylaminopropane chloride (DLin-TAP.Cl), 1,2-dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), or 3-(N,N-dilinoleylamino)-1,2-propanediol (DLinAP), 3-(N,N-dioleylamino)-1,2-propanediol (DOAP), 1,2-dilinoleyloxo-3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), 1,2-dilinolenyloxy-N,N-dimethylaminopropane (DLinDMA), 2,2-dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxolane (DLin-K-DMA) or similar compounds. (3aR,5s,6aS)-N,N-dimethyl-2,2-di((9Z,12Z)-octadeca-9,12-dienyltetrahydro-3aH-cyclopenta[d][1,3]dioxol-5-amine (ALN100), (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl4-(dimethylamino)butanoate (MC3), 1,1'-(2-(4-(2-((2-(bis(2-hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl)amino)ethyl)piperazine-1-ylethylazandiyldidodecane-2-ol (Tech Examples include G1), or mixtures thereof. Cationic lipids may constitute, for example, about 20 mol% to about 50 mol% or about 40 mol% of the total lipids present in the particles.

[0043] Ionizable / noncationic lipids may be anionic or neutral lipids, but are not limited to distearoyl phosphatidylcholine (DSPC), dioleoyl phosphatidylcholine (DOPC), dipalmitoyl phosphatidylcholine (DPPC), dioleoyl phosphatidylglycerol (DOPG), dipalmitoyl phosphatidylglycerol (DPPG), dioleoyl-phosphatidylethanolamine (DOPE), palmitoyloleoyl phosphatidylcholine (POPC), and palmitoyloleoyl phosphatidylethanolamine. Examples include amines (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidylethanolamine (DSPE), 16-O-monomethylPE, 16-O-dimethylPE, 18-1-transPE, 1-stearoyl-2-oleoyl-phosphatidylethanolamine (SOPE), cholesterol, or mixtures thereof. Noncationic lipids may constitute about 5 mol% to about 90 mol%, about 10 mol%, or about 60 mol% of the total lipids present in the particles, for example, if cholesterol is included.

[0044] The conjugate lipids that inhibit particle aggregation may be, for example, polyethylene glycol (PEG) lipids, including, but not limited to, PEG-diacylglycerol (DAG), PEG-dialkyloxypropyl (DAA), PEG-phospholipids, PEG-ceramide (Cer), or mixtures thereof. The PEG-DAA conjugate may be, for example, PEG-dilauryloxypropyl (C) 12 ), PEG-Dimyristyloxypropyl (C 14 ), PEG-Dipalmityloxypropyl (C 16 ), or PEG-distearyloxypropyl (C 18) may be the case. The complex lipids that prevent particle aggregation may be, for example, 0 mol% to about 20 mol% or about 2 mol% of the total lipids present in the particles.

[0045] Lipids may have carbon chains of 4 to 22 lengths and neutral, cationic, or anionic head groups. In some embodiments, the particles further contain sterols (e.g., cholesterol) in an amount of about 10 mol% to about 60 mol% or about 50 mol% of the total lipids present in the particles.

[0046] Micelle A micelle is a specific type of molecular assembly in which amphiphilic molecules are arranged in a spherical structure such that all hydrophobic parts of the molecule face inward, while the hydrophilic parts remain in contact with the surrounding aqueous phase. Micelles can be made of lipids. The micelle phase is caused by the packing behavior of single-tail lipids in a bilayer. The difficulty of filling all the internal volume of the bilayer while corresponding to the region per head group forced on the molecule by the hydration of the lipid head group leads to micelle formation. This type of micelle is known as a normal-phase micelle (oil-in-water micelle). An inverse micelle has a head group in the center and tails extending outward (water-in-oil micelle).

[0047] Micelles are nearly spherical. Other phases are possible, including ellipsoidal, cylindrical, and bilayer shapes. The shape and size of a micelle are functions of the molecular structure of its surfactant molecule and solution conditions such as surfactant concentration, temperature, pH, and ionic strength. The process of micelle formation is known as micellation and, depending on its polymorphism, forms part of the phase behavior of many lipids.

[0048] Phospholipids The lipid-polymer composite particles described herein may contain one or more phospholipids. Phospholipids generally consist of two hydrophobic fatty acid tails and a hydrophilic head having a phosphate group. The two components are usually linked by a glycerol molecule. The phosphate group may be modified with organic molecules such as choline, ethanolamine, or serine. Suitable phospholipids that may be used in the compositions described herein include, for example, phosphatidylcholine, phosphatidylserine, phosphatidylglycerol, phosphatidylethanolamine, and phosphatidylinositol. The concentration of phospholipids in the lipid-polymer composite particles may be about 2% to about 20% (v / v) (e.g., about 4% to about 18%, about 5% to about 15%, for example, about 10%).

[0049] Block copolymer The lipid-polymer composite particles described herein may include block copolymers. A block copolymer refers to a linear polymer having regions or blocks along its main chain characterized by similar hydrophilic, hydrophobic, or chemical properties. A block copolymer may contain, for example, two, three, four, or more blocks (e.g., diblock or triblock copolymers). A multiblock copolymer contains multiple blocks.

[0050] Diblock copolymer The compositions described herein may include diblock copolymers comprising two distinct blocks of repeating polymer units. An example of a diblock copolymer described herein is an amphiphilic copolymer, for example, a copolymer comprising a region containing a hydrophilic chain of a repeating unit, connected to a region containing a hydrophobic chain of a repeating unit with or without a linker. Such a diblock copolymer may include a hydrophilic chain of a polyoxyethylene (PEO) subunit connected to a hydrophobic chain of a polyoxypropylene (PPO) subunit. A diblock copolymer of PEO and PPO subunits can be represented by the following formula: X1(C2H4O) m -L-(C3H6O) nX2. X1 and X2 may be any chemical part. L may be an optional linker. In some embodiments, the PEO and PPO subunit blocks are directly covalently bonded. In some embodiments, X1 and X2 are H and OH, respectively. Other diblock copolymers include, for example, poly(ethylene glycol)-poly(γ-benzyl L-glutamate)PEG-PBLA, poly(ethylene glycol)-poly(D,L-lactic acid)PEG-PDLLA, poly(ethylene glycol)-poly(L-lactic acid)PEG-PLLA, poly(ethylene glycol)-poly(ε-caprolactone)PEG-PCL, poly(ethylene glycol)-poly(D,L-lactide-co-glycolide)PEG-PLGA, poly(ethylene glycol)-poly(γ-benzyl L-glutamate)PEG-PBLG, poly(ethylene glycol)-poly(β-benzyl L-aspartate)PEG-PBLA, poly(ethylene glycol)-poly(α-benzylcarboxylate-ε-caprolactone)PEG-PBCL, and poly(ethylene glycol)-poly(δ-valerolactone)PEG-PVL. For clarity, as used herein, X1-[PEO]-L-[PPO]-X2 refers to the following structure:

[0051] [ka]

[0052] The length of the polymer block can be customized. As a result, many different diblock copolymers exist. Diblock copolymers suitable for use in conjunction with the compositions and methods of this disclosure include those having an average molecular weight of about 5 kDa to about 30 kDa (e.g., 6 kDa, 7 kDa, 8 kDa, 9 kDa, 10 kDa, 11 kDa, 12 kDa, 13 kDa, 14 kDa, 15 kDa, 16 kDa, 17 kDa, 18 kDa, 19 kDa, 20 kDa, 21 kDa, 22 kDa, 23 kDa, 24 kDa, 25 kDa, 26 kDa, 27 kDa, 28 kDa, 29 kDa, or 30 kDa). Because the synthesis of diblock copolymers is related to the natural degree of change from one batch to another, the values ​​cited above (and those used herein to characterize a given diblock copolymer) may not be precisely attainable during synthesis, and the average values ​​will vary to a certain extent. Therefore, as used herein, the term “diblock copolymer” may be used interchangeably with the term “diblock copolymers” (representing several diblock copolymer entities, also called mixtures of diblock copolymers) unless otherwise explicitly stated. The term “average” as used herein, relating to the number of monomer units or molecular weight of a diblock copolymer (plural), is a result of the fact that it is technically impossible to produce diblock copolymers that are all identical in composition and therefore have the same molecular weight. Diblock copolymers prepared according to state-of-the-art methods are presented as mixtures of diblock copolymers, each exhibiting variability with respect to molecular weight, but the mixture as a whole averaging the molecular weights specified herein.

[0053] Poloxamer An example of a triblock copolymer is a poloxamer. A poloxamer refers to a nonionic triblock copolymer composed of a central hydrophobic chain of polyoxypropylene with two adjacent hydrophilic chains of polyoxyethylene. Poloxamers are also known by the trade names "PLURONIC®" or "SYNPERONIC®" (BASF). The block copolymer can be represented by the following formula: HO(C2H4O) x (C3H6O) y (C2H4O) z H. The lengths of the polymer blocks can be customized. As a result, many different poloxamers exist. The synthesis of block copolymers is associated with a natural variability from batch to batch, so the numerical values used herein to characterize a given poloxamer may not be exactly achievable during synthesis, and the average values may vary to some extent. Therefore, the term "poloxamer" as used herein can be used in the same sense as the term "poloxamers" (representing the components of several poloxamers, also called mixtures of poloxamers) if not explicitly stated otherwise. The term "average" as related to the number of monomer units or molecular weight of the poloxamer(s) used herein results from the fact that it is not technically possible to produce poloxamers that all have the same composition and thus the same molecular weight. Poloxamers made according to state-of-the-art methods each exhibit variability with respect to molecular weight but are presented as mixtures of poloxamers that, as a whole, average the molecular weights specified herein. Poloxamers suitable for use in the compositions described herein are disclosed in Alexandridis and Bodratti, Journal of Functional Materials 9(1):11(2018), the disclosure of which is hereby incorporated by reference in its entirety.

[0054] Poloxamers that can be used in combination with the compositions and methods of this disclosure include those with an average molar mass of polyoxypropylene subunits greater than 2,050 g / mol (e.g., approximately 2,055 g / mol, 2,060 g / mol, 2,075 g / mol, 2,080 g / mol, 2,085 g / mol, 2,090 g / mol, 2,095 g / mol, 2,100 g / mol, 2,200 g / mol, 2,300 g / mol, 2,400 g / mol, 2,500 g / mol, 2,600 g / mol, 2,700 g / mol, 2,800 g / mol, 2,900 g / mol, 3,000 g / mol, 3 Examples include poloxamers having an average molar mass of polyoxypropylene subunits of 100 g / mol, 3,200 g / mol, 3,300 g / mol, 3,400 g / mol, 3,500 g / mol, 3,600 g / mol, 3,700 g / mol, 3,800 g / mol, 3,900 g / mol, 4,000 g / mol, 4,100 g / mol, 4,200 g / mol, 4,300 g / mol, 4,400 g / mol, 4,500 g / mol, 4,600 g / mol, 4,700 g / mol, 4,800 g / mol, 4,900 g / mol, or 5,000 g / mol.

[0055] In some embodiments, the poloxamer has an average molar mass of polyoxypropylene subunits greater than 2,250 g / mol (e.g., about 2,300 g / mol, 2,400 g / mol, 2,500 g / mol, 2,600 g / mol, 2,700 g / mol, 2,800 g / mol, 2,900 g / mol, 3,000 g / mol, 3,100 g / mol, 3,200 g / mol, 3,300 g / mol, 3,400 g / mol, 3,500 g / mol). It has an average molar mass of polyoxypropylene subunits of 1 mol, 3,600 g / mol, 3,700 g / mol, 3,800 g / mol, 3,900 g / mol, 4,000 g / mol, 4,100 g / mol, 4,200 g / mol, 4,300 g / mol, 4,400 g / mol, 4,500 g / mol, 4,600 g / mol, 4,700 g / mol, 4,800 g / mol, 4,900 g / mol, or 5,000 g / mol.

[0056] In some embodiments, the poloxamer has an average molar mass of polyoxypropylene subunits greater than 2,750 g / mol (e.g., about 2,800 g / mol, 2,900 g / mol, 3,000 g / mol, 3,100 g / mol, 3,200 g / mol, 3,300 g / mol, 3,400 g / mol, 3,500 g / mol, 3,600 g / mol, 3,700 g / mol, 3, It has an average molar mass of polyoxypropylene subunits of 800 g / mol, 3,900 g / mol, 4,000 g / mol, 4,100 g / mol, 4,200 g / mol, 4,300 g / mol, 4,400 g / mol, 4,500 g / mol, 4,600 g / mol, 4,700 g / mol, 4,800 g / mol, 4,900 g / mol, or 5,000 g / mol.

[0057] In some embodiments, the poloxamer has an average molar mass of polyoxypropylene subunits greater than 3,250 g / mol (for example, an average molar mass of polyoxypropylene subunits of about 3,300 g / mol, 3,400 g / mol, 3,500 g / mol, 3,600 g / mol, 3,700 g / mol, 3,800 g / mol, 3,900 g / mol, 4,000 g / mol, 4,100 g / mol, 4,200 g / mol, 4,300 g / mol, 4,400 g / mol, 4,500 g / mol, 4,600 g / mol, 4,700 g / mol, 4,800 g / mol, 4,900 g / mol, or 5,000 g / mol).

[0058] In some embodiments, the poloxamer has an average molar mass of polyoxypropylene subunits greater than 3,625 g / mol (for example, an average molar mass of polyoxypropylene subunits of about 3,700 g / mol, 3,800 g / mol, 3,900 g / mol, 4,000 g / mol, 4,100 g / mol, 4,200 g / mol, 4,300 g / mol, 4,400 g / mol, 4,500 g / mol, 4,600 g / mol, 4,700 g / mol, 4,800 g / mol, 4,900 g / mol, or 5,000 g / mol).

[0059] In some embodiments, the poloxamer has a molecular weight of from about 2,050 g / mol to about 4,000 g / mol (e.g., about 2,050 g / mol, 2,055 g / mol, 2,060 g / mol, 2,065 g / mol, 2,070 g / mol, 2,075 g / mol, 2,080 g / mol, 2,085 g / mol, 2,090 g / mol, 2,095 g / mol, 2,100 g / mol, 2,105 g / mol, 2,110 g / mol, 2,115 g / mol, 2,120 g / mol, 2,125 g / mol, 2,130 g / mol, 2,135 g / mol, 2,140 g / mol, 2,145 g / mol, 2,150 g / mol, 2,155 g / mol, 2,160 g / mol, 2,165 g / mol, 2,170 g / mol, 2,175 g / mol, 2,180 g / mol, 2,185 g / mol, 2,190 g / mol, 2,195 g / mol, 2,200 g / mol, 2,205 g / mol, 2,210 g / mol, 2,215 g / mol, 2,220 g / mol, 2,225 g / mol, 2,230 g / mol, 2,235 g / mol, 2,240 g / mol, 2,245 g / mol, 2,250 g / mol, 2,255 g / mol, 2,260 g / mol, 2,265 g / mol, 2,270 g / mol, 2,275 g / mol, 2,280 g / mol, 2,285 g / mol, 2,290 g / mol, 2,295 g / mol, 2,300 g / mol, 2,305 g / mol, 2,310 g / mol, 2,315 g / mol, 2,320 g / mol, 2,325 g / mol, 2,330 g / mol, 2,335 g / mol, 2,340 g / mol, 2,345 g / mol, 2,350 g / mol, 2,355 g / mol, 2,360 g / mol, 2,365 g / mol, 2,370 g / mol, 2,375 g / mol, 2,380 g / mol, 2,385 g / mol, 2,390 g / mol, 2,395 g / mol, 2,400 g / mol, 2,405 g / mol, 2,410 g / mol, 2,415 g / mol, 2,420 g / mol, 2,425 g / mol, 2,430 g / mol, 2,435 g / mol, 2,440 g / mol, 2,445 g / mol, 2,450 g / mol, 2,455 g / mol, 2,460 g / mol, 2,465 g / mol, 2,470 g / mol, 2,475 g / mol, 2,480g / mol、2,485g / mol、2,490g / mol、2,495g / mol、2,500g / mol、2,505g / mol、2,510g / mol、2,515g / mol、2,520g / mol、2,525g / mol、2,530g / mol、2,535g / mol、2,540g / mol、2,545g / mol、2,550g / mol、2,555g / mol、2,560g / mol、2,565g / mol、2,570g / mol、2,575g / mol、2,580g / mol、2,585g / mol、2,590g / mol、2,595g / mol、2,600g / mol、2,605g / mol、2,610g / mol、2,615g / mol、2,620g / mol、2,625g / mol、2,630g / mol、2,635g / mol、2,640g / mol、2,645g / mol、2,650g / mol、2,655g / mol、2,660g / mol、2,665g / mol、2,670g / mol、2,675g / mol、2,680g / mol、2,685g / mol、2,690g / mol、2,695g / mol、2,700g / mol、2,705g / mol、2,710g / mol、2,715g / mol、2,720g / mol、2,725g / mol、2,730g / mol、2,735g / mol、2,740g / mol、2,745g / mol、2,750g / mol、2,755g / mol、2,760g / mol、2,765g / mol、2,770g / mol、2,775g / mol、2,780g / mol、2,785g / mol、2,790g / mol、2,795g / mol、2,800g / mol、2,805g / mol、2,810g / mol、2,815g / mol、2,820g / mol、2,825g / mol、2,830g / mol、2,835g / mol、2,840g / mol、2,845g / mol、2,850g / mol、2,855g / mol、2,860g / mol、2,865g / mol、2,870g / mol、2,875g / mol、2,880g / mol、2,885g / mol、2,890g / mol、2,895g / mol、2,900g / mol、2,905g / mol、2,910g / mol、2,915g / mol、2,920g / mol、2,925g / mol、2,930g / mol、2,935g / mol、2,940g / mol、2,945g / mol、2,950g / mol、2,955g / mol、2,960g / mol、2,965g / mol、2,970g / mol、2,975g / mol、2,980g / mol、2,985g / mol、2,990g / mol、2,995g / mol、3,000g / mol、3,005g / mol、3,010g / mol、3,015g / mol、3,020g / mol、3,025g / mol、3,030g / mol、3,035g / mol、3,040g / mol、3,045g / mol、3,050g / mol、3,055g / mol、3,060g / mol、3,065g / mol、3,070g / mol、3,075g / mol、3,080g / mol、3,085g / mol、3,090g / mol、3,095g / mol、3,100g / mol、3,105g / mol、3,110g / mol、3,115g / mol、3,120g / mol、3,125g / mol、3,130g / mol、3,135g / mol、3,140g / mol、3,145g / mol、3,150g / mol、3,155g / mol、3,160g / mol、3,165g / mol、3,170g / mol、3,175g / mol、3,180g / mol、3,185g / mol、3,190g / mol、3,195g / mol、3,200g / mol、3,205g / mol、3,210g / mol、3,215g / mol、3,220g / mol、3,225g / mol、3,230g / mol、3,235g / mol、3,240g / mol、3,245g / mol、3,250g / mol、3,255g / mol、3,260g / mol、3,265g / mol、3,270g / mol、3,275g / mol、3,280g / mol、3,285g / mol、3,290g / mol、3,295g / mol、3,300g / mol、3,305g / mol、3,310g / mol、3,315g / mol、3,320g / mol、3,325g / mol、3,330g / mol、3,335g / mol、3,340g / mol、3,345g / mol、3,350g / mol、3,355g / mol、3,360g / mol、3,365g / mol、3,370g / mol、3,375g / mol、3,380g / mol、3,385g / mol、3,390g / mol、3,395g / mol、3,400g / mol、3,405g / mol、3,410g / mol、3,415g / mol、3,420g / mol、3,425g / mol、3,430g / mol、3,435g / mol、3,440g / mol、3,445g / mol、3,450g / mol、3,455g / mol、3,460g / mol、3,465g / mol、3,470g / mol、3,475g / mol、3,480g / mol、3,485g / mol、3,490g / mol、3,495g / mol、3,500g / mol、3,505g / mol、3,510g / mol、3,515g / mol、3,520g / mol、3,525g / mol、3,530g / mol、3,535g / mol、3,540g / mol、3,545g / mol、3,550g / mol、3,555g / mol、3,560g / mol、3,565g / mol、3,570g / mol、3,575g / mol、3,580g / mol、3,585g / mol、3,590g / mol、3,595g / mol、3,600g / mol、3,605g / mol、3,610g / mol、3,615g / mol、3,620g / mol、3,625g / mol、3,630g / mol、3,635g / mol、3,640g / mol、3,645g / mol、3,650g / mol、3,655g / mol、3,660g / mol、3,665g / mol、3,670g / mol、3,675g / mol、3,680g / mol、3,685g / mol、3,690g / mol、3,695g / mol、3,700g / mol、3,705g / mol、3,710g / mol、3,715g / mol、3,720g / mol、3,725g / mol、3,730g / mol、3,735g / mol、3,740g / mol、3,745g / mol、3,750g / mol、3,755g / mol、3,760g / mol、3,765g / mol、3,770g / mol、3,775g / mol、3,780g / mol、3,785g / mol、3,790g / mol、3,795g / mol、3,800g / mol、3,805g / mol、3,810g / mol、3,815g / mol、3,820g / mol、3,825g / mol、3,830g / mol、3,835g / mol、3,840g / mol、3,845g / mol, 3,850g / mol, 3,855g / mol, 3,860g / mol, 3,865g / mol, 3,870g / mol, 3,875g / mol, 3,880g / mol, 3,885g / mol l, 3,890g / mol, 3,895g / mol, 3,900g / mol, 3,905g / mol, 3,910g / mol, 3,915g / mol, 3,920g / mol, 3,925g / mol, 3,930g It has an average molar mass of polyoxypropylene subunits of 3,935 g / mol, 3,940 g / mol, 3,945 g / mol, 3,950 g / mol, 3,955 g / mol, 3,960 g / mol, 3,965 g / mol, 3,970 g / mol, 3,975 g / mol, 3,980 g / mol, 3,985 g / mol, 3,990 g / mol, 3,995 g / mol, or 4,000 g / mol).

[0060] In some embodiments, the poloxamer has a molecular weight of from about 2,750 g / mol to about 4,000 g / mol (e.g., about 2,750 g / mol, 2,755 g / mol, 2,760 g / mol, 2,765 g / mol, 2,770 g / mol, 2,775 g / mol, 2,780 g / mol, 2,785 g / mol, 2,790 g / mol, 2,795 g / mol, 2,800 g / mol, 2,805 g / mol, 2,810 g / mol, 2,815 g / mol, 2,820 g / mol, 2,825 g / mol, 2,830 g / mol, 2,835 g / mol, 2,840 g / mol, 2,845 g / mol, 2,850 g / mol, 2,855 g / mol, 2,860 g / mol, 2,865 g / mol, 2,870 g / mol, 2,875 g / mol, 2,880 g / mol, 2,885 g / mol, 2,890 g / mol, 2,895 g / mol, 2,900 g / mol, 2,905 g / mol, 2,910 g / mol, 2,915 g / mol, 2,920 g / mol, 2,925 g / mol, 2,930 g / mol, 2,935 g / mol, 2,940 g / mol, 2,945 g / mol, 2,950 g / mol, 2,955 g / mol, 2,960 g / mol, 2,965 g / mol, 2,970 g / mol, 2,975 g / mol, 2,980 g / mol, 2,985 g / mol, 2,990 g / mol, 2,995 g / mol, 3,000 g / mol, 3,005 g / mol, 3,010 g / mol, 3,015 g / mol, 3,020 g / mol, 3,025 g / mol, 3,030 g / mol, 3,035 g / mol, 3,040 g / mol, 3,045 g / mol, 3,050 g / mol, 3,055 g / mol, 3,060 g / mol, 3,065 g / mol, 3,070 g / mol, 3,075 g / mol, 3,080 g / mol, 3,085 g / mol, 3,090 g / mol, 3,095 g / mol, 3,100 g / mol, 3,105 g / mol, 3,110 g / mol, 3,115 g / mol, 3,120 g / mol, 3,125 g / mol, 3,130 g / mol, 3,135 g / mol, 3,140 g / mol, 3,145 g / mol, 3,150 g / mol, 3,155 g / mol, 3,160 g / mol, 3,165 g / mol, 3,170 g / mol, 3,175 g / mol, 3,180g / mol、3,185g / mol、3,190g / mol、3,195g / mol、3,200g / mol、3,205g / mol、3,210g / mol、3,215g / mol、3,220g / mol、3,225g / mol、3,230g / mol、3,235g / mol、3,240g / mol、3,245g / mol、3,250g / mol、3,255g / mol、3,260g / mol、3,265g / mol、3,270g / mol、3,275g / mol、3,280g / mol、3,285g / mol、3,290g / mol、3,295g / mol、3,300g / mol、3,305g / mol、3,310g / mol、3,315g / mol、3,320g / mol、3,325g / mol、3,330g / mol、3,335g / mol、3,340g / mol、3,345g / mol、3,350g / mol、3,355g / mol、3,360g / mol、3,365g / mol、3,370g / mol、3,375g / mol、3,380g / mol、3,385g / mol、3,390g / mol、3,395g / mol、3,400g / mol、3,405g / mol、3,410g / mol、3,415g / mol、3,420g / mol、3,425g / mol、3,430g / mol、3,435g / mol、3,440g / mol、3,445g / mol、3,450g / mol、3,455g / mol、3,460g / mol、3,465g / mol、3,470g / mol、3,475g / mol、3,480g / mol、3,485g / mol、3,490g / mol、3,495g / mol、3,500g / mol、3,505g / mol、3,510g / mol、3,515g / mol、3,520g / mol、3,525g / mol、3,530g / mol、3,535g / mol、3,540g / mol、3,545g / mol、3,550g / mol、3,555g / mol、3,560g / mol、3,565g / mol、3,570g / mol、3,575g / mol、3,580g / mol、3,585g / mol、3,590g / mol、3,595g / mol、3,600g / mol、3,605g / mol、3,610g / mol、3,615g / mol、3,620g / mol、3,625g / mol、3,630g / mol、3,635g / mol, 3,640g / mol, 3,645g / mol, 3,650g / mol, 3,655g / mol, 3,660g / mol, 3,665g / mol, 3,670g / mol, 3,675g / mol, 3,680g / m ol, 3,685g / mol, 3,690g / mol, 3,695g / mol, 3,700g / mol, 3,705g / mol, 3,710g / mol, 3,715g / mol, 3,720g / mol, 3,725g / mol, 3,73 0g / mol, 3,735g / mol, 3,740g / mol, 3,745g / mol, 3,750g / mol, 3,755g / mol, 3,760g / mol, 3,765g / mol, 3,770g / mol, 3,775g / mol , 3,780g / mol, 3,785g / mol, 3,790g / mol, 3,795g / mol, 3,800g / mol, 3,805g / mol, 3,810g / mol, 3,815g / mol, 3,820g / mol, 3,825g / mol, 3,830g / mol, 3,835g / mol, 3,840g / mol, 3,845g / mol, 3,850g / mol, 3,855g / mol, 3,860g / mol, 3,865g / mol, 3,870g / mol, 3 ,875g / mol, 3,880g / mol, 3,885g / mol, 3,890g / mol, 3,895g / mol, 3,900g / mol, 3,905g / mol, 3,910g / mol, 3,915g / mol, 3,920g / m It has an average molar mass of polyoxypropylene subunits of ol, 3,925 g / mol, 3,930 g / mol, 3,935 g / mol, 3,940 g / mol, 3,945 g / mol, 3,950 g / mol, 3,955 g / mol, 3,960 g / mol, 3,965 g / mol, 3,970 g / mol, 3,975 g / mol, 3,980 g / mol, 3,985 g / mol, 3,990 g / mol, 3,995 g / mol, or 4,000 g / mol).

[0061] In some embodiments, the poloxamer has a molecular weight of from about 3,250 g / mol to about 4,000 g / mol (e.g., about 3,250 g / mol, 3,255 g / mol, 3,260 g / mol, 3,265 g / mol, 3,270 g / mol, 3,275 g / mol, 3,280 g / mol, 3,285 g / mol, 3,290 g / mol, 3,295 g / mol, 3,300 g / mol, 3,305 g / mol, 3,310 g / mol, 3,315 g / mol, 3,320 g / mol, 3,325 g / mol, 3,330 g / mol, 3,335 g / mol, 3,340 g / mol, 3,345 g / mol, 3,350 g / mol, 3,355 g / mol, 3,360 g / mol, 3,365 g / mol, 3,370 g / mol, 3,375 g / mol, 3,380 g / mol, 3,385 g / mol, 3,390 g / mol, 3,395 g / mol, 3,400 g / mol, 3,405 g / mol, 3,410 g / mol, 3,415 g / mol, 3,420 g / mol, 3,425 g / mol, 3,430 g / mol, 3,435 g / mol, 3,440 g / mol, 3,445 g / mol, 3,450 g / mol, 3,455 g / mol, 3,460 g / mol, 3,465 g / mol, 3,470 g / mol, 3,475 g / mol, 3,480 g / mol, 3,485 g / mol, 3,490 g / mol, 3,495 g / mol, 3,500 g / mol, 3,505 g / mol, 3,510 g / mol, 3,515 g / mol, 3,520 g / mol, 3,525 g / mol, 3,530 g / mol, 3,535 g / mol, 3,540 g / mol, 3,545 g / mol, 3,550 g / mol, 3,555 g / mol, 3,560 g / mol, 3,565 g / mol, 3,570 g / mol, 3,575 g / mol, 3,580 g / mol, 3,585 g / mol, 3,590 g / mol, 3,595 g / mol, 3,600 g / mol, 3,605 g / mol, 3,610 g / mol, 3,615 g / mol, 3,620 g / mol, 3,625 g / mol, 3,630 g / mol, 3,635 g / mol, 3,640 g / mol, 3,645 g / mol, 3,650 g / mol, 3,655 g / mol, 3,660 g / mol, 3,665 g / mol, 3,670 g / mol, 3,675 g / mol, 3,680g / mol, 3,685g / mol, 3,690g / mol, 3,695g / mol, 3,700g / mol, 3,705g / mol, 3,710g / mol, 3,715g / mol, 3,720g / mol, 3,725g / mol, 3,730g / mol, 3,735g / mol, 3,740g / mol, 3,745g / mol, 3,750g / mol, 3,755g / mol, 3,760g / mol, 3 ,765g / mol, 3,770g / mol, 3,775g / mol, 3,780g / mol, 3,785g / mol, 3,790g / mol, 3,795g / mol, 3,800g / mol, 3,805g / mol, 3,810g / mol, 3,815g / mol, 3,820g / mol, 3,825g / mol, 3,830g / mol, 3,835g / mol, 3,840g / mol, 3,845g / mol, 3 ,850g / mol, 3,855g / mol, 3,860g / mol, 3,865g / mol, 3,870g / mol, 3,875g / mol, 3,880g / mol, 3,885g / mol, 3,890g / mol, 3,895g / mol, 3,900g / mol, 3,905g / mol, 3,910g / mol, 3,915g / mol, 3,920g / mol, 3,925g / mol, 3,930g / mol, It has an average molar mass of polyoxypropylene subunits of 3,935 g / mol, 3,940 g / mol, 3,945 g / mol, 3,950 g / mol, 3,955 g / mol, 3,960 g / mol, 3,965 g / mol, 3,970 g / mol, 3,975 g / mol, 3,980 g / mol, 3,985 g / mol, 3,990 g / mol, 3,995 g / mol, or 4,000 g / mol.

[0062] In some embodiments, poloxamer is present in concentrations of approximately 3,625 g / mol to approximately 4,000 g / mol (for example, approximately 3,625 g / mol, 3,630 g / mol, 3,635 g / mol, 3,640 g / mol, 3,645 g / mol, 3,650 g / mol, 3,655 g / mol, 3,660 g / mol, 3,665 g / mol, 3,670 g / mol, 3,675 g / mol, 3,680 g / mol, 3,685 g / mol, 3,690 g / mol, 3,695 g / mol, 3,70700 g / mol, 3,680 g / mol, 3,685 g / mol, 3,690 g / mol, 3,695 g / mol, 3,600 g / mol, 3,600 g / mol, 3,600 g / mol, 3,600 g / mol, 3,600 g / mol, 3,600 g / mol, 3,600 g / mol, 3,600 g / mol, 3,600 g / mol, 3,600 g / mol, 3,600 g / mol, 3,600 g / mol, 3,600 g / mol, 5g / mol, 3,710g / mol, 3,715g / mol, 3,720g / mol, 3,725g / mol, 3,730g / mol, 3,735g / mol, 3,740g / mol, 3,745g / mol, 3,750g / mol, 3,755g / mol l, 3,760g / mol, 3,765g / mol, 3,770g / mol, 3,775g / mol, 3,780g / mol, 3,785g / mol, 3,790g / mol, 3,795g / mol, 3,800g / mol, 3,805g / mol, 3,8 10g / mol, 3,815g / mol, 3,820g / mol, 3,825g / mol, 3,830g / mol, 3,835g / mol, 3,840g / mol, 3,845g / mol, 3,850g / mol, 3,855g / mol, 3,860g / mol mol, 3,865g / mol, 3,870g / mol, 3,875g / mol, 3,880g / mol, 3,885g / mol, 3,890g / mol, 3,895g / mol, 3,900g / mol, 3,905g / mol, 3,910g / mol, 3 It has an average molar mass of polyoxypropylene subunits of 915 g / mol, 3,920 g / mol, 3,925 g / mol, 3,930 g / mol, 3,935 g / mol, 3,940 g / mol, 3,945 g / mol, 3,950 g / mol, 3,955 g / mol, 3,960 g / mol, 3,965 g / mol, 3,970 g / mol, 3,975 g / mol, 3,980 g / mol, 3,985 g / mol, 3,990 g / mol, 3,995 g / mol, or 4,000 g / mol).

[0063] In some embodiments, poloxamer is present in amounts exceeding 40% by mass (for example, about 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 6%). It has an average ethylene oxide content of 8%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, or greater.

[0064] In some embodiments, the poloxamer has an average ethylene oxide content of more than 50% by mass (for example, about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, or greater).

[0065] In some embodiments, the poloxamer has an average ethylene oxide content of more than 60% by mass (for example, about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, or greater).

[0066] In some embodiments, the poloxamer has an average ethylene oxide content of more than 70% by mass (for example, about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, or greater).

[0067] In some embodiments, the poloxamer has an average ethylene oxide content of about 40% to about 90% (for example, about 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, or 90%).

[0068] In some embodiments, the poloxamer has an average ethylene oxide content of about 50% to about 85% (for example, about 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, or 85%).

[0069] In some embodiments, the poloxamer has an average ethylene oxide content of about 60% to about 80% (for example, about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%).

[0070] In some embodiments, poloxamer is present in concentrations greater than 10,000 g / mol (e.g., approximately 10,100 g / mol, 10,200 g / mol, 10,300 g / mol, 10,400 g / mol, 10,500 g / mol, 10,600 g / mol, 10,700 g / mol, 10,800 g / mol, 10,900 g / mol, 11,000 g / mol, 11,100 g / mol). 0g / mol, 11,200g / mol, 11,300g / mol, 11,400g / mol, 11,500g / mol, 11,600g / mol, 11,700g / mol, 11 ,800g / mol, 11,900g / mol, 12,000g / mol, 12,100g / mol, 12,200g / mol, 12,300g / mol, 12,400g / mol, 12,500g / mol, 12,600g / mol, 12,700g / mol, 12,800g / mol, 12,900g / mol, 13,000g / mol, 13,100g / m ol, 13,200g / mol, 13,300g / mol, 13,400g / mol, 13,500g / mol, 13,600g / mol, 13,700g / mol, 13,800g It has an average molar mass of 1 / mol, 13,900 g / mol, 14,000 g / mol, 14,100 g / mol, 14,200 g / mol, 14,300 g / mol, 14,400 g / mol, 14,500 g / mol, 14,600 g / mol, 14,700 g / mol, 14,800 g / mol, 14,900 g / mol, or 15,000 g / mol.

[0071] In some embodiments, the poloxamer is present in concentrations greater than 11,000 g / mol (for example, about 11,100 g / mol, 11,200 g / mol, 11,300 g / mol, 11,400 g / mol, 11,500 g / mol, 11,600 g / mol, 11,700 g / mol, 11,800 g / mol, 11,900 g / mol, 12,000 g / mol, 12,100 g / mol, 12,200 g / mol, 12,300 g / mol, 12,400 g / mol, 12,500 g / mol, 12,600 g / mol, 12,700 g / mol, 12,800 g / mol, 12,900 g / mol). It has an average molar mass of 13,000 g / mol, 13,100 g / mol, 13,200 g / mol, 13,300 g / mol, 13,400 g / mol, 13,500 g / mol, 13,600 g / mol, 13,700 g / mol, 13,800 g / mol, 13,900 g / mol, 14,000 g / mol, 14,100 g / mol, 14,200 g / mol, 14,300 g / mol, 14,400 g / mol, 14,500 g / mol, 14,600 g / mol, 14,700 g / mol, 14,800 g / mol, 14,900 g / mol, or 15,000 g / mol.

[0072] In some embodiments, poloxamers are present in concentrations greater than 12,000 g / mol (for example, about 12,100 g / mol, 12,200 g / mol, 12,300 g / mol, 12,400 g / mol, 12,500 g / mol, 12,600 g / mol, 12,700 g / mol, 12,800 g / mol, 12,900 g / mol, 13,000 g / mol, 13,100 g / mol, 13,200 g / mol, 13,300 g / mol, 13,400 g / mol, It has an average molar mass of 13,500 g / mol, 13,600 g / mol, 13,700 g / mol, 13,800 g / mol, 13,900 g / mol, 14,000 g / mol, 14,100 g / mol, 14,200 g / mol, 14,300 g / mol, 14,400 g / mol, 14,500 g / mol, 14,600 g / mol, 14,700 g / mol, 14,800 g / mol, 14,900 g / mol, or 15,000 g / mol.

[0073] In some embodiments, poloxamer is present in concentrations greater than 12,500 g / mol (e.g., approximately 12,600 g / mol, 12,700 g / mol, 12,800 g / mol, 12,900 g / mol, 13,000 g / mol, 13,100 g / mol, 13,200 g / mol, 13,300 g / mol, 13,400 g / mol, 13,500 g / mol, 13,600 g / mol, 13,700 g / mol). It has an average molar mass of g / mol, 13,800 g / mol, 13,900 g / mol, 14,000 g / mol, 14,100 g / mol, 14,200 g / mol, 14,300 g / mol, 14,400 g / mol, 14,500 g / mol, 14,600 g / mol, 14,700 g / mol, 14,800 g / mol, 14,900 g / mol, or 15,000 g / mol.

[0074] In some embodiments, poloxamer is present in concentrations of approximately 10,000 g / mol to approximately 15,000 g / mol (for example, approximately 10,000 g / mol, 10,100 g / mol, 10,200 g / mol, 10,300 g / mol, 10,400 g / mol, 10,500 g / mol, 10,600 g / mol, 10,700 g / mol, 10,800 g / mol, 10,900 g / mol). l, 11,000g / mol, 11,100g / mol, 11,200g / mol, 11,300g / mol, 11,400g / mol, 11,500g / mol, 11,600g / mol , 11,700g / mol, 11,800g / mol, 11,900g / mol, 12,000g / mol, 12,100g / mol, 12,200g / mol, 12,300g / mol, 12,400g / mol, 12,500g / mol, 12,600g / mol, 12,700g / mol, 12,800g / mol, 12,900g / mol, 13,000g / mol, 13,100g / mol, 13,200g / mol, 13,300g / mol, 13,400g / mol, 13,500g / mol, 13,600g / mol, 13,700g / mol, 1 It has an average molar mass of 3,800 g / mol, 13,900 g / mol, 14,000 g / mol, 14,100 g / mol, 14,200 g / mol, 14,300 g / mol, 14,400 g / mol, 14,500 g / mol, 14,600 g / mol, 14,700 g / mol, 14,800 g / mol, 14,900 g / mol, or 15,000 g / mol.

[0075] In some embodiments, poloxamer is present in concentrations of approximately 11,000 g / mol to approximately 15,000 g / mol (for example, approximately 11,000 g / mol, 11,100 g / mol, 11,200 g / mol, 11,300 g / mol, 11,400 g / mol, 11,500 g / mol, 11,600 g / mol, 11,700 g / mol, 11,800 g / mol, 11,900 g / mol, 12,000 g / mol, 12,100 g / mol, 12,200 g / mol, 12,300 g / mol, 12,400 g / mol, 12,500 g / mol, 12,600 g / mol, 12,700 g / mol, 12,800 g / mol). It has an average molar mass of 12,900 g / mol, 13,000 g / mol, 13,100 g / mol, 13,200 g / mol, 13,300 g / mol, 13,400 g / mol, 13,500 g / mol, 13,600 g / mol, 13,700 g / mol, 13,800 g / mol, 13,900 g / mol, 14,000 g / mol, 14,100 g / mol, 14,200 g / mol, 14,300 g / mol, 14,400 g / mol, 14,500 g / mol, 14,600 g / mol, 14,700 g / mol, 14,800 g / mol, 14,900 g / mol, or 15,000 g / mol.

[0076] In some embodiments, poloxamer is present in concentrations of approximately 11,500 g / mol to approximately 15,000 g / mol (for example, approximately 11,500 g / mol, 11,600 g / mol, 11,700 g / mol, 11,800 g / mol, 11,900 g / mol, 12,000 g / mol, 12,100 g / mol, 12,200 g / mol, 12,300 g / mol, 12,400 g / mol, 12,500 g / mol, 12,600 g / mol, 12,700 g / mol, 12,800 g / mol, 12,900 g / mol, 13,000 g / mol, 13,100 g / mol). It has an average molar mass of g / mol, 13,200 g / mol, 13,300 g / mol, 13,400 g / mol, 13,500 g / mol, 13,600 g / mol, 13,700 g / mol, 13,800 g / mol, 13,900 g / mol, 14,000 g / mol, 14,100 g / mol, 14,200 g / mol, 14,300 g / mol, 14,400 g / mol, 14,500 g / mol, 14,600 g / mol, 14,700 g / mol, 14,800 g / mol, 14,900 g / mol, or 15,000 g / mol.

[0077] In some embodiments, poloxamer is present in concentrations of approximately 12,000 g / mol to approximately 15,000 g / mol (for example, approximately 12,000 g / mol, 12,100 g / mol, 12,200 g / mol, 12,300 g / mol, 12,400 g / mol, 12,500 g / mol, 12,600 g / mol, 12,700 g / mol, 12,800 g / mol, 12,900 g / mol, 13,000 g / mol, 13,100 g / mol, 13,200 g / mol, 13,300 g / mol, It has an average molar mass of 13,400 g / mol, 13,500 g / mol, 13,600 g / mol, 13,700 g / mol, 13,800 g / mol, 13,900 g / mol, 14,000 g / mol, 14,100 g / mol, 14,200 g / mol, 14,300 g / mol, 14,400 g / mol, 14,500 g / mol, 14,600 g / mol, 14,700 g / mol, 14,800 g / mol, 14,900 g / mol, or 15,000 g / mol.

[0078] In some embodiments, poloxamer is present in concentrations of approximately 12,500 g / mol to approximately 15,000 g / mol (for example, approximately 12,500 g / mol, 12,600 g / mol, 12,700 g / mol, 12,800 g / mol, 12,900 g / mol, 13,000 g / mol, 13,100 g / mol, 13,200 g / mol, 13,300 g / mol, 13,400 g / mol, 13,500 g / mol, 13,600 g / mol). It has an average molar mass of g / mol, 13,700 g / mol, 13,800 g / mol, 13,900 g / mol, 14,000 g / mol, 14,100 g / mol, 14,200 g / mol, 14,300 g / mol, 14,400 g / mol, 14,500 g / mol, 14,600 g / mol, 14,700 g / mol, 14,800 g / mol, 14,900 g / mol, or 15,000 g / mol.

[0079] Poloxamer P288, P335, P338, and P407 Suitable poloxamers that can be used in combination with the compositions and methods disclosed herein include those with the chemical formula HO(C2H4O) x (C3H6O) y (C2H4O) z One example is "poloxamer 288" (also known in the art as "P288" and poloxamer "F98"), which has H [wherein the formula, the sum of x and y is approximately 236.36 and z is approximately 44.83]. The average molecular weight of P288 is approximately 13,000 g / mol.

[0080] In some embodiments, poloxamer is of the formula HO(C2H4O) x (C3H6O) y (C2H4O) z Variants of P288, such as a variant of H [wherein the formula, the sum of x and y is approximately 220 to approximately 250, and z is approximately 40 to approximately 50]. In some embodiments, the average molecular weight of poloxamers is approximately 12,000 g / mol to approximately 14,000 g / mol.

[0081] Suitable poloxamers that can be used in combination with the compositions and methods disclosed herein include those with the chemical formula HO(C2H4O) x (C3H6O) y (C2H4O) z Further examples include "poloxamer 335" (also known in the art as "P335" and poloxamer "P105") having H [wherein the formula, the sum of x and y is approximately 73.86 and z is approximately 56.03]. The average molecular weight of P335 is approximately 6,500 g / mol.

[0082] In some embodiments, poloxamer is of the formula HO(C2H4O) x (C3H6O) y (C2H4O) z Variants of P335, such as a variant of H [wherein the formula, the sum of x and y is approximately 60 to approximately 80, and z is approximately 50 to approximately 60]. In some embodiments, the average molecular weight of poloxamers is approximately 6,000 g / mol to approximately 7,000 g / mol.

[0083] Suitable poloxamers that can be used in combination with the compositions and methods disclosed herein include those with the chemical formula HO(C2H4O) x (C3H6O) y (C2H4O) z Further examples include "poloxamer 338" (also known in the art as "P338" and poloxamer "F108") having H [wherein the formula, the sum of x and y is approximately 265.45 and z is approximately 50.34]. The average molecular weight of P335 is approximately 14,600 g / mol.

[0084] In some embodiments, poloxamer is of the formula HO(C2H4O) x (C3H6O) y (C2H4O) z Variants of P338, such as a variant of H [wherein the formula, the sum of x and y is approximately 260 to approximately 270, and z is approximately 45 to approximately 55]. In some embodiments, the average molecular weight of poloxamer is approximately 14,000 g / mol to approximately 15,000 g / mol.

[0085] Suitable poloxamers that can be used in combination with the compositions and methods disclosed herein include those with the chemical formula HO(C2H4O) x (C3H6O) y (C2H4O) z Further examples include "Poloxamer 407" (also known in the art as "P407" and "Poloxamer F127"), which has H [wherein the formula, the sum of x and y is approximately 200.45 and z is approximately 65.17]. Its average molecular weight is approximately 12,600 g / mol.

[0086] In some embodiments, poloxamer is of the formula HO(C2H4O) x (C3H6O) y (C2H4O) z Variants of P407, such as a variant of H [wherein the formula, the sum of x and y is approximately 190 to approximately 210, and z is approximately 60 to approximately 70]. In some embodiments, the average molecular weight of poloxamers is approximately 12,000 g / mol to approximately 13,000 g / mol.

[0087] For clarity, the terms “average molar mass” and “average molecular weight” are used interchangeably in this specification and refer to the same quantity. The average molar mass, ethylene oxide content, and propylene oxide content of poloxamers described herein may be measured using the methods disclosed in Alexandridis and Hatton, Colloids and Surfaces A: Physicochemical and Engineering Aspects 96:1-46 (1995), the entire disclosure of which is incorporated herein by reference.

[0088] Sterols The lipid-polymer composite particles described herein may further contain one or more sterols. Sterols are lipids commonly found naturally in plants, animals, and fungi. Phytosterols refer to a collection of plant sterol molecules, naturally occurring compounds found in plant cell membranes. Phytosterols include both plant sterols and stanols. Phytosterols can be extracted from any common plant source, such as soybeans, wood, tall oil, and vegetable oil. Phytosterols include β-sitosterol, campesterol, stigmasterol, stigmamanol, campestanol, brassicasterol, ergosterol, lupeol, and cycloartenol. The sterols described herein may be any cholesterol or derivative thereof that alters the fluidity of the lipid layer. The sterols may be naturally occurring sterols, e.g., sterols derived from or found in natural sources. Alternatively, the sterols may be synthetic sterols, e.g., sterol analogs or derivatives that do not exist in nature. In some preferred embodiments, the sterol is cholesterol or an analogue of cholesterol (e.g., thiocholesterol, epicholesterol, β-sitosterol (Si-Lip), stigmasterol (St-Lip), or lanosterol (La-Lip)). The concentration of sterol in the compositions described herein may be, for example, about 1% to about 50% of the total lipid composition (e.g., about 5% to about 45%, about 10% to about 40%).

[0089] Sterols and phospholipids may be present in particles in a sterol:phospholipid weight ratio of, for example, about 0.01 to about 0.5. For example, the sterol:phospholipid weight ratio may be, for example, about 0.01 to about 0.1, about 0.1 to about 0.2, about 0.2 to about 0.3, about 0.3 to about 0.4, about 0.4 to about 0.5, about 0.01 to about 0.2, about 0.01 to about 0.3, about 0.01 to about 0.4, about 0.1 to about 0.15, and about 0.1 to about 0.25. For example, the sterol:phospholipid weight ratio may be approximately 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, 0.4, 0.45, or 0.5.

[0090] Sterols also include their esterified derivatives, sometimes called sterol esters or stanol esters. Sterol esters are long-chain (e.g., C6-C6) sterols such as octanoic acid, decanoic acid, undecanoic acid, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid. 24 For example, C 10 ~C 24 For example, C 14 ~C 24 These are sterols esterified with fatty acids, such as sterols. The sterols and their esters may be completely saturated (e.g., hydrogenated). A pharmaceutical composition containing sterols or their esters may contain one or more of the aforementioned components or a mixture thereof.

[0091] Bioactive agents The lipid-polymer composite particles described herein can encapsulate bioactive agents. These bioactive agents may include therapeutic agents, nutritional supplements, or stimulants. In some embodiments, the bioactive agent is a cannabinoid or a cannabinoid derivative. In more preferred embodiments, the cannabinoid or its derivative is cannabigerol acid (CBGA), cannabigerol monomethyl ether (CBGAM), cannabigerol (CBG), cannabigerol monomethyl ether (CBGM), cannabigerovalic acid (CBGVA), cannabigerovaline (CBGV), cannabichromeneic acid (CBCA), cannabichromene (CBC), cannabichromevalic acid (CBCVA), cannabichromevaline (CBCV), cannabidiolic acid (CBDA), delta-9-tetrahydrocannabinolic acid A (THCA-A), delta-9-tetrahydrocannabinolic acid Cannabinolate B (THCA-B), Delta-9-tetrahydrocannabinol (THC), Delta-9-tetrahydrocannabinolate-C4 (THCA-C4), Delta-9-tetrahydrocannabinol-C4 (THC-C4), Delta-9-tetrahydrocannabivaric acid (THCVA), Delta-9-tetrahydrocannabivarin (THCV), Delta-9-tetrahydrocannabiolcholic acid (THCA-C1), Delta-9-tetrahydrocannabiolcholic acid (THC-C1), Delta-7-cis-iso-tetrahydrocannabivarin, Delta-8-tetrahydrocannabinolate (Δ 8 -THCA), delta-8-tetrahydrocannabinol (Δ 8-THC), Cannabicycloalic acid (CBLA), Cannabicyclol (CBL), Cannabicyclovaline (CBLV), Cannabielsonic acid A (CBEA-A), Cannabielsonic acid B (CBEA-B), Cannabielsoin (CBE), Cannabinolic acid (CBNA), Cannabinol (CBN), Cannabinol methyl ether (CBNM), Cannabinol-C4 (CBN-C4), Cannabivarin (CBV), Cannabinol-C2 (CBN-C2), Cannabiolcol (CBN-C1), Cannabinodivaline (CBND), Cannabinodivaline (CBVD), Cannabitriol (CBT), 10-Ethoxy-9-Hydroxy-Delta-6α-Tetrahydrocannabinol, 8,9-Dihydroxy-Delta-6α-Tetrahydrocannabinol The cannabinoid derivatives are one or more of the following: cannabinol, cannabitriolvaline (CBTV), ethoxycannabitriolvaline (CBTVE), dehydrocannabifuran (DCBF), cannabifuran (CBF), cannabichromanone (CBCN), cannabicitran (CBT), 10-oxo-delta-6a-tetrahydrocannabinol (OTHC), delta-9-cis-tetrahydrocannabinol (cis-THC), 3,4,5,6-tetrahydro-7-hydroxy-alpha-alpha-2-trimethyl-9-n-propyl-2,6-methano-2H-1-benzoxosin-5-methanol (OH-iso-HHCV), cannabilipsol (CBR), and trihydroxy-delta-9-tetrahydrocannabinol (triOH-THC). In some embodiments, the cannabinoid derivatives are naturally occurring. In some embodiments, the cannabinoid derivatives are of non-natural origin, including those synthesized chemically or enzymatically. In some embodiments, the bioactive agent is a terpene, flavonoid, antibiotic, antiseptic, antifungal, antimicrobial, analgesic, anti-inflammatory, antiprotozoan, steroid, antiviral, lipophilic drug (e.g., with a solubility of less than 1 mg / ml), anti-VEGF agent, anti-glaucoma agent, essential oil, nicotine or nicotine analogue, cyclosporine A, tacrolimus, isotretinoin, propofol, griseofulvin, or any combination thereof.

[0092] In some embodiments, the bioactive agent includes an immunogen (e.g., a vaccine component, e.g., a DNA vaccine, an RNA vaccine, or a polypeptide vaccine). The immunogen may be, for example, a polypeptide, or a fragment, variant, or analog thereof, a nucleic acid (e.g., DNA or RNA), or another component of a cell. In some embodiments, the present invention features a vaccine comprising a composition such as those described herein.

[0093] In some embodiments, the essential oils include tea tree oil, myrrh oil, eucalyptus oil, clove oil, lavender oil, peppermint oil, chamomile oil, Roman chamomile oil, German chamomile oil, frankincense oil, helichrysum oil, cypress oil, angelica oil, labdanum oil, petitgrain bigarade oil, orange bigarade oil, bergamot oil, sweet orange oil, palmarosa oil, lemon iron bark oil, may chang oil, basil oil, sweet marjoram oil, geranium oil, patchouli oil, valerian oil, sandalwood oil, neroli bigarade oil, grapefruit oil, coriander oil, Includes citronella oil, black peppermint oil, garly gum oil, juniper yig oil, spearmint oil, scotch pine oil, rosemary oil, clary oil, ginger oil, lemon oil, mandarin oil, cumin oil, juniper berry oil, lemon balm oil, myrtle oil, ravensara oil, sweet thyme oil, everlasting oil, manuka oil, dwarf pine oil, oregano oil, vetiver oil, melissa oil, white fir oil, cassia oil, lemongrass oil, lime oil, wintergreen oil, fennel oil, ylang-ylang oil, or a combination thereof.

[0094] In some embodiments, the weight ratio between poloxamer and bioactive agent is about 4 to about 8 (e.g., about 4.1 to about 7.9, or about 4.2 to about 7.8, about 4.3 to about 7.7, about 4.4 to about 7.6, about 4.5 to about 7.5, about 4.6 to about 7.4, about 4.7 to about 7.3, about 4.8 to about 7.2, about 4.9 to about 7.1, about 5.0 to about 7.0, about 5.1 to about 6.9, about 5.2 to about 6.8, about 5.3 to about 6.7, about 5.4 to about 6.6, about 5.5 to about 6.5, about 5.6 to about 6.4, about 5.7 to about 6.3, about 5.8 to about 6.2, or about 5.9 to about 6.1). In some embodiments, the bioactive agent (e.g., cannabinoid or its derivative) is present in amounts of about 0.01% to about 10% by weight of the composition (e.g., about 0.05% to about 9.5%, about 0.1% to about 9%, about 0.2% to about 8.5%, about 0.4% to about 8%, about 0.5% to about 7.5%, about 1% to about 7%, about 1.5% to about 6.5%, about 2% to about 6%, about 2.5% to about 5.5%, about 3% to about 5%, about 3.5% to about 4.5%, about 4% to about 4.49%, for example, approximately 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.30%, 0.31% %, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.38%, 0.39%, 0.40%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.50%, 0.60%, 0.61%, 0.62%, 0.63%, 0.64%, 0.65%, 0.66%, 0.67%, 0.68%, 0.69%, 0.70%, 0.71%, 0 0.72%, 0.73%, 0.74%, 0.75%, 0.76%, 0.77%, 0.78%, 0.79%, 0.80%, 0.81%, 0.82%, 0.83%, 0.84%, 0.85%, 0.86%, 0.87%, 0.88%, 0.89%, 0.90%, 0.91%, 0.92%, 0.93%, 0.94%, 0.95%, 0.96%, 0.97%, 0.98%, 0.99%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5. It is present in the formulation at concentrations of 9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, or 10.0%. In some embodiments, the bioactive agent (e.g., essential oil) is present in a composition of approximately 0.01% to approximately 95% by weight (e.g., approximately 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.11%, 0.12%, 0.13%, 0.14%, 0.15%). %, 0.16%, 0.17%, 0.18%, 0.19%, 0.20%, 0.21%, 0.22%, 0.23%, 0.24%, 0.25%, 0.26%, 0.27%, 0.28%, 0.29%, 0.30%, 0.31%, 0.32%, 0.33%, 0.34%, 0.35%, 0.36%, 0.37%, 0.3 8%, 0.39%, 0.40%, 0.41%, 0.42%, 0.43%, 0.44%, 0.45%, 0.46%, 0.47%, 0.48%, 0.49%, 0.50%, 0.60%, 0.61%, 0.62%, 0.63%, 0.64%, 0.65%, 0.66%, 0.67%, 0.68%, 0.69%, 0. 70%, 0.71%, 0.72%, 0.73%, 0.74%, 0.75%, 0.76%, 0.77%, 0.78%, 0.79%, 0.80%, 0.81%, 0.82%, 0.83%, 0.84%, 0.85%, 0.86%, 0.87%, 0.88%, 0.89%, 0.90%, 0.91%, 0.92%, 0.93%, 0.94%, 0.95%, 0.96%, 0.97%, 0.98%, 0.99%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3.0%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4.0%, 4.1%, 4.2%, 4.3%, 4.4 %, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5.0%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6.0%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7.0%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8.0%, 8.1%, 8.2%, 8.3%, 8.4%, 8.5%, 8.6%, 8 0.7%, 8.8%, 8.9%, 9.0%, 9.1%, 9.2%, 9.3%, 9.4%, 9.5%, 9.6%, 9.7%, 9.8%, 9.9%, 10.0%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46% It is present in the composition at concentrations of 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, or 95%.

[0095] The bioactive agents encapsulated by the lipid-polymer composite particles described herein may also be terpenes, flavonoids, antibiotics, preservatives, antifungal agents, antimicrobial agents, analgesics, anti-inflammatory agents, antiprotozoal agents, steroids, antiviral agents, lipophilic drugs, anti-VEGF agents, or antiglaucoma agents. In some embodiments, the terpenes are myrcene, beta-caryophyllene, linalool, alpha-pinene, beta-pinene, ocimene, terpinolene, ocimene, terpinolene, alpha-tepinol, alpha-terpinene, gamma-terpinene, alpha-phellandrene, cymene, camphene, delta-3-carene, fencol, 1,8-cineole, nerolidol, borneol, eucalyptol, camphene, or limonene. In some embodiments, the flavonoid is selected from the group consisting of cannaflavin A, cannaflavin B, phenolic acids, stilbenoids, phytochemicals, dihydroflavonols, anthocyanins, anthocyanidins, polyphenols, tannins, flavones, flavonols, flavan-3-ol, flavan-4-ol, flavan-3,4-diol, homoisoflavonoids, phenylpropanoids, phloroglucinol, coumarin, phenolic acids, naphthodianthrone, steroid glycosides, bioflavonoids, or isoflavonoids.

[0096] In some embodiments, the bioactive agent is a vaccine component, for example, a protein or polypeptide fragment that induces immunity, prevents an infection, and / or reduces the risk of infection.

[0097] In some embodiments, the bioactive compound is nicotine, a nicotine analog, or a nicotine derivative. In other embodiments, the bioactive agent is cyclosporine A, tacrolimus, isotretinoin, propofol, griseofulvin, azithromycin, or a nonsteroidal anti-inflammatory drug (NSAID).

[0098] Pharmaceutical composition The compositions described herein are preferably formulated into pharmaceutical compositions for administration to human subjects in a biocompatible form suitable for in vivo administration. The pharmaceutical compositions may be formulated together with pharmaceutically acceptable carriers or excipients. A pharmaceutically acceptable carrier or excipient is a carrier (e.g., carrier, medium, diluent, solvent, vehicle, etc.) that does not significantly interfere with the biological activity or efficacy of the active ingredient(s) of the pharmaceutical composition and is not excessively toxic to the host at the concentrations used or administered. Other pharmaceutically acceptable components may similarly be present in the composition. Suitable substances for the formulation of pharmaceutically active compounds and their use are well known in the art (for example, see Remington: The Science and Practice of Pharmacy. 21st Edition. Philadelphia, PA. Lippincott Williams & Wilkins, 2005 for further discussion of pharmaceutically acceptable substances and methods for preparing various types of pharmaceutical compositions).

[0099] Pharmaceutical compositions are typically formulated to suit their intended route of administration. For oral administration, drugs can be formulated by combining a bioactive agent with a pharmaceutically acceptable carrier known in the art. Such carriers enable the formulation of the drugs of the present invention as powders, tablets, pills, capsules, lozenges, liquids, gels, syrups, slurries, suspensions, etc. When administered orally, it is recognized that some pharmaceutical compositions must be protected from digestion. This is typically achieved either by complexing a protein with the composition to make it resistant to acidic and enzymatic hydrolysis, or by packaging the protein in a suitable resistant carrier such as liposomes. Suitable excipients for oral dosage forms include, for example, sugars such as lactose, sucrose, mannitol, or sorbitol; and cellulose preparations such as starch, gelatin, tragacanth gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP). For example, disintegrants such as cross-linked polyvinylpyrrolidone, agar, or alginic acid or sodium alginate, or their salts, may be added. Optionally, the oral formulation may be formulated with physiological saline or buffer to neutralize the internal acidity, or it may be administered without a carrier.

[0100] For administration by inhalation, the pharmaceutical composition may be formulated in the form of an aerosol spray from a pressurized container or dispenser containing a suitable propellant, such as carbon dioxide, fluorocarbon, or a gas such as a nebulizer. Liquids or dry aerosols (e.g., dry powder, large porous particles, etc.) may also be used. For topical application, the pharmaceutical composition may be formulated in a suitable ointment, lotion, gel, or cream containing the active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers suitable for use in such compositions. Compositions formulated for intraocular administration may be formulated with, for example, hyaluronic acid.

[0101] The compositions described herein may be administered to a target in various forms depending on the selected route of administration, as will be understood by those skilled in the art. The compositions described herein may be administered, for example, by any route that allows the composition (e.g., lipid-polymer composite particles, e.g., micelles or LNPs) to reach target cells. The compositions may be administered, for example, orally, topically, parenterally, subarachnoidally, intraventricularly, intraparenchymically, intraorally, sublingually, transnasally, rectally, by patch, pump, transdermally, sublingually, vaginally, orally, ocularly, or transnasally, and by compounded pharmaceutical compositions as appropriate. The compositions may be administered by inhalation or spray. Parenteral administration includes intravenously, intraperitoneally, subcutaneously, intramuscularly, transepithelially, transnasally, intrapulmonaryly, rectally, and by topical administration, sublingually, intraorally, rectally, transvaginally, via the route of the eye, via the route of the ear, or via the route of the nose.

[0102] In some embodiments, the compositions described herein are formulated, for example, as food or as part of a food carrier that can be administered with a meal. In some embodiments, the compositions described herein include one or more essential oils, for example, tea tree oil, myrrh oil, eucalyptus oil, clove oil, lavender oil, peppermint oil, Roman chamomile oil, German chamomile oil, frankincense oil, helichrysum oil, cypress oil, angelica oil, labdanum oil, petitgrain bigarade oil, orange bigarade oil, bergamot oil, sweet orange oil, palmarosa oil, lemon iron bark oil, may chang oil, basil oil, sweet marjoram oil, geranium oil, patchouli oil, valerian oil, sandalwood oil, neroli bigarade oil, grapefruit oil, etc. This composition includes leander oil, citronella oil, black peppermint oil, garly gum oil, juniper yig oil, spearmint oil, scotch pine oil, rosemary oil, clary oil, ginger oil, lemon oil, mandarin oil, cumin oil, juniper berry oil, lemon balm oil, myrtle oil, ravensara oil, sweet thyme oil, everlasting oil, manuka oil, dwarf pine oil, oregano oil, vetiver oil, melissa oil, white fir oil, cassia oil, lemongrass oil, lime oil, wintergreen oil, fennel oil, ylang-ylang oil, or a combination thereof. In some embodiments, the composition includes a plurality of essential oils (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 essential oils). In some embodiments, the concentration of essential oils in the composition is 0.01% to 95% by weight of the composition (for example, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%).

[0103] In some embodiments, the compositions described herein are formulated as eye drops. In some embodiments, the eye drops contain one or more essential oils in an amount of, for example, 0.01% to 95% by weight (e.g., 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) of the composition.

[0104] Generally, the dosage of a pharmaceutical composition (e.g., a bioactive agent) is approximately 1 ng to 1 g (e.g., approximately 1 ng to 10 ng, e.g., 2 ng, 3 ng, 4 ng, 5 ng, 6 ng, 7 ng, 8 ng, 9 ng, 10 ng, e.g., 10 ng to 100 ng, e.g., 20 ng, 30 ng, 40 ng, 50 ng, 60 ng, 70 ng, 80 ng, 90 ng, 100 ng, e.g., For example, approximately 100ng to approximately 1μg, for example, 200ng, 300ng, 400ng, 500ng, 600ng, 700ng, 800ng, 900ng, 1μg, for example, approximately 1μg to approximately 10μg, for example, 1μg, 2μg, 3μg, 4μg, 5μg, 6μg, 7μg, 8μg, 9μg, 10μg, for example, approximately 10μg to approximately 100μg, for example, 20μg. 30μg, 40μg, 50μg, 60μg, 70μg, 80μg, 90μg, 100μg, for example 100μg to 1mg, for example 200μg, 300μg, 400μg, 500μg, 600μg, 700μg, 800μg, 900μg, 1mg, for example approximately 1mg to approximately 10mg, for example 2mg, 3mg, 4mg, 5mg, 6mg, 7mg, 8mg, The dosage may be in the range of 9 mg, 10 mg, for example, approximately 10 mg to approximately 100 mg, for example, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, for example, approximately 100 mg to approximately 1 g, for example, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 700 mg, 800 mg, 900 mg, or 1 g).

[0105] The dosage of a pharmaceutical composition (e.g., a bioactive agent) can be administered per kg of body weight of the subject. For example, the dosage can range from approximately 0.01 mg / kg to approximately 100 mg / kg, for example, approximately 0.01 mg / kg to approximately 30 mg / kg, approximately 0.01 mg / kg to approximately 10 mg / kg, approximately 0.1 mg / kg to approximately 1 mg / kg, for example, approximately 0.02 mg / kg, 0.03 mg / kg, 0.03 mg / g, 0.04 mg / kg, 0.05 mg / kg, 0.06 mg / kg, 0.07 mg / kg, 0.08 mg / kg, 0.08 mg / kg, 0.09 mg / kg, 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 0 The dose may be 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, 10 mg / kg, 15 mg / kg, 20 mg / kg, 25 mg / kg, 30 mg / kg, 35 mg / kg, 40 mg / kg, 45 mg / kg, 50 mg / kg, 60 mg / kg, 65 mg / kg, 70 mg / kg, 75 mg / kg, 80 mg / kg, 85 mg / kg, 90 mg / kg, 95 mg / kg, or 100 mg / kg. The aforementioned doses may be administered once a day, once a week, once a month, or once a year.

[0106] The dosage of the compositions described herein (e.g., compositions containing bioactive agents) may vary depending on many factors, such as the pharmacodynamic properties of the drug, the mode of administration, the age, health status, and weight of the recipient, the nature and severity of the symptoms, the frequency of treatment, and the type of concurrent treatment (if any), as well as the clearance rate of the composition in the treated animal. The compositions described herein may be initially administered in a suitable dosage that can be adjusted as needed depending on the clinical response. In some embodiments, the dosage of the composition (e.g., compositions containing bioactive agents) is a preventive or therapeutically effective amount. Furthermore, it is understood that all dosages may be administered continuously or divided into predetermined dosages at predetermined time intervals. The composition may be administered, for example, hourly, daily, weekly, monthly, or yearly. In some embodiments, the composition may be administered continuously or systemically.

[0107] The compositions described herein may be prepared using a multi-step process. In some embodiments, a series of steps in the process provide optimal particle size, polydispersity, solution clarity, pH, isotonicity, size distribution, stability, and loading of the bioactive agent. In some embodiments, the bioactive agent is homogenized with the polymer in a first step, for example, at a temperature of about 50°C to about 70°C, for example, about 60°C. In a second step, a solution containing lipids and sterols is added to the homogenized bioactive agent and polymer suspension using immersion injection (e.g., ethanol injection).

[0108] How to use The compositions described herein are formulated to treat diseases or conditions (for example, eye conditions such as inflammation, eye pain, conjunctivitis, dark circles under the eyes, red eyes, bacterial eye infections, fungal eye infections, viral eye infections, swelling, angiogenesis, malnutrition, macular degeneration, glaucoma, or elevated intraocular pressure, pain, bacterial infection, fungal infection, protozoan infection, anxiety, agitation, stress, fatigue, insomnia, mental fatigue, memory loss, organ rejection, eczema, acne, and skin infections). In some embodiments, the compositions are used to supplement nutrition. In some embodiments, the compositions are used for pleasure.

[0109] The compositions described herein may be formulated as eye drops for treating eye conditions such as dry eye disease, inflammation, eye pain, conjunctivitis, dark circles under the eyes, red eyes, bacterial eye infections, fungal eye infections, viral eye infections, nutrient deficiencies, macular degeneration, glaucoma, or elevated intraocular pressure.

[0110] In some embodiments, the composition may be used to treat a disease or condition selected from inflammation, pain, bacterial infection, fungal infection, protozoan infection, anxiety, agitation, stress, fatigue, insomnia, mental fatigue, memory loss, organ rejection, eczema, acne, and skin infections (e.g., athlete's foot, tinea, or jock itch).

[0111] In some embodiments, the compositions described herein may be administered to a subject as an immunogenic composition (e.g., a vaccine). The bioactive agent may include an immunogen (e.g., a vaccine component, e.g., a DNA vaccine, RNA vaccine, or polypeptide vaccine). The immunogen may be, for example, a polypeptide or its fragments, variants, or analogs, nucleic acids (e.g., DNA or RNA), or another component of a cell. The composition may be administered to a subject to prevent the onset of a disease or condition, or to reduce the risk of acquiring a disease or condition. The immunogenic composition may be administered, for example, topically, orally, by injection, sublingually, intraorally, rectally, vaginally, via the ocular route, via the ear route, via the nasal route, by inhalation, by spray, or percutaneously. In some embodiments, the immunogenic composition is administered intravenously, subcutaneously, or intramuscularly.

[0112] This composition may be administered to the target eye, the area surrounding the target eye, the external area of ​​the eye, the eyelid (for example, the outer area of ​​the eyelid, the inner area of ​​the eyelid), or the lacrimal duct. This composition may be administered as eye drops to each affected eye (for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 eye drops per day).

[0113] In some embodiments, the composition may be administered to a subject for pleasurable use. The composition may be administered to a subject for pleasurable use, as needed, for example, orally, by inhalation, parenterally, or topically.

[0114] This composition may be administered once, twice, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three, or twenty-four times a day. This composition may be administered weekly, bi-weekly, monthly, or every other month. This composition may be administered before or after bedtime.

[0115] In some embodiments, the composition is administered to a subject topically, orally, by injection, sublingually, orally, rectally, vaginally, through the eye, through the ear, through the nose, by inhalation, by spray, or percutaneously. [Examples]

[0116] The following examples are provided to those skilled in the art to illustrate how the compositions and methods described herein may be used, manufactured, and evaluated, and are intended purely to illustrate the present invention and not to limit the scope of what the inventors consider to be their invention.

[0117] Example 1. Preparation of phospholipid pre-liposomes Phospholipid liposome compositions were prepared and visually characterized. Three variant formulations, Fa1, Fa2, and Fa3, were prepared using 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cannabidiol (CBD), and optionally cholesterol. Fa1 was prepared by mixing DSPC (1 mg / ml), CBD (30 mg / ml), and cholesterol (0.37 mg / ml) in ethanol and subsequently injecting into DI water (Table 1). Fa2 was prepared by mixing DSPC (1 mg / ml) and CBD (30 mg / ml) in ethanol and subsequently injecting into DI water. Fa3 was prepared by mixing DSPC (2 mg / ml), CBD (30 mg / ml), and cholesterol (0.73 mg / ml) in ethanol and subsequently injecting into DI water.

[0118] [Table 1]

[0119] Visual characterization of the liposome suspensions Fa1, Fa2, and Fa3 revealed that all suspensions had a cloudy appearance with some precipitate (Table 2). The relative abundance of liposomes was qualitatively characterized using an optical microscope (Figure 1), and it was observed that the process of formulating Fa3 produced the highest concentration of liposomes.

[0120] [Table 2]

[0121] Example 2. Effect of temperature on phospholipid liposomes To overcome the precipitation observed in the Fa formulation experiments, liposome suspensions were prepared under different temperature conditions. The effect of the aqueous solvent temperature on the solubility and visual appearance of the liposome suspensions was evaluated. Three formulations, Fb1, Fb2, and Fb3, were prepared as shown in Table 3. Fb1 was prepared in deionized water at room temperature (20°C) and ethanol at 4°C. An ethanol solution containing DSPC (2 mg / ml), CBD (30 mg / ml), and cholesterol (0.73 mg / ml, 15 mol% of DSPC) was introduced into the water using the immersion injection technique. The resulting suspension had a cloudy appearance with precipitates of CBD and non-dispersed lipids. Fb2 was prepared by air injection of DSPC (2 mg / ml), CBD (30 mg / ml), and cholesterol (0.73 mg / ml, 15 mol% of DSPC) using deionized water at 20°C. The resulting suspension also had a cloudy appearance with precipitates of CBD and non-dispersed lipids. Fb3 was prepared by immersion injection of CBD and non-dispersed lipids using deionized water at 20°C. Fb3 had a turbid appearance, and precipitates of CBD and non-dispersed lipids were observed. (Figure 2)

[0122] [Table 3]

[0123] Example 3. Effects of CBD concentration and preparation method The preparation of lipid nanoparticle formulations (Fc1-Fc9, Table 4) was performed to investigate the effects of cannabidiol (CBD) concentration, temperature, and homogenization method on the appearance, particle size, and zeta potential of the nanoparticles in the suspension.

[0124] [Table 4]

[0125] The formulations used two concentrations of CBD (2 mg / ml and 30 mg / ml), while the concentration of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) was kept constant at 2 mg / ml in all formulations. The cholesterol concentration was also kept constant at 0.73 mg / ml (15 mol% of DSPC). Formulations Fc1 and Fc4 were formulations that directly compared CBD concentrations while maintaining consistent preparation conditions, such as temperature and concentration of DSPC and cholesterol. The effect of temperature was also evaluated. For Fc1 to Fc4, differences in the appearance, precipitate, particle size, and zeta potential of the suspensions were examined at either room temperature (20°C) or high temperature (60°C) (Table 5 and Figures 3-4). Homogenization of the suspensions was performed at either 20°C (Fc5) or 60°C (Fc8) for formulations Fc5 and Fc8. Visual examination of the appearance, precipitate, particle size, and zeta potential of the suspension at either room temperature (20°C) or high temperature (60°C) (Table 5 and Figures 3-4). For formulations Fc6 and Fc9, the effect of CBD concentration (2 mg / ml and 30 mg / ml) was examined in relation to the appearance of the suspension, precipitate, particle size, and zeta potential of nanoparticles while the suspension was kept at 60°C and CBD was added dropwise (Table 5 and Figures 3-4). Formulation Fc7 was a control formulation containing only the drug.

[0126] [Table 5]

[0127] The particle size of formulations Fc1 to Fc9 was evaluated as mean effective diameter (nm) and mean polydispersity with a standard deviation margin (Figure 3). The smallest nanoparticle size was observed starting with formulation Fc1. The preparation methods for formulations Fc1 and Fc4 differed only in the concentration of CBD used. Higher CBD concentrations resulted in larger particles, likely due to uncaptured CBD particles, which coincided with the largest particle size observed in formulation Fc7. The preparation method for formulation Fc1 also resulted in a mixture of smaller lipid nanoparticles and larger uncaptured CBD microparticles, which was reflected in the high polydispersity observed in Fc1. Formulation Fc4 demonstrated the optimal method for proceeding with a predetermined minimum nanoparticle size and low polydispersity value (Figure 3).

[0128] Zeta potentials were also measured for formulations Fc1 to Fc9 (Figure 4). The average zeta potential in mV was recorded along with the standard deviation (SD) from three measurements, where each recorded measurement is the average of 10 runs. The zeta potential value for formulation Fc4 was -0.05mV with an SD of 0.09, which is consistent with selecting Fc4 as the optimal formulation, as it was confirmed that the CBD was loaded into nanoparticles and the surface charge of the nanoparticles was close to neutral.

[0129] Example 4. Effect of poloxamer on polymer particle composition The effects of poloxamer concentration on suspension clarity, precipitate volume, particle size, suspension pH, and suspension viscosity were investigated using PLURONIC® F127 as the poloxamer. Micelle formulations Fd1 to Fd6 were prepared using 0.5 g / 100 ml of CBD and homogenized for 2 minutes at the lowest speed on a homogenizer. The concentration of PLURONIC® F127 varied from 1% to 10% (w / v) for formulations Fd1 to Fd6, as shown in Table 6.

[0130] [Table 6]

[0131] Formulations Fd1 to Fd6 all had a translucent appearance, in contrast to the cloudy appearance observed with lipid-only nanoparticles (Figures 5-6). Various amounts of precipitate were observed, correlating with the concentration of PLURONIC® F127 used (Table 7).

[0132] [Table 7]

[0133] Suspensions containing higher levels of PLURONIC® F127 exhibited greater clarity and lower precipitate levels, demonstrating the effect of PLURONIC® F127 on CBD dissolution (Figures 5-6). Particle size was characterized, and all formulations yielded micelles of similar size (less than 50 nm) (Figure 7). The pH and viscosity of formulations Fd1-Fd6 were measured and compared to two commercially available eye drops, THEALOZ® DUO and HYABAK® (Figures 8-9). All PLURONIC® F127 formulations except Fd3 had pH values ​​close to neutral. Fd3 showed a lower pH, close to 6.2 (Figure 8). Viscosity was also lower for all Fd formulations except one compared to the commercially available eye drops. Fd10 had a significantly higher viscosity, which may be due to it being the formulation with the highest CBD concentration (Figure 9). The stability of Fd1-Fd6 was also characterized on the day of preparation and after storage at room temperature (20°C) or 4°C for 30 days (Figure 10).

[0134] Example 5. Formulation of lipid-polymer composite particles Lipid-polymer composite particles were prepared into formulations Fe1 to Fe9 using PLURONIC® F127 at concentrations of 1%, 3%, and 5% (w / v), a 0.5% CBD concentration, 2 mg / ml of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and 0.73 mg / ml of cholesterol (Table 8). Characterization of the Fe formulations included visual inspection of clarity and presence of precipitates, particle size and polydispersity, suspension pH, suspension tonicity, size distribution, and size stability after 19 days of storage. Formulations Fe1, Fe4, and Fe7 were homogenized with DSPC at 60°C, and cholesterol was added to the suspension mixture by ethanol injection. Formulations Fe2, Fe5, and Fe8 were prepared in two steps. First, PLURONIC® F127 was homogenized with CBD, followed by the addition of DSPC and cholesterol by ethanol injection. Formulations Fe3, Fe6, and Fe9 were also prepared in two separate steps. First, PLURONIC® F127 was homogenized with CBD. Then, a powder mixture of DSPC and cholesterol was directly added to the PLURONIC® F127-CBD suspension at 60°C. Visual inspection revealed that all suspensions were translucent (Table 9 and Figure 11). Formulations Fe1, Fe4, and Fe7 had a milky appearance, but this milky appearance decreased as the PLURONIC® F127 concentration increased.

[0135] [Table 8]

[0136] Formulations Fe2 and Fe5 also showed a decrease in precipitate and milky appearance as the concentration of PLURONIC® F127 increased. Fe8 appeared transparent with no observable precipitate (Table 9 and Figure 11). Formulations Fe3, Fe6, and Fe9 also had a milky appearance (Figure 11), and the soft / turbid precipitate observed in Fe6 and Fe9 was present.

[0137] [Table 9]

[0138] In formulations Fe1, Fe4, and Fe7, particle size and polydispersity decreased with increasing PLURONIC® concentration. This trend was also observed in formulations Fe2, Fe5, and Fe8. In formulations Fe3, Fe6, and Fe9, PLURONIC® concentration did not correlate with particle size. This may be due to reduced solubility of DSPC and cholesterol due to the direct addition of DSPC and cholesterol in powder form (Figure 12). The particle span values ​​for all formulations ranged from approximately 1.00 to 1.60 (Figure 12), with polydispersity being lowest in the particles of formulation Fe5. Generally, formulations Fe2, Fe5, and Fe8 had the smallest average particle size. The pH was close to neutral across all formulations (Figure 13). The isotonicity of the formulations was also measured and compared with a physiological saline control solution, a suspension of PLURONIC® F127 and CBD alone, and commercially available eye drops HYABAK® 0.15% and THEALOZ® DUO (Figure 14). The size distribution was characterized for all formulations exhibiting two particle size populations. Formulations Fe5 and Fe8 showed a decrease in the abundance of the larger particle population compared to the smaller particle population. The stability of Fd1-Fd6 was also characterized on the day of preparation and after 19 days of storage at either room temperature (20°C) or 4°C (Figure 15). All formulations, except Fe1 and Fe9, showed stable size stability on day 19 at either room temperature (20°C) or 4°C. Visual inspection of all formulations was performed on day 19 from preparation, and all formulations stored at room temperature showed minimal to moderate color change to orange / pink. The formulations stored at 4°C showed no observable changes in color or appearance from the date of preparation (Table 10).

[0139] [Table 10]

[0140] Example 6. Method optimization of the selected formulation The selection of formulations was further optimized (Table 11). Replicas of each formulation, Fd5, Fe5, Fe8, and Fe2, were prepared and characterized for particle size and polydispersity (Figure 16). Optimal particle size and polydispersity were obtained for formulations Fd5, Fe5, and Fe8.

[0141] [Table 11]

[0142] Example 7. Effect of PLURONIC® F127 and lipid concentration on CBD solubility Due to its very low solubility in water and high lipophilicity, CBD is classified as Class 2 by the Biopharmaceutical Classification System (BCS). This low water solubility presents challenges to the bioavailability of CBD during pharmaceutical administration. In fasting individuals, the oral bioavailability of CBD is only about 6%. The aim of this study was to investigate the effect on CBD solubility of increasing the concentration of PLURONIC® F127, as well as increasing the concentration of lipids DSPC and cholesterol, along with a combination of PLURONIC® F127, DSPC, and cholesterol.

[0143] PLURONIC® F127, 18:0 DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine) and cholesterol (wool) were obtained from Sigma Aldrich. Cannabidiol (CBD) was supplied by MaxBiotech. Formulations containing PLURONIC® F127 at concentrations of 1, 3, 5, 7, 9, 11, 13, and 15% (w / w) were prepared by dissolving PLURONIC® F127 in distilled water. To prepare PLURONIC® F127 formulations at these concentrations, the formulations were stirred with a heated magnetic stirrer at 34°C. Lipid stock formulations for ethanol injection were prepared using DSPC at concentrations of 2, 4, 8, and 16 mg / ml in ethanol, and cholesterol at concentrations of 0.73, 1.46, 2.92, and 5.84 mg / ml in ethanol.

[0144] method CBD saturation solubility To determine the saturation solubility of CBD in the presence of PLURONIC® F127, PLURONIC® formulations were prepared as described above. Excess CBD was added to each formulation and stirred overnight at room temperature using a magnetic stirrer (Figure 17). To confirm how the addition of lipids affects saturation solubility, lipids were added to 5% PLURONIC® F127 containing excess CBD using the ethanol injection method. For this purpose, a 19G needle was immersed in the stirred formulation and 8.11% (volume of the final sample volume) of the lipid stock formulation was injected at a rate of 750 μl / 7 sec. Next, each formulation was stirred overnight as described above. The following day, the formulations were filtered using a 0.45 μm syringe filter, and the amount of dissolved CBD was quantified using high-pressure liquid chromatography.

[0145] Determining the effects of increasing lipid concentration When investigating the effect of lipids in the system, a volume of 20 ml (v1) was used as the target final volume. Two methods were used to investigate the effect of increasing lipid concentration on the micelle system. In the first method, CBD was added before ethanol injection, while in the second method, CBD was added after ethanol injection and after the evaporation of ethanol from the system.

[0146] Different lipid stocks were prepared in ethanol as described above. Excess CBD was added to 20 ml (v1) of 5% PLURONIC® F127 and stirred with a magnetic stirrer to bring the formulation to a temperature of 45-60°C. Lipids were added to the PLURONIC® formulation using the ethanol injection method while stirring on a hot plate. A 19G needle was immersed in the stirring formulation and 1.62 ml (8.11% (v / v) (v2)) of the lipid stock formulation was injected at a rate of approximately 750 μl / 7 seconds. Here, v2 = (v1 / 100) × 8.11. To evaporate the ethanol, the formulation was left uncovered and stirred on a hot plate at 45°C for 2 hours, and then stirred overnight at room temperature without a cover.

[0147] Another method to examine the effects of incorporating higher concentrations of lipids was to incorporate CBD after ethanol infusion. This method was similar to the one described in the previous section, but the difference was that the ethanol infusion for lipid infusion was performed before the addition of CBD. After ethanol infusion, the formulation was stirred at 45°C for 2 hours with the cover removed, and then stirred overnight at room temperature to evaporate the ethanol. The following day, the excess CBD was added to the formulation and stirred.

[0148] CBD content was determined by diluting the sample with acetonitrile (ACN) (if necessary) and then analyzing it using HPLC. Calibration curves and sample quantification were performed using an Agilent Infinity 1260 system. An Eclipse Plus C18, 4.6 × 150 mm, 3.5 μm column was purchased from Phenomenex. The mobile phase used was ACN:water (82:18 (v / v)). HPLC analysis was performed at room temperature, at a flow rate of 1 ml / min, and with an injection volume of 20 μl. The UV detector for HPLC analysis was set to 220 nm. Particle size and polydispersity were determined using a Brookhaven dynamic light scattering system.

[0149] result PLURONIC® F127 significantly increases the solubility of CBD in water, which enhances permeability and thereby increases the bioavailability of orally administered CBD. A peak CBD concentration of approximately 2% (w / v) was observed in formulations containing 7% to 11% PLURONIC® F127 (Figure 18). During TEM-based visual characterization of particles generated in the formulation, densely packed micelles ranging in size from 20 to 30 μm were observed in a 5% PLURONIC® F127 suspension (Figure 19). In a lipid-only suspension (DSPC:Chol; (0.21:0.19Mm)), the presence of two distinct structural types—small, diffused spherical structures and large aggregated structures—was observed (Figure 19). In a suspension of 5% PLURONIC(registered trademark) F127 + DSPC:Chol (0.21:0.19 mM), two distinct characteristics were observed: dense structures with particle sizes of 20-30 μm and monolayer and multilayer structures in which small micelles were encapsulated.

[0150] Characterization of formulations to which CBD has been added before ethanol injection. Formulations to which CBD was added before ethanol injection were characterized. The appearance of the formulations was recorded (Table 12). Three formulations in which PLURONIC® F127 was absent, CBD was added before ethanol injection, and the lipid concentration in the water was increased, showed a cloudy appearance.

[0151] [Table 12]

[0152] When PLURONIC® F127 was included in the formulations at a 5% (w / v) concentration, all three different formulations (Table 12) were cloudy, showed precipitates, and had a pinkish hue. The particle size and polydispersity of the formulations (Figure 20) were also characterized. Particle size was larger in the absence of PLURONIC® F127, and in formulations containing PLURONIC® F127, particle size increased with increasing lipid concentration (Figure 20). The pH of the formulations was also measured, and there was no substantial difference in pH in the presence or absence of PLURONIC® F127 (Figure 21).

[0153] The amount of CBD dissolved in formulations containing PLURONIC® F127 and formulations without PLURONIC® F127 was characterized (Figure 22). There was a significant difference in the amount of dissolved CBD when PLURONIC® F127 was included in the formulation. The concentration of lipids in the formulation did not have an effect on the amount of CBD solubility in the pure aqueous solution (Figure 22).

[0154] Characterization of formulations to which CBD has been added after ethanol injection. Preparations in which CBD was added after ethanol injection. The appearance of the preparations was recorded (Table 13). Three preparations in which PLURONIC® F127 was absent, CBD was added before ethanol injection, and the lipid concentration in the water was increased, showed a cloudy appearance.

[0155] [Table 13]

[0156] When PLURONIC® F127 was included in the formulation at 5% (w / v), all three different formulations (Table 13) were cloudy in appearance and contained observable precipitates, but the pink hue observed when ethanol infusion was performed after CBD addition was absent. The particle size and polydispersity of the formulations (Figure 23) were also characterized. Particle size was larger without PLURONIC® F127, and in formulations containing PLURONIC® F127, particle size increased with increasing lipid concentration (Figure 23). The pH of the formulations was also measured, and there was no substantial difference in pH in the presence or absence of PLURONIC® F127 (Figure 24).

[0157] The amount of CBD dissolved in formulations containing PLURONIC® F127 and formulations without PLURONIC® F127 was characterized (Figure 25). There was a significant difference in the amount of dissolved CBD when PLURONIC® F127 was included in the formulation. The concentration of lipids in the formulation did not have an effect on the amount of CBD solubility in the pure aqueous solution (Figure 25).

[0158] Consideration As demonstrated in this experiment, PLURONIC® F127 was able to significantly increase the solubility of CBD in water. TEM images show that incorporating lipids into the CBD-containing PLURONIC® formulation results in the formation of monolayer and multilayer liposomes, which appear to contain micelles. These hybrid systems lead to enhanced absorption of orally administered CBD. While the increased lipid concentration may have resulted in increased concentrations of these multiparticle systems, CBD solubility is only slightly improved compared to PLURONIC® alone. However, the hybrid particles containing lipid-coated micelles may act by shielding the micelles internally when the formulation is diluted, as well as at administration, thereby maintaining micelle integrity. Another interesting observation at a DSPC:Chol concentration of 1:64:1.51 was that substantially larger (twice the size) particles were formed by injecting ethanol after incorporating CBD into the system. This may mean that this sequence of CBD incorporation and ethanol injection leads to the encapsulation of more micelles within the liposomes and / or more drug within the hybrid composite particles.

[0159] Other Embodiments While the present invention has been described in relation to its specific embodiments, further modifications are possible, and this application aims to cover any modifications, uses, or adaptations of the present invention, including any deviations from the present invention, which are applicable to the essential features described herein and are generally based on the principles of the present invention and are within the scope of known or conventional practices in the art to which the invention relates. It will be understood that other embodiments are subject to the claims.

Claims

1. A composition comprising a plurality of particles encapsulating a hydrophobic bioactive agent, wherein the plurality of particles comprises liposome particles containing poloxamer 407, lipids selected from the group consisting of neutral lipids, cationic lipids, and anionic lipids, and cholesterol, wherein the plurality of particles have an average particle size between 10 and 1000 nanometers, the poloxamer 407 forms micelles encapsulating the bioactive agent, the liposome particles encapsulate the micelles, and the weight ratio of the poloxamer 407 to the bioactive agent is 2 to 15.

2. The composition according to claim 1, wherein the bioactive agent is a therapeutic agent, a nutritional supplement, or a pleasure agent.

3. The composition according to claim 1, wherein the bioactive agent is cannabinoid or cannabinoid derivative, terpene, flavonoid, antibiotic, preservative, antifungal agent, antibacterial agent, analgesic, anti-inflammatory agent, antiprotozoan agent, steroid, antiviral agent, lipophilic drug, anti-VEGF agent, anti-glaucoma agent, essential oil, vaccine, nicotine or nicotine analog, cyclosporine A, tacrolimus, isotretinoin, propofol, griseofulvin, or any combination thereof.

4. The essential oils mentioned above include tea tree oil, myrrh oil, eucalyptus oil, clove oil, lavender oil, peppermint oil, Roman chamomile oil, German chamomile oil, frankincense oil, helichrysum oil, cypress oil, angelica oil, labdanum oil, petitgrain bigarade oil, orange bigarade oil, bergamot oil, sweet orange oil, palmarosa oil, lemon iron bark oil, may chang oil, basil oil, sweet marjoram oil, geranium oil, patchouli oil, valerian oil, sandalwood oil, neroli bigarade oil, grapefruit oil, coriander oil, citronella oil, and black pepper. The composition according to claim 3, comprising mint oil, garlic gum oil, juniper yig oil, spearmint oil, scotch pine oil, rosemary oil, clary oil, ginger oil, lemon oil, mandarin oil, cumin oil, juniper berry oil, lemon balm oil, myrtle oil, ravensara oil, sweet thyme oil, everlasting oil, manuka oil, dwarf pine oil, oregano oil, vetiver oil, melissa oil, white fir oil, cassia oil, lemongrass oil, lime oil, wintergreen oil, fennel oil, ylang-ylang oil, or a combination thereof.

5. The composition according to claim 4, wherein the concentration of the essential oil is 0.01% by weight to 95% by weight.

6. The composition according to any one of claims 1 to 5, wherein the composition is formulated as an eye drop formulation.

7. The composition according to any one of claims 1 to 6, wherein the lipid comprises a carbon chain of length 4 to 22 and a neutral, cationic, or anionic head group.

8. The composition according to claim 7, wherein the lipid is phosphatidylcholine, phosphatidylserine, phosphatidylglycerol, phosphatidylethanolamine, or phosphatidylinositol.

9. The composition according to any one of claims 1 to 8, wherein the concentration of the lipid is about 0.1 mol% to about 10 mol%.

10. The composition according to any one of claims 1 to 9, wherein the concentration of the cholesterol is about 5 mol% to about 50 mol% of the total lipid composition.

11. The composition according to any one of claims 1 to 10, wherein the weight ratio of cholesterol to lipids is about 0.01 to about 0.

50.

12. The composition according to any one of claims 1 to 11, wherein the plurality of particles have a polydispersity index of 0.3 or less.

13. The composition according to any one of claims 1 to 12, wherein the composition does not contain an organic solvent for solubilizing the biological agent.

14. A composition according to any one of claims 1 to 13, for use in providing a biologically active agent to a target.

15. The composition according to claim 14, formulated for local, oral, or injectable administration, sublingually, intraorally, rectally, vaginally, via the ocular route, via the ear route, via the nasal route, by inhalation, by spray, or transdermally.

16. (a) A step of homogenizing a hydrophobic bioactive agent with poloxamer 407 to produce a homogenized solution, and after step (a) (b) by the step of injecting cholesterol and lipids, selected from the group consisting of neutral lipids, cationic lipids, and anionic lipids, into the homogenized solution, A method for preparing a composition comprising a plurality of particles, wherein a plurality of particles encapsulate a hydrophobic bioactive agent, the plurality of particles comprising liposome particles containing poloxamer 407, lipids, and cholesterol, having an average particle size between 10 and 1000 nanometers, the poloxamer 407 forming micelles that encapsulate the bioactive agent, the liposome particles encapsulating the micelles, and the weight ratio of the poloxamer 407 to the bioactive agent being 2 to 15.

17. The method according to claim 16, wherein step (b) comprises immersion injection of the cholesterol and the lipids.

18. The method according to claim 17, wherein step (b) comprises immersion ethanol injection of the cholesterol and the lipids.

19. The method according to any one of claims 16 to 18, wherein step (a) is carried out at 50°C to 70°C.

20. The method according to any one of claims 16 to 19, wherein the composition does not contain an organic solvent for solubilizing the biological agent.

21. The method according to any one of claims 16 to 20, wherein the plurality of particles have a polydispersity index of 0.3 or less.

22. A composition according to any one of claims 1 to 13 for pleasure purposes.

23. A composition according to any one of claims 1 to 13 for use in treating a disease or condition selected from inflammation, pain, bacterial infection, fungal infection, protozoan infection, anxiety, agitation, stress, fatigue, insomnia, mental fatigue, memory loss, organ rejection, eczema, acne, skin infection, dry eye disease, eye pain, contagious conjunctivitis, dark circles under the eyes, red eyes, bacterial eye infection, fungal eye infection, viral eye infection, swelling, neovascularization, nutritional deficiency, macular degeneration, glaucoma, or elevated intraocular pressure.

24. An immunogenic composition comprising the composition according to any one of claims 1 to 13.

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