Lipid-polymer compositions and methods of use

Abstract

A composition comprising a plurality of lipid-polymer composite particles encapsulating a biologically active 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, wherein the plurality of lipid-polymer composite particles have an average particle size of between 10 and 1000 nanometers. A method of making a plurality of lipid-polymer composite particles encapsulating a biologically active 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, wherein the plurality of lipid-polymer composite particles have an average particle size of between 10 and 1000 nanometers, the method comprising homogenizing the biologically active agent with the polymer to produce a homogenized solution and injecting the lipid and the sterol into the homogenized solution.

Description

Background Technology

[0001] Delivering bioactive agents with low solubility in aqueous media has always been a challenge in drug delivery. One approach to targeted delivery of insoluble bioactive agents is to use liposomes and / or micellar nanoparticles as carrier systems. However, challenges related to stability, degradation, and bioavailability have hindered the widespread implementation of such delivery systems. Therefore, novel formulations using nanoparticle delivery systems are needed to effectively deliver bioactive agents. Summary of the Invention

[0002] In one aspect, the present invention provides a composition comprising a plurality of lipid-polymer composite particles encapsulating a bioactive agent. The lipid-polymer composite particles may comprise block copolymers, 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, about 500 nm to about 1000 nm).

[0003] In some embodiments, the bioactive agent is a therapeutic agent.

[0004] In some embodiments, the bioactive agent is a terpene, flavonoid, antibiotic, preservative, antifungal agent, antibacterial agent, analgesic, anti-inflammatory agent, antiprotozoal agent, steroid, antiviral agent, lipophilic drug, antiVEGF agent, antiglaucoma agent, essential oil, immunogen (e.g., vaccine component), nicotine or nicotine analogue, 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, immortelle oil, cypress oil, angelica oil, rockrose oil, lime leaf oil, orange leaf oil, bergamot oil, sweet orange oil, palmarosa oil, lemon bark oil, litsea oil, basil oil, sweet marjoram oil, geranium oil, patchouli oil, valerian oil, sandalwood oil, bitter orange oil, grapefruit oil, coriander oil, lemongrass oil, black peppermint oil, gully gum oil, juniper twig oil, spearmint oil, Scots pine oil, rosemary oil, clary oil, ginger oil, lemon oil, citrus oil, cumin oil, juniper oil, lemon balm, myrtle oil, and ravensara oil. The essential oils include sweet thyme oil, immortelle oil, manuka oil, dwarf pine oil, oregano oil, vetiver oil, bee pollen oil, white fir oil, cinnamon oil, lemongrass oil, lime oil, wintergreen oil, fennel oil, ylang-ylang oil, or combinations thereof. In some embodiments, the concentration of the essential oils is from 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 include 2 to 10 essential oils. In some embodiments, the concentration of the plurality of essential oils is from 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 is formulated as an eye drop preparation.

[0005] In some embodiments, the block copolymer is poloxamer (e.g., poloxamer 407).

[0006] In some embodiments, the weight ratio of poloxamer to the bioactive agent is about 2 to about 15.

[0007] In some embodiments, the lipid comprises a carbon chain of length 4 to 22 and a head group having a neutral, cationic, or anionic head group. In some embodiments, the lipid is phosphatidylcholine, phosphatidylserine, phosphatidylglycerol, phosphatidylethanolamine, or phosphatidylinositol.

[0008] In some embodiments, the concentration of the lipid is from about 0.1 mol% to about 10 mol%.

[0009] In some embodiments, the sterol is a phytosterol, a synthetic sterol, cholesterol, or a cholesterol analogue.

[0010] In some embodiments, the concentration of the sterol is from about 5 mol% to about 50 mol% of the total lipid composition.

[0011] In some embodiments, the weight ratio of the sterol to the lipid is from about 0.01 to about 0.50.

[0012] In another aspect, the present invention is characterized by immunogenic compositions comprising compositions as described herein.

[0013] In another aspect, the present invention is characterized by a method of providing a bioactive agent to a subject by administering to the subject any of the compositions described in the above embodiments.

[0014] In some embodiments, the dosage of the bioactive agent is from about 0.01 mg / kg to about 30 mg / kg.

[0015] In some implementation methods, the administration can be local, oral, by injection, sublingual, buccal, rectal, vaginal, ocular, audible, nasal, inhaled, nebulized, or transdermal.

[0016] In another aspect, the present invention is characterized by a method for preparing the compositions described in any of the above embodiments. In some embodiments, the preparation of any of the compositions described in the above embodiments comprises a multi-step process. In some embodiments, the multi-step process comprises a first step comprising homogenizing a bioactive agent with any of the polymers described in the above embodiments to produce a homogeneous solution, and a second step comprising injecting (e.g., immersion injection) any of the lipids and sterols described in the above embodiments into the homogeneous solution of the first step.

[0017] In another aspect, the present invention provides a method for treating dry eye, inflammation, eye pain, acute conjunctivitis (pink eye), dark circles, red eye, bacterial eye infection, fungal eye infection, viral eye infection, nutritional deficiency, macular degeneration, glaucoma, or elevated intraocular pressure, the method comprising administering to the eye of a subject a composition of lipid-polymer composite particles having a plurality of encapsulated bioactive agents, wherein the lipid-polymer composite particles comprise block copolymers, lipids selected from the group consisting of neutral lipids, cationic lipids, and anionic lipids, and sterols, wherein the plurality of lipid-polymer composite particles have an average particle size between 10 and 1000 nanometers.

[0018] In another aspect, the present invention is characterized by a method for treating diseases and symptoms 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, the method comprising administering to a subject a composition of lipid-polymer composite particles having a plurality of encapsulated bioactive agents, wherein the lipid-polymer composite particles comprise block copolymers, lipids selected from the group consisting of neutral lipids, cationic lipids, and anionic lipids, and sterols, wherein the plurality of lipid-polymer composite particles have an average particle size between 10 and 1000 nanometers.

[0019] In some embodiments of any of the foregoing aspects, the bioactive agent is a terpene, flavonoid, antibiotic, preservative, antifungal agent, antibacterial agent, analgesic, anti-inflammatory agent, antiprotozoal agent, steroid, antiviral agent, lipophilic drug, antiVEGF agent, antiglaucoma agent, essential oil, immunogen (e.g., vaccine component), nicotine or nicotine analogue, cyclosporine A, tacrolimus, isotretinoin, propofol, griseofulvin, or any combination thereof.

[0020] In some embodiments, the block copolymer is poloxamer. In some embodiments, the weight ratio of the poloxamer to the bioactive agent is between 2 and 15.

[0021] 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 a phytosterol, or a synthetic sterol, or cholesterol or a cholesterol analogue. 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 the sterol to the lipid is from about 0.01 to about 0.50.

[0022] In any of the embodiments described above, 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, immortelle oil, cypress oil, angelica oil, rockrose oil, lime leaf oil, petitgrain oil, bergamot oil, sweet orange oil, palmarosa oil, lemon bark oil, litsea cubeba oil, basil oil, marjoram oil, geranium oil, patchouli oil, valerian oil, sandalwood oil, bitter orange oil, etc. Grapefruit oil, coriander oil, lemongrass oil, black pepper oil, gum oil, juniper twig oil, spearmint oil, Scotch pine oil, rosemary oil, sage oil, ginger oil, lemon oil, citrus oil, cumin oil, juniper oil, lemon balm, myrtle oil, ravensa leaf oil, sweet thyme oil, immortelle oil, manuka oil, dwarf pine oil, oregano oil, vetiver oil, bee pollen oil, white fir oil, cinnamon oil, lemongrass oil, pearberry oil, wintergreen oil, fennel oil, ylang-ylang oil, or combinations thereof. In some embodiments, the concentration of the essential oil is from 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 multiple essential oils are from 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, the method comprises formulating the composition into an eye drop preparation.

[0023] 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 a phytosterol, or a synthetic sterol, or cholesterol or a cholesterol analogue. 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 the sterol to the lipid is from about 0.01 to about 0.50.

[0024] definition

[0025] To facilitate understanding of the invention, several terms are defined below. The terms defined herein have meanings commonly understood by one of ordinary skill in the art related to this invention. Terms such as “a,” “an,” and “the” are not intended to refer to a single entity, but rather to encompass general categories that can be illustrated using specific examples. The terms herein are used to describe specific embodiments of the invention, but their use does not limit the invention unless set forth in the claims.

[0026] As used in this article, any value provided within the numerical range includes the upper and lower limits, as well as any value contained within the upper and lower limits.

[0027] As used in this article, the term “about” means ±10% of the listed values.

[0028] The term "application" refers to introducing the formulation of the present invention into a patient in need to treat a disease or symptom.

[0029] As used herein, the term "bioactive agent" refers to any synthetic or naturally occurring compound (in free, salt, solvated, or hydrated form) having the desired biological or physiological effect, such as proteins, pharmaceuticals, antigens, nutrients, cosmetics, flavorings, taste agents, diagnostic agents, drugs, vitamins, or dietary agents, and will be formulated at a level sufficient to provide a functional level in vivo (including local concentrations in topical compositions). In some cases, one or more components of the lipid matrix i) (e.g., components (a), (b), (c), and / or (d)) may also be active agents, although it is preferred that the optional bioactive agent (iii) should not be one of these components (e.g., should not be a component of the lipid matrix). The most preferred active agents are pharmaceuticals (e.g., APIs), including drugs, vaccines, and diagnostic agents.

[0030] As used herein, the term "block copolymer" refers to a linear polymer having regions or blocks along its main chain that are characterized by similar hydrophilic, hydrophobic, or chemical properties.

[0031] The term "diblock copolymer" refers to a block copolymer that contains two blocks.

[0032] The term "triblock copolymer" refers to a block copolymer that contains three blocks.

[0033] The term "multiblock copolymer" refers to a block copolymer that contains multiple blocks.

[0034] As used herein, the terms “encapsulate,” “encapsulated,” or “encapsulating” refer to encapsulating a portion (e.g., a bioactive agent as defined herein) within a closed polymer assembly structure (e.g., micelles). The encapsulated bioactive agent is encapsulated within the polymer assembly structure, for example, the encapsulated portion being located within the hydrophobic interior of the polymer assembly structure (e.g., the lumen of a micelle).

[0035] As used in this article, the term "anionic head group" refers to a lipid head group that carries a net negative charge at physiological pH.

[0036] As used in this article, the term "cationic head group" refers to a lipid head group that carries a net positive charge at physiological pH.

[0037] As used in this article, the term "neutral head group" refers to a lipid head group that exists in an uncharged form at physiological pH.

[0038] As used herein, "lipid nanoparticles" or "LNPs" are vesicles containing a lipid layer encapsulating a substantially solid lipid core; the lipid core may contain pharmaceutically active molecules. LNPs typically contain cationic lipids, non-cationic lipids, and lipids that prevent particle aggregation (e.g., PEG-lipid conjugates).

[0039] As used herein, the term "lipid-polymer composite particle" refers to a molecular complex 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 can include macromolecular complexes forming, for example, spherical structures. Lipid-polymer composite particles include, for example, lipid nanoparticles and micelles.

[0040] As used herein, the term “micelle” or variations thereof refers to a polymer assembly consisting of a hydrophilic shell (or crown) and a hydrophobic shell (or crown) and / or ionic interior. Furthermore, the term micelle can refer to any polyionic complex assembly consisting of a multiblock copolymer with a net positive charge and a suitable negatively charged polynucleotide.

[0041] As used herein, the term “nanoparticle” refers to polymer-based particles having a diameter in the nanometer range (e.g., 1 nm–1000 nm).

[0042] As used in this article, the term “multiple” means more than one, such as at least 2, 20, 50, 100, 1000, 10000, 100000, 100000, 1000000, 1000000, or even more.

[0043] As used herein, the term “subject” can refer to a human, a non-human primate or other mammal, such as, but not limited to, dogs, cats, horses, cattle, pigs, turkeys, goats, fish, monkeys, chickens, rats, mice and sheep.

[0044] As used herein, the term "sterol" includes all sterols but is not limited to, for example:

[0045] Sitosterol, campesterol, stigmasterol, brassosterol (including dihydrobrassosterol), sterols, chalinosterol, porous sterol, perforated sponge sterol, ergosterol, codostellarol, pine sterol, isoflavosterol, fucosterol, cinnamesterol, nervisterol, 7-enylcholine, stellasterol, spinachsterol, peposterol, alfalfa sterol, isoalfalfa sterol, codostellarol, pollinastasterol, 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 this invention.

[0046] The term "therapeutic agent" refers to any substance that has therapeutic properties that produce a desired, generally beneficial effect. For example, a therapeutic agent can treat, improve, and / or prevent disease. Such agents can be synthetic or naturally occurring non-peptides, proteins or peptides, oligonucleotides or nucleotides, polysaccharides or sugars.

[0047] A “vesicle” is defined herein as a lipid-polymer composite particle in which amphiphilic molecules (e.g., lipids) collectively define a volume, for example, a substantially spherical volume. Amphiphilic molecules (e.g., lipids) typically constitute at least one shell of the vesicle. Within this shell, the amphiphilic molecules are arranged in a bilayer, with the hydrophilic portions of the molecules facing outwards relative to the bilayer plane, and the hydrophobic portions of the molecules predominantly located within the bilayer. If the surrounding medium is hydrophobic, the opposite arrangement exists. Attached Figure Description

[0048] Figure 1 Optical micrographs of the primary liposome formulations Fa1, Fa2, and Fa3 are shown. Scale bar is 50 μm.

[0049] Figure 2 Optical micrographs of primary liposome formulations Fb1, Fb2, and Fb3, prepared to evaluate the effect of temperature on particulate formulations, are shown. Scale bar is 10 μm.

[0050] Figure 3This is a table showing the effective diameter and polydispersity of formulations Fc1-Fc7. Formulations Fc1, Fc2, and Fc9 are loaded with 0.02% w / w CBD. Formulations Fc3-Fc8 are loaded with 0.25% w / w CBD. Report the mean and standard deviation values.

[0051] Figure 4 This is a table showing the average and standard deviation values ​​of the measured zeta potential of particles in formulations Fc1-Fc9.

[0052] Figure 5 This is a photographic image showing the optical transparency of formulations Fd1-Fd6. Transparency varies with poloxamer. The concentration of CBD increased with increasing F127 concentration. The CBD concentration in all suspensions was kept constant at 0.5% w / w, while the concentrations in formulations Fd1, Fd2, Fd3, Fd4, Fd5, and Fd6 increased. The F127 concentrations were 1%, 2%, 3%, 4%, 5%, and 10%, respectively.

[0053] Figure 6 These are photographic images showing the effect of poloxamer concentration on the observable precipitation amounts in formulations Fd6, Fd5, Fd4, Fd2, and Fd1. The CBD concentration in all suspensions was kept constant at 0.5% w / w, while the concentrations in formulations Fd1, Fd2, Fd4, Fd5, and Fd6 were... The F127 concentrations were 1%, 2%, 4%, 5%, and 10%, respectively.

[0054] Figure 7 This is a graph showing the effective diameter and polydispersity of formulations Fd1-Fd6. All formulations produced micelles of similar size, less than 50 μm.

[0055] Figure 8 This shows the pH of formulations Fd1-Fd6 compared to commercially available eye drops. DUO and A graph comparing pH values.

[0056] Figure 9 This shows the viscosity of formulations Fd1-Fd6 compared to water and two commercially available eye drops. DUO and The graph compares the viscosity of the two.

[0057] Figure 10 This is a graph showing the dimensional stability of formulations Fd1-Fd6. The effective diameter and polydispersity of all formulations were calculated on the day of preparation (left bar) and after preparation and storage at room temperature (20°C) or 4°C (right bar) for 30 days.

[0058] Figure 11These are photographic images showing the optical transparency of formulations Fe1-Fe9. Formulations Fe1, Fe4, and Fe7 were prepared by homogenization with poloxamer, followed by the addition of 0.5% CBD w / w, phospholipids, and cholesterol via ethanol infusion. Formulations Fe2, Fe5, and Fe8 were prepared by homogenization with poloxamer, followed by the addition of 0.5% CBD w / w via ethanol infusion, and then the addition of phospholipids and cholesterol. Formulations Fe3, Fe6, and Fe9 were prepared by homogenization with poloxamer, with 0.5% CBD w / w in powder form, followed by the addition of phospholipids and cholesterol.

[0059] Figure 12 This is a graph showing the particle size, polydispersity, and span values ​​of the lipid-polymer composite particles in formulation Fe1-Fe9.

[0060] Figure 13 This is a graph showing the pH values ​​of formulations Fe1-Fe9. The pH values ​​of all formulations are close to neutral.

[0061] Figure 14 This demonstrates the tonicity of formulations Fe1-Fe9 compared to saline, control poloxamer and CBD suspensions, and three commercially available eye drops. 0.15% DUO New and A table comparing the tension of DUO.

[0062] Figure 15 This is a graph showing the dimensional stability of formulations Fe1-Fe9. The effective diameter and polydispersity of all formulations were calculated on the day of preparation (left bar) and after preparation and storage at room temperature (20°C) or 4°C (right bar) for 19 days.

[0063] Figure 16 This is a graph showing the particle size, polydispersity, and span values ​​of the optimized formulations Fd5, Fe5, Fe8, and Fe2.

[0064] Figure 17 This shows the stirring to concentrations of 1, 3, 5, 7, 9, 11, 13, and 15% w / v. Photograph of an excessive amount of CBD in the formulation of F127. The leftmost vial does not contain CBD. F127 was used as a control. The viscosity of the formulation increased with... The concentration of F127 increases with the increase of F127 concentration.

[0065] Figure 18 The table shows CBD concentrations in water (0%), 1%, 3%, 5%, 7%, 9%, 11%, 13%, and 15%. F127, lipid-only (0% + lipid) and 5% containing lipid (5% + lipid) A graph showing the saturated solubility of F127. The lipid concentrations used were 0.21 mM DSPC and 0.19 mM cholesterol. The solubility of CBD increased with... The concentration of F127 increases with increasing concentration, ranging from 7-11%. The solubility of CBD peaks near F127. At the lipid concentrations used, the solubility of CBD does not differ significantly with lipid incorporation.

[0066] Figure 19 This is a series of transmission electron microscopy (TEM) images of three different formulations containing CBD. At 5% In the F127 formulation (top row), closely packed spherical structures with diameters ranging from 20 to 30 μm were observed. These are likely micelles. In the lipid-only formulation (middle row) (DSPC:Chol; (0.21:0.19 Mm)), two distinct characteristics were observed: smaller, diffuse spherical structures and larger, bulky structures. In formulations containing lipids and... F127 formulation (bottom row) (5%) Two distinct features were also observed in F127+DSPC:Chol(0.21:0.19mM). Closely packed 20-30 μm micelle structures and larger monolayer and multilayer structures containing smaller micelles were observed.

[0067] Figure 20 This is a graph showing the particle size and polydispersity of formulations containing high concentrations of DSPC and cholesterol. (Compared to...) Compared to when F127 binds to DSPC:Chol at a concentration of 1.64:1.51 mM, in the absence of... Higher particle size is typically observed in the case of F127. Furthermore, it is noteworthy that particle size increases with the presence of... The increase is due to the increase in lipid concentration in the formulation of F127.

[0068] Figure 21 It indicates whether it is present or absent. The pH of the F127 formulations did not differ significantly, and the pH did not change significantly with increasing lipid (DSPC and cholesterol) concentrations, as shown in the graph.

[0069] Figure 22 This shows the dissolution of lipids in the absence of (left) and the presence of (right) lipids with increased concentrations. A diagram of CBD in the formulation of F127 (right).

[0070] Figure 23 This is a graph showing the particle size and polydispersity of formulations containing high concentrations of lipids (DSPC and cholesterol). Similarly, in the absence of... In the case of F127, the particle size is higher, and with The increase is due to the increase in lipid concentration in the F127 formulation.

[0071] Figure 24 It indicates whether it is present or absent. The pH of the F127 formulations did not differ significantly, and the pH did not change significantly with increasing lipid concentration, as shown in the graph.

[0072] Figure 25 The diagram shows the difference between deficiency (left) and presence (right) of lipids (DSPC and cholesterol) with increased concentrations of these substances. A diagram of CBD in the formulation of F127. Detailed Implementation

[0073] The present invention is characterized by novel lipid-polymer composite particles that can be used in the formulation of bioactive agents for administration to subjects, such as human subjects. The lipid-polymer composite particles comprise multiple (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) nanoparticles encapsulating a bioactive agent. The nanoparticles comprise block copolymers, lipids such as phospholipids, and sterols. Formulation of the bioactive agents (e.g., therapeutic agents) described herein provides for easier loading of lipid-polymer composite particles with higher drug loading capacity, formulation stability, and lower surface tension, allowing for lipid coating and retention. Such improved processes for preparing lipid-polymer composite particles (e.g., micelles) with lipid coatings allow for aqueous loading of hydrophobic bioactive agents and controlled drug release. Furthermore, the compositions and methods described herein avoid the use of organic solvents such as ethanol to dissolve hydrophobic bioactive agents. The components of the formulation are described in more detail below.

[0074] Lipid-polymer composite particles

[0075] Lipid-polymer composite particles can be used to formulate bioactive agents (e.g., therapeutic agents) for delivery. Lipid-polymer composite particles comprise defined molecular complexes (e.g., lipids and polymers) held together by non-covalent bonds (hydrogen-injected bonds, van der Waals forces, electrostatic interactions, hydrophobic effects, and Pi-Pi interactions). Lipid-polymer composite particles can include macromolecular complexes forming globular, rod-like, or sheet-like structures. Lipid-polymer composite particles include, for example, micelles and LNPs. Lipid-polymer composite particles can have predetermined sizes. The size of the structure can vary depending on the size or number of molecules of the components enclosed within the structure (e.g., the size or number of bioactive agent molecules).

[0076] The size of lipid-polymer composite particles can vary from, for example, from about 10 nm to about 1,000 nm. Non-limiting examples of Z-average particle size 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 may have, for example, about 10 nm, about 15 nm, about 20 nm, about 25 nm, about 30 nm, about 35 nm, about 40 nm, about 45 nm, about 50 nm, about 55 nm, about 60 nm, about 65 nm, about 70 nm, about 75 nm, about 80 nm, about 85 nm, about 90 nm, about 95 nm, about 100 nm, about 105 nm, about 110 nm, about 115 nm, about 120 nm, about 125 nm, about 130 nm, about 135 nm, about 140 nm, about 145 nm, about 150 nm, about 155 nm, about 160 nm, about 165 nm, and so on. 170nm, approximately 175nm, approximately 180nm, approximately 185nm, approximately 190nm, approximately 195nm, approximately 200nm, approximately 205nm, approximately 210nm, approximately 215nm, approximately 220nm, approximately 225nm, approximately 230nm, approximately 235nm, approximately 240nm, approximately 245nm, approximately 250nm, approximately 255nm, approximately 260nm, approximately 265nm, approximately 270nm, approximately 275nm, approximately 280nm, approximately 285nm, approximately 290nm, approximately 295nm, approximately 300nm, approximately 305nm, approximately 310nm, approximately 315nm, approximately 320nm, approximately 325nm, approximately 33 0nm, approximately 335nm, approximately 340nm, approximately 345nm, approximately 350nm, approximately 355nm, approximately 360nm, approximately 365nm, approximately 370nm, approximately 375nm, approximately 380nm, approximately 385nm, approximately 390nm, approximately 395nm, approximately 400nm, approximately 405nm, approximately 410nm, approximately 415nm, approximately 420nm, approximately 425nm, approximately 430nm, approximately 435nm, approximately 440nm, approximately 445nm, approximately 450nm, approximately 455nm, approximately 460nm, approximately 465nm, approximately 470nm, approximately 475nm, approximately 480nm, approximately 485nm, approximately 490nm m, approximately 495nm, approximately 500nm, approximately 505nm, approximately 510nm, approximately 515nm, approximately 520nm, approximately 525nm, approximately 530nm, approximately 535nm, approximately 540nm, approximately 545nm, approximately 550nm, approximately 555nm, approximately 560nm, approximately 565nm, approximately 570nm, approximately 575nm, approximately 580nm, approximately 585nm, approximately 590nm, approximately 595nm, approximately 600nm, approximately 605nm, approximately 610nm, approximately 615nm, approximately 620nm, approximately 625nm, approximately 630nm, approximately 635nm, approximately 640nm, approximately 645nm, approximately 650nm,Approximately 655nm, approximately 660nm, approximately 665nm, approximately 670nm, approximately 675nm, approximately 680nm, approximately 685nm, approximately 690nm, approximately 695nm, approximately 700nm, approximately 705nm, approximately 710nm, approximately 715nm, approximately 720nm, approximately 725nm, approximately 730nm, approximately 735nm, approximately 740nm, approximately 745nm, approximately 750nm, approximately 755nm, approximately 760nm, approximately 765nm, approximately 770nm, approximately 775nm, approximately 780nm, approximately 785nm, approximately 790nm, approximately 795nm, approximately 800nm, approximately 805nm, approximately 810nm, approximately 815nm, approximately 820nm, approximately 825nm, approximately 830nm Z-average particle size at approximately 835 nm, 840 nm, 850 nm, 855 nm, 860 nm, 865 nm, 870 nm, 875 nm, 880 nm, 885 nm, 890 nm, 895 nm, 900 nm, 905 nm, 910 nm, 915 nm, 920 nm, 925 nm, 930 nm, 935 nm, 940 nm, 945 nm, 950 nm, 955 nm, 960 nm, 965 nm, 970 nm, 975 nm, 980 nm, 985 nm, 990 nm, 995 nm, or approximately 1000 nm.

[0077] 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 a particular embodiment, the lipid-polymer composite particles contain a Z-mean average 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) can have a range of Z-mean average particle sizes within a population. Therefore, the population may be polydisperse. 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).

[0078] lipid nanoparticles

[0079] The bioactive agents of the present invention can be completely encapsulated in lipid formulations, such as LNPs or other lipid-polymer complex particles. LNPs are extremely useful for systemic application because they exhibit prolonged circulation life after intravenous (iv) injection and accumulate at distant sites (e.g., sites physically separated from the application site). LNPs include “pSPLP”, which comprises encapsulated condenser-nucleic acid complexes as listed in PCT Publication No. WO 2000 / 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 present in the LNPs of the present invention, the bioactive agents are resistant to degradation by nucleases in aqueous solutions. Nucleic acid-lipid particles and methods for their preparation are disclosed, for example, in U.S. Patent Nos. 5,976,567; 5,981,501; 6,534,484; 6,586,410; 6,815,432; U.S. Publication No. 2010 / 0324120 and PCT Publication No. WO96 / 40964.

[0080] In one embodiment, the ratio of lipids to the drug (mass / mass ratio) (e.g., the ratio of lipids to the bioactive agent) will be in the range of about 1:1 to about 50:1, about 1:1 to about 25:1, about 3:1 to about 15:1, about 4:1 to about 10:1, about 5:1 to about 9:1, or about 6:1 to about 9:1. The intermediate ranges above are also considered part of the invention.

[0081] Non-limiting examples of cationic lipids include N,N-dioleoyl-N,N-dimethylammonium chloride (DODAC), N,N-distearate-N,N-dimethylammonium bromide (DDAB), N-(I-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP), N-(I-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA), N,N-dimethyl-2,3-dioleoyloxy)propylamine (DODMA), 1,2-dilinoleoyloxy-N,N-dimethylaminopropane (DLinDMA), and 1,2-dilinoleenoyloxy-N,N-dimethylaminopropane (DLenDMA). DMA), 1,2-dilinoleoylcarbamoyloxy-3-dimethylaminopropane (DLin-C-DAP), 1,2-dilinoleoyloxy-3-(dimethylamino)acetoxypropane (DLin-DAC), 1,2-dilinoleoyloxy-3-morpholinopropane (DLin-MA), 1,2-dilinoleoyl-3-dimethylaminopropane (DLinDAP), 1,2-dilinoleoylthio-3-dimethylaminopropane (DLin-S-DMA), 1-linoleoyl-2-linoleoyloxy-3-dimethylaminopropane (DLin-2-DMAP), 1,2-dilinoleoyloxy-3-trimethylaminopropane chloride (DLin-TMA) .Cl), 1,2-dilinoleoyl-3-trimethylaminopropane chloride (DLin-TAP.Cl), 1,2-dilinoleoyloxy-3-(N-methylpiperazine)propane (DLin-MPZ), or 3-(N,N-dilinoleoylamino)-1,2-propanediol (DLinAP), 3-(N,N-dioleoylamino)-1,2-propanediol (DOAP), 1,2-dilinoleoyloxy-3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), 1,2-dilinoleoyloxy-N,N-dimethylaminopropane (DLinDMA), 2,2-dilinoleoyl-4-dimethylaminomethyl-[1,3] Dioxane (DLin-K-DMA) or its analogues, (3aR,5s,6aS)-N,N-dimethyl-2,2-di((9Z,12Z)-octadecyl-9,12-diynetetrahydro-3aH-cyclopentadieno[d][1,3]dioxin-5-amine (ALN100), (6Z,9Z,28Z,31Z)-heptahepta-6,9,28,31-tetraen-19-yl4-(dimethylamino)butyric acid (MC3), 1,1'-(2-(4-(2-((2-(bis(2-hydroxydodecyl)amino)ethyl)(2-hydroxydodecyl)amino)ethyl)piperazin-1-ylethylazabicyclododecane-2-ol (Tech G1), or mixtures thereof. The cationic lipids may comprise, for example, about 20 mol% to about 50 mol% or about 40 mol% of the total lipids present in the particles.

[0082] Ionizable / non-cationic lipids can be anionic or neutral lipids, including but not limited to distearylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dioleoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoylphosphatidylcholine (POPC), palmitoylphosphatidylethanolamine (POPE), dioleoylphosphatidylethanolamine 4-(N-maleimidemethyl)cyclohexane-1-carboxylic acid ester (DOPE-mal), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), distearylphosphatidylethanolamine (DSPE), 16-O-monomethylPE, 16-O-dimethylPE, 18-1-transPE, 1-stearoyl-2-oleoylphosphatidylethanolamine (SOPE), cholesterol, and mixtures thereof. If cholesterol is included, noncationic lipids may be, for example, about 5 mol% to about 90 mol%, about 10 mol%, or about 60 mol% of the total lipids present in the particle.

[0083] Conjugated lipids that inhibit particle aggregation can be, for example, polyethylene glycol (PEG) lipids, including but not limited to PEG-diacylglycerol (DAG), PEG-dialkoxypropyl (DAA), PEG-phospholipids, PEG-ceramides (Cer), or mixtures thereof. PEG-DAA conjugates can be, for example, PEG-dilauroyloxypropyl (Cer)... 12 ), PEG-dimyristoyloxypropyl (C 14 ), PEG-dipalmitoyloxypropyl (C 16 ) or PEG-distearatepropyl (C 18 Conjugated lipids that prevent particle aggregation may account for, for example, about 0 mol% to about 20 mol% or about 2 mol% of the total lipids present in the particles.

[0084] Lipids can have carbon chains of 4 to 22 in length and neutral, cationic or anionic head groups.

[0085] In some embodiments, the particles also contain, for example, about 10 mol% to about 60 mol% or about 50 mol% of the total lipids present in the particles, sterols (e.g., cholesterol).

[0086] micelles

[0087] Micelles are a special type of molecular assembly in which amphiphilic molecules are arranged in a spherical structure, with all hydrophobic portions of the molecules facing inwards, while the hydrophilic portions are in contact with the surrounding aqueous phase. Micelles can be made of lipids. The micellar phase is caused by the stacking behavior of single-tailed lipids in a bilayer. The inability to fill all the volumes within the bilayer, coupled with the regulation of the area of ​​each head group through hydration of the lipid head groups, leads to the formation of micelles. Such micelles are called normal-phase micelles (oil-in-water micelles). Inverse micelles have a head group at the center, while the tails extend outwards (water-in-oil micelles).

[0088] Micelles are generally spherical. Other shapes, including ellipsoids, cylinders, and bilayers, are also possible. The shape and size of micelles are a function of the molecular geometry of their surfactant molecules and solution conditions such as surfactant concentration, temperature, pH, and ionic strength. The process of micelle formation is called micellization and, according to their polymorphism, is part of the phase behavior of many lipids.

[0089] Phospholipids

[0090] The lipid-polymer composite particles described herein may contain one or more phospholipids. Phospholipids typically consist of two hydrophobic fatty acid tails and a hydrophilic head with a phosphate group. These two components are typically linked together 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 from about 2% to about 20% v / v (e.g., from about 4% to about 18%, from about 5% to about 15%, for example, about 10%).

[0091] Block copolymers

[0092] The lipid-polymer composite particles described herein may comprise block copolymers. A block copolymer is a linear polymer having regions or blocks along its backbone that are characterized by similar hydrophilic, hydrophobic, or chemical properties. Block copolymers may comprise, for example, two, three, four, or more blocks (e.g., diblock or triblock copolymers). Multiblock copolymers comprise multiple blocks.

[0093] Diblock copolymer

[0094] The compositions described herein may comprise diblock copolymers containing two distinct repeating polymer unit blocks. An example of a diblock copolymer as described herein includes amphiphilic copolymers, such as those having a region containing a hydrophilic chain comprising repeating units linked to a region containing a hydrophobic chain comprising repeating units with or without a connector. Such diblock copolymers may comprise hydrophilic chains of polyoxyethylene (PEO) subunits linked to hydrophobic chains of polyoxypropylene (PPO) subunits. A diblock copolymer of PEO and PPO subunits may be represented by the following formula: X1(C2H4O) m -L-(C3H6O) n X2. X1 and X2 can be any chemical motif. L can be a linker that may be present optionally. In some embodiments, the PEO and PPO subunit blocks are directly covalently linked. In some embodiments, X1 and X2 are H and OH, respectively. Other diblock copolymers include, for example, poly(ethylene glycol)-poly(γ-benzyl L-glutamic acid) 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-glutamic acid) PEG-PBLG, poly(ethylene glycol)-poly(β-benzyl L-aspartic acid) PEG-PBLA, poly(ethylene glycol)-poly(α-carboxylic acid benzyl ester-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 structure:

[0095]

[0096] The length of the polymer blocks can be customized. Therefore, 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 degree of natural variation between batches, the above values ​​(and those used herein to characterize a given diblock copolymer) may not be precisely achievable during synthesis, and the average values ​​may vary to some extent. Therefore, unless otherwise explicitly stated, the term "diblock copolymer" as used herein may be used interchangeably with the term "multiple diblock copolymers" (representing entities of several diblock copolymers, also referred to as mixtures of diblock copolymers). As used herein, the term "average," relating to the number of monomer units or molecular weight of one or more diblock copolymers, is a result of the technical impossibility of producing all diblock copolymers having the same composition and therefore the same molecular weight. Diblock copolymers produced according to methods prior art in the field will exist as mixtures of diblock copolymers, each exhibiting variability in its molecular weight, but the mixture as a whole will have the average molecular weight specified herein.

[0097] Polosham

[0098] One example of a triblock copolymer is poloxamer. Poloxamer refers to a nonionic triblock copolymer composed of a central hydrophobic chain of polyoxypropylene flanked by two hydrophilic chains of polyethylene oxide. Poloxamer is also known by trade name... or (BAS F) is known. Block copolymers can be represented by the following formula: HO(C2H4O) x (C3H6O) y (C2H4O) zH. The length of the polymer blocks can be customized. Therefore, many different poloxamers exist. Because the synthesis of block copolymers is related to the degree of natural variation between batches, the numerical values ​​used herein to characterize a given poloxamer may not be precisely achievable during synthesis, and the average value will vary to some extent. Therefore, unless otherwise explicitly stated, the term "poloxam" as used herein may be used interchangeably with the term "multiple poloxamers" (representing entities of several poloxamers, also known as mixtures of poloxamers). As used herein, the term "average," related to the number of monomer units or molecular weight of one or more poloxamers, is a result of the technical impossibility of producing all poloxamers having the same composition and therefore the same molecular weight. Poloxamers produced according to methods prior art in the field will exist as mixtures of poloxamers, each exhibiting variability in its molecular weight, but the mixture as a whole will have the average molecular weight specified herein. Poloxamer suitable for use with the compositions described herein is disclosed in Alexandridis and Bodratti, Journal of Functional Materials 9(1):11(2018), the disclosure of which is incorporated herein by reference in its entirety.

[0099] Poloxamers that can be used in conjunction with the compositions and methods of this disclosure include polyoxypropylene subunits with an average molar mass greater than 2,050 g / mol (e.g., the average molar mass of the polyoxypropylene subunits is about 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). Those poloxamers (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, 3,800 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).

[0100] In some embodiments, the poloxamer has an average molar mass of polyoxypropylene subunits greater than 2,250 g / mol (e.g., the average molar mass of the polyoxypropylene subunits is 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, 400g / mol, 3,500g / mol, 3,600g / mol, 3,700g / mol, 3,800g / mol, 3,900g / mol, 4,000g / mol, 4,100g / mol, 4,200g / mol, 4,300g / mol, 4,400g / mol, 4,500g / mol, 4,600g / mol, 4,700g / mol, 4,800g / mol, 4,900g / mol, or 5,000g / mol).

[0101] In some embodiments, the poloxamer has an average molar mass of polyoxypropylene subunits greater than 2,750 g / mol (e.g., the average molar mass of the polyoxypropylene subunits is 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). mol, 3,700g / mol, 3,800g / mol, 3,900g / mol, 4,000g / mol, 4,100g / mol, 4,200g / mol, 4,300g / mol , 4,400g / mol, 4,500g / mol, 4,600g / mol, 4,700g / mol, 4,800g / mol, 4,900g / mol, or 5,000g / mol).

[0102] In some embodiments, the poloxamer has an average molar mass of polyoxypropylene subunits greater than 3,250 g / mol (e.g., the average molar mass of the polyoxypropylene subunits is 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).

[0103] In some embodiments, the poloxamer has an average molar mass of polyoxypropylene subunits greater than 3,625 g / mol (e.g., the average molar mass of the polyoxypropylene subunits is 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).

[0104] In some embodiments, the poloxamer has an average molar mass of polyoxypropylene subunits of 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, 2,0 ... ,135g / mol, 2,140g / mol, 2,145g / mol, 2,150g / mol, 2,155g / mol, 2,160g / mol, 2,165g / mol, 2,170g / mol, 2,175g / mol, 2,180g / mol, 2,185g / mol, 2,190 g / mol, 2,195g / mol, 2,200g / mol, 2,205g / mol, 2,210g / mol, 2,215g / mol, 2,220g / mol, 2,225g / mol, 2,230g / mol, 2,235g / mol, 2,240g / mol, 2,245g / mo l, 2,250g / mol, 2,255g / mol, 2,260g / mol, 2,265g / mol, 2,270g / mol, 2,275g / mol, 2,280g / mol, 2,285g / mol, 2,290g / mol, 2,295g / mol, 2,300g / mol, 2 ,305g / mol, 2,310g / mol, 2,315g / mol, 2,320g / mol, 2,325g / mol, 2,330g / mol, 2,335g / mol, 2,340g / mol, 2,345g / mol, 2,350g / mol, 2,355g / mol, 2,360 g / mol, 2,365g / mol, 2,370g / mol, 2,375g / mol, 2,380g / mol, 2,385g / mol, 2,390g / mol, 2,395g / mol, 2,400g / mol, 2,405g / mol, 2,410g / mol, 2,415g / mo l, 2,420g / mol, 2,425g / mol, 2,430g / mol, 2,435g / mol, 2,440g / mol, 2,445g / mol, 2,450g / mol, 2,455g / mol, 2,460g / mol, 2,465g / mol, 2,470g / mol, 2,475g / 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、3,840 g / mol, 3,845 g / mol, 3,850 g / mol, 3,855 g / mol, 3,860 g / mol, 3,865 g / mol, 3,870 g / mol, 3,875 g / mol, 3,880 g / mol, 3,885 g / mol, 3,890 g / mol, 3,895 g / mol, 3,900 g / mol, 3,905 g / mol, 3,910 g / mol, 3,915 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).

[0105] In some embodiments, the poloxamer has an average molar mass of polyoxypropylene subunits of 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, 2,0 ... ,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, 2,890 g / 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 / mo l, 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, 3,060 g / 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 / mo l, 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、3,630 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, 3,700 g / mol, 3,705 g / mol, 3,710 g / mol, 3,715 g / mol, 3,720 g / mol, 3,725 g / mol, 3,730 g / mol, 3,735 g / mol, 3,740 g / mol, 3,745 g / mol, 3,750 g / mol, 3,755 g / mol, 3,760 g / mol, 3,765 g / mol, 3,770 g / mol, 3,775 g / mol, 3,780 g / mol, 3,785 g / mol, 3,790 g / mol, 3,795 g / mol, 3,800 g / mol, 3,805 g / mol, 3,810 g / mol, 3,815 g / mol, 3,820 g / mol, 3,825 g / mol, 3,830 g / mol, 3,835 g / mol, 3,840 g / mol, 3,845 g / mol, 3,850 g / mol, 3,855 g / mol, 3,860 g / mol, 3,865 g / mol, 3,870 g / mol, 3,875 g / mol, 3,880 g / mol, 3,885 g / mol, 3,890 g / mol, 3,895 g / mol, 3,900 g / mol, 3,905 g / mol, 3,910 g / mol, 3,915 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). ,

[0106] In some embodiments, the poloxamer has an average molar mass of polyoxypropylene subunits of 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, 3,3 ... ,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, 3,390 g / 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 / mo l, 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, 3,560 g / 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 / mo l, 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, 3,675 g / mol, 3,680 g / mol, 3,685 g / mol, 3,690 g / mol, 3,695 g / mol, 3,700 g / mol, 3,705 g / mol, 3,710 g / mol, 3,715 g / mol, 3,720 g / mol, 3,725 g / mol, 3,730 g / mol, 3,735 g / mol, 3,740 g / mol, 3,745 g / mol, 3,750 g / mol, 3,755 g / mol, 3,760 g / mol, 3,765 g / mol, 3,770 g / mol, 3,775 g / mol, 3,780 g / mol, 3,785 g / mol, 3,790 g / mol, 3,795 g / mol, 3,800 g / mol, 3,805 g / mol, 3,810 g / mol, 3,815 g / mol, 3,820 g / mol, 3,825 g / mol, 3,830 g / mol, 3,835 g / mol, 3,840 g / mol, 3,845 g / mol, 3,850 g / mol, 3,855 g / mol, 3,860 g / mol, 3,865 g / mol, 3,870 g / mol, 3,875 g / mol, 3,880 g / mol, 3,885 g / mol, 3,890 g / mol, 3,895 g / mol, 3,900 g / mol, 3,905 g / mol, 3,910 g / mol, 3,915 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). ,

[0107] In some embodiments, the poloxamer has an average molar mass of polyoxypropylene subunits of about 3,625 g / mol to about 4,000 g / mol (e.g., about 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). l, 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, 3,8 00g / 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, 3,850 g / 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 / m ol, 3,905g / mol, 3,910g / mol, 3,915g / mol, 3,920g / mol, 3,925g / mol, 3,930g / mol, 3,935g / mol, 3,940g / mol, 3,945g / mol, 3,950g / mol , 3,955g / mol, 3,960g / mol, 3,965g / mol, 3,970g / mol, 3,975g / mol, 3,980g / mol, 3,985g / mol, 3,990g / mol, 3,995g / mol, or 4,000g / mol).

[0108] In some embodiments, the poloxamer has an average ethylene oxide content greater than 40% by mass (e.g., 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%, 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).

[0109] In some embodiments, the poloxamer has an average ethylene oxide content greater than 50% by mass (e.g., 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).

[0110] In some embodiments, the poloxamer has an average ethylene oxide content of more than 60% by mass (e.g., 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).

[0111] In some embodiments, the poloxamer has an average ethylene oxide content greater than 70% by mass (e.g., 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).

[0112] In some embodiments, the poloxamer has an average ethylene oxide content of about 40% to about 90% by mass (e.g., 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%).

[0113] In some embodiments, the poloxamer has an average ethylene oxide content of about 50% to about 85% by mass (e.g., 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%).

[0114] In some embodiments, the poloxamer has an average ethylene oxide content of about 60% to about 80% by mass (e.g., about 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%).

[0115] In some embodiments, the poloxamer has an average molar mass greater than 10,000 g / mol (e.g., about 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). ,11,100g / 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, 12,4 00g / 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 / m ol, 13,800g / mol, 13,900g / mol, 14,000g / mol, 14,100g / mol, 14,200g / mol, 14,300g / mol, 14,40 0g / mol, 14,500g / mol, 14,600g / mol, 14,700g / mol, 14,800g / mol, 14,900g / mol, or 15,000g / mol).

[0116] In some embodiments, the poloxamer has an average molar mass greater than 11,000 g / mol (e.g., 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, 12,9 ... 00g / 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, 13,800g / mol, 13,900g / mol, 14, 000g / mol, 14,100g / mol, 14,200g / mol, 14,300g / mol, 14,400g / mol, 14,500g / mol, 14,600g / mol, 14,700g / mol, 14,800g / mol, 14,900g / mol, or 15,000g / mol).

[0117] In some embodiments, the poloxamer has an average molar mass greater than 12,000 g / mol (e.g., 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). 00g / mol, 13,500g / mol, 13,600g / mol, 13,700g / mol, 13,800g / mol, 13,900g / mol, 14,000g / mol, 14,100g / mol, 14,200g / m ol, 14,300g / mol, 14,400g / mol, 14,500g / mol, 14,600g / mol, 14,700g / mol, 14,800g / mol, 14,900g / mol, or 15,000g / mol).

[0118] In some embodiments, the poloxamer has an average molar mass greater than 12,500 g / mol (e.g., about 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). l, 13,700g / mol, 13,800g / mol, 13,900g / mol, 14,000g / mol, 14,100g / mol, 14,200g / mol, 14,300g / mol, 14,400g / mol, 14,500g / mol, 14,600g / mol, 14,700g / mol, 14,800g / mol, 14,900g / mol, or 15,000g / mol).

[0119] In some embodiments, the poloxamer has an average molar mass of about 10,000 g / mol to about 15,000 g / mol (e.g., about 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, etc.). 00g / mol, 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, 13,800g / mol, 13,900g / mol, 14,000g / mol, 14,100g / mol, 14,200g / mol, 14,300g / mol, 14 ,400g / mol, 14,500g / mol, 14,600g / mol, 14,700g / mol, 14,800g / mol, 14,900g / mol, or 15,000g / mol).

[0120] In some embodiments, the poloxamer has an average molar mass of about 11,000 g / mol to about 15,000 g / mol (e.g., about 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, 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, 13,800g / mol, 13,900g / mol, 14,000g / mol, 14,100g / mol, 14,200g / mol, 14,300g / mol, 14,400g / mol, 14,500g / mol, 14,600g / mol, 14,700g / mol, 14,800g / mol, 14,900g / mol, or 15,000g / mol).

[0121] In some embodiments, the poloxamer has an average molar mass of about 11,500 g / mol to about 15,000 g / mol (e.g., about 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). ol, 13,100g / mol, 13,200g / mol, 13,300g / mol, 13,400g / mol, 13,500g / mol, 13,600g / mol, 13,700g / mol, 13,800g / mol, 13,900g / mol, 14,000g / mol , 14,100g / mol, 14,200g / mol, 14,300g / mol, 14,400g / mol, 14,500g / mol, 14,600g / mol, 14,700g / mol, 14,800g / mol, 14,900g / mol, or 15,000g / mol).

[0122] In some embodiments, the poloxamer has an average molar mass of about 12,000 g / mol to about 15,000 g / mol (e.g., about 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, 300g / mol, 13,400g / mol, 13,500g / mol, 13,600g / mol, 13,700g / mol, 13,800g / mol, 13,900g / mol, 14,000g / mol, 14,100g / mol, 14 ,200g / mol, 14,300g / mol, 14,400g / mol, 14,500g / mol, 14,600g / mol, 14,700g / mol, 14,800g / mol, 14,900g / mol, or 15,000g / mol).

[0123] In some embodiments, the poloxamer has an average molar mass of about 12,500 g / mol to about 15,000 g / mol (e.g., about 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). ol, 13,600g / mol, 13,700g / mol, 13,800g / mol, 13,900g / mol, 14,000g / mol, 14,100g / mol, 14,200g / mol, 14,30 0g / mol, 14,400g / mol, 14,500g / mol, 14,600g / mol, 14,700g / mol, 14,800g / mol, 14,900g / mol, or 15,000g / mol).

[0124] Polosham P288, P335, P338 and P407

[0125] Poloxamers that can be used in conjunction with the compositions and methods disclosed herein include those having the approximate chemical formula HO(C2H4O). x (C3H6O) y (C2H4O) z H is “Poloxamer 288” (also known in the art as “P288” and poloxamer “F98”), where the sum of x and y is about 236.36 and z is about 44.83. The average molecular weight of P288 is about 13,000 g / mol.

[0126] In some implementations, poloxamer is a variant of P288, such as HO(C2H4O). x (C3H6O) y (C2H4O) z Variants of H, wherein the sum of x and y is about 220 to about 250, and z is about 40 to about 50. In some embodiments, the average molecular weight of poloxamer is about 12,000 g / mol to about 14,000 g / mol.

[0127] Poloxamer, which can be used in conjunction with the compositions and methods of this disclosure, also includes those having the approximate chemical formula HO(C2H4O). x (C3H6O) y (C2H4O) zH is “Poloxamer 335” (also known in the art as “P335” and poloxamer “P105”), where the sum of x and y is about 73.86 and z is about 56.03. The average molecular weight of P335 is about 6,500 g / mol.

[0128] In some implementations, poloxamer is a variant of P335, such as HO(C2H4O). x (C3H6O) y (C2H4O) z Variants of H, wherein the sum of x and y is about 60 to about 80, and z is about 50 to about 60. In some embodiments, the average molecular weight of poloxamer is about 6,000 g / mol to about 7,000 g / mol.

[0129] Poloxamer, which can be used in conjunction with the compositions and methods of this disclosure, also includes those having the approximate chemical formula HO(C2H4O). x (C3H6O) y (C2H4O) z H is “Poloxamer 338” (also known in the art as “P338” and poloxamer “F108”), where the sum of x and y is about 265.45 and z is about 50.34. The average molecular weight of P335 is about 14,600 g / mol.

[0130] In some implementations, poloxamer is a variant of P338, such as HO(C2H4O). x (C3H6O) y (C2H4O) z Variants of H, wherein the sum of x and y is about 260 to about 270, and z is about 45 to about 55. In some embodiments, the average molecular weight of poloxamer is about 14,000 g / mol to about 15,000 g / mol.

[0131] Poloxamer, which can be used in conjunction with the compositions and methods of this disclosure, also includes those having the approximate chemical formula HO(C2H4O). x (C3H6O) y (C2H4O) z H is “Poloxamer 407” (also known in the art as “P407” and poloxamer “F127”), where the sum of x and y is about 200.45 and z is about 65.17. The average molecular weight is about 12,600 g / mol.

[0132] In some implementations, poloxamer is a variant of P407, such as HO(C2H4O). x (C3H6O) y (C2H4O) zVariants of H, wherein the sum of x and y is about 190 to about 210, and z is about 60 to about 70. In some embodiments, the average molecular weight of poloxamer is about 12,000 g / mol to about 13,000 g / mol.

[0133] For clarity, the terms “mean molar mass” and “mean molecular weight” are used interchangeably herein to refer to the same quantity. As described herein, the mean molar mass, ethylene oxide content, and propylene oxide content of poloxamer can be determined using the methods disclosed in Alexandridis and Hatton, Colloids and Surfaces A: Physicochemical and Engineering Aspects 96:1-46 (1995), the disclosure of which is incorporated herein by reference in its entirety.

[0134] Sterols

[0135] The lipid-polymer composite particles described herein may also contain one or more sterols. Sterols are lipids that are commonly found naturally in plants, animals, and fungi. Phytosterols refer to a class of plant sterol molecules, which are naturally occurring compounds found in plant cell membranes. Phytosterols include phytosterols and steranols. Phytosterols can be derived from any common plant source, such as soybeans, wood, tall oil, vegetable oils, etc. Phytosterols include β-sitosterol, campesterol, stigmasterol, stigmasterol, campesterol, brassosterol, ergosterol, lupeol, cycloartenol, etc. Sterols as described herein can be any cholesterol or its derivatives that alter the fluidity of the lipid layer. Sterols can be naturally occurring sterols, such as those derived from or present in natural sources. Alternatively, sterols can be synthetic sterols, such as non-naturally occurring sterol analogs or derivatives. In some preferred embodiments, the sterol is cholesterol or an analogue thereof (e.g., thiocholesterol, epicholesterol, β-sitosterol (Si-Lip), stigmasterol (St-Lip), or lanosterol (La-Lip)). The concentration of the sterol in the compositions described herein may be, for example, from about 1% to about 50% of the total lipid composition (e.g., from about 5% to about 45%, from about 10% to about 40%).

[0136] Sterols and phospholipids may be present in the particles in, for example, a sterol:phospholipid weight ratio of 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, or about 0.1 to about 0.25. For example, the sterol:phospholipid weight ratio may be about 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.

[0137] Sterols also encompass their esterified derivatives, sometimes referred to as sterol esters or steraneol esters. Sterol esters are sterols esterified with fatty acids, such as long-chain (e.g., C6-C) sterols. 24 For example, C 10 -C 24 For example, C 14 -C 24 Fatty acids, such as caprylic acid, capric acid, undecanoic acid, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid. Sterols and their esters may be fully saturated (e.g., hydrogenated). Pharmaceutical compositions containing sterols or their esters may comprise one or more of the foregoing components or mixtures thereof.

[0138] Bioactive agents

[0139] The lipid-polymer composite particles described herein can encapsulate bioactive agents. Bioactive agents may include therapeutic agents. In some embodiments, the bioactive agent is a terpene, flavonoid, antibiotic, preservative, antifungal agent, antibacterial agent, analgesic, anti-inflammatory agent, antiprotozoal agent, steroid, antiviral agent, lipophilic drug (e.g., with a solubility less than 1 mg / ml), antiVEGF agent, antiglaucoma agent, essential oil, nicotine or nicotine analogues, cyclosporine A, tacrolimus, isotretinoin, propofol, griseofulvin, or any combination thereof.

[0140] In some embodiments, the bioactive agent includes an immunogen (e.g., a vaccine component, such as a DNA vaccine, RNA vaccine, or peptide vaccine). An immunogen can be, for example, a peptide or fragment thereof, a variant thereof, or an analogue thereof, a nucleic acid (e.g., DNA or RNA), or another component of a cell. In some embodiments, the present invention is characterized by a vaccine comprising a composition as described herein.

[0141] In some implementations, 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, immortelle oil, cypress oil, angelica oil, rockrose oil, lime leaf oil, petitgrain oil, bergamot oil, sweet orange oil, palmarosa oil, lemon bark oil, litsea oil, basil oil, marjoram oil, geranium oil, patchouli oil, valerian oil, sandalwood oil, bitter orange oil, and grapefruit oil. Coriander oil, lemongrass oil, black pepper oil, gum oil, juniper twig oil, spearmint oil, Scotch pine oil, rosemary oil, sage oil, ginger oil, lemon oil, citrus oil, cumin oil, juniper oil, lemon balm, myrtle oil, ravensa leaf oil, sweet thyme oil, immortelle oil, manuka oil, dwarf pine oil, oregano oil, vetiver oil, bee pollen oil, white fir oil, cinnamon oil, lemongrass oil, pear berry oil, wintergreen oil, fennel oil, ylang-ylang oil, or combinations thereof.

[0142] In some embodiments, the weight ratio of poloxamer to the 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 is present in amounts from 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%, e.g., about 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%, 0.72%, 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. The concentrations of 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% are present in the formulation. In some embodiments, the bioactive agent (e.g., essential oil) is present in amounts from about 0.01% to about 95% (e.g., about 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%, 0.01%) based on the weight of the composition. 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%, 0.72%, 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%, 8.7%, 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%, 47%, 48%, 4 The formulation is present at concentrations of 9%, 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%.

[0143] The bioactive agents encapsulated in the lipid-polymer composite particles described herein may also be terpenes, flavonoids, antibiotics, preservatives, antifungal agents, antibacterial 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, β-caryophyllene, galactoside, α-pinene, β-pinene, ocimene, terpinene oil, ocimene, terpinene oil, α-terpineol, α-terpinene, γ-terpinene, α-phellandrene, umbelliferone, camphene, δ-3-carene, fentanyl alcohol, 1,8-cineole, nerolidol, borneol, eucalyptol, camphene, or limonene. In some implementations, the flavonoids are selected from the group consisting of: cannabinoid A, cannabinoid B, phenolic acids, stilbenes, phytochemicals, dihydroflavonols, anthocyanins, anthocyanin aglycones, polyphenols, tannins, flavones, flavonols, flavan-3-ols, flavan-4-ols, flavan-3,4-diols, isoflavones, phenylpropanoids, phloroglucinol, coumarins, phenolic acids, benzodiazepines, steroidal glycosides, bioflavonoids, or isoflavones.

[0144] In some implementations, the bioactive agent is a vaccine component, such as a protein or polypeptide fragment that induces immunity, prevents infectious diseases, and / or reduces the risk of infectious diseases.

[0145] In some embodiments, the bioactive compound is nicotine, a nicotine analogue, 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).

[0146] Pharmaceutical Composition

[0147] Preferably, the compositions described herein are formulated into pharmaceutical compositions for administration to human subjects in a biocompatible form suitable for in vivo administration. The pharmaceutical compositions may be formulated using pharmaceutically acceptable carriers or excipients. Pharmaceutically acceptable carriers or excipients are those that do not significantly interfere with the biological activity or efficacy of the active ingredient in the pharmaceutical composition and are not excessively toxic to the host at their concentrations used or administered. Other pharmaceutically acceptable components may also be present in the composition. Suitable substances and their use in formulations of pharmaceutically active compounds are well known in the art (see, for example, 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).

[0148] Pharmaceutical compositions are typically formulated to be compatible with their intended route of administration. For oral administration, formulations can be made by combining a bioactive agent with a pharmaceutically acceptable carrier well known in the art. Such carriers enable the agents of the present invention to be formulated as powders, tablets, pills, capsules, lozenges, liquids, gels, syrups, slurries, suspensions, etc. It has been recognized that, if administered orally, some pharmaceutical compositions must be prevented from being digested. This is typically achieved by conjugating proteins to the composition to make it resistant to acid and enzymatic hydrolysis or by packaging the proteins in a suitable resistant carrier such as liposomes. Excipients suitable for oral dosage forms include, for example, fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, starch, gelatin, tragacanth gum, methylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, and / or polyvinylpyrrolidone (PVP). Disintegrants such as croscarmellose, agar, or alginate or its salts (such as sodium alginate) may be added. Alternatively, oral formulations may be prepared in saline or buffer solutions to neutralize internal acidic conditions, or may be administered without any carrier.

[0149] For inhalation, the pharmaceutical composition can be formulated as an aerosol spray from a pressurized container or dispenser, containing a suitable propellant (e.g., a gas such as carbon dioxide, a fluorocarbon) or in the form of a nebulized inhaler. Liquid or dry aerosols (e.g., dry powder, large porous particles, etc.) can also be used. For topical application, the pharmaceutical composition can be formulated as a suitable ointment, lotion, gel, or cream containing an active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers used in such compositions. Compositions for ocular application can be formulated, for example, with hyaluronic acid.

[0150] As those skilled in the art will understand, the compositions described herein can be administered to a subject in a variety of forms, depending on the chosen route of administration. The compositions described herein can be administered, for example, via any route that allows the composition (e.g., lipid-polymer complex particles, such as micelles or LNPs) to reach target cells. The compositions can be administered, for example, orally, topically, parenterally, intrathecally, intraventricularly, intraparenchymally, buccally, sublingually, nasally, rectally, via patch, pump, transdermally, sublingually, vaginally, ocularly, ocularly, or nasally, and the pharmaceutical compositions are formulated accordingly. The compositions can be administered via inhalation or nebulization. Parenterally administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, transnasal, intrapulmonary, transrectal, and topically administration modalities. Sublingual, buccal, transrectal, transvaginal, ocular, ocular, ocular, or nasal routes are also possible.

[0151] In some embodiments, the compositions described herein are formulated as part of food delivery, for example, they may be administered as food or with a meal.

[0152] In some embodiments, the compositions described herein comprise one or more essential oils, such as tea tree oil, myrrh oil, eucalyptus oil, clove oil, lavender oil, peppermint oil, Roman chamomile oil, German chamomile oil, frankincense oil, immortelle oil, cypress oil, angelica oil, rockrose oil, lime leaf oil, petitgrain oil, bergamot oil, sweet orange oil, palmarosa oil, lemon bark oil, litsea oil, basil oil, marjoram oil, geranium oil, patchouli oil, valerian oil, sandalwood oil, etc. Bitter orange oil, grapefruit oil, coriander oil, lemongrass oil, black pepper oil, gum oil, juniper twig oil, spearmint oil, Scots pine oil, rosemary oil, sage oil, ginger oil, lemon oil, citrus oil, cumin oil, juniper oil, lemon balm, myrtle oil, ravensa leaf oil, sweet thyme oil, immortelle oil, manuka oil, dwarf pine oil, oregano oil, vetiver oil, bee pollen oil, white fir oil, cinnamon oil, lemongrass oil, pearberry oil, wintergreen oil, fennel oil, ylang-ylang oil, or combinations thereof. In some embodiments, the composition comprises multiple essential oils (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 essential oils). In some embodiments, the concentration of essential oil in the composition is from 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%).

[0153] In some embodiments, the compositions described herein are formulated as eye drops. In some embodiments, the eye drops containing one or more essential oils are present in an amount from 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%).

[0154] Generally, the dosage of a pharmaceutical composition (e.g., a bioactive agent) can be from about 1 ng to about 1 g (e.g., from about 1 ng to about 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-100 ng, e.g., 20 ng, 30 ng, 40 ng, 50 ng, 60 ng, 70 ng, 80 ng, 90 ng, 100 ng, e.g., from about 100 ng to about 1 μg, e.g., 200 ng, 300 ng, 400 ng, 500 ng, 600 ng, 700 ng, 800 ng, 900 ng, 1 μg, e.g., from about 1 μg to about 10 μg, e.g., 1 μg, 2 μg, 3 μg, 4 μg, 5 μg, 6 μg, 7 μg, 8 μg, 9 μg, 10 μg, e.g., from about 10 μg to about 100 μg, e.g., 20 μg). 30μg, 40μg, 50μg, 60μg, 70μg, 80μg, 90μg, 100μg, for example, 100μg–1mg, for example, 200μg, 300μg, 400μg, 500μg, 600μg, 700μg, 800μg, 900μg, 1mg, for example, about 1mg to about 10mg, for example, 2mg, 3mg, 4mg, 5mg, 6mg, 7mg, The range is 8mg, 9mg, 10mg, for example, about 10mg to about 100mg, for example, 20mg, 30mg, 40mg, 50mg, 60mg, 70mg, 80mg, 90mg, 100mg, for example, about 100mg to about 1g, for example, 200mg, 300mg, 400mg, 500mg, 600mg, 700mg, 800mg, 900mg, or 1g).

[0155] The dosage of the pharmaceutical composition (e.g., a bioactive agent) may be administered per kg of the subject's body weight. For example, the dosage may be from about 0.01 mg / kg to about 100 mg / kg, for example, from about 0.01 mg / kg to about 30 mg / kg, for example, from about 0.01 mg / kg to about 10 mg / kg, from about 0.1 mg / kg to about 1 mg / kg, for example, from about 0.02 mg / kg, 0.03 mg / kg, 0.03 mg / kg, 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, 0.6 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 daily, weekly, monthly, or annually.

[0156] The dosage of the compositions described herein can vary depending on many factors, such as the pharmacodynamic properties of the agent; the route of administration; the recipient's age, health status, and weight; the nature and severity of symptoms; the frequency of treatment and the type of concurrent treatment (if any); and the clearance rate of the composition in the treated animals. The compositions described herein can be initially administered at a suitable dosage, which may be adjusted as needed based on clinical response. In some embodiments, the dosage of the composition (e.g., a composition containing a bioactive agent) is a preventative or therapeutically effective amount. Furthermore, it should be understood that all doses can be administered continuously or divided into doses administered for each given time interval. For example, the composition can be administered hourly, daily, weekly, monthly, or yearly. In some embodiments, the composition can be administered continuously or systemically.

[0157] The compositions described herein can be prepared using a multi-step process. In some embodiments, the sequence of steps in this process provides optimal particle size, polydispersity, solution transparency, pH, tension, size distribution, stability, and loading of the bioactive agent. In some embodiments, the bioactive agent is homogenized with the polymer in the first step, for example at a temperature of about 50°C to about 70°C, such as about 60°C. In the second step, a solution containing lipids and sterols is added to the homogenized suspension of bioactive agent and polymer using an immersion injection (e.g., ethanol injection).

[0158] How to use

[0159] The compositions described herein are formulated to treat diseases or conditions (e.g., ocular conditions such as inflammation, eye pain, acute conjunctivitis, dark circles, red eyes, bacterial eye infections, fungal eye infections, viral eye infections, swelling, neovascularization, nutritional deficiencies, macular degeneration, glaucoma or elevated intraocular pressure, pain, bacterial infections, fungal infections, protozoan infections, anxiety, agitation, stress, fatigue, insomnia, mental exhaustion, memory loss, organ rejection, eczema, acne, and skin infections). In some embodiments, the compositions are used as nutritional supplements.

[0160] The compositions described herein can be formulated as eye drops to treat eye conditions such as dry eye, inflammation, eye pain, acute conjunctivitis, dark circles, red eyes, bacterial eye infections, fungal eye infections, viral eye infections, nutritional deficiencies, macular degeneration, glaucoma, or elevated intraocular pressure.

[0161] In some embodiments, the composition can be used to treat diseases and symptoms selected from inflammation, pain, bacterial infection, fungal infection, protozoan infection, anxiety, agitation, stress, fatigue, insomnia, mental exhaustion, memory loss, organ rejection, eczema, acne, and skin infections (e.g., athlete's foot, tinea cruris, or tinea pedis).

[0162] In some embodiments, the compositions described herein can 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, such as a DNA vaccine, RNA vaccine, or peptide vaccine). An immunogen may be, for example, a peptide or fragment thereof, a variant thereof, or an analogue thereof, a nucleic acid (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 symptom or to reduce the risk of developing a disease or symptom. The immunogenic composition may be administered, for example, topically, orally, by injection, sublingually, buccally, rectally, vaginally, ocularly, auricularly, nasally, by inhalation, by nebulization, or transdermally. In some embodiments, the immunogenic composition is administered intravenously, subcutaneously, or intramuscularly.

[0163] The composition can be applied to the subject's eye, the area around the subject's eye, the outer area of ​​the eye, the eyelid (e.g., the outer area of ​​the eyelid, the inner area of ​​the eyelid), or the lacrimal duct.

[0164] The composition can be applied as an eye drop (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 drops per day) to each affected eye.

[0165] The composition can be applied once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times, eleven times, twelve times, thirteen times, fourteen times, fifteen times, sixteen times, eighteen times, nineteen times, twenty times, twenty-one times, twenty-two times, twenty-three times, or twenty-four times a day. The composition can also be applied weekly, bi-weekly, monthly, or bi-monthly. The composition can be applied before or after sleep.

[0166] In some embodiments, the composition is administered to the subject topically, orally, by injection, sublingually, buccally, rectally, vaginally, via the eye, via the ear, via the nose, by inhalation, by nebulization, or transdermally.

[0167] Example

[0168] The following examples are provided to provide those skilled in the art with a description of how to use, prepare, and evaluate the compositions and methods described herein, and these examples are intended only as examples of the invention and not to limit the scope of what the inventors consider to be their invention.

[0169] Example 1. Preparation of primary phospholipid liposomes

[0170] Phospholipid liposome compositions were prepared and visually characterized. Three variant formulations, Fa1, Fa2, and Fa3, were prepared using 1,2-distearatel-sn-glycerol-3-phosphocholine (DSPC), cannabidiol (CBD), and optional cholesterol. Fa1 was prepared by mixing DSPC (1 mg / ml), CBD (30 mg / ml), and cholesterol (0.37 mg / ml) in ethanol and then infusing it into dissolved water (Table 1). Fa2 was prepared by mixing DSPC (1 mg / ml) and CBD (30 mg / ml) in ethanol and then infusing it into dissolved water. Fa3 was prepared by mixing DSPC (2 mg / ml), CBD (30 mg / ml), and cholesterol (0.73 mg / ml) in ethanol and then infusing it into dissolved water.

[0171] Table 1. Primary liposome formulations Fa.

[0172]

[0173] Visual characterization of liposome suspensions, Fa1, Fa2, and Fa3 yielded all suspensions with a turbid appearance and some precipitation (Table 2). Qualitative characterization of the relative abundance of liposomes was performed using optical microscopy. Figure 1 Furthermore, it was observed that the process of preparing Fa3 produced the highest concentration of liposomes.

[0174] Table 2. Visual appearance of primary liposome formulation Fa.

[0175] Fa1 Turbid suspension with sediment Fa2 Turbid suspension with sediment Fa3 Turbid suspension with sediment

[0176] Example 2. Effect of temperature on primary phospholipid liposomes

[0177] To overcome the precipitation observed in the Fa formulation experiments, liposome suspensions were prepared under different temperature conditions. The effect of temperature of the aqueous solvent 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 in 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 water using an immersion infusion technique. The resulting suspension had a turbid appearance, with CBD precipitation and non-dispersed lipids. Fb2 was prepared using deionized water at 20°C, followed by air infusion of DSPC (2 mg / ml), CBD (30 mg / ml), and cholesterol (0.73 mg / ml, 15 mol% of DSPC). The resulting suspension also had a turbid appearance, with CBD precipitation 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 with observable CBD precipitation and non-dispersed lipids. Figure 2 )

[0178] Table 3. Water temperature and ethanol injection method for Fb formulations.

[0179] Fb1 Water (20°C), ethanol (4°C), immersion injection Fb2 Air injection, water (20℃) Fb3 Immersion injection, water (20℃)

[0180] Example 3. Effects of CBD concentration and preparation method

[0181] Lipid nanoparticle formulations (Fc1-Fc9, Table 4) were prepared to examine the effects of cannabidiol (CBD) concentration, temperature, and homogenization method on the visual appearance, particle size, and zeta potential of nanoparticles in suspension.

[0182] Table 4. Formulations of CBD encapsulated in lipid nanoparticles prepared at room temperature (20°C) or 60°C, c(Fc).

[0183]

[0184] Two concentrations of CBD (2 mg / ml and 30 mg / ml) were used in the formulations, while the concentration of 1,2-distearate-sn-glycerol-3-phosphocholine (DSPC) was constant at 2 mg / ml for all formulations. Cholesterol concentrations were also kept constant at 0.73 mg / ml (15 mol% of DSPC). Fc1 and Fc4 are formulations for which CBD concentrations were directly compared when preparation conditions were consistent (e.g., temperature and concentrations of DSPC and cholesterol). The effect of temperature was also evaluated. The differences in suspension appearance, precipitation, particle size, and zeta potential at room temperature (20°C) or high temperature (60°C) were examined for Fc1–Fc4 (Table 5 and 10). Figures 3 to 4 Homogenization of the suspensions was performed in formulations Fc5 and Fc8 at 20°C (Fc5) or 60°C (Fc8). Visual inspection of the appearance, sedimentation, particle size, and zeta potential of the suspensions at room temperature (20°C) or high temperature (60°C) (Table 5 and 1). Figures 3 to 4 Formulations Fc6 and Fc9 were used to examine the effects of CBD concentrations (2 mg / ml and 30 mg / ml) on suspension appearance, precipitation, particle size, and nanoparticle zeta potential when the suspension was maintained at 60 °C and CBD was added dropwise (Tables 5 and 6). Figures 3 to 4 Formulation Fc7 is a drug control formulation only.

[0185] Table 5. Visual appearance and presence of precipitates in formulations Fc1-Fc9.

[0186] Fc1 Opaque / turbid suspension with sediment Fc2 Opaque / turbid suspension with sediment Fc3 A slightly turbid suspension that is translucent / milky white / with a small amount of sediment. Fc4 A slightly turbid suspension that is translucent / milky white / with a small amount of sediment. Fc5 A slightly turbid suspension that is translucent / milky white / with a small amount of sediment. Fc6 A slightly turbid suspension that is translucent / milky white / with a small amount of sediment. Fc7 A slightly turbid suspension that is translucent or contains crystalline precipitates. Fc8 A slightly turbid suspension that is translucent / milky white / with a small amount of sediment. Fc9 Turbid suspension with sediment

[0187] The particle size of formulations Fc1-Fc9 was assessed as the average effective diameter (nm) and the average polydispersity with standard deviation limits. Figure 3 The smallest nanoparticles were observed in formulation Fc1. The preparation methods of formulations Fc1 and Fc4 differed only in the concentration of CBD used. The higher CBD concentration resulting in larger particles is likely due to unencapsulated CBD particles consistent with the largest particles observed in formulation Fc7. The preparation method of formulation Fc1 also produced a mixture of smaller lipid nanoparticles and larger unencapsulated CBD particles, reflecting the large polydispersity seen in Fc1. Formulation Fc4 demonstrates the optimal method for a given minimum nanoparticle size and low polydispersity value. Figure 3 ).

[0188] The formulation Fc1-Fc9 was also measured. Figure 4The zeta potential of the nanoparticles was measured. The average zeta potential, in mV, was recorded along with the standard deviation (SD) of three measurements, where each measurement was the average of 10 runs. The zeta potential of formulation Fc4 was -0.05 mV with an SD of 0.09, consistent with the selection of Fc4 as the optimal formulation, as it confirmed that CBD was loaded into the nanoparticles and that the surface charge of the nanoparticles was close to neutral.

[0189] Example 4. Effect of poloxamer on polymer particle composition

[0190] use F127 was used as poloxamer, and the effects of poloxamer concentration on suspension transparency, sedimentation, particle size, suspension pH, and suspension viscosity were investigated. Micellar formulations Fd1-Fd6 were prepared using 0.5 g / 100 ml CBD and homogenized at the lowest speed of a homogenizer for 2 minutes. The concentrations of F127 from formulations Fd1 to Fd6 varied from 1% to 10% w / v, as shown in Table 6.

[0191] Table 6. Those with CBD Formulation of F127 micelles.

[0192] Fd1 1% F127 + 0.5% CBD Homogenize for 2 minutes (speed: lowest). Fd2 2% F127 + 0.5% CBD Homogenize for 2 minutes (speed: lowest). Fd3 3% F127 + 0.5% CBD Homogenize for 2 minutes (speed: lowest). Fd4 4% F127 + 0.5% CBD Homogenize for 2 minutes (speed: lowest). Fd5 5% F127 + 0.5% CBD Homogenize for 2 minutes (speed: lowest). Fd6 10% F127 + 0.5% CBD Homogenize for 2 minutes (speed: lowest).

[0193] Formulations Fd1-Fd6 all have a translucent appearance. Figures 5 to 6 This contrasts with the turbid appearance observed in lipid nanoparticles alone. The observed difference is consistent with the use of... Different amounts of precipitate related to F127 concentration (Table 7).

[0194] Table 7. Observed transparency and presence of sediment in micelle suspensions.

[0195]

[0196]

[0197] High The F127 suspension showed high transparency and minimal sedimentation, indicating that... The effect of F127 on CBD dissolution Figures 5 to 6 The particle size was characterized, and all formulations produced micelles of similar size, less than 50 nm. Figure 7 The pH and viscosity of formulations Fd1-Fd6 were measured and compared with two commercially available eye drops. DUO and ( Figures 8 to 9 (Compare). Except for Fd3, all The F127 formulation has a near-neutral pH. Fd3 shows a lower pH value close to 6.2. Figure 8 For all formulations except one Fd formulation, the viscosity was also lower than that of commercially available eye drops. Fd10 had a significantly higher viscosity, which can be attributed 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 ).

[0198] Example 5. Formulation of lipid-polymer composite particles

[0199] Using concentrations of 1%, 3%, and 5% w / v F127, 0.5% CBD, 2 mg / ml 1,2-distearate-sn-glycerol-3-phosphocholine (DSPC), and 0.73 mg / ml cholesterol were used to prepare lipid-polymer composite particles into formulations Fe1-Fe9 (Table 8). Characterization of the Fe formulations included visual inspection of transparency and precipitation presence after 19 days of storage, particle size and polydispersity, suspension pH, suspension tension, size distribution, and dimensional stability. Formulations Fe1, Fe4, and Fe7 were homogenized with all components at 60°C, and DSPC and cholesterol were added to the suspension mixture via ethanol injection. Formulations Fe2, Fe5, and Fe8 were prepared in two steps. First, [the following steps were performed]. F127 was homogenized with CBD, and then DSPC and cholesterol were added via ethanol injection. Formulations Fe3, Fe6, and Fe9 were also prepared in two separate steps. First, [the following steps were performed]... F127 was homogenized with CBD. Then, the powder mixture of DSPC and cholesterol was added directly at 60°C. In F127-CBD suspensions. Visually inspected, all suspensions were translucent (Table 9 and...). Figure 11 The formulations Fe1, Fe4, and Fe7 have a milky white appearance, and as... As the concentration of F127 increases, the milky white appearance becomes lighter.

[0200] Table 8. Formulation of lipid-polymer composite particles.

[0201]

[0202] along with With increasing F127 concentration, formulations Fe2 and Fe5 also showed reduced precipitation and a milky white appearance. Fe8 appeared transparent and no observable precipitation was observed (Table 9 and). Figure 11 The formulations Fe3, Fe6, and Fe9 also have a milky white appearance. Figure 11Soft / cloudy precipitates were observed in Fe6 and Fe9.

[0203] Table 9. Observed transparency and presence of precipitation in Fe preparations.

[0204]

[0205]

[0206] In formulations Fe1, Fe4, and Fe7, particle size and polydispersity increased with... The concentration increases and then decreases. This trend was also observed in formulations Fe2, Fe5, and Fe8. In formulations Fe3, Fe6, and Fe9, The concentration is independent of particle size. This may be due to reduced solubility caused by directly adding DSPC and cholesterol in powder form. Figure 12 The particle span values ​​for all formulations range from approximately 1.00 to 1.60. Figure 12 The Fe5 formulation exhibited the lowest polydispersity of particles. Generally, the Fe2, Fe5, and Fe8 formulations had the lowest average particle size. All formulations had a pH close to neutral. Figure 13 The tension of the formulation was also measured and compared with a saline control solution, with only... F127 and CBD suspensions and commercially available eye drops 0.15% and DUO comparison ( Figure 14 The size distribution of all formulations was characterized, revealing two particle size populations. Formulations Fe5 and Fe8 showed a reduced abundance of larger particle sizes compared to the smaller particle population. The stability of Fe1–Fe6 was also characterized on the day of preparation and after 19 days of storage at room temperature (20°C) or 4°C. Figure 15 Except for Fe1 and Fe9, all formulations showed stable dimensional stability for 19 days at room temperature (20°C) or 4°C. Visual inspection of all formulations was performed on day 19 from the start of preparation, and all formulations stored at room temperature showed minimal to moderate color change of orange / pink hues. Formulations stored at 4°C showed no observable color or appearance changes from the date of preparation (Table 10).

[0207] Table 10. Color changes in Fe in the formulation.

[0208]

[0209]

[0210] Example 6. Method optimization of the selected formulation

[0211] A series of formulations were further optimized (Table 11). Reproducibles of each formulation for Fd5, Fe5, Fe8, and Fe2 were prepared, and particle size and polydispersity were characterized. Figure 16 Formulations Fd5, Fe5, and Fe8 produced optimal particle size and polydispersity.

[0212] Table 11. Optimized formulations of lipid-polymer composite particles.

[0213]

[0214] Example 7. Effects of F127 and lipid concentration on CBD solubility

[0215] CBD's very low solubility and high lipophilicity in water classify it as a Category 2 drug in the Biomedical Classification System (BCS). This low water solubility poses a challenge to the bioavailability of CBD during drug administration. For fasting individuals, the oral bioavailability of CBD is only about 6%. The aim of this study was to investigate... Increased F127 concentration, along with increased lipid DSPC and cholesterol concentrations, and The effect of the combination of F127, DSPC and cholesterol on CBD solubility.

[0216] F127, 18:0DSPC (1,2-distearyl-sn-glycerol-3-phosphocholine), and cholesterol (lanolin) were purchased from Sigma Aldrich. Cannabidiol (CBD) was supplied by MaxBiotech. Contains concentrations of 1, 3, 5, 7, 9, 11, 13, and 15% w / v. F127 formulation through F127 is prepared by dissolving it in distilled water. To prepare these concentrations... For formulation F127, the formulation was stirred at 34°C on a heated magnetic stirrer. Lipid reserve formulations for ethanol infusion 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.

[0217] method

[0218] CBD saturation solubility

[0219] In order to determine in F127 has a saturated solubility for CBD, as prepared above. Formulations. Add excess CBD to each formulation and stir overnight at room temperature on a magnetic stirrer. Figure 17To understand how the addition of lipids affected saturated solubility, lipids were added to 5% of a solution containing excess CBD using an ethanol infusion method. In F127, a 19G needle was immersed in the stirred formulation, and 8.11% (of the final sample volume) of the lipid stock formulation was injected at a rate of 750 μl / 7 seconds. Each formulation was then stirred overnight as described above. The next day, the formulation was filtered using a 0.45 μm syringe filter, and the amount of dissolved CBD was quantified using high-performance liquid chromatography (HPLC).

[0220] Determine the effect of increasing lipid concentration

[0221] When studying the effects 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 ethanol evaporated from the system.

[0222] Different lipid stock solutions were prepared in ethanol as described above. Excess CBD was added to 20 ml (v1) of 5% [amount missing]. The mixture was stirred in F127 on a magnetic stirrer until the formulation reached a temperature of 45-60°C. While continuing stirring on the hot plate, lipids were added using an ethanol infusion method. In the formulation, a 19G needle was immersed in the stirred 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) x 8.11. To evaporate the ethanol, the formulation was left uncovered and stirred on a hot plate at 45°C for 2 hours, then left uncovered and stirred overnight at room temperature.

[0223] Another method for examining the effects of incorporating higher concentrations of lipids involves incorporating CBD after ethanol infusion. This method is similar to that described in the previous section, except that the ethanol infusion for lipid incorporation is performed before the addition of CBD. After ethanol infusion, the formulation is stirred uncovered at 45°C for 2 hours, then left overnight at room temperature to evaporate the ethanol. The next day, excess CBD is added to the formulation and stirred.

[0224] CBD content was determined by diluting samples in acetonitrile (ACN) as needed and then analyzing them using HPLC. Calibration curves and sample quantification were performed using an Agilent Infinity 1260 system. An Eclipse Plus C18, 4.6 x 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 with a flow rate of 1 mL / min and an injection volume of 20 μL. The UV detector used for HPLC analysis was set to 220 nm. Particle size and polydispersity were determined using a Brookhaven dynamic light scattering system.

[0225] result

[0226] F127 can significantly increase the solubility of CBD in water, which will increase permeability and thus improve the bioavailability of orally administered CBD. In solutions containing 7% to 11% CBD... A peak CBD concentration of approximately 2% w / v was observed in the formulation of F127. Figure 18 After visual characterization of the particles generated in the formulation using TEM, it was observed that at 5%... F127 suspension contains closely packed micelles with a size of 20-30 μm. Figure 19 Two different types of structures were observed in the lipid-only suspension (DSPC:Chol; (0.21:0.19 Mm)): small, diffuse spherical structures and large agglomerate structures. Figure 19 ). At 5% In a suspension of F127+DSPC:Chol (0.21:0.19mM), two distinct characteristics were observed: a close-packed structure with a particle size of 20-30 μm and a single-layer and multi-layer structure in which smaller micelles were encapsulated.

[0227] Characterization of formulations in which CBD is added prior to ethanol injection

[0228] The formulations in which CBD was added prior to ethanol injection were characterized. The visual appearance of the formulations was recorded (Table 12). In the absence of... F127, along with the addition of CBD prior to ethanol injection, and three formulations containing increased lipid concentrations in water, exhibited a turbid appearance.

[0229] Table 12. Visual appearance of formulations with CBD added before ethanol injection

[0230]

[0231] when When F127 was included in the formulation at 5% w / v, the appearance of all three different formulations (Table 12) was turbid with observable precipitation and a pinkish tint. The particle size and polydispersity of the formulations were also characterized. Figure 20 ).No F127 has a larger particle size, while containing In F127 formulations, particle size increases with increasing lipid concentration. Figure 20 (Whether it exists or not) In the case of F127, the pH of the formulation was also measured, and there was no significant difference in pH value. Figure 21 ).

[0232] Characterized the solubility in F127 and none The amount of CBD in the formulation of F127 ( Figure 22 When the formulation contains At F127, there were significant differences in the amount of CBD dissolved. The lipid concentration in the formulation had no effect on the solubility of CBD in pure aqueous solution. Figure 22 ).

[0233] Characterization of formulations in which CBD is added after ethanol injection

[0234] Formulations containing CBD were added after ethanol injection. The visual appearance of the formulations was recorded (Table 13). In the absence of... F127, along with the addition of CBD prior to ethanol injection, and three formulations containing increased lipid concentrations in water, exhibited a turbid appearance.

[0235] Table 13. Visual appearance of formulations with CBD added after ethanol injection

[0236]

[0237] when When F127 was included in the formulations at 5% w / v, all three different formulations appeared cloudy with observable precipitation, but no pink color was observed when ethanol infusion was performed after the addition of CBD. The particle size and polydispersity of the formulations were also characterized. Figure 23 ).No F127 has a larger particle size, while containing In F127 formulations, particle size increases with increasing lipid concentration. Figure 23 (Whether it exists or not) In the case of F127, the pH of the formulation was also measured, and there was no significant difference in pH value. Figure 24 ).

[0238] Characterized the solubility in F127 and none The amount of CBD in the formulation of F127 ( Figure 25 When the formulation contains At F127, there were significant differences in the amount of CBD dissolved. The lipid concentration in the formulation had no effect on the solubility of CBD in pure aqueous solution. Figure 25 ).

[0239] discuss

[0240] As shown in this experiment, F127 can significantly improve the solubility of CBD in water. TEM images show that in water containing CBD... Incorporating lipids into formulations creates monolayer and multilayer liposomes, which appear to contain micelles. These hybrid systems translate to enhanced absorption of orally administered CBD. While increasing lipid concentration may lead to increased concentrations of these multi-particulate systems, compared to... Compared to its counterpart, CBD solubility was slightly improved. However, the mixed particles containing lipid-coated micelles function by shielding the micelles within them, thus maintaining micellar integrity when the formulation is diluted (as may happen at administration). Another interesting observation with DSPC:Chol at a concentration of 1:64:1.51 was the formation of significantly larger (twice the size) particles using a method of injecting ethanol after CBD incorporation into the system. This may mean that this sequence of CBD incorporation and ethanol injection results in greater micellar retention within the liposomes and / or greater drug retention within the mixed composite particles.

[0241] Other implementation plans

[0242] Although the invention has been described in conjunction with its specific embodiments, it should be understood that further modifications can be made to it, and this application is intended to cover any variations, uses, or adaptations of the invention based on the principles thereof, including known or conventional practices that, while not part of the invention, fall within the scope of the invention, and changes that fall within the essential features described above and are within the scope of the claims. Other embodiments are defined in the claims.

Claims

1. A composition comprising a plurality of lipid-polymer composite particles encapsulating a hydrophobic bioactive agent, wherein the lipid-polymer composite particles are prepared by a method comprising the steps of: (a) Homogenize the bioactive agent with poloxamer 407 to prepare a homogeneous solution; and after step (a) (b) Cholesterol and lipids selected from neutral lipids, cationic lipids and anionic lipids are injected into the homogeneous solution to prepare multiple lipid-polymer composite particles encapsulating bioactive agents; The plurality of lipid-polymer composite particles have an average particle size between 10 and 1000 nanometers, the lipid-polymer composite particles comprise a single-layer or multi-layer structure encapsulating one or more micelles, the weight ratio of poloxamer 407 to the bioactive agent is between 2 and 15, and the composition does not contain an organic solvent for dissolving the bioactive agent.

2. The composition of claim 1, wherein the bioactive agent is a therapeutic agent.

3. The composition of claim 1, wherein the bioactive agent is cannabidiol.

4. The composition according to any one of claims 1-3, wherein the composition is formulated into an eye drop formulation.

5. The composition of any one of claims 1-3, wherein the lipid comprises a carbon chain of length 4 to 22 and a neutral, cationic or anionic head group.

6. The composition of claim 5, wherein the lipid is phosphatidylcholine, phosphatidylserine, phosphatidylglycerol, phosphatidylethanolamine, or phosphatidylinositol.

7. The composition according to any one of claims 1-3, wherein the concentration of the lipid is from 0.1 mol% to 10 mol%.

8. The composition according to any one of claims 1-3, wherein the concentration of cholesterol is from 5 mol% to 50 mol% of the total lipid composition.

9. The composition of any one of claims 1-3, wherein the weight ratio of cholesterol to lipid is 0.01 to 0.

50.

10. Use of the composition of any one of claims 1-9 in the preparation of a medicament for providing a bioactive agent to a subject.

11. The use as described in claim 10, wherein the bioactive agent is a therapeutic agent.

12. The use as described in claim 11, wherein the bioactive agent is cannabidiol.

13. The use as described in any one of claims 10-12, wherein the composition is formulated for topical application.

14. The use according to any one of claims 10-12, wherein the composition is formulated for administration via oral, sublingual, buccal, rectal, vaginal, ocular, auditory, or nasal route.

15. The use as described in any one of claims 10-12, wherein the composition is formulated for administration by injection, inhalation, or transdermal application.

16. The use as described in any one of claims 10-12, wherein the composition is formulated for atomized application.

17. A method for preparing a composition comprising a plurality of encapsulated hydrophobic bioactive agents in lipid-polymer composite particles, the method comprising: (a) Homogenize the bioactive agent with poloxamer 407 to prepare a homogeneous solution; And after step (a) (b) Cholesterol and lipids selected from neutral lipids, cationic lipids and anionic lipids are injected into the homogenized solution. This allows for the preparation of multiple lipid-polymer composite particles encapsulating the bioactive agent, wherein the multiple lipid-polymer composite particles comprise a single-layer or multi-layer structure encapsulating one or more micelles, wherein the multiple lipid-polymer composite particles have an average particle size between 10 and 1000 nanometers, wherein the weight ratio of poloxamer 407 to the bioactive agent is between 2 and 15, and wherein the composition does not contain an organic solvent for dissolving the bioactive agent.

18. The method of claim 17, wherein the bioactive agent is a therapeutic agent.

19. The method of claim 17 or 18, wherein the bioactive agent is cannabidiol.

20. The method of claim 17 or 18, wherein the lipid comprises a carbon chain of length 4 to 22 and a neutral, cationic or anionic head group.

21. The method of claim 17 or 18, wherein the lipid is phosphatidylcholine, phosphatidylserine, phosphatidylglycerol, phosphatidylethanolamine, or phosphatidylinositol.

22. The method of claim 17 or 18, wherein the concentration of the lipid is from 0.1 mol% to 10 mol%.

23. The method of claim 17 or 18, wherein the concentration of cholesterol is from 5 mol% to 50 mol% of the total lipid composition.

24. The method of claim 17 or 18, wherein the weight ratio of cholesterol to lipid is 0.01 to 0.

50.

25. The method of claim 17 or 18, wherein step (b) comprises immersion injection of cholesterol and lipids.

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

27. The method of claim 17 or 18, wherein step (a) is performed at 50°C to 70°C.

28. A composition prepared by the method of any one of claims 17-27.

29. Use of the composition of any one of claims 1-9 in the preparation of a medicament for treating dry eye, inflammation, eye pain, acute conjunctivitis (pink eye), dark circles, red eyes, bacterial eye infections, fungal eye infections, viral eye infections, swelling, neovascularization, nutritional deficiencies, macular degeneration, glaucoma, or elevated intraocular pressure.

30. Use of the composition of any one of claims 1-9 in the preparation of a medicament for treating diseases and symptoms 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.

31. The use as described in claim 29 or 30, wherein the composition is formulated into an eye drop formulation.

32. An immunogenic composition comprising the composition as described in any one of claims 1-9.

Images