Nanoparticle compositions for oxygen delivery

Abstract

The present disclosure relates, in some aspects, generally to nanoparticle compositions for oxygen delivery, methods of using the same, and methods of making the same. The present disclosure relates, in some aspects, generally to topical formulations capable of delivering oxygen to tissue and methods for their use in the treatment of diseases, such as hypoxemia.

Description

[0001] Cross-references to related applications

[0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 462,207, filed April 26, 2023, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] This disclosure relates in several aspects to nanoparticle compositions for oxygen delivery, methods of using them, and methods of preparation thereof. In other aspects, this disclosure relates in several aspects to formulations capable of delivering oxygen to tissues and methods for treating diseases such as hypoxemia. Background Technology

[0004] Acute respiratory distress syndrome (ARDS) is a life-threatening condition characterized by the failure of oxygen and CO2 transalveolar-capillary membrane movement (gas exchange). Damage caused by ARDS in the lungs leads to the formation of fluid within the alveolar units, hindering the transport of oxygen and carbon dioxide. This fluid accumulation impedes the rate of oxygen and carbon dioxide exchange at the alveolar epithelial cells and capillaries, resulting in a lack of oxygenated blood in the system.

[0005] ARDS has a high incidence and often leads to long-term health complications. Traumatic injury, direct exposure to chemical and / or biological warfare agents, or respiratory complications from worldwide respiratory viral diseases pose a serious threat to surgical success, especially in resource-constrained settings. Clinical management of ARDS is supportive and involves the use of supportive measures to correct critical hypoxemia, such as mechanical ventilation to treat respiratory failure, systemic corticosteroids to reduce inflammation, and, where feasible, extracorporeal life support (ECLS) to deliver O2 and remove CO2 directly from the bloodstream. While these supportive measures exist, mechanical ventilation can cause cellular trauma as alveoli are stretched and deformed under positive pressure, resulting in further lung injury; corticosteroids may reduce inflammation, but the underlying inflammatory processes that contribute to ARDS remain; and most hospitals do not offer ECLS services. Therefore, there is an urgent need for novel or improved treatment options for ARDS-related refractory hypoxemia and hypercapnia, especially when the lungs are no longer able to effectively facilitate normal O2 and CO2 transport.

[0006] To develop more effective treatment options for hypoxemia, oxygen delivery compositions, particularly those containing perfluorocarbons (PFCs), have been developed. PFCs are a class of molecules primarily composed of fluorine and carbon. They are characterized by high gas solubility, rapid release, high volumetric mass, moderate volatility, good tissue compatibility, and non-absorption and metabolism in vivo. Due to their ability to dissolve large amounts of physiologically important gases (primarily oxygen and carbon dioxide), numerous artificial oxygen carriers based on PFC emulsions have been developed over the past century. However, there remains a need to develop more effective, stable, and safer PFC-based compositions for efficient O2 delivery and the treatment of hypoxemia. This application addresses this need, along with other requirements. Summary of the Invention

[0007] In some aspects, this document provides a nanoparticle composition for oxygen delivery comprising nanoparticles, wherein the nanoparticles comprise a perfluorocarbon component and a lipid component, wherein the perfluorocarbon component comprises perfluoronaphthene (PFD) and perfluorotripropylamine (PFTPA), and optionally wherein the nanoparticle composition comprises no more than about 2% (w / w) of a polymeric surfactant in the total nanoparticle composition.

[0008] In some embodiments, the perfluorocarbon component comprises (e.g., the perfluorocarbon component is) a mixture of perfluoronaphthylene (PFD) and perfluorotripropylamine (PFTPA) optionally in a weight ratio of about 2:1 to about 3:1. In some embodiments, the lipid component comprises (e.g., the lipid component is) a mixture of DSPC and DPPC optionally in a weight ratio of about 1:1 to about 4:1. In some embodiments, the lipid component comprises no more than about 2% (w / w) of hydrogenated soybean phosphatidylcholine (HSPC) of the total nanoparticle composition. In some embodiments, the nanoparticle composition further comprises a buffer component. In some embodiments, the nanoparticle composition comprises a buffer comprising a buffer component and water. In some embodiments, the buffer component comprises sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is about 5:1 to about 20:1, optionally 12:1. In some embodiments, the weight ratio of the perfluorocarbon component to the buffer component is about 3.5:1 to about 4.2:1, such as about 3.8:1. In some embodiments, the buffer comprises about 20 mM to about 100 mM citrate, optionally at a weight percentage of about 40% to about 60% (w / w) of the total nanoparticle composition. In some embodiments, the nanoparticle composition comprises no more than about 2% (w / w) of phosphate buffer of the total nanoparticle composition. In some embodiments, the perfluorocarbon component comprises about 40% to about 60% (w / w) of the total nanoparticle composition. In some embodiments, the lipid component comprises about 1.5% to about 3.5% (w / w) of the total nanoparticle composition. In some embodiments, the nanoparticle composition has a pH of about 3.5 to about 8.0. In some embodiments, the nanoparticle composition has a viscosity of no more than about 100 cp. In some embodiments, the average particle size of the nanoparticles is no more than about 3 μm. In some embodiments, when diluted with 5 × (v / v) citrate buffer, the nanoparticles have an average particle size of no more than about 500 nm. In some embodiments, the nanoparticles have a multi-lamellar structure comprising at least two lamellar layers, wherein each lamellar layer includes perfluorocarbon sandwiched between two lipid layers. In some embodiments, the nanoparticle composition is enriched with at least about 5 ppm of dissolved oxygen. In some embodiments, the nanoparticle composition is stable at room temperature for at least 4 days without phase separation.

[0009] In some embodiments, the nanoparticle composition comprises no more than about 2% (w / w) of centrimonium bromide (CTAB) of the total nanoparticle composition. In some embodiments, the nanoparticle composition comprises no more than about 2% (w / w) of Tween 80 of the total nanoparticle composition. In some embodiments, the nanoparticle composition comprises no more than about 2% (w / w) of soybean lecithin of the total nanoparticle composition. In some embodiments, the nanoparticle composition comprises no more than about 2% (w / w) of poloxamer of the total nanoparticle composition. In some embodiments, the nanoparticle composition comprises no more than about 2% (w / w) of surfactant or lipids other than DSPC and DPPC. In some embodiments, the nanoparticle composition is suitable for inhalation, topical application, or enema administration.

[0010] In some embodiments, the nanoparticle composition comprises (i) PFD, which is about 35.5% by weight of the total nanoparticle composition; (ii) PFTPA, which is about 14.5% by weight of the total nanoparticle composition; (iii) DPPC, which is about 0.6% by weight of the total nanoparticle composition; (iv) DSPC, which is about 1.7% by weight of the total nanoparticle composition; and (v) a citrate buffer (50 mM) at pH 6.0, which is about 47.7% by weight of the total nanoparticle composition.

[0011] In some embodiments, the composition comprises (i) PFD, which is about 35.5% by weight of the total nanoparticle composition; (ii) PFTPA, which is about 14.5% by weight of the total nanoparticle composition; (iii) DPPC, which is about 0.6% by weight of the total nanoparticle composition; (iv) DSPC, which is about 1.7% by weight of the total nanoparticle composition; (v) citric acid, which is about 0.1% by weight of the total nanoparticle composition; (vi) sodium citrate dihydrate, which is about 1.2% by weight of the total nanoparticle composition; and (viii) water, which is about 46.4% by weight of the total nanoparticle composition.

[0012] In some aspects, this document provides a wound healing composition comprising the nanoparticle composition described herein, silver dioxide, and collagen. In some aspects, this document provides an enema gel comprising the nanoparticle composition described herein, poloxamer, and a PBS solution. In some aspects, this document provides a cream composition comprising the nanoparticle composition described herein and pharmaceutically acceptable excipients.

[0013] In some aspects, this document provides a method for delivering to a subject in need tissue experiencing a reduction in normal oxygen levels (as determined by comparison with a healthy state, as measured by techniques known in the art), comprising applying to the subject a nanoparticle composition described herein, a wound healing composition described herein, an enema gel described herein, or a cream composition described herein. In some embodiments, the tissue is in the skin, in the colon or gastrointestinal tract, or in the lungs. In some embodiments, this document provides a method for treating a condition in a subject in need, comprising applying to the subject topically a nanoparticle composition described herein, a wound healing composition described herein, an enema gel described herein, or a cream composition described herein. In some embodiments, the condition is a wound, open injury, burn, colonic or gastrointestinal condition, or lung condition. In some embodiments, the lung condition is COVID, chronic obstructive pulmonary disease (COPD), asthma, or lung injury. In some embodiments, the condition is ARDS. In some embodiments, ARDS is caused by a bacterial infection. Attached Figure Description

[0014] The accompanying drawings illustrate some embodiments of the features and advantages of this disclosure. These embodiments are not intended to limit the scope of the appended claims in any way.

[0015] Figure 1 shows an exemplary scheme of a gas exchange process achieved through nanoparticles in a nanoparticle composition.

[0016] Figure 2 shows a microscopic image of the nanoparticles in the nanoparticle composition.

[0017] Figure 3A shows a microscopic image of the nanoparticles in the nanoparticle composition NF-62. Figure 3B shows a microscopic image of the nanoparticles in the nanoparticle composition NF-66.

[0018] Figure 4 shows the low-energy method (high-pressure homogenization / ultrasonic treatment) and the high-energy method (microfluidization) process used to prepare nanoparticle compositions.

[0019] Figure 5A shows a microscopic image of lung tissue from mice with ARDS treated with the nanoparticle composition NF-83. Figure 5B shows a microscopic image of untreated lung tissue from mice with ARDS.

[0020] Figure 6 shows microscopic images of lung tissue from mice suffering from ARDS induced by bacterial infection. The top image shows lung tissue from untreated and normal mice. The bottom left image shows lung tissue from untreated and diseased mice. The bottom right image shows an image of lung tissue from diseased mice treated with the nanoparticle composition NF-83.

[0021] Figure 7A shows the trajectory of nanoparticles in the nanoparticle composition NF-83 through the microchannel. Figures 7B and 7C show the trajectories of individual nanoparticles in the nanoparticle composition NF-83 during 0 to 10 seconds and 0 to 30 seconds, respectively.

[0022] Figure 8 shows a sample image of the nanoparticles.

[0023] Figure 9A shows the level of CFTR1 expression in the lung tissue of mice treated with LPS. The saline group served as a control, and the formulation group was administered the nanoparticle composition NF-83. Figure 9B shows the level of TRPV1 expression in the lung tissue of mice treated with LPS. The saline group served as a control, and the formulation group was administered the nanoparticle composition NF-83. Figures 9C and 9D show fluorescence (representing COX-PTGS2 expression levels) in the lung tissue of mice treated with LPS. The control group refers to untreated and normal mice. The untreated group refers to untreated and diseased mice. The treated group refers to diseased mice treated with the nanoparticle composition NF-83.

[0024] Figure 10 shows the conformational changes of collagen induced by the exemplary nanoparticle composition.

[0025] Figure 11 shows an image of lung tissue with a distribution of the nanoparticle composition stained with Texas Red.

[0026] Figure 12 shows the quenching of fluorescein (FITC) using an exemplary nanoparticle composition. Detailed Implementation

[0027] The following description is presented to enable those skilled in the art to make and use various embodiments. The description of specific apparatuses, techniques, and applications is provided only as examples. Various modifications to the embodiments described herein will be apparent to those skilled in the art, and the general principles defined herein can be applied to other embodiments and applications without departing from the spirit and scope of the various embodiments. Therefore, the various embodiments are not intended to be limited to the embodiments described and shown herein, but are accorded the scope consistent with the claims.

[0028] I. Definition

[0029] As used in this specification, the following words and phrases are generally intended to have the meanings described below, unless the context in which they are used indicates otherwise.

[0030] As used herein and in the appended claims, unless the context clearly indicates otherwise, the singular forms “a”, “an” and “the” include the plural forms.

[0031] As used herein, and unless otherwise specified, the terms “about” and “approximately” when used in conjunction with a dose, amount, or weight percentage of an ingredient in a composition or dosage form mean a dose, amount, or weight percentage recognized by one of ordinary skill in the art to provide a pharmacological effect equivalent to that obtained from a specified dose, amount, or weight percentage. Specifically, where applicable, the terms “about” and “approximately” when used in the context herein contemplate a dose, amount, or weight percentage within 15% of a specified dose, amount, or weight percentage.

[0032] The terms “patient,” “subject,” “individual,” etc., are used interchangeably herein and refer to any animal, in some embodiments a mammal, and in some embodiments a human, including those requiring treatment or susceptible to a disease or its sequelae. An individual may include, for example, dogs, cats, pigs, cows, sheep, goats, horses, rats, rabbits, hamsters, guinea pigs, monkeys, mice, and humans. In some embodiments, the individual is a human. In some embodiments, the individual is an animal.

[0033] The term "treatment" generally refers to a clinical intervention aimed at altering the natural processes of the treated individual or cells during the course of a clinical lesion. Desired therapeutic effects include slowing the rate of disease progression, modifying or alleviating the disease state, and mitigating or improving prognosis. For example, an individual is successfully "treated" if one or more symptoms associated with a disease or disorder are reduced or eliminated, including but not limited to a decrease in the frequency and / or severity of signs and / or symptoms caused by the disease, an improvement in the quality of life of individuals with the disease, a reduction in the dosage of other medications required to treat the disease, and / or an extension of the individual's survival. Treatment can be preventative (to prevent or delay the onset of the disease, or to prevent the manifestation of its clinical or subclinical symptoms), or therapeutically suppress or alleviate symptoms after the disease has manifested.

[0034] The term "therapeutic effective amount" or "effective amount" refers to the amount of a compound or preparation described herein that, when administered to a patient requiring such treatment, is sufficient to affect the treatment as defined herein. A therapeutically effective amount of a composition or preparation may be an amount sufficient to treat a hypoxia-related disease. Therapeuticly effective amounts will vary depending on factors such as the subject and the disease condition being treated, the subject's weight and age, the severity of the disease condition, the specific compound, the dosing regimen to be followed, the time of administration, and the method of administration, all of which can be readily determined by one of ordinary skill in the art. Therapeuticly effective amounts can be determined experimentally, for example, by measuring the blood concentration of the chemical entity, or theoretically by calculating bioavailability.

[0035] In some embodiments, as used herein, "polymeric" refers to structural features similar to those of a polymer. "Polymer" generally refers to very large molecules, such as macromolecules, that contain multiple repeating subunits. In some embodiments, a polymer refers to a molecule having one or more repeating units and having a molecular weight of at least about 2,500 Daltons, at least about 3,000 Daltons, at least about 4,000 Daltons, or at least about 5,000 Daltons. In some embodiments, the polymer does not include lipids. In some embodiments, the polymer surfactant does not include lipid surfactants.

[0036] In some embodiments, this disclosure also includes any or all stereochemical forms of the compounds provided herein (e.g., perfluorocarbons and lipids), including any enantiomers or diastereomers of the described compounds, as well as any tautomers or other forms. In some embodiments, unless the stereochemistry is explicitly indicated in the chemical structure or name, the structure or name is intended to include all possible stereoisomers of the depicted compound. Compositions containing mixtures of the compounds described herein in any ratio are also included in this disclosure as mixtures of two or more stereochemical forms of the compounds in any ratio, thereby including racemic, non-racemic, enantiomer-rich, and proportionate mixtures of the compounds. N-oxides are also provided and described in the presence of one or more tertiary amine moieties in the compound.

[0037] In some embodiments, this disclosure also includes additional isotopic labeling and / or isotopic enrichment forms of the compounds described herein (e.g., perfluorocarbons and lipids). The compounds described herein may contain atomic isotopes other than deuterium in non-natural proportions at one or more of the atoms constituting such compounds. Exemplary additional isotopes that may be incorporated into the compounds described herein include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, and chlorine, such as... 2 H, 3 H, 11 C 13 C 14 C 13 N、 15 O、 17 O、 35 S, 18 F, 36 Cl. As used herein, each case of enriching, substituting, or replacing an atom with its corresponding isotope covers approximately one of the following isotope enrichment levels: 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.6%, 99.7%, 99.8%, 99.9%, or 100%, or a range between any two of the aforementioned percentages.

[0038] As used herein, the term “substantially free of” means that the composition contains not more than 15%, not more than 10%, not more than 5%, not more than 4%, not more than 3%, not more than 2% or not more than 1% by weight of one or more of the specified substances.

[0039] It should be understood that the implementation schemes described herein as "comprising" include implementation schemes that are "composed of" and "substantially composed of".

[0040] II. Nanoparticle Composition

[0041] In one aspect, this document provides a nanoparticle (e.g., nanoemulsion) composition for oxygen delivery comprising nanoparticles, wherein the nanoparticles comprise a perfluorocarbon component and a lipid component. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition comprises no more than about 2% (w / w) of a polymeric surfactant. For example, in some embodiments, the nanoparticle composition is substantially free of (e.g., completely free of) polymeric surfactants. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition comprises nanoparticles comprising perfluoronaphthyl (PFD), perfluorotripropylamine (PFTPA), DSPC, DPPC, and a buffering agent, wherein the nanoparticle (e.g., nanoemulsion) composition is substantially free of surfactants other than lipids, and the nanoparticle (e.g., nanoemulsion) composition is stable at room temperature (e.g., does not undergo phase separation) for at least four days. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition comprises nanoparticles, wherein the nanoparticles comprise perfluoronaphthylene (PFD), perfluorotripropylamine (PFTPA), DSPC, DPPC, and the nanoparticle composition comprises a buffer component, optionally wherein the buffer component comprises sodium citrate hydrate and citric acid. In some embodiments, the buffer component comprises sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1, such as any one of about 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, or any value between the foregoing values. In some embodiments, the weight ratio of PFD to PFTPA is from about 2:1 to about 3:1. In some embodiments, the weight ratio of DSPC to DPPC is from about 1:1 to about 4:1. In some embodiments, the perfluorocarbon component comprises about 40% to about 60% (w / w) of the total nanoparticle (e.g., nanoemulsion) composition, and the lipid component comprises about 1.5% to about 3.5% (w / w) of the total nanoparticle (e.g., nanoemulsion) composition. In some embodiments, the buffer comprises about 50 mM citrate, optionally at a weight percentage of about 40% to about 60% (w / w) of the total nanoparticle (e.g., nanoemulsion) composition. In some embodiments, the weight ratio of the perfluorocarbon component to the buffer component in the nanoparticle composition is about 2.5:1 to about 5, for example, about 3:1 to about 4.5:1, about 3.5:1 to about 4.5:1, or about 3.8:1. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition has a pH of about 5.0 to about 8.0, such as about 6.0 to about 7.0. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition has a viscosity of not more than about 100 cp.In some embodiments, when diluted with 5 × (v / v) citrate buffer, the nanoparticles have an average particle size of not more than about 500 nm, such as about 200 nm to about 500 nm. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is enriched with at least about 5 ppm of dissolved oxygen. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is stable at room temperature for at least 4 days, such as at least 7 days or 10 days without phase separation. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is substantially free of hydrogenated soybean phosphatidylcholine (HSPC), cetrimonium bromide (CTAB), Tween 80, soybean lecithin, or other surfactants or lipids other than DSPC and DPPC. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is suitable for topical application, such as inhalation.

[0042] In some embodiments, the nanoparticle (e.g., nanoemulsion) compositions provided herein are effective for oxygen delivery and offer at least several key advantages over known perfluorocarbon emulsions. First, known perfluorocarbon emulsions exhibit poor stability, especially at room temperature. For example, although perfluorocarbon concentrations are low, artificial blood fluorosols can only be stored frozen, severely hindering their use, storage, and transport. In contrast, the nanoparticle (e.g., nanoemulsion) compositions provided herein are stable at room temperature (e.g., without phase separation) for at least one day. Furthermore, to improve stability, current perfluorocarbon emulsions typically contain polymeric surfactants, such as poloxamer, which can induce complement activation. The nanoparticle compositions provided herein are essentially free of polymeric surfactants, thus being safer and less irritating. Additionally, known perfluorocarbon emulsions are typically administered intravenously, which increases operational complexity and cost. Therefore, the nanoparticle (e.g., nanoemulsion) compositions described herein are crucial because they can effectively deliver O2 while remaining stable and non-toxic, and can be administered in a non-invasive and convenient manner, such as topical application.

[0043] Perfluorocarbon

[0044] In some embodiments, the perfluorocarbon component includes a perfluoroalkyl group. In some embodiments, the perfluoroalkyl group is an alkyl group in which all hydrogens are independently substituted by F or Br. In some embodiments, the alkyl group is C4-C. 20 In some embodiments, the alkyl group is cyclic, acyclic, branched, or unbranched. In some embodiments, the perfluorocarbon component comprises a trifluoroalkylamine, wherein the perfluoroalkyl group is cyclic, acyclic, branched, or unbranched. In some embodiments, the perfluoroalkyl group is C1-C. 15In some embodiments, the trifluoroalkylamine may be protonated. In some embodiments, the perfluorocarbon component includes perfluorobutane, perfluoropentane, perfluorotert-butylcyclohexane, perfluoro-2-methylhexane, perfluoronaphthane (PFD), perfluorotripropylamine (PFTPA), perfluorotributylamine, perfluorotripentylamine, perfluorotrihexylamine, perfluoro-N-(4-methylcyclohexyl)piperidine, perfluoro-N-methylpiperidine, perfluoro-dicyclohexyl-methylamine, perfluorooctyl bromide, perfluorodecyl bromide, or any combination thereof. In some embodiments, the perfluorocarbon component includes perfluoronaphthane (PFD), perfluorotripropylamine (PFTPA), perfluorotributylamine, perfluorotripentylamine, perfluorobutane, or any combination thereof. In some embodiments, the perfluorocarbon component includes PFD. In some embodiments, the perfluorocarbon component includes PFTPA. In some embodiments, the perfluorocarbon component includes both PFD and PFTPA. In some embodiments, the perfluorocarbon component is substantially composed of PFD and PFTPA. In some embodiments, the perfluorocarbon component includes no more than about 2% (w / w), for example, no more than any one of about 1.5% (w / w), 1.0% (w / w), 0.5% (w / w), 0.1% (w / w), 0.01% (w / w), or 0.001% (w / w) of perfluorocarbon other than PFD and PFTPA. In some embodiments, the perfluorocarbon component is composed of PFD and PFTPA. In some embodiments, the weight ratio of PFD to PFTPA in the nanoparticle (e.g., nanoemulsion) composition is about 2:1 to about 3:1, for example, any one of about 2:1 to about 2.8:1, about 2:1 to about 2.6:1, about 2.2:1 to about 2.6:1, or about 2.4:1.

[0045] In some embodiments, the weight percentage of the perfluorocarbon component in the total nanoparticle (e.g., nanoemulsion) composition is at least about 20% (w / w), for example, at least about 24% (w / w), 26% (w / w), 28% (w / w), 30% (w / w), 32% (w / w), 34% (w / w), 36% (w / w), 40% (w / w), 42% (w / w), 44% (w / w), 46% (w / w), 48% (w / w), 50% (w / w), or 55% (w / w). In some embodiments, the weight percentage of the perfluorocarbon component in the total nanoparticle (e.g., nanoemulsion) composition is no more than about 90% (w / w), for example, no more than any one of about 88% (w / w), 86% (w / w), 84% (w / w), 82% (w / w), 80% (w / w), 78% (w / w), 76% (w / w), 72% (w / w), 70% (w / w), 68% (w / w), 66% (w / w), 64% (w / w), 62% (w / w), or 60% (w / w). In some embodiments, the weight percentage of the perfluorocarbon component in the total nanoparticle (e.g., nanoemulsion) composition is from about 20% to about 90% (w / w), for example, from about 25% to about 85% (w / w), from about 30% to about 70% (w / w), from about 40% to about 50% (w / w), or about 50% (w / w).

[0046] lipids

[0047] In some embodiments, the lipid component includes phospholipids, for example, more than one type of phospholipid. In some embodiments, the phospholipids may be of naturally occurring or synthetic origin. In some embodiments, the lipid component includes phosphatidic acid, phosphatidylethanolamine, phosphatidylcholine, phosphatidylserine, phosphatidylinositol, distearylphosphatidylcholine (DSPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylcholine, dimyristoylphosphatidylcholine (DMPC), palmitoyloleoylphosphatidylcholine (POPC), phosphatidylethanolamine, sphingomyelin, phosphatidylserine and phosphatidylinositol, and a mixture of fats containing glycerophosphatidylinosides such as soy lecithin, soy phosphatidylcholine (HSPC), 1-oleoyl-2-palmitoyl-sn-glycerol-3-phosphocholine (OPPC), 1,2-dioleoyl-sn-glycerol-3-phosphocholine (DOPC), 2-disorcinol-sn-glycerol-3-phosphocholine (DEPC) or any combination thereof.

[0048] In some embodiments, the lipid component includes DSPC and DPPC. In some embodiments, the lipid component consists essentially of DSPC and DPPC. In some embodiments, the lipid component includes no more than about 2% (w / w), for example, no more than about 1.5% (w / w), 1.0% (w / w), 0.5% (w / w), 0.1% (w / w), 0.01% (w / w), or 0.001% (w / w) of lipids other than DSPC or DPPC. In some embodiments, the lipid component includes no more than about 2% (w / w), for example, no more than about 1.5% (w / w), 1.0% (w / w), 0.5% (w / w), 0.1% (w / w), 0.01% (w / w), or 0.001% (w / w) of soy lecithin. In some embodiments, the total nanoparticle (e.g., nanoemulsion) composition comprises no more than about 2% (w / w) of the total nanoparticle (e.g., nanoemulsion) composition, for example, no more than about 1.5% (w / w), 1.0% (w / w), 0.5% (w / w), 0.1% (w / w), 0.01% (w / w), or 0.001% (w / w). In some embodiments, the lipid component comprises no more than about 2% (w / w), for example, no more than about 1.5% (w / w), 1.0% (w / w), 0.5% (w / w), 0.1% (w / w), 0.01% (w / w), or 0.001% (w / w). In some embodiments, the total nanoparticle (e.g., nanoemulsion) composition contains no more than about 2% (w / w) of the total nanoparticle (e.g., nanoemulsion) composition, for example, no more than any one of about 1.5% (w / w), 1.0% (w / w), 0.5% (w / w), 0.1% (w / w), 0.01% (w / w), or 0.001% (w / w) of HSPC.

[0049] In some embodiments, the lipid component comprises (e.g., the lipid component is) a mixture of DSPC and DPPC, wherein the weight ratio of DSPC to DPPC is from about 1:1 to about 5:1, such as from about 1:1 to about 4.5:1, from about 1:1 to about 4:1, from about 1.5:1 to about 4:1, from about 2:1 to about 4:1, from about 2:1 to about 3.5:1, or from about 2.8:1. In some embodiments, the lipid component comprises (e.g., the lipid component is) a mixture of DSPC and DPPC in a weight ratio of about 2.8:1.

[0050] In some embodiments, the lipid component in the total nanoparticle (e.g., nanoemulsion) composition is at least about 0.5% (w / w), for example, at least about 0.8% (w / w), 1.0% (w / w), 1.2% (w / w), 1.4% (w / w), 1.6% (w / w), 2% (w / w), 2.2% (w / w), 2.4% (w / w), 2.6% (w / w), 2.8% (w / w), or 3% (w / w). In some embodiments, the weight percentage of the lipid component in the total nanoparticle (e.g., nanoemulsion) composition is not greater than 4.5% (w / w), for example, at least about 4.2% (w / w), 4% (w / w), 3.8% (w / w), 3.5% (w / w), 3.2% (w / w), 3% (w / w), 2.8% (w / w), 2.6% (w / w), or 2.5% (w / w). In some embodiments, the weight percentage of the lipid component in the total nanoparticle (e.g., nanoemulsion) composition is about 1.5% to about 3.5% (w / w), for example, 2.0% to about 3.5% (w / w), 2.0% to about 3.0% (w / w), 2.0% to about 2.5% (w / w), or about 2.3%.

[0051] In some embodiments, the weight ratio of the perfluorocarbon component to the lipid component in the nanoparticle composition is at least about 5:1, for example, at least about 6:1, 7:1, 8:1, 9:1, 10:1, 12:1, 14:1, 16:1, 18:1, 20:1, or 22:1. In some embodiments, the weight ratio of the perfluorocarbon component to the lipid component is not greater than about 50:1, for example, not greater than about 45:1, 40:1, 35:1, 30:1, 25:1, or 23:1. In some embodiments, the weight ratio of the perfluorocarbon component to the lipid component in the nanoparticle composition is from about 5:1 to about 40:1, for example, from about 10:1 to about 30:1, from about 15:1 to about 23:1, or about 23:1.

[0052] Buffers and Buffer Components

[0053] In some embodiments, the nanoparticle composition further comprises a buffering component. In some embodiments, the nanoparticle composition further comprises a buffering agent comprising a buffering component and water. In some embodiments, the buffering agent can maintain a stable pH and / or control stress. In some embodiments, the buffering agent includes an acetate buffer, a formate buffer, a citrate buffer, a glutamate buffer, or a phosphate buffer, or any combination thereof. In some embodiments, the buffering agent includes a citrate buffer. In some embodiments, the buffering agent comprises about 20 mM to about 100 mM, such as about 30 mM to 70 mM or 50 mM of citrate buffer. In some embodiments, the buffering agent comprises 50 mM of citrate buffer. In some embodiments, the pH of the citrate buffer is 6.0. In some embodiments, the buffer component comprises sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1, such as any one of about 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, or any value between the foregoing values. In some embodiments, the buffer component comprises sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid results in the buffer having a pH of about 4 to about 8, such as about 6.

[0054] In some embodiments, the buffer comprises a phosphate buffer with a pH of 6.8. In some embodiments, the buffer does not include a phosphate buffer. In some embodiments, the buffer comprises no more than about 2% (w / w), for example, no more than about 1.5% (w / w), 1.0% (w / w), 0.5% (w / w), 0.1% (w / w), 0.01% (w / w), or 0.001% (w / w) of a phosphate buffer. In some embodiments, the total nanoparticle (e.g., nanoemulsion) composition comprises no more than about 2% (w / w), for example, no more than about 1.5% (w / w), 1.0% (w / w), 0.5% (w / w), 0.1% (w / w), 0.01% (w / w), or 0.001% (w / w) of a phosphate buffer.

[0055] In some embodiments, the buffer is at least about 20% (w / w) of the total nanoparticle (e.g., nanoemulsion) composition, for example, at least about 22% (w / w), 25% (w / w), 27% (w / w), 30% (w / w), 32% (w / w), 35% (w / w), 37% (w / w), 40% (w / w), 42% (w / w), 44% (w / w), or 46% (w / w). In some embodiments, the buffer is no more than 70% (w / w) of the total nanoparticle (e.g., nanoemulsion) composition, for example, no more than 68% (w / w), 65% (w / w), 63% (w / w), 60% (w / w), 57% (w / w), 55% (w / w), 52% (w / w), 50% (w / w), or 48% (w / w). In some embodiments, the buffer is about 20% to about 70% (w / w) of the total nanoparticle (e.g., nanoemulsion) composition, for example, about 30% to about 60% (w / w), about 30% to about 50% (w / w), about 40% to about 60% (w / w), or about 48% (w / w).

[0056] In some embodiments, the buffer component comprises sodium citrate dihydrate, and the weight percentage of sodium citrate dihydrate in the total nanoparticle (e.g., nanoemulsion) composition is at least about 5% (w / w), for example, at least about 6% (w / w), 7% (w / w), 8% (w / w), 9% (w / w), 10% (w / w), 11% (w / w), or 12% (w / w). In some embodiments, the buffer component comprises sodium citrate dihydrate, and the weight percentage of sodium citrate dihydrate in the total nanoparticle (e.g., nanoemulsion) composition is not greater than 10% (w / w), for example, not greater than 11% (w / w), 12% (w / w), 13% (w / w), 14% (w / w), 15% (w / w), 16% (w / w), or 20% (w / w). In some embodiments, the buffer component includes sodium citrate dihydrate, and the sodium citrate dihydrate in the total nanoparticle (e.g., nanoemulsion) composition is present in a weight percentage of about 5% to about 20% (w / w), for example, about 7% to about 15% (w / w) or about 12% (w / w).

[0057] In some embodiments, the buffer component includes citric acid, and the weight percentage of citric acid in the total nanoparticle (e.g., nanoemulsion) composition is at least about 0.001% (w / w), for example, at least about 0.003% (w / w), 0.005% (w / w), 0.007% (w / w), 0.009% (w / w), 0.1% (w / w), 0.12% (w / w), or 0.15% (w / w). In some embodiments, the buffer component includes citric acid, and the weight percentage of citric acid in the total nanoparticle (e.g., nanoemulsion) composition is not greater than 0.2% (w / w), for example, not greater than 0.18% (w / w), 0.16% (w / w), 0.14% (w / w), 0.12% (w / w), or 0.1% (w / w). In some embodiments, the buffer component includes citric acid, and the weight percentage of citric acid in the total nanoparticle (e.g., nanoemulsion) composition is from about 0.01% to about 0.2% (w / w), for example, from about 0.05% to about 0.15% (w / w) or about 0.1% (w / w).

[0058] In some embodiments, the weight ratio of the perfluorocarbon component to the buffer is at least about 0.5:1, for example, at least about 0.75:1, 0.9:1, 1:1, 9:1, or 1.1:1. In some embodiments, the weight ratio of the perfluorocarbon component to the buffer is not greater than about 2:1, for example, not greater than about 1.8:1, 1.7:1, 1.5:1, 1.4:1, 1.3:1, 1.1:1, or 1:1. In some embodiments, the weight ratio of the perfluorocarbon component to the lipid component is from about 0.8:1 to about 2:1, for example, from about 0.8:1 to about 1.5:1, from about 0.8:1 to about 1.3:1, or about 1.1:1.

[0059] In some embodiments, the buffer comprises a buffer component and water. In some embodiments, the buffer component comprises sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1, such as any one of about 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, or any value between the foregoing values. In some embodiments, the buffer component comprises sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid results in the buffer having a pH of about 4 to about 8, such as about 6. In some embodiments, the weight ratio of the perfluorocarbon component to the buffer component is at least about 1:1, for example, at least any one of about 1.5:1, 2:1, 2.5:1, 3:1, or 3.5:1. In some embodiments, the weight ratio of the perfluorocarbon component to the buffer component is no greater than about 6:1, for example, no greater than any of about 5.5:1, 5:1, 4.5:1, 4:1, or 3.5:1. In some embodiments, the weight ratio of the perfluorocarbon component to the buffer component is from about 2.5:1 to about 5, for example, from about 3:1 to about 4.5:1, from about 3.5:1 to about 4.5:1, or about 3.8:1.

[0060] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition described herein may be diluted with a buffer (e.g., citrate buffer), wherein the diluted nanoparticle (e.g., nanoemulsion) composition is also covered in this invention. In some embodiments, the nanoparticle composition may be diluted with a citrate buffer of at least 1× (v / v) (e.g., any one of about 1× (v / v), 2× (v / v), 3× (v / v), 4× (v / v), 5× (v / v), 6× (v / v), or 7× (v / v). In some embodiments, a composition comprising the nanoparticle (e.g., nanoemulsion) composition described herein and an additional citrate buffer is provided, wherein the volume ratio of the additional citrate buffer to the nanoparticle (e.g., nanoemulsion) composition is at least 0.5:1 (e.g., any one of about 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, or 8:1).

[0061] feature

[0062] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition has a pH of at least about 4, for example, at least about 4, 4.5, 5, or 5.5. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition has a pH of not more than about 9, for example, not more than about 8.5, 8, 7.5, 7, 6.5, or 6. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition has a pH of about 3.5 to about 8, for example, about 4 to 8, about 5 to about 8, about 5 to about 7.5, or about 6 to about 7.5.

[0063] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition has a viscosity of not more than about 500 cp, for example, not more than any of about 400 cp, 300 cp, 200 cp, 100 cp, 80 cp, 60 cp, 40 cp or 20 cp, as determined by a viscometer.

[0064] In some embodiments, the average particle size of the nanoparticles is not greater than about 5 μm (e.g., not greater than any of about 4 μm, 3 μm, 2 μm, 1 μm, or 500 nm). In some embodiments, the average particle size of the nanoparticles in the nanoparticle (e.g., nanoemulsion) composition can be further reduced by diluting the nanoparticle (e.g., nanoemulsion) composition with a buffer. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition can be further diluted with at least about 0.5 × (e.g., at least about 1 ×, 2 ×, 3 ×, 4 ×, 5 ×, or 6 ×) citrate buffer. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition can be diluted with 5 × citrate buffer, and the average particle size of the nanoparticles in the diluted nanoparticle (e.g., nanoemulsion) composition is not greater than about 1 μm (e.g., about 50 nm to about 900 nm, 50 nm to about 700 nm, 50 nm to about 600 nm, 50 nm to about 400 nm, or 200 nm to about 400 nm). In some embodiments, the polydispersity index (PDI) of the nanoparticles in the 5× citrate-diluted nanoparticle (e.g., nanoemulsion) composition is about 0.5 to about 1 (e.g., any one of about 0.6 to 1, 0.7 to 1, 0.8 to 1, 0.9 to 1, or 1). In some embodiments, the zeta potential of the nanoparticles in the 5× citrate-diluted nanoparticle (e.g., nanoemulsion) composition is about 0.02 mV to about 0.8 mV (e.g., any one of about 0.02 mV to 0.8 mV, 0.02 mV to 0.7 mV, 0.02 mV to 0.2 mV, or 0.5 mV to 0.7 mV). In some embodiments, the average particle size of the nanoparticles in the nanoparticle (e.g., nanoemulsion) composition, whether before dilution, after dilution, or both, is suitable for atomization and inhalation. In some embodiments, the average particle size of the nanoparticles in the nanoparticle (e.g., nanoemulsion) composition is determined by dynamic light scattering. In some embodiments, the zeta potential of the nanoparticles in the nanoparticle (e.g., nanoemulsion) composition is determined by a zeta potential meter.

[0065] In some embodiments, the nanoparticles in the nanoparticle (e.g., nanoemulsion) composition have a multi-lamellar structure comprising at least two lamellar layers. Figure 2 shows a CryoEM image of a representative nanoparticle in the nanoparticle composition. In some embodiments, each lamellar layer comprises perfluorocarbon sandwiched between two lipid layers.

[0066] In some embodiments, the nanoparticle (e.g., nanoemulsion) compositions described herein are capable of dissolving oxygen. In some embodiments, the nanoparticle (e.g., nanoemulsion) compositions are enriched with dissolved oxygen. In some embodiments, the nanoparticle (e.g., nanoemulsion) compositions are enriched with dissolved oxygen at least about 1 ppm, for example, at least about 1 ppm, 2 ppm, 3 ppm, 4 ppm, 5 ppm, 6 ppm, 7 ppm, or 8 ppm. In some embodiments, the nanoparticle (e.g., nanoemulsion) compositions are enriched with dissolved oxygen from about 1 ppm to about 20 ppm, for example, about 3 ppm to about 17 ppm, about 5 ppm to about 15 ppm, about 7 ppm to about 13 ppm, about 7 ppm to about 10 ppm, about 7 ppm, about 8 ppm, about 9 ppm, or about 10 ppm. In some embodiments, the dissolved oxygen concentration is determined by an oxygen probe.

[0067] In some embodiments, the nanoparticle (e.g., nanoemulsion) compositions described herein are capable of binding with CO2. In some embodiments, the nanoparticles in the nanoparticle (e.g., nanoemulsion) compositions provided herein are capable of binding with both O2 and CO2. In some embodiments, the nanoparticles in the nanoparticle (e.g., nanoemulsion) compositions provided herein are capable of selectively binding with O2 or CO2, depending on the concentration of O2 or CO2, or the relative concentration between O2 and CO2. In some embodiments, as shown in FIG1, when applied to tissue, the nanoparticles in the nanoparticle (e.g., nanoemulsion) compositions provided herein are capable of promoting CO2 and O2 exchange. In some embodiments, the tissue is in the lungs, on the skin, in the oral cavity, in the colon, or in the gastrointestinal tract. In some embodiments, the nanoparticles in the nanoparticle (e.g., nanoemulsion) compositions provided herein are capable of promoting CO2 and O2 exchange in the lungs, as shown in Figure 1, wherein the nanoparticle (e.g., nanoemulsion) composition is enriched with O2 before the nanoparticles reach the alveolar epithelial cells, and enriched with CO2 after contact with the alveolar epithelial cells.

[0068] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is physically stable, for example, not exhibiting phase separation at temperatures above 10°C (e.g., room temperature). In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is physically stable at temperatures above 10°C (e.g., room temperature) for at least about 3 days, for example, at least about 4, 5, 6, 7, 8, 9, 10, or 12 days. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is chemically stable, for example, not exhibiting oxidation or degradation at temperatures above 10°C (e.g., room temperature). In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is chemically stable at temperatures above 10°C (e.g., room temperature) for at least 3 days, for example, at least about 4, 5, 6, 7, 8, 9, 10, or 12 days. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is stable at room temperature for at least 10 days without phase separation or chemical oxidation.

[0069] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition contains no more than 5% (w / w), for example, no more than 2% (w / w), 1% (w / w), 0.5% (w / w), 0.1% (w / w), or 0.01% (w / w). In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is substantially free of cetrimonium bromide (CTAB). CTAB is a quaternary ammonium surfactant and has known toxicological effects, such as respiratory irritation or skin irritation. In some embodiments, by removing the CTAB surfactant, the nanoparticles (e.g., nanoemulsions) of the present invention reduce side effects while maintaining high stability.

[0070] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition contains no more than 5% (w / w), for example, no more than 2% (w / w), 1% (w / w), 0.5% (w / w), 0.1% (w / w), or 0.01% (w / w) of Tween 80. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is substantially free of Tween 80. Tween 80 is a polyether nonionic surfactant and has known toxicological effects, such as respiratory irritation or skin irritation. In some embodiments, by removing Tween 80, the nanoparticles (e.g., nanoemulsions) of the present invention reduce side effects while maintaining high stability.

[0071] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition contains no more than 5% (w / w), for example, no more than 2% (w / w), 1% (w / w), 0.5% (w / w), 0.1% (w / w), or 0.01% (w / w) of soy lecithin. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is substantially free of soy lecithin. Lecithin is a mixture of fats containing glycerophospholipids and can have toxic effects, such as undesirable irritation, when inhaled or applied to wounds. In some embodiments, by removing soy lecithin, the nanoparticles (e.g., nanoemulsions) of the present invention reduce side effects while maintaining high stability.

[0072] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition contains no more than 5% (w / w), for example, no more than 2% (w / w), 1% (w / w), 0.5% (w / w), 0.1% (w / w), or 0.01% (w / w) of a polymeric surfactant. In some embodiments, the polymeric surfactant includes poloxamer, and the nanoparticle (e.g., nanoemulsion) composition contains no more than 5% (w / w), for example, no more than 2% (w / w), 1% (w / w), 0.5% (w / w), 0.1% (w / w), or 0.01% (w / w) of a poloxamer (such as Pluronic®). Poloxamer is a nonionic triblock copolymer composed of a central hydrophobic chain of polyoxypropylene (poly(propylene oxide)) with two hydrophilic chains of polyoxyethylene (poly(ethylene oxide)) side-attached. Studies have shown that poloxamer exhibits various toxicities when administered to animals, including causing hypercholesterolemia, hypertriglyceridemia, and tissue irritation. In some embodiments, by removing poloxamer, the nanoparticles (e.g., nanoemulsions) of the present invention reduce side effects while maintaining high stability. In some embodiments, by removing polymeric surfactants, the nanoparticles (e.g., nanoemulsions) of the present invention reduce side effects while maintaining high stability. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition comprises no more than 5% (w / w), for example, no more than 2% (w / w), 1% (w / w), 0.5% (w / w), 0.1% (w / w), or 0.01% (w / w) of any surfactant other than lipids.

[0073] In some embodiments, this document provides a nanoparticle (e.g., nanoemulsion) composition prepared by mixing about 25% to 40% (e.g., about 35.5%) (w / w) of PFD, about 10% to 20% (e.g., about 14.5%) (w / w) of PFTPA, about 0.1% to 1% (e.g., about 0.6%) (w / w) of DPPC, about 1.5% to 2.5% (e.g., about 1.7%) (w / w) of DSPC, and about 40% to 60% (e.g., about 47.7%) (w / w) of a citrate buffer (50 mM) at pH 6.0. In some of the foregoing embodiments, mixing is carried out in a microfluidic apparatus.

[0074] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is suitable for application to the surface of tissue. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is suitable for application to the surface of the alveoli, optionally administered by inhalation. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is suitable for application to the surface of the skin, such as wounds or injuries. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is suitable for application to the surface of the gastrointestinal tract, for example, administered by enema. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is administered in a non-invasive manner. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is not administered by injection (including intravenous and intraperitoneal injection).

[0075] Exemplary nanoparticle (e.g., nanoemulsion) compositions

[0076] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition comprises:

[0077] (i) PFD, comprising about 25% to about 40% (e.g., about 35.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight; (ii) PFTPA, comprising about 10% to 20% (e.g., about 14.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight; (iii) DPPC, comprising about 0.1% to 1% (e.g., about 0.6%) of the total nanoparticle (e.g., nanoemulsion) composition by weight; (iv) DSPC, comprising about 1.5% to 2.5% (e.g., about 1.7%) of the total nanoparticle (e.g., nanoemulsion) composition by weight; and (v) a citrate buffer (50 mM) at pH 6.0, comprising 40% to 60% (e.g., about 47.7%) of the total nanoparticle (e.g., nanoemulsion) composition by weight. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is contained within the atomizer. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is included in a gel composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is included in a cream composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition contains no more than about 2% (w / w) of a polymeric surfactant of the total nanoparticle (e.g., nanoemulsion) composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition has a viscosity of no more than about 100 cp. In some embodiments, the average particle size of the nanoparticles is no more than about 3 μm.

[0078] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition comprises:

[0079] (i) PFD, comprising about 35% to about 55% (e.g., about 44.6%) of the total nanoparticle (e.g., nanoemulsion) composition by weight; (ii) PFTPA, comprising about 10% to 20% (e.g., about 14.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight; (iii) DPPC, comprising about 0.1% to 1% (e.g., about 0.6%) of the total nanoparticle (e.g., nanoemulsion) composition by weight; (iv) DSPC, comprising about 1.5% to 2.5% (e.g., about 1.7%) of the total nanoparticle (e.g., nanoemulsion) composition by weight; and (v) a citrate buffer (50 mM) at pH 6.0, comprising about 25% to about 45% (e.g., about 35.4%) of the total nanoparticle (e.g., nanoemulsion) composition by weight. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is contained within the atomizer. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is included in a gel composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is included in a cream composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition contains no more than about 2% (w / w) of a polymeric surfactant of the total nanoparticle (e.g., nanoemulsion) composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition has a viscosity of no more than about 100 cp. In some embodiments, the average particle size of the nanoparticles is no more than about 3 μm.

[0080] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition comprises:

[0081] (i) PFD, comprising about 40% to about 60% (e.g., about 50%) of the total nanoparticle (e.g., nanoemulsion) composition by weight; (ii) PFTPA, comprising about 10% to about 30% (e.g., about 20%) of the total nanoparticle (e.g., nanoemulsion) composition by weight; (iii) DPPC, comprising 0.1% to 1% (e.g., about 0.7%) of the total nanoparticle (e.g., nanoemulsion) composition by weight; (iv) DSPC, comprising about 1% to 3% (e.g., about 2%) of the total nanoparticle (e.g., nanoemulsion) composition by weight; and (v) a phosphate buffer with a pH of 6.8, comprising about 15% to about 40% (e.g., about 27.3%) of the total nanoparticle (e.g., nanoemulsion) composition by weight. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is contained in an atomizer. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is contained in a gel composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is included in the cream composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition comprises no more than about 2% (w / w) of a polymeric surfactant in the total nanoparticle (e.g., nanoemulsion) composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition has a viscosity of no more than about 100 cp. In some embodiments, the average particle size of the nanoparticles is no more than about 3 μm.

[0082] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition comprises:

[0083] (i) PFD, which is about 25% to about 40% by weight of the total nanoparticle composition (e.g., about 30% to 40%, or about 35.5%).

[0084] (ii) PFTPA, which is about 10% to 20% by weight of the total nanoparticle composition (e.g., about 12% to 18%, or about 14.5%).

[0085] (iii) DPPC, which is about 0.1% to 1% by weight of the total nanoparticle (e.g., nanoemulsion) composition (e.g., about 0.2% to 0.8%, or about 0.6%).

[0086] (iv) DSPC, which is about 1.5% to 2.5% by weight of the total nanoparticle composition (e.g., about 1.5% to about 2%, or about 1.7%).

[0087] (v) Citric acid, in a weight percentage of about 0.01% to 0.15% of the total nanoparticle composition (e.g., about 0.05% to 0.15%, or about 0.1%).

[0088] (vi) Sodium citrate dihydrate, comprising, by weight, about 0.5% to about 2% of the total nanoparticle composition (e.g., about 0.5% to about 1.5%, or about 1.2%); and

[0089] (viii) Water, which is about 30% to about 60% by weight of the total nanoparticle composition (e.g., about 40% to about 50%, or about 46.4%).

[0090] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is contained in an atomizer. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is contained in a gel composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is contained in a cream composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition contains no more than about 2% (w / w) of a polymeric surfactant of the total nanoparticle (e.g., nanoemulsion) composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition has a viscosity of no more than about 100 cp. In some embodiments, the average particle size of the nanoparticles is no more than about 3 μm.

[0091] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition comprises:

[0092] (i) A perfluorocarbon component comprising PFD and PFTPA, wherein the weight ratio of PFD to PFTPA is from about 2:1 to about 3:1 (e.g., from about 2:1 to about 2.6:1, or about 2.4:1);

[0093] (ii) A lipid component comprising DSPC and DPPC, wherein the weight ratio of DSPC to DPPC is from about 1:1 to about 5:1 (e.g., from about 2:1 to about 3.5:1, or about 2.8:1);

[0094] (iii) A buffer component comprising sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1 (e.g., from about 7:1 to about 15:1, or about 12:1); wherein

[0095] The weight ratio between the perfluorocarbon component and the lipid component is from about 10:1 to about 30:1 (e.g., from about 15:1 to about 23:1, or about 23:1); and

[0096] The weight ratio between the perfluorocarbon component and the buffer component is from about 2.5:1 to about 5 (e.g., from about 3.5:1 to about 4.5:1, or about 3.8:1).

[0097] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is contained in an atomizer. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is contained in a gel composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is contained in a cream composition. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition contains no more than about 2% (w / w) of a polymeric surfactant. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition has a viscosity of no more than about 100 cp. In some embodiments, the average particle size of the nanoparticles is no more than about 3 μm.

[0098] In some embodiments, any of the aforementioned nanoparticle (e.g., nanoemulsion) compositions can be used as an inhalable formulation. In some embodiments, any of the aforementioned nanoparticle (e.g., nanoemulsion) compositions can be mixed with pharmaceutically acceptable excipients to prepare wound healing compositions, enema gels, or topical creams, as described in detail herein.

[0099] Preparation method

[0100] In some respects, methods for preparing the nanoparticle compositions described herein are provided.

[0101] Typically, in some embodiments, methods for preparing the nanoparticle compositions described herein involve blending a lipid and a perfluorocarbon component in a solvent (such as a water-miscible solvent, e.g., ethanol), subjecting the mixture to high shear conditions, and then removing the solvent. In some embodiments, the lipid component is first blended in a solvent, and then the perfluorocarbon component is added. In some embodiments, the solvent is an organic solvent, such as ketones, esters, ethers, chlorinated solvents, and other solvents known in the art. In some embodiments, the high shear conditions are provided by one or more of acoustic treatment, homogenization, or microfluidization.

[0102] Techniques for subjecting mixtures to high shear conditions are known in the art. In some embodiments, high shear conditions are provided through acoustic treatment (including probe acoustic treatment or ultrasonic treatment), homogenization, or microfluidization. For example, in some embodiments, the mixture is subjected to high-pressure homogenization (e.g., using Avestin, APV Gaulin, Microfluidizer™ (such as the Microfluidizer™ processor ML 10EH from Microfluidics, Stansted), or an Ultra Turrax homogenizer). In some embodiments, subjecting the mixture to high shear conditions includes one or more cycles of high shear conditions. For example, the mixture may be cycled through a high-pressure homogenizer for about 2 to about 100 cycles, such as about 5 to about 50 cycles or about 8 to about 20 cycles (e.g., any one of about 8, 10, 12, 14, 16, 18, or 20 cycles). Solvents (such as ethanol) can be removed by evaporation using suitable equipment known for this purpose, which can operate in batch or continuous mode. This equipment includes, but is not limited to, rotary evaporators, falling film evaporators, wiped film evaporators, spray dryers, and the like. In some embodiments, the solvent can be removed by exposure to atmospheric pressure or by applying a reduced pressure (such as any one of about 25 mmHg, 30 mmHg, 40 mmHg, 50 mmHg, 100 mmHg, 200 mmHg, or 300 mmHg). The amount of time used to remove the solvent can be adjusted according to the volume of the formulation and the amount of solvent present.

[0103] In one aspect, this document provides a method for preparing a nanoparticle (e.g., nanoemulsion) composition comprising nanoparticles, wherein the nanoparticles comprise a perfluorocarbon component and a lipid component, the method comprising: (i) mixing the lipid with a solvent; (ii) evaporating the solvent from the mixture of (i); and (iii) adding the perfluorocarbon to the mixture of (ii). In some embodiments, the solvent is an alcohol. In some embodiments, the solvent is ethanol. In some embodiments, the solvent is evaporated by rotary evaporation. In some embodiments, the mixture of (iii) is subjected to continuous acoustic treatment for a period of time, optionally from about 30 minutes to about 1 hour.

[0104] In one aspect, this document provides a method for preparing a nanoparticle (e.g., nanoemulsion) composition comprising nanoparticles, wherein the nanoparticles comprise a perfluorocarbon component and a lipid component, the method comprising: (i) mixing the lipid with a solvent; (ii) adding the perfluorocarbon to the mixture of (i); (iii) homogenizing the mixture of (ii); and (iv) evaporating the solvent. In some embodiments, the solvent is a water-miscible solvent, such as an alcohol. In some embodiments, the solvent is ethanol. In some embodiments, the solvent is evaporated by rotary evaporation. In some embodiments, in step (i), the lipid is mixed with the solvent in the presence of a surfactant. In some embodiments, in step (i), the lipid is mixed with the solvent in the absence of a surfactant. In some embodiments, the method further comprises (v) dispersing the mixture of (iv) in water or a buffer to form a nanoparticle composition, optionally under probe acoustic treatment.

[0105] In one aspect, this document provides a method for preparing a nanoparticle (e.g., nanoemulsion) composition comprising nanoparticles, wherein the nanoparticles comprise a perfluorocarbon component and a lipid component, the method comprising: (i) combining the lipid with the perfluorocarbon to form a mixture; and (ii) homogenizing the mixture. In some embodiments, homogenization is performed by probe homogenization. In some embodiments, the method further comprises (iii) adding water or a buffer to the mixture to obtain a second mixture; and (iv) homogenizing the second mixture. In some embodiments, homogenization may be performed at about 1 rpm to about 5 rpm, such as at about 3.4 rpm. In some embodiments, the second mixture in (iv) may be homogenized for at least 1 minute, such as any one of about 1 minute to 60 minutes, 10 minutes to 40 minutes, 10 minutes to 30 minutes, or about 20 minutes.

[0106] In one aspect, this document provides a method for preparing a nanoparticle (e.g., nanoemulsion) composition comprising nanoparticles, wherein the nanoparticles comprise a perfluorocarbon component and a lipid component, the method comprising: (i) adding a lipid to a solvent to form a first mixture; (ii) adding a perfluorocarbon to the first mixture to form a second mixture; and (iii) subjecting the second mixture to probe acoustic treatment. In some embodiments, in step (i), the lipid is added to the solvent along with about 30% to 70% (e.g., about 50%) of a surfactant required in the composition. In some embodiments, the first mixture is clear. In some embodiments, the solvent is a water-miscible solvent, such as an alcohol. In some embodiments, the solvent is ethanol. In some embodiments, the second mixture is subjected to probe acoustic treatment for at least about 1 minute, such as any one of about 1 minute to 30 minutes, 5 minutes to 20 minutes, or about 10 minutes. In some embodiments, the method further comprises adding an aqueous solution to the first mixture, wherein the aqueous solution comprises a surfactant. In some embodiments, the method further comprises dispersing the probe acoustic-treated second mixture in an aqueous surfactant solution. In some embodiments, the surfactant (e.g., a non-lipid or polymeric surfactant) is about 30% to 70% (e.g., about 50%) of the desired surfactant in the composition.

[0107] In one aspect, this document provides a method for preparing a nanoparticle (e.g., nanoemulsion) composition comprising nanoparticles, wherein the nanoparticles comprise a perfluorocarbon component and a lipid component, the method comprising: (i) mixing the lipid with a solvent; (ii) adding the perfluorocarbon to the mixture of (i); (iii) subjecting the mixture of (ii) to probe acoustic treatment; and (iv) slowly adding water or a buffer to the mixture of (iii). In some embodiments, the solvent is a water-miscible solvent, such as an alcohol. In some embodiments, the solvent is ethanol. In some embodiments, in step (iv), an aqueous solution comprising a surfactant and optionally a buffer is added to the mixture of (iii). In some embodiments, the method further comprises subjecting the mixture to probe acoustic treatment after step (iv). In some embodiments, the mixture of (iv) is subjecting the mixture to probe acoustic treatment for about 30 minutes to about 1 hour.

[0108] In one aspect, this document provides a method for preparing a nanoparticle (e.g., nanoemulsion) composition comprising nanoparticles, wherein the nanoparticles comprise a perfluorocarbon component and a lipid component, the method comprising: (i) mixing the lipid with a solvent; (ii) evaporating the solvent in the mixture of (i) to form a thin film; (iii) hydrating the thin film by adding water or a buffer; and (iv) adding the perfluorocarbon to the mixture of (iii). In some embodiments, the solvent is a water-miscible solvent, such as an alcohol. In some embodiments, the solvent is ethanol. In some embodiments, the solvent is evaporated by rotary evaporation. In some embodiments, in step (iii), the thin film is hydrated by an aqueous solution comprising a surfactant and optionally a buffer. In some embodiments, the mixture of (iv) is subjected to continuous acoustic treatment (e.g., probe acoustic treatment) for a period of time, optionally from about 5 minutes to about 30 minutes.

[0109] In some embodiments, this document provides a method for preparing a nanoparticle (e.g., nanoemulsion) composition comprising nanoparticles, wherein the nanoparticles comprise a perfluorocarbon component and a lipid component, the method comprising: (i) mixing the lipid with the perfluorocarbon; (ii) slowly adding water or a buffer to the mixture in (i); and (iii) microfluidizing the mixture in (ii). In some embodiments of (ii), a citrate buffer is added to the mixture in (i). In some embodiments, step (i) can be performed by probe homogenization. In some embodiments, step (ii) can be performed by probe homogenization. In some embodiments of step (ii), an aqueous solution comprising a surfactant and optionally a buffer is added to the mixture in (i). In some embodiments, the aqueous solution does not contain (e.g., contains less than 5 wt%, 4 wt%, 3 wt%, 2 wt%, 1 wt%, or 0.1 wt%) a surfactant. In some embodiments, the mixture in (ii) is microfluidized at about 10,000 PSI to about 30,000 PSI, optionally at about 20,000 PSI or 25,000 PSI. In some embodiments, the mixture in (ii) is microfluidized for at least two cycles, for example, two, three, four, or five cycles. In some embodiments, the composition is cooled between each microfluidization cycle. In some embodiments, the composition is further sterilized by autoclaving. In some embodiments, the composition is sterilized at a temperature of at least 100°C, such as about 105°C, 110°C, 115°C, or 120°C.

[0110] In some embodiments, the lipid comprises DSPC and DPPC. In some embodiments, the lipid is a mixture of DSPC and DPPC. In some embodiments, the perfluorocarbon comprises PFD and PFTA. In some embodiments, the perfluorocarbon is a mixture of PFD and PFTA.

[0111] In some aspects, nanoparticle compositions are provided that are produced using the methods taught herein. For example, in some embodiments, a nanoparticle composition is provided that is produced by mixing a lipid and a perfluorocarbon component in a solvent (such as a water-miscible solvent, e.g., ethanol), subjecting the mixture to high shear conditions, and removing the solvent to produce the nanoparticle composition.

[0112] III. Topical preparations

[0113] Inhalable preparations

[0114] In one aspect, this document provides an inhalable formulation comprising the nanoparticle (e.g., nanoemulsion) composition described herein. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition described herein is a composition suitable for direct atomization and inhalation. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition described herein is an inhalable formulation.

[0115] In some embodiments, the inhalable formulation comprising a nanoparticle (e.g., nanoemulsion) composition is capable of binding with O2. In some embodiments, the inhalable formulation comprising a nanoparticle (e.g., nanoemulsion) composition is capable of binding with CO2. In some embodiments, the inhalable formulation comprising a nanoparticle (e.g., nanoemulsion) composition is capable of selectively binding with O2 or CO2, depending on the concentration of O2 or CO2, or the relative concentration between O2 and CO2. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is enriched with O2 before the nanoparticles reach the alveolar epithelial cells, and enriched with CO2 after contact with the alveolar epithelial cells. In some embodiments, the inhalable formulation comprising a nanoparticle (e.g., nanoemulsion) composition can be exhaled. In some embodiments, the inhalable formulation comprising a nanoparticle (e.g., nanoemulsion) composition is capable of delivering O2 to the lungs.

[0116] This article also provides devices (such as nebulizers) that can be used to administer inhalable formulations.

[0117] Wound healing composition

[0118] In one aspect, this article provides a wound healing composition comprising a nanoparticle (e.g., nanoemulsion) composition as described herein and a pharmaceutically acceptable excipient.

[0119] In some embodiments, the wound healing composition comprises the nanoparticle (e.g., nanoemulsion) composition described herein, silver dioxide, and collagen. In some embodiments, the collagen is bovine collagen. In some embodiments, the bovine collagen is hydrolyzed type I bovine collagen or porous type I bovine collagen. In some embodiments, hydrolyzed type I bovine collagen comprises 10% of the collagen contents processed to have a collagen structure similar to fetal skin. In some embodiments, porous type I bovine collagen is processed to have a collagen structure similar to fetal skin. In some embodiments, the collagen is human collagen.

[0120] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is about 30% to about 90% (w / w) of the total wound healing composition, such as any one of about 40% to 90% (w / w), 40% to 85% (w / w), 50% to 85% (w / w), 55% (w / w), or 82% (w / w).

[0121] In some embodiments, silver dioxide is about 0.1% to about 5% (w / w) of the total wound healing composition, such as any one of about 0.5% to 3% (w / w), 0.5% to 2.5% (w / w), 0.5% to 2% (w / w), 1% (w / w), or 1.5% (w / w).

[0122] In some embodiments, collagen (e.g., bovine collagen) comprises about 10% to about 65% (w / w) of the total wound healing composition, such as any one of about 10% to 60% (w / w), 10% to 55% (w / w), 10% to 50% (w / w), 15% to 50% (w / w), 15% (w / w), or 44% (w / w). In some embodiments, the collagen is porous type 1 bovine collagen, comprising about 15% by weight of the total wound healing composition. In some embodiments, the collagen is hydrolyzed type 1 bovine collagen, comprising about 44% by weight of the total wound healing composition.

[0123] In some embodiments, the wound healing composition comprises:

[0124] (a) A nanoparticle (e.g., nanoemulsion) composition comprising, by weight percentage, about 40% to about 70% (e.g., about 55%) of the total wound healing composition, wherein the nanoparticle (e.g., nanoemulsion) composition contains

[0125] (i) PFD, which is about 25% to about 40% (e.g., about 35.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0126] (ii) PFTPA, which is about 10% to 20% (e.g., about 14.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0127] (iii) DPPC, which is about 0.1% to 1% (e.g., about 0.6%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0128] (iv) DSPC, comprising about 1.5% to 2.5% (e.g., about 1.7%) by weight of the total nanoparticle (e.g., nanoemulsion) composition; and

[0129] (v) A citrate buffer (50 mM) with a pH of 6.0, comprising about 40% to 60% (e.g., about 47.7%) of the total nanoparticle (e.g., nanoemulsion) composition by weight.

[0130] (b) Silver dioxide, comprising about 0.5% to about 1.5% by weight (e.g., about 1%) of the total wound healing composition, and

[0131] (c) Hydrolyzed bovine collagen type 1, comprising about 35% to about 55% (e.g., about 44%) of the total wound healing composition by weight, wherein the hydrolyzed bovine collagen type 1 comprises 10% of the collagen contents processed to have a collagen structure similar to that of fetal skin.

[0132] In some embodiments, the wound healing composition comprises:

[0133] (a) A nanoparticle (e.g., nanoemulsion) composition comprising, by weight percentage, about 70% to about 90% (e.g., about 82%) of the total wound healing composition, wherein the nanoparticle (e.g., nanoemulsion) composition contains

[0134] (i) PFDS, which are about 25% to about 40% (e.g., about 35.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0135] (ii) PFTPA, which is about 10% to 20% (e.g., about 14.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0136] (iii) DPPC, which is about 0.1% to 1% (e.g., about 0.6%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0137] (iv) DSPC, comprising about 1.5% to 2.5% (e.g., about 1.7%) by weight of the total nanoparticle (e.g., nanoemulsion) composition; and

[0138] (v) A citrate buffer (50 mM) with a pH of 6.0, comprising about 40% to 60% (e.g., about 47.7%) of the total nanoparticle (e.g., nanoemulsion) composition by weight.

[0139] (b) Silver dioxide, comprising about 1% to about 2% (e.g., 1.5%) of the total wound healing composition by weight, and

[0140] (c) Porous type 1 bovine collagen, comprising about 10% to about 20% (e.g., about 15%) of the total wound healing composition by weight, wherein the porous type 1 bovine collagen is processed to have a collagen structure similar to fetal skin.

[0141] In some embodiments, the wound healing composition comprises:

[0142] (a) A nanoparticle (e.g., nanoemulsion) composition comprising, by weight percentage, about 70% to about 90% (e.g., about 82%) of the total wound healing composition, wherein the nanoparticle (e.g., nanoemulsion) composition contains

[0143] (i) PFD, which is about 25% to about 40% by weight of the total nanoparticle composition (e.g., about 30% to 40%, or about 35.5%).

[0144] (ii) PFTPA, which is about 10% to 20% by weight of the total nanoparticle composition (e.g., about 12% to 18%, or about 14.5%).

[0145] (iii) DPPC, which is about 0.1% to 1% by weight of the total nanoparticle (e.g., nanoemulsion) composition (e.g., about 0.2% to 0.8%, or about 0.6%).

[0146] (iv) DSPC, which is about 1.5% to 2.5% by weight of the total nanoparticle composition (e.g., about 1.5% to about 2%, or about 1.7%).

[0147] (v) Citric acid, in a weight percentage of about 0.01% to 0.15% of the total nanoparticle composition (e.g., about 0.05% to 0.15%, or about 0.1%).

[0148] (vi) Sodium citrate dihydrate, comprising, by weight, about 0.5% to about 2% of the total nanoparticle composition (e.g., about 0.5% to about 1.5%, or about 1.2%); and

[0149] (viii) Water, which is about 30% to about 60% by weight of the total nanoparticle composition (e.g., about 40% to about 50%, or about 46.4%).

[0150] (b) Silver dioxide, comprising about 1% to about 2% (e.g., 1.5%) of the total wound healing composition by weight, and

[0151] (c) Porous type 1 bovine collagen, comprising about 10% to about 20% (e.g., about 15%) of the total wound healing composition by weight, wherein the porous type 1 bovine collagen is processed to have a collagen structure similar to fetal skin.

[0152] In some embodiments, the wound healing composition comprises:

[0153] (a) A nanoparticle (e.g., nanoemulsion) composition comprising, by weight percentage, about 70% to about 90% (e.g., about 82%) of the total wound healing composition, wherein the nanoparticle (e.g., nanoemulsion) composition contains

[0154] (i) A perfluorocarbon component comprising PFD and PFTPA, wherein the weight ratio of PFD to PFTPA is from about 2:1 to about 3:1 (e.g., from about 2:1 to about 2.6:1, or about 2.4:1);

[0155] (ii) A lipid component comprising DSPC and DPPC, wherein the weight ratio of DSPC to DPPC is from about 1:1 to about 5:1 (e.g., from about 2:1 to about 3.5:1, or about 2.8:1); and

[0156] (iii) A buffer component comprising sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1 (e.g., from about 7:1 to about 15:1, such as about 12:1); wherein

[0157] The weight ratio of the perfluorocarbon component to the lipid component in the nanoparticle composition is from about 10:1 to about 30:1 (e.g., from about 15:1 to about 23:1, or about 23:1); and

[0158] The weight ratio of the perfluorocarbon component to the buffer component in the nanoparticle composition is from about 2.5:1 to about 5 (e.g., from about 3.5:1 to about 4.5:1, or about 3.8:1).

[0159] (b) Silver dioxide, comprising about 1% to about 2% (e.g., 1.5%) of the total wound healing composition by weight, and

[0160] (c) Porous type 1 bovine collagen, comprising about 10% to about 20% (e.g., about 15%) of the total wound healing composition by weight, wherein the porous type 1 bovine collagen is processed to have a collagen structure similar to fetal skin.

[0161] In some embodiments, this document provides a wound healing composition prepared by:

[0162] (1) Preparing a nanoparticle composition comprising about 70% to about 90% (by weight%) (e.g., about 82%) of the total wound healing composition, wherein the nanoparticle composition is prepared by mixing:

[0163] (i) PFD, which is about 25% to about 40% by weight of the total nanoparticle composition (e.g., about 30% to 40%, or about 35.5%).

[0164] (ii) PFTPA, which is about 10% to 20% by weight of the total nanoparticle composition (e.g., about 12% to 18%, or about 14.5%).

[0165] (iii) DPPC, which is about 0.1% to 1% by weight of the total nanoparticle (e.g., nanoemulsion) composition (e.g., about 0.2% to 0.8%, or about 0.6%).

[0166] (iv) DSPC, which is about 1.5% to 2.5% by weight of the total nanoparticle composition (e.g., about 1.5% to about 2%, or about 1.7%).

[0167] (v) Citric acid, in a weight percentage of about 0.01% to 0.15% of the total nanoparticle composition (e.g., about 0.05% to 0.15%, or about 0.1%).

[0168] (vi) Sodium citrate dihydrate, comprising, by weight, about 0.5% to about 2% of the total nanoparticle composition (e.g., about 0.5% to about 1.5%, or about 1.2%); and

[0169] (viii) Water, which is about 30% to about 60% by weight of the total nanoparticle composition (e.g., about 40% to about 50%, or about 46.4%).

[0170] (2) Silver dioxide is added to the nanoparticle composition, wherein the silver oxide comprises about 1% to about 2% (wt%) (e.g., 1.5%) of the total wound healing composition, and

[0171] (3) Porous type 1 bovine collagen is added to the nanoparticle composition, wherein the collagen accounts for about 10% to about 20% (by weight%) (e.g., about 15%) of the total wound healing composition to form a wound healing composition, wherein the porous type 1 bovine collagen is processed to have a collagen structure similar to fetal skin.

[0172] In some embodiments, this document provides a wound healing composition prepared by:

[0173] (1) Prepare a nanoparticle composition comprising about 70% to about 90% (by weight%) (e.g., about 82%) of the total wound healing composition, wherein the nanoparticle composition is prepared by mixing.

[0174] (i) A perfluorocarbon component comprising PFD and PFTPA, wherein the weight ratio of PFD to PFTPA is from about 2:1 to about 3:1 (e.g., from about 2:1 to about 2.6:1, or about 2.4:1);

[0175] (ii) A lipid component comprising DSPC and DPPC, wherein the weight ratio of DSPC to DPPC is from about 1:1 to about 5:1 (e.g., from about 2:1 to about 3.5:1, or about 2.8:1); and

[0176] (iii) A buffer component comprising sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1 (e.g., from about 7:1 to about 15:1, such as about 12:1); wherein

[0177] The weight ratio of the perfluorocarbon component to the lipid component in the nanoparticle composition is from about 10:1 to about 30:1 (e.g., from about 15:1 to about 23:1, or about 23:1); and

[0178] The weight ratio of the perfluorocarbon component to the buffer component in the nanoparticle composition is from about 2.5:1 to about 5 (e.g., from about 3.5:1 to about 4.5:1, or about 3.8:1).

[0179] (2) Silver dioxide is added to the nanoparticle composition, wherein the silver oxide comprises about 1% to about 2% (wt%) (e.g., 1.5%) of the total wound healing composition, and

[0180] (3) Porous type 1 bovine collagen is added to the nanoparticle composition, wherein the collagen accounts for about 10% to about 20% (by weight%) (e.g., about 15%) of the total wound healing composition to form a wound healing composition, wherein the porous type 1 bovine collagen is processed to have a collagen structure similar to fetal skin.

[0181] Enema gel

[0182] In one aspect, this document provides an enema gel comprising the nanoparticle (e.g., nanoemulsion) composition described herein and a pharmaceutically acceptable excipient. In some embodiments, the enema gel comprises the nanoparticle (e.g., nanoemulsion) composition described herein, poloxamer, and a PBS solution.

[0183] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is about 20% to about 70% (w / w) of the total enema gel, such as any one of about 20% to 60% (w / w), 25% to 55% (w / w), 25% to 50% (w / w), 35% to 50% (w / w), or 40% (w / w).

[0184] In some embodiments, poloxamer is P407 poloxamer or P188 poloxamer. In some embodiments, poloxamer is P407 poloxamer, comprising about 5% to about 50% by weight of the total enema gel, such as any one of about 10% to 40%, 15% to 35%, or 17% to 25%. In some embodiments, poloxamer is P407 poloxamer, comprising about 25% by weight of the total enema gel. In some embodiments, the PBS solution comprises about 35% of the total enema gel.

[0185] In some embodiments, poloxamer is P188 poloxamer, comprising about 5% to about 70% by weight of the total enema gel, such as any one of about 10% to 60%, 15% to 55%, or 20% to 50%. In some embodiments, poloxamer is P188 poloxamer, comprising about 50% by weight of the total enema gel. In some embodiments, the PBS solution comprises about 10% of the total enema gel.

[0186] In some implementations, the enema gel comprises:

[0187] (a) A nanoparticle (e.g., nanoemulsion) composition comprising, by weight percentage, about 30% to about 50% (e.g., about 40%) of the total enema gel, containing

[0188] (i) PFD, which is about 25% to about 40% (e.g., about 35.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0189] (ii) PFTPA, which is about 10% to 20% (e.g., about 14.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0190] (iii) DPPC, which is about 0.1% to 1% (e.g., about 0.6%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0191] (iv) DSPC, comprising about 1.5% to 2.5% (e.g., about 1.7%) by weight of the total nanoparticle (e.g., nanoemulsion) composition; and

[0192] (v) A citrate buffer (50 mM) with a pH of 6.0, comprising about 40% to 60% (e.g., about 47.7%) of the total nanoparticle (e.g., nanoemulsion) composition by weight.

[0193] (b) P407 poloxamer, comprising approximately 20% to approximately 30% (e.g., approximately 25%) of the total enema gel by weight, and

[0194] (c) PBS solution, which is about 30% to about 40% (e.g., about 35%) of the total enema gel by weight.

[0195] In some implementations, the enema gel comprises:

[0196] (a) A nanoparticle (e.g., nanoemulsion) composition comprising, by weight percentage, about 30% to about 50% (e.g., about 40%) of the total enema gel, containing

[0197] (i) PFD, which is about 25% to about 40% (e.g., about 35.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0198] (ii) PFTPA, which is about 10% to 20% (e.g., about 14.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0199] (iii) DPPC, which is about 0.1% to 1% (e.g., about 0.6%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0200] (iv) DSPC, comprising about 1.5% to 2.5% (e.g., about 1.7%) by weight of the total nanoparticle (e.g., nanoemulsion) composition; and

[0201] (v) A citrate buffer (50 mM) with a pH of 6.0, comprising about 40% to 60% (e.g., about 47.7%) of the total nanoparticle (e.g., nanoemulsion) composition by weight.

[0202] (b) P188 poloxamer, comprising approximately 40% to approximately 60% (e.g., approximately 50%) of the total enema gel by weight, and

[0203] (c) PBS solution, which is about 5% to about 15% (e.g., about 10%) of the total enema gel by weight.

[0204] In some implementations, the enema gel comprises:

[0205] (a) A nanoparticle (e.g., nanoemulsion) composition comprising, by weight percentage, about 70% to about 90% (e.g., about 82%) of the total enema gel, containing

[0206] (i) PFD, which is about 25% to about 40% by weight of the total nanoparticle composition (e.g., about 30% to 40%, or about 35.5%).

[0207] (ii) PFTPA, which is about 10% to 20% by weight of the total nanoparticle composition (e.g., about 12% to 18%, or about 14.5%).

[0208] (iii) DPPC, which is about 0.1% to 1% by weight of the total nanoparticle (e.g., nanoemulsion) composition (e.g., about 0.2% to 0.8%, or about 0.6%).

[0209] (iv) DSPC, which is about 1.5% to 2.5% by weight of the total nanoparticle composition (e.g., about 1.5% to about 2%, or about 1.7%).

[0210] (v) Citric acid, in a weight percentage of about 0.01% to 0.15% of the total nanoparticle composition (e.g., about 0.05% to 0.15%, or about 0.1%).

[0211] (vi) Sodium citrate dihydrate, comprising, by weight, about 0.5% to about 2% of the total nanoparticle composition (e.g., about 0.5% to about 1.5%, or about 1.2%); and

[0212] (viii) Water, which is about 30% to about 60% by weight of the total nanoparticle composition (e.g., about 40% to about 50%, or about 46.4%).

[0213] (b) P407 poloxamer, comprising approximately 20% to approximately 30% (e.g., approximately 25%) of the total enema gel by weight, and

[0214] (c) PBS solution, which is about 30% to about 40% (e.g., about 35%) of the total enema gel by weight.

[0215] In some implementations, the enema gel comprises:

[0216] (a) A nanoparticle (e.g., nanoemulsion) composition comprising, by weight percentage, about 70% to about 90% (e.g., about 82%) of the total enema gel, containing

[0217] (i) A perfluorocarbon component comprising PFD and PFTPA, wherein the weight ratio of PFD to PFTPA is from about 2:1 to about 3:1 (e.g., from about 2:1 to about 2.6:1, or about 2.4:1);

[0218] (ii) A lipid component comprising DSPC and DPPC, wherein the weight ratio of DSPC to DPPC is from about 1:1 to about 5:1 (e.g., from about 2:1 to about 3.5:1, or about 2.8:1); and

[0219] (iii) A buffer component comprising sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1 (e.g., from about 7:1 to about 15:1, such as about 12:1); wherein

[0220] The weight ratio of the perfluorocarbon component to the lipid component in the nanoparticle composition is from about 10:1 to about 30:1 (e.g., from about 15:1 to about 23:1, or about 23:1); and

[0221] The weight ratio of the perfluorocarbon component to the buffer component in the nanoparticle composition is from about 2.5:1 to about 5 (e.g., from about 3.5:1 to about 4.5:1, or about 3.8:1).

[0222] (b) P188 poloxamer, comprising approximately 40% to approximately 60% (e.g., approximately 50%) of the total enema gel by weight, and

[0223] (c) PBS solution, which is about 5% to about 15% (e.g., about 10%) of the total enema gel by weight.

[0224] In some embodiments, this document provides an enema gel prepared by:

[0225] (1) A nanoparticle composition is prepared, comprising about 70% to about 90% (by weight%) of the total enema gel (e.g., about 82%), wherein the nanoparticle composition is prepared by mixing.

[0226] (i) PFD, which is about 25% to about 40% by weight of the total nanoparticle composition (e.g., about 30% to 40%, or about 35.5%).

[0227] (ii) PFTPA, which is about 10% to 20% by weight of the total nanoparticle composition (e.g., about 12% to 18%, or about 14.5%).

[0228] (iii) DPPC, which is about 0.1% to 1% by weight of the total nanoparticle (e.g., nanoemulsion) composition (e.g., about 0.2% to 0.8%, or about 0.6%).

[0229] (iv) DSPC, which is about 1.5% to 2.5% by weight of the total nanoparticle composition (e.g., about 1.5% to about 2%, or about 1.7%).

[0230] (v) Citric acid, in a weight percentage of about 0.01% to 0.15% of the total nanoparticle composition (e.g., about 0.05% to 0.15%, or about 0.1%).

[0231] (vi) Sodium citrate dihydrate, comprising, by weight, about 0.5% to about 2% of the total nanoparticle composition (e.g., about 0.5% to about 1.5%, or about 1.2%); and

[0232] (viii) Water, which is about 30% to about 60% by weight of the total nanoparticle composition (e.g., about 40% to about 50%, or about 46.4%).

[0233] (b) Adding P188 poloxamer to the nanoparticle composition, wherein P188 poloxamer constitutes approximately 40% to approximately 60% (e.g., approximately 50%) by weight of the total enema gel, and

[0234] (c) Adding a PBS solution to the nanoparticle composition to form an enema gel, wherein the PBS solution accounts for about 5% to about 15% (e.g., about 10%) of the total enema gel by weight.

[0235] In some embodiments, this document provides an enema gel prepared by:

[0236] (1) A nanoparticle composition is prepared, comprising about 70% to about 90% (by weight%) of the total enema gel (e.g., about 82%), wherein the nanoparticle composition is prepared by mixing.

[0237] (i) A perfluorocarbon component comprising PFD and PFTPA, wherein the weight ratio of PFD to PFTPA is from about 2:1 to about 3:1 (e.g., from about 2:1 to about 2.6:1, or about 2.4:1);

[0238] (ii) A lipid component comprising DSPC and DPPC, wherein the weight ratio of DSPC to DPPC is from about 1:1 to about 5:1 (e.g., from about 2:1 to about 3.5:1, or about 2.8:1); and

[0239] (iii) A buffer component comprising sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1 (e.g., from about 7:1 to about 15:1, such as about 12:1); wherein

[0240] The weight ratio of the perfluorocarbon component to the lipid component in the nanoparticle composition is from about 10:1 to about 30:1 (e.g., from about 15:1 to about 23:1, or about 23:1); and

[0241] The weight ratio of the perfluorocarbon component to the buffer component in the nanoparticle composition is from about 2.5:1 to about 5 (e.g., from about 3.5:1 to about 4.5:1, or about 3.8:1).

[0242] (b) Adding P188 poloxamer to the nanoparticle composition, wherein P188 poloxamer constitutes approximately 40% to approximately 60% (e.g., approximately 50%) by weight of the total enema gel, and

[0243] (c) Adding a PBS solution to the nanoparticle composition to form an enema gel, wherein the PBS solution accounts for about 5% to about 15% (e.g., about 10%) of the total enema gel by weight.

[0244] Topical cream

[0245] In one aspect, this document provides a cream composition comprising the nanoparticle (e.g., nanoemulsion) composition described herein and a pharmaceutically acceptable excipient. In some embodiments, the excipient includes a thickener, an antioxidant, or a combination thereof.

[0246] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is provided as a weight percentage of about 20% to about 70% (w / w) of the total cream composition, such as about 20% to 70% (w / w), 25% to 55% (w / w), 25% to 50% (w / w), 40% to 60% (w / w), or 40% (w / w).

[0247] In some embodiments, pharmaceutically acceptable excipients include thickeners. In some embodiments, thickeners include petroleum jelly. In some embodiments, petroleum jelly is present in the total cream composition at about 20% to about 80% (w / w), such as any one of about 30% to 70% (w / w), 40% to 70% (w / w), 50% to 70% (w / w), 55% to 75% (w / w), or 60% (w / w). In some embodiments, thickeners include colloidal oat flakes and purified coconut oil. In some embodiments, colloidal oat flakes are present in the total cream composition at about 20% (w / w) to about 30% (w / w). In some embodiments, purified coconut oil is present in the total cream composition at about 5% to about 50%, such as any one of about 10% to 40%, 15% to 35%, or 20% (w / w) to 30% (w / w).

[0248] In some embodiments, pharmaceutically acceptable excipients include antioxidants. In some embodiments, antioxidants include colloidal chaga extract, *Fomitopsis pinnatifida* extract, or combinations thereof. In some embodiments, the antioxidant includes colloidal chaga extract, and the colloidal chaga extract is present in the form of about 5% to about 50% of the total cream composition, such as any one of about 10% to 40%, 15% to 35%, or 20% (w / w) to 30% (w / w). In some embodiments, the antioxidant includes *Fomitopsis pinnatifida* extract, and the *Fomitopsis pinnatifida* extract is present in the form of about 5% to about 50% of the total cream composition, such as any one of about 10% to 40%, 15% to 35%, or 20% (w / w) to 30% (w / w).

[0249] In some embodiments, the cream composition comprises:

[0250] (a) A nanoparticle (e.g., nanoemulsion) composition, comprising about 40% by weight of the total cream composition, containing

[0251] (i) PFD, which is about 25% to about 40% (e.g., about 35.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0252] (ii) PFTPA, which is about 10% to 20% (e.g., about 14.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0253] (iii) DPPC, which is about 0.1% to 1% (e.g., about 0.6%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0254] (iv) DSPC, comprising about 1.5% to 2.5% (e.g., about 1.7%) by weight of the total nanoparticle (e.g., nanoemulsion) composition; and

[0255] (v) a citrate buffer (50 mM) at pH 6.0, comprising approximately 40% to 60% (e.g., approximately 47.7%) by weight of the total nanoparticle (e.g., nanoemulsion) composition; and

[0256] (b) Petroleum gel, which comprises about 50% to about 70% (e.g., about 60%) of the total cream composition by weight.

[0257] In some embodiments, the cream composition comprises:

[0258] (a) A nanoparticle (e.g., nanoemulsion) composition comprising, by weight percentage, about 70% to about 90% (e.g., about 82%) of the total cream composition, containing

[0259] (i) PFD, which is about 25% to about 40% by weight of the total nanoparticle composition (e.g., about 30% to 40%, or about 35.5%).

[0260] (ii) PFTPA, which is about 10% to 20% by weight of the total nanoparticle composition (e.g., about 12% to 18%, or about 14.5%).

[0261] (iii) DPPC, which is about 0.1% to 1% by weight of the total nanoparticle (e.g., nanoemulsion) composition (e.g., about 0.2% to 0.8%, or about 0.6%).

[0262] (iv) DSPC, which is about 1.5% to 2.5% by weight of the total nanoparticle composition (e.g., about 1.5% to about 2%, or about 1.7%).

[0263] (v) Citric acid, in a weight percentage of about 0.01% to 0.15% of the total nanoparticle composition (e.g., about 0.05% to 0.15%, or about 0.1%).

[0264] (vi) Sodium citrate dihydrate, comprising, by weight, about 0.5% to about 2% of the total nanoparticle composition (e.g., about 0.5% to about 1.5%, or about 1.2%); and

[0265] (viii) Water, comprising about 30% to about 60% by weight of the total nanoparticle composition (e.g., about 40% to about 50%, or about 46.4%); and

[0266] (b) Petroleum gel, which comprises about 50% to about 70% (e.g., about 60%) of the total cream composition by weight.

[0267] In some embodiments, the cream composition comprises:

[0268] (a) A nanoparticle (e.g., nanoemulsion) composition comprising, by weight percentage, about 70% to about 90% (e.g., about 82%) of the total cream composition, containing

[0269] (i) A perfluorocarbon component comprising PFD and PFTPA, wherein the weight ratio of PFD to PFTPA is from about 2:1 to about 3:1 (e.g., from about 2:1 to about 2.6:1, or about 2.4:1);

[0270] (ii) A lipid component comprising DSPC and DPPC, wherein the weight ratio of DSPC to DPPC is from about 1:1 to about 5:1 (e.g., from about 2:1 to about 3.5:1, or about 2.8:1); and

[0271] (iii) A buffer component comprising sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1 (e.g., from about 7:1 to about 15:1, such as about 12:1); wherein

[0272] The weight ratio of the perfluorocarbon component to the lipid component in the nanoparticle composition is from about 10:1 to about 30:1 (e.g., from about 15:1 to about 23:1, or about 23:1); and

[0273] The weight ratio of the perfluorocarbon component to the buffer component in the nanoparticle composition is from about 2.5:1 to about 5 (e.g., from about 3.5:1 to about 4.5:1, or about 3.8:1); and

[0274] (b) Petroleum gel, which comprises about 50% to about 70% (e.g., about 60%) of the total cream composition by weight.

[0275] In some embodiments, the cream composition comprises:

[0276] (a) A nanoparticle (e.g., nanoemulsion) composition, comprising about 40% to about 60% by weight of the total cream composition, wherein the composition contains

[0277] (i) PFD, which is about 25% to about 40% (e.g., about 35.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0278] (ii) PFTPA, which is about 10% to 20% (e.g., about 14.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0279] (iii) DPPC, which is about 0.1% to 1% (e.g., about 0.6%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0280] (iv) DSPC, comprising about 1.5% to 2.5% (e.g., about 1.7%) by weight of the total nanoparticle (e.g., nanoemulsion) composition; and

[0281] (v) A citrate buffer (50 mM) with a pH of 6.0, comprising about 40% to 60% (e.g., about 47.7%) of the total nanoparticle (e.g., nanoemulsion) composition by weight.

[0282] (b) Colloidal oat flakes, comprising about 20% to about 30% by weight of the total cream composition, and

[0283] (c) Purified coconut oil, comprising about 20% to about 30% by weight of the total cream composition. Structure.

[0284] In some embodiments, the cream composition comprises:

[0285] (a) A nanoparticle (e.g., nanoemulsion) composition, comprising about 40% to about 60% by weight of the total cream composition, wherein the composition contains

[0286] (i) PFD, which is about 25% to about 40% by weight of the total nanoparticle composition (e.g., about 30% to 40%, or about 35.5%).

[0287] (ii) PFTPA, which is about 10% to 20% by weight of the total nanoparticle composition (e.g., about 12% to 18%, or about 14.5%).

[0288] (iii) DPPC, which is about 0.1% to 1% by weight of the total nanoparticle (e.g., nanoemulsion) composition (e.g., about 0.2% to 0.8%, or about 0.6%).

[0289] (iv) DSPC, which is about 1.5% to 2.5% by weight of the total nanoparticle composition (e.g., about 1.5% to about 2%, or about 1.7%).

[0290] (v) Citric acid, in a weight percentage of about 0.01% to 0.15% of the total nanoparticle composition (e.g., about 0.05% to 0.15%, or about 0.1%).

[0291] (vi) Sodium citrate dihydrate, comprising, by weight, about 0.5% to about 2% of the total nanoparticle composition (e.g., about 0.5% to about 1.5%, or about 1.2%); and

[0292] (viii) Water, which is about 30% to about 60% by weight of the total nanoparticle composition (e.g., about 40% to about 50%, or about 46.4%).

[0293] (b) Colloidal oat flakes, comprising about 20% to about 30% by weight of the total cream composition, and

[0294] (c) Purified coconut oil, comprising about 20% to about 30% by weight of the total cream composition. Structure.

[0295] In some embodiments, the cream composition comprises:

[0296] (a) A nanoparticle (e.g., nanoemulsion) composition, comprising about 40% to about 60% by weight of the total cream composition, wherein the composition contains

[0297] (i) A perfluorocarbon component comprising PFD and PFTPA, wherein the weight ratio of PFD to PFTPA is from about 2:1 to about 3:1 (e.g., from about 2:1 to about 2.6:1, or about 2.4:1);

[0298] (ii) A lipid component comprising DSPC and DPPC, wherein the weight ratio of DSPC to DPPC is from about 1:1 to about 5:1 (e.g., from about 2:1 to about 3.5:1, or about 2.8:1); and

[0299] (iii) A buffer component comprising sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1 (e.g., from about 7:1 to about 15:1, such as about 12:1); wherein

[0300] The weight ratio of the perfluorocarbon component to the lipid component in the nanoparticle composition is from about 10:1 to about 30:1 (e.g., from about 15:1 to about 23:1, or about 23:1); and

[0301] The weight ratio of the perfluorocarbon component to the buffer component in the nanoparticle composition is from about 2.5:1 to about 5 (e.g., from about 3.5:1 to about 4.5:1, or about 3.8:1).

[0302] (b) Colloidal oat flakes, comprising about 20% to about 30% by weight of the total cream composition, and

[0303] (c) Purified coconut oil, comprising about 20% to about 30% by weight of the total cream composition. Structure.

[0304] In some embodiments, the cream composition comprises:

[0305] (a) A nanoparticle (e.g., nanoemulsion) composition, comprising about 40% to about 60% by weight of the total cream composition, wherein the composition contains

[0306] (i) PFD, which is about 25% to about 40% (e.g., about 35.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0307] (ii) PFTPA, which is about 10% to 20% (e.g., about 14.5%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0308] (iii) DPPC, which is about 0.1% to 1% (e.g., about 0.6%) of the total nanoparticle (e.g., nanoemulsion) composition by weight;

[0309] (iv) DSPC, comprising about 1.5% to 2.5% (e.g., about 1.7%) by weight of the total nanoparticle (e.g., nanoemulsion) composition; and

[0310] (v) A citrate buffer (50 mM) with a pH of 6.0, comprising about 40% to 60% (e.g., about 47.7%) of the total nanoparticle (e.g., nanoemulsion) composition by weight.

[0311] (b) Colloidal Chaga mushroom extract, comprising about 20% to about 30% by weight of the total cream composition, and

[0312] (c) Extract of *Phellinus tiliaceus*, which is about 20% to about 30% by weight of the total cream composition.

[0313] In some embodiments, the cream composition comprises:

[0314] (a) A nanoparticle (e.g., nanoemulsion) composition, comprising about 40% to about 60% by weight of the total cream composition, wherein the composition contains

[0315] (i) PFD, which is about 25% to about 40% by weight of the total nanoparticle composition (e.g., about 30% to 40%, or about 35.5%).

[0316] (ii) PFTPA, which is about 10% to 20% by weight of the total nanoparticle composition (e.g., about 12% to 18%, or about 14.5%).

[0317] (iii) DPPC, which is about 0.1% to 1% by weight of the total nanoparticle (e.g., nanoemulsion) composition (e.g., about 0.2% to 0.8%, or about 0.6%).

[0318] (iv) DSPC, which is about 1.5% to 2.5% by weight of the total nanoparticle composition (e.g., about 1.5% to about 2%, or about 1.7%).

[0319] (v) Citric acid, in a weight percentage of about 0.01% to 0.15% of the total nanoparticle composition (e.g., about 0.05% to 0.15%, or about 0.1%).

[0320] (vi) Sodium citrate dihydrate, comprising, by weight, about 0.5% to about 2% of the total nanoparticle composition (e.g., about 0.5% to about 1.5%, or about 1.2%); and

[0321] (viii) Water, which is about 30% to about 60% by weight of the total nanoparticle composition (e.g., about 40% to about 50%, or about 46.4%).

[0322] (b) Colloidal Chaga mushroom extract, comprising about 20% to about 30% by weight of the total cream composition, and

[0323] (c) Extract of *Phellinus tiliaceus*, which is about 20% to about 30% by weight of the total cream composition.

[0324] In some embodiments, the cream composition comprises:

[0325] (a) A nanoparticle (e.g., nanoemulsion) composition, comprising about 40% to about 60% by weight of the total cream composition, wherein the composition contains

[0326] (i) A perfluorocarbon component comprising PFD and PFTPA, wherein the weight ratio of PFD to PFTPA is from about 2:1 to about 3:1 (e.g., from about 2:1 to about 2.6:1, or about 2.4:1);

[0327] (ii) A lipid component comprising DSPC and DPPC, wherein the weight ratio of DSPC to DPPC is from about 1:1 to about 5:1 (e.g., from about 2:1 to about 3.5:1, or about 2.8:1); and

[0328] (iii) A buffer component comprising sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1 (e.g., from about 7:1 to about 15:1, such as about 12:1); wherein

[0329] The weight ratio of the perfluorocarbon component to the lipid component in the nanoparticle composition is from about 10:1 to about 30:1 (e.g., from about 15:1 to about 23:1, or about 23:1); and

[0330] The weight ratio of the perfluorocarbon component to the buffer component in the nanoparticle composition is from about 2.5:1 to about 5 (e.g., from about 3.5:1 to about 4.5:1, or about 3.8:1).

[0331] (b) Colloidal Chaga mushroom extract, comprising about 20% to about 30% by weight of the total cream composition, and

[0332] (c) Extract of *Phellinus tiliaceus*, which is about 20% to about 30% by weight of the total cream composition.

[0333] IV. Treatment Methods

[0334] In one aspect, this document provides methods of using any of the compositions and formulations provided herein. In some embodiments, this document provides a method of delivering oxygen to the tissues of a subject in need, comprising administering (e.g., administering a pharmaceutically effective amount) a nanoparticle (e.g., nanoemulsion) composition to the subject. In some embodiments, the tissue is damaged tissue, such as oxygen-required tissue.

[0335] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is applied topically, i.e., applied to the surface of a tissue. In some embodiments, the advantages of topical application include, but are not limited to, avoiding the risks and inconveniences of parenteral treatment; avoiding the variability in absorption and metabolism associated with intravenous or intraperitoneal injection; and the continuity of drug administration. In some embodiments, suitable routes of topical application include, but are not limited to: buccal application, skin application, endotracheal application, enteral application, nasal application, nasogastric application, rectal application, respiratory tract application (inhalation), sublingual application, transdermal application, or transmucosal application. In some embodiments, the methods provided herein include topical application of the nanoparticle (e.g., nanoemulsion) composition described herein. In some embodiments, topical application includes applying the nanoparticle (e.g., nanoemulsion) composition topically to a specific treatment site (e.g., the digestive tract, gastrointestinal tract (“GI”), lungs, eyes, joints, or skin) to provide topical application to an area requiring treatment (e.g., oral cavity, gastrointestinal tract, lungs, colon, eyes, or skin). In some embodiments, the nanoparticle (e.g., nanoemulsion) composition can penetrate the nail and can be applied directly topically to the nail. In some embodiments, systemic exposure to the chemical entity is minimized during the local application. In some embodiments, suitable routes of local application include, but are not limited to, rectal administration, sublingual administration, transmucosal administration, skin administration, or inhalation. In some embodiments, the composition is not intended for intravenous or peritoneal injection.

[0336] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is applied in the form of a wound healing composition, enema gel, or cream composition provided herein. In some embodiments, the method includes topically applying the nanoparticle (e.g., nanoemulsion) composition to the surface of a tissue. In some embodiments, the tissue is in the skin. In some embodiments, the tissue is in the colon or gastrointestinal tract. In some embodiments, the tissue is in the lungs.

[0337] In some embodiments, the nanoparticle (e.g., nanoemulsion) composition, optionally in the form of the wound healing composition, enema gel, or cream composition provided herein, can be applied immediately as needed, such as in emergency situations. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition, optionally in the form of the wound healing composition, enema gel, or cream composition provided herein, can be applied according to a suitable schedule, such as approximately daily or approximately monthly. The dosage of the nanoparticle (e.g., nanoemulsion) composition can be determined by those skilled in the art. Furthermore, it is noteworthy that the clinician or attending physician will know how and when to interrupt, adjust, or terminate the therapy based on the individual's response. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition can be repeatedly applied until the condition to be treated is reduced or eliminated.

[0338] In some embodiments, the method treats or prevents hypoxemia in a subject in need, comprising administering (e.g., a pharmaceutically effective amount) of a nanoparticle (e.g., nanoemulsion) composition to the subject. In some embodiments, this document provides a method for treating or preventing hypoxemia-related symptoms in a subject in need, comprising administering (e.g., a pharmaceutically effective amount) of a nanoparticle (e.g., nanoemulsion) composition to the subject. In some embodiments, this document provides a method for alleviating or preventing symptoms related to hypoxemia-related symptoms in a subject in need, comprising administering (e.g., a pharmaceutically effective amount) of a nanoparticle (e.g., nanoemulsion) composition to the subject. In some implementations, conditions include, but are not limited to: acute respiratory distress syndrome (ARDS), acute lung injury, pulmonary fibrosis (idiopathic pulmonary fibrosis), mechanical ventilator-induced lung injury, acute implant dysfunction following lung transplantation and post-transplant obliterative bronchiolitis, bronchial asthma, acute bronchitis, emphysema, chronic obstructive emphysema, chronic obstructive pulmonary disease, centrilobular emphysema, panaminiferal emphysema, chronic obstructive bronchitis, smoker's disease, reactive airway disease, cystic fibrosis, black lung disease, bronchiectasis, acquired bronchiectasis, and kartaagener's disease. Symptoms include: pulmonary atelectasis, acute atelectasis, chronic atelectasis, pneumonia, idiopathic thrombocytosis, Legionnaires' disease, psittacosis, fibrotic dust disease, hypersensitivity of the lungs, idiopathic infiltrative lung disease, chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome, lung tumors, pulmonary hypertension, and diseases caused by organic dust, cyanide poisoning, nicotine, insulin, irritant gases, Alzheimer's disease, nasal diseases (such as allergic rhinitis), sinusitis and chemicals (such as cyanide, ozone), lung or sinus infections, cancer, sleep apnea, migraine, COVID, and diseases caused by chronic obstructive pulmonary disease (COPD) or asthma. In some implementations, the symptoms are COVID, chronic obstructive pulmonary disease (COPD), asthma, or lung injury.

[0339] In some embodiments, this document provides a method of treating a local condition in a subject of need, comprising administering to the subject a nanoparticle (e.g., nanoemulsion) composition described herein. In some embodiments, the nanoparticle (e.g., nanoemulsion) composition is administered in the form of a wound healing composition, enema gel, or cream composition provided herein. In some embodiments, the local condition includes a wound, open injury, burn, colonic or gastrointestinal condition, or lung condition. In some embodiments, the lung condition is COVID, chronic obstructive pulmonary disease (COPD), asthma, or lung injury. In some embodiments, the condition is ARDS. In some embodiments, ARDS is caused by a bacterial infection.

[0340] In some embodiments, this document provides a method of treating a lung condition in a subject of need, comprising administering to the subject any of the nanoparticle compositions described herein. In some embodiments, the lung condition is COVID, chronic obstructive pulmonary disease (COPD), asthma, or lung injury. In some embodiments, the lung condition is acute respiratory distress syndrome (ARDS). In some embodiments, ARDS is caused by a bacterial infection. In some embodiments, the nanoparticle composition may be nebulized prior to administration. In some embodiments, the nanoparticle composition is administered by inhalation.

[0341] In some embodiments, this document provides a method of treating a skin condition in a subject in need, comprising applying to the subject any of the nanoparticle composition, wound healing composition, or cream described herein. In some embodiments, the skin condition is a wound, open injury, or burn. In some embodiments, the nanoparticle composition is applied topically, such as topically onto the skin. In some embodiments, the nanoparticle composition may be atomized prior to application. In some embodiments, the nanoparticle composition is applied as a cream or aerosol.

[0342] In some embodiments, this document provides a method for treating a gastrointestinal condition in a subject of need, comprising administering to the subject either the nanoparticle composition described herein or an enema gel. In some embodiments, the gastrointestinal condition is a gastrointestinal wound. In some embodiments, the nanoparticle composition is applied topically, such as by applying it topically to the surface of gastrointestinal tissue.

[0343] In some embodiments, this document provides a method for treating ARDS, comprising administering to a subject any of the nanoparticle compositions provided herein. In some embodiments, the amount of the nanoparticle composition administered is effective in delaying the destruction of lung tissue in the subject, such as slowing the reduction of intact alveoli in the subject by at least 5%, such as at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 95%, 99%, or 100%. Therefore, in some embodiments, this document provides a method for delaying the destruction of lung tissue in a subject, comprising administering to the subject any of the nanoparticle compositions provided herein. In some embodiments, the subject suffers from ARDS. In some embodiments, the amount of the applied nanoparticle composition is effective in reducing alveolar structure degradation in a subject, such as by at least 5%, or by any of at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 95%, 99%, or 100%. Therefore, in some embodiments, this document provides a method for reducing alveolar structure degradation in a subject, comprising administering to the subject any of the nanoparticle compositions provided herein. In some embodiments, the subject suffers from ARDS. In some embodiments, the amount of the applied nanoparticle composition is effective in increasing CFTR expression in the lungs of the subject, such as by at least 5%, or by any of at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 95%, 99%, or 100%. In some embodiments, the amount of the applied nanoparticle composition is effective in reducing TRPV1 expression in a subject, such as by at least 5%, or by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 95%, 99%, or 100%. Therefore, in some embodiments, this document provides a method for reducing TRPV1 expression in a subject, comprising administering to the subject any of the nanoparticle compositions provided herein. In some embodiments, the subject suffers from ARDS. In some embodiments, the amount of the applied nanoparticle composition is effective in reducing COX-PTGS2 expression in a subject, such as by at least 5%, or by at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 95%, 99%, or 100%.Therefore, in some embodiments, this document provides a method for reducing COX-PTGS2 expression in a subject, comprising administering to the subject any of the nanoparticle compositions provided herein. In some embodiments, the subject suffers from ARDS. In some embodiments, the nanoparticle composition comprises (i) a perfluorocarbon component comprising PFD and PFTPA, wherein the weight ratio of PFD to PFTPA is from about 2:1 to about 3:1 (e.g., from about 2:1 to about 2.6:1, or about 2.4:1); (ii) a lipid component comprising DSPC and DPPC, wherein the weight ratio of DSPC to DPPC is from about 1:1 to about 5:1 (e.g., from about 2:1 to about 3.5:1, or about 2.8:1); and (iii) a buffer component comprising sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1 (e.g., from about 7:1 to about 15:1, such as about 12:1); wherein the weight ratio of the perfluorocarbon component to the lipid component in the nanoparticle composition is from about 10:1 to about 30:1. (e.g., about 15:1 to about 23:1, or about 23:1); and the weight ratio of the perfluorocarbon component to the buffer component in the nanoparticle composition is about 2.5:1 to about 5 (e.g., about 3.5:1 to about 4.5:1, or about 3.8:1). In some embodiments, the nanoparticle composition is contained in an atomizer. In some embodiments, the nanoparticle composition contains no more than about 2% (w / w) of a polymeric surfactant of the total nanoparticle composition. In some embodiments, the nanoparticle composition has a viscosity of no more than about 100 cp. In some embodiments, the average particle size of the nanoparticles is no more than about 3 μm.

[0344] In some embodiments, this document provides a method for restoring oxyhemoglobin in the tissues of a subject, comprising administering to the subject any of the nanoparticle compositions provided herein. In some embodiments, the nanoparticle composition is applied topically. In some embodiments, the nanoparticle composition can penetrate the skin. In some embodiments, the amount of nanoparticle composition applied is sufficient to effectively restore at least 5% of oxyhemoglobin in the tissue (e.g., an increase in the amount of oxyhemoglobin of at least about 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 95%, 99%, or 100%). In some embodiments, the amount of oxyhemoglobin can be determined by blood oxygenation methods, such as pulse oximetry. In some embodiments, the nanoparticle composition comprises (i) a perfluorocarbon component comprising PFD and PFTPA, wherein the weight ratio of PFD to PFTPA is from about 2:1 to about 3:1 (e.g., from about 2:1 to about 2.6:1, or about 2.4:1); (ii) a lipid component comprising DSPC and DPPC, wherein the weight ratio of DSPC to DPPC is from about 1:1 to about 5:1 (e.g., from about 2:1 to about 3.5:1, or about 2.8:1); and (iii) a buffer component comprising sodium citrate hydrate and citric acid, wherein the weight ratio of sodium citrate hydrate to citric acid is from about 5:1 to about 20:1. (e.g., about 7:1 to about 15:1, such as about 12:1); wherein the weight ratio of the perfluorocarbon component to the lipid component in the nanoparticle composition is about 10:1 to about 30:1 (e.g., about 15:1 to about 23:1, or about 23:1); and the weight ratio of the perfluorocarbon component to the buffer component in the nanoparticle composition is about 2.5:1 to about 5 (e.g., about 3.5:1 to about 4.5:1, or about 3.8:1). In some embodiments, the nanoparticle composition is contained in an atomizer. In some embodiments, the nanoparticle composition is contained in a gel composition. In some embodiments, the nanoparticle composition is contained in a cream composition. In some embodiments, the nanoparticle composition contains no more than about 2% (w / w) of a polymeric surfactant of the total nanoparticle composition. In some embodiments, the nanoparticle composition has a viscosity of no more than about 100 cp. In some embodiments, the average particle size of the nanoparticles is no more than about 3 μm.

[0345] V. Medicine box

[0346] Articles and / or kits containing any of the nanoparticle (e.g., nanoemulsion) compositions described herein are also provided. In some embodiments, the articles and / or kits contain any of the wound healing compositions, enema gels, or cream compositions provided herein. In some embodiments, the articles may include a labeled container. In some embodiments, suitable containers include, for example, bottles, vials, and test tubes. Containers may be formed from a variety of materials, such as glass or plastic. In some embodiments, the container may contain the nanoparticle (e.g., nanoemulsion) compositions provided herein. In some embodiments, the container is a nebulizer. In some embodiments, the nebulizer contains the nanoparticle (e.g., nanoemulsion) compositions described herein and may atomize the nanoparticle (e.g., nanoemulsion) compositions prior to use. In some embodiments, a label on the container may indicate that the pharmaceutical composition is for the prevention, treatment, or suppression of the conditions described herein, and may also indicate instructions for use.

[0347] In one aspect, this document provides a kit containing any of the nanoparticle (e.g., nanoemulsion) compositions described herein and instructions for use. The kit may contain instructions for use to treat any of the diseases or conditions described herein for an individual in need. The kit may additionally contain any materials or devices that can be used to administer the compound or composition, such as a nebulizer.

[0348] Example

[0349] The subject matter disclosed herein will be better understood by referring to the following embodiments, which are provided as examples of the invention and not as limitations.

[0350] Example 1. Method for preparing emulsions by rotary evaporation

[0351] NF-1

[0352] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC) and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) were added to ethanol, and the mixture was stirred until the solution became clear. The solution was then transferred to a round-bottom flask (RBF) and rotary evaporated at 40°C to evaporate the ethanol. The mixture was then kept in a vacuum oven overnight to remove the remaining ethanol at room temperature. PFD and PFTA were added to the membrane, and the membrane was subjected to continuous acoustic treatment for approximately 30 minutes to obtain NF-1. Table 1 summarizes the amounts of each component.

[0353] Example 2. Method for preparing emulsions by homogenization

[0354] NF-2, NF-3, NF-4

[0355] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC) and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) were added to ethanol and stirred until the solution became clear. A mixture of PFD and PFTA was then slowly added to the above mixture, and homogenized at 3.4 rpm for 10 minutes. After homogenization, the ethanol was removed from the mixture, and the vials containing the lipid and perfluorocarbon mixture were redispersed with water to form NF2, NF-3, and NF-4. Table 1 summarizes the quantities of each component.

[0356] Example 3. Method for preparing emulsions via probe acoustic processing

[0357] NF-5, NF-5A, NF-6, NF-7

[0358] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC) and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) were added to ethanol, and the mixture was stirred until the solution became clear. A portion of the non-lipid surfactant (if any) was added, and the mixture was stirred until clear. This solution was added to a mixture of PFD and PFTA, and the mixture was then probe-treated with acoustic waves at an amplitude of 40 for approximately 10 minutes. After the probe-treatment, the mixture was slowly added to water containing the remaining non-lipid surfactant (if any) to form NF-5, NF-5A, NF-6, and NF-7. Table 1 summarizes the amounts of each component.

[0359] NF-8, NF-9, NF-10

[0360] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC) and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), along with a non-lipid surfactant (if present), were mixed with a perfluorocarbon (PFD+PFTA) mixture, and the mixture was then probe-treated for approximately 5 minutes. Water was slowly added to the above mixture, and the new mixture was probe-treated at an amplitude of 20 for approximately 15 minutes to obtain NF-8, NF-9, and NF-10. Table 1 summarizes the quantities of each component.

[0361] NF-11, NF-12, NF-13

[0362] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC) and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), along with a non-lipid surfactant, were mixed with an API (PFD+PFTA) mixture, and the new mixture was then probe-treated for approximately 5 minutes. Water was slowly added to the mixture, and the mixture was probe-treated at an amplitude of 20 for approximately 15 minutes to obtain NF-11, NF-12, and NF-13. Table 1 summarizes the quantities of each component.

[0363] Example 4. Method for preparing emulsions by homogenization and probe acoustic treatment

[0364] NF-14, NF-16, NF-17

[0365] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), and a non-lipid surfactant (if present) were added to ethanol, and the mixture was stirred until the solution became clear. A mixture of PFD and PFTA was then slowly added to the above mixture, and the resulting mixture was homogenized at 3.4 rpm for 10 minutes. After homogenization, the ethanol was removed from the mixture, and the remaining lipid and perfluorocarbon mixture was redispersed with water. The resulting solution was subjected to probe acoustic treatment at an amplitude of 20 for approximately 15 minutes. Table 1 summarizes the quantities of each component.

[0366] Example 5. Method for preparing emulsions by homogenization

[0367] NF-15

[0368] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), and a non-lipid surfactant (if present) were added to ethanol, and the mixture was stirred until the solution became clear. After homogenization, a mixture of PFD and PFTA was slowly added to the above mixture. Water was added to the above mixture, and homogenization was carried out at 8.4 rpm for 30 minutes. The resulting solution was kept at room temperature overnight, and the ethanol was removed to obtain NF-15. Table 1 summarizes the amount of each component.

[0369] Example 6. A method for preparing emulsions by homogenization (e.g., probe homogenization) and rotary evaporation.

[0370] NF-18

[0371] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), and a non-lipid surfactant (if present) were added to ethanol, and the mixture was stirred until the solution became clear. After homogenization, a mixture of PFD and PFTA was slowly added to the above mixture. The above mixture was transferred to an RBF for ethanol evaporation. The RBF containing the lipid and perfluorocarbon mixture was redispersed with water, and the resulting solution was subjected to probe acoustic treatment at an amplitude of 20 for approximately 10 minutes. Table 1 summarizes the quantities of each component.

[0372] Example 7. Method for preparing emulsions by direct addition

[0373] NF-19, NF-20, NF-24

[0374] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC) and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), along with a non-lipid surfactant (if present), were mixed with a perfluorocarbon (PFD+PFTA) mixture, and the resulting solution was then probe-treated for approximately 5 minutes. Water was slowly added to the mixture, and the new mixture was probe-treated at an amplitude of 40 for approximately 15 minutes to obtain NF-19, NF-20, and NF-24. Table 1 summarizes the quantities of each component.

[0375] Example 8. Method for preparing emulsions via thin-film hydration

[0376] NF-21, NF-22, NF-23

[0377] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), and a non-lipid surfactant (if present) were added to ethanol, and the mixture was stirred until clear. As the film formed, the solution was transferred to an RBF and evaporated. The film was then hydrated with water. A mixture of PFD and PFTA was slowly added to the above mixture, and the probe was acoustically treated at 20 amplitude for 10 minutes to obtain NF-21, NF-22, and NF-23. Table 1 summarizes the quantities of each component.

[0378] NF-25

[0379] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), and HSPC were added to ethanol, and the solution was stirred until clear. The solution was transferred to RBF and rotary evaporated to form a film. The film was then hydrated with water. A mixture of PFD and PFTA was slowly added to the above mixture, and the probe was acoustically treated at 20 amplitude for 10 minutes to obtain NF-25. Table 1 summarizes the amount of each component.

[0380] NF-26, NF-27, NF-28, NF-29, NF-30

[0381] 1,2-Dipalmitoyl-sn-glycerol-3-phosphate choline (DPPC), 1,2-distearatel-sn-glycerol-3-phosphate choline (DSPC), and a non-lipid surfactant (e.g., cetrimonium bromide (CTAB), if present) were added to ethanol, and the solution was stirred until clear. As the film formed, the solution was transferred to an RBF and evaporated. The film was then hydrated with water. A mixture of PFD and PFTA was slowly added to the above mixture, and the mixture was subjected to probe acoustic treatment at 20 amplitude for 10 minutes to obtain the title composition. Table 1 summarizes the quantities of each component.

[0382] Example 9. Method for preparing emulsions by direct addition

[0383] NF-31, NF-3, NF-33, NF-34, NF-35, NF-36,

[0384] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC) and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) were mixed with PFD and PFTA, and the mixture was then subjected to probe acoustic treatment for approximately 2 minutes. Water or an aqueous buffer and a non-lipid surfactant (e.g., cetrimonium bromide (CTAB), Tween 80, if available) were slowly added to the above mixture, and the resulting solution was subjected to probe acoustic treatment at an amplitude of 20 (e.g., at 45°C) for 8 minutes to obtain the title composition. Table 1 summarizes the quantities of each component.

[0385] Example 10. Method for preparing emulsions via thin-film hydration

[0386] NF-44

[0387] 1,2-Dipalmitoyl-sn-glycerol-3-phosphate choline (DPPC), 1,2-distearatel-sn-glycerol-3-phosphate choline (DSPC), and a non-lipid surfactant (e.g., Tween 80, if available) were added to ethanol, and the solution was stirred until clear. As the film formed, the solution was transferred to an RBF and evaporated. The film was then hydrated with water. A mixture of PFD and PFTA was slowly added to the above mixture, and the mixture was subjected to probe acoustic treatment at 20 amplitude for 10 minutes to obtain the title composition. Table 1 summarizes the quantities of each component.

[0388] NF-45

[0389] 1,2-Dipalmitoyl-sn-glycerol-3-phosphate choline (DPPC) and 1,2-distearatel-sn-glycerol-3-phosphate choline (DSPC) were added to ethanol, and the solution was stirred until clear. The solution was transferred to an RBF and rotary evaporated to form a film. The film was then hydrated with the desired amount of water and a surfactant (e.g., soy lecithin, if available). A mixture of PFD and PFTA was slowly added to the above mixture, and the mixture was subjected to probe acoustic treatment at 20 amplitude for 10 minutes to obtain the title composition. Table 1 summarizes the quantities of each component.

[0390] Example 11. Method for preparing emulsions by direct addition

[0391] NF-46

[0392] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC) and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) were mixed with perfluorocarbon (PFD+PFTA), and the mixture was vortexed. Water was slowly added to the mixture, and the resulting mixture was probe-treated at an amplitude of 20 for 10 minutes to obtain NF-46. Table 1 summarizes the quantities of each component.

[0393] NF-47, NF-48

[0394] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC) and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) were mixed with water, and the mixture was then vortexed. PFD and PFTA were slowly added to the mixture, and the resulting mixture was probe-treated at an amplitude of 20 for 10 minutes. Table 1 summarizes the quantities of each component.

[0395] Example 12. Method for preparing emulsions via thin-film hydration

[0396] NF-49, NF-50, NF-51, NF-53 and NF-54

[0397] Weigh out amounts of 1,2-dipalmitoyl-sn-glycerol-3-phosphate choline (DPPC), 1,2-distearatel-sn-glycerol-3-phosphate choline (DSPC), and HSPC (if present) and add them to ethanol, stirring the mixture until it becomes clear. Transfer the resulting solution to RBF and rotary evaporate it to form a film. Then hydrate the film with water. Slowly add a mixture of PFD and PFTA to the above mixture and perform probe acoustic treatment at 20 amplitude for 10 minutes. Table 1 summarizes the amounts of each component.

[0398] NF-52

[0399] 1,2-Dipalmitoyl-sn-glycerol-3-phosphate choline (DPPC) and 1,2-distearatel-sn-glycerol-3-phosphate choline (DSPC) were added to ethanol, and the mixture was stirred until clear. The solution was transferred to an RBF and rotary evaporated to form a thin film. The film was then hydrated with water by tank acoustic treatment at 65°C. PFD and PFTA were slowly added to the mixture, and probe acoustic treatment was performed at 20 amplitude for 10 minutes. Table 1 summarizes the quantities of each component.

[0400] Example 13. Method for preparing emulsions by direct addition

[0401] NF-54

[0402] 1,2-Distearate-sn-glycerol-3-phosphate choline (DSPC), 1,2-dipalmitoyl-sn-glycerol-3-phosphate choline (DPPC), and La-hydrogenated phosphatidylcholine (soybean) (HSPC) were mixed and vortexed. The desired amount of perfluorocarbon (PFD+PFTA) mixture was slowly added to the above mixture, and the probe was acoustically treated at 20 amplitude for 10 minutes. Table 1 summarizes the amount of each component.

[0403] Example 14. Method for preparing emulsions by direct addition using a microfluidic analyzer

[0404] NF-55, NF-57

[0405] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC), HSPC, and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) were mixed with PFD and PFTA, and the mixture was vortexed. Water was slowly added to the mixture and vortexed, then the mixture was microfluidized for two cycles at 25,000 PSI to obtain NF-55. Table 1 summarizes the quantities of each component.

[0406] NF-56

[0407] 1,2-Distearayoyl-sn-glycerol-3-phosphocholine (DSPC), HSPC, and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) were mixed with PFD and PFTA and vortexed. Water and a non-lipid surfactant (e.g., soy lecithin) were slowly added to the mixture, and microfluidization was performed at 25,000 PSI for 2 and 4 cycles. Table 1 summarizes the quantities of each component.

[0408] NF-58

[0409] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC), HSPC, and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) were mixed with PFD and PFTA and top-stirred at 3.5 rpm. Water was slowly added to the mixture and the solution was stirred for 30 minutes. The resulting mixture was then microfluidized at 25,000 PSI for 2, 3, and / or 4 cycles. Table 1 summarizes the quantities of each component.

[0410] NF-59

[0411] 1,2-Distearatel-sn-glycerol-3-phosphocholine (DSPC) and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) were mixed with PFD+PFTA. The resulting mixture was homogenized at 3.5 rpm for 1 minute. Water was then slowly added to the mixture, and homogenization was performed at 3.4 rpm for 19 minutes. Table 1 summarizes the quantities of each component.

[0412] NF-60 to NF-79

[0413] 1,2-Distearyl-sn-glycerol-3-phosphocholine (DSPC) and 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) were mixed with PFD and PFTA, and the mixture was homogenized for 1 minute. Water or a buffer was added to the mixture, and homogenization was carried out at 3.4 rpm for 1 minute. The resulting mixture was then microfluidized at 25,000 PSI for 3, 4, or 5 cycles to obtain the title composition. Optionally, the microfluidized mixture was cooled between cycles. The composition was filled into Type 1 glass vials fitted with rubber stoppers and sealed. The mixture was autoclaved at 121°C for 15 minutes. Sterilize the vials with psi for 20 minutes. Table 1 summarizes the quantity of each component.

[0414] NF-74

[0415] PFD and PFTPA were added to the manufacturing vessel. 1,2-Dipalmitoyl-sn-glycerol-3-phosphate choline (DPPC) and (1,2-distearatel-sn-glycerol-3-phosphate choline) (DSPC) were added to the perfluorocarbon mixture at 3.5 rpm using a probe homogenizer. A buffer was added to the above solution, and the mixture was stirred at 3.5 rpm using a probe homogenizer to obtain a homogenized solution. The solution was then microfluidized at 25,000 psi for 3 cycles at room temperature. The obtained bulk solution was filled into 5.0 mL amber glass vials and sealed with coated rubber stoppers. The bulk solution in the vials was terminally sterilized at 121°C for 20 minutes.

[0416] NF-83

[0417] PFD and PFTPA were added to a stainless steel container. 1,2-Dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC) and (1,2-distearate-sn-glycerol-3-phosphocholine) (DSPC) were added to perfluorocarbon and homogenized at 3.5 rpm for 2 minutes. A buffer was added to the solution, and the mixture was stirred at 3.5 rpm to obtain a homogenized solution until no further phase separation occurred. The solution was then microfluidized at 25,000 psi for 3 cycles. The resulting bulk solution was filled into clear glass vials and sealed with coated rubber stoppers. The solution was then autoclaved at 121°C for 15 minutes. Sterilize the vials with psi for 20 minutes.

[0418] For NF-83, the ratios between the various components are summarized in Table A below:

[0419] Table A

[0420] Ingredient Concentration (g / g) Perfluoronaphthalene (PFD) 0.355 Perfluorotri-n-propylamine (PFTPA) 0.145 Distearoylphosphatidylcholine (DSPC) 0.006 Dipalmitoylphosphatidylcholine (DPPC) 0.017 Sodium citrate dihydrate 0.012 Citric acid 0.001 Water 0.464

[0421]

[0422]

[0423]

[0424]

[0425]

[0426]

[0427]

[0428]

[0429]

[0430] Example 15. Characterization of the prepared nanoparticle composition

[0431] The phase stability, pH, viscosity, dissolved oxygen, particle size, PDI, and Zeta potential of the nanoparticle compositions prepared in Examples 1 through 14 were further tested. Table 2 summarizes the observations on the phase stability of each nanoparticle composition.

[0432] Particle size and zeta potential measurement

[0433] The particle size, polydispersity index, and surface potential of the prepared nanoparticle composition were investigated using dynamic light scattering (FLSR). The particle size was determined using a measuring dish (10 mm in diameter) at 25 °C. Prior to the size measurement studies, the prepared nanoparticle composition was diluted (1:5) with a citrate buffer at pH 6.0 to ensure free Brownian motion of the droplets.

[0434] Viscosity measurement

[0435] The viscosity of the prepared nanoparticle compositions was determined at 25 ± 0.5 °C using a Brookfield cone-plate rheometer (manufacturer: Brookfield Engineering Laboratory, model: DV2T) with a rotor # CPE40 at a constant speed of 80 rpm. The software used for calculations was Rheocalc V2.6. All experiments were performed three times.

[0436] pH measurement

[0437] The pH of the prepared nanoparticle composition was determined using a Mettler Toledo pH meter. The device was calibrated with buffer solutions at pH 4.00 ± 0.05, pH 7.00 ± 0, and pH 10.00 ± 0.05 prior to measurement. pH measurements were performed three times at room temperature.

[0438] osmolar concentration

[0439] The osmolality was determined using a osmometer (manufacturer: Vapro, model: 5600) via freezing point measurement technology. The instrument was calibrated using a standardized solution from the osmometer supplier. A 100 μL sample of the evaluated nanoparticles was placed in a dedicated microtube and then inserted into the measurement head.

[0440] Cryo-TEM

[0441] The shape, size, and integrity of nanoparticles in their diluted state were accurately characterized using Cryo-TEM by freezing samples. Nanoparticles were diluted with lung fluid at a 1:5 ratio to minimize droplet overlap, enabling subsequent particle size measurements. A current of 25 mA / 50 s was applied to a gold grid with a 300-mesh Lacey-type carbon film. The grid was then loaded into a Vitrobot Mark IV sample vitrification robot. Samples diluted with lung fluid at 1:5 and 1:10 ratios were placed into the grid, and excess sample was removed using an 8.5 s duration and -15 force imprint. The grid was immediately frozen in liquid ethane and preserved in liquid nitrogen until microscopic examination. Images were captured using a Talos Arctica TEM model operating at 200 kV. The microscope was equipped with a camera for acquiring digital images.

[0442] Stability of nanoparticle compositions

[0443] Accelerated stability testing was performed on the prepared compositions in accordance with the recommendations of the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH). The perfluorocarbon-loaded compositions were stored for 24 weeks under long-term and accelerated conditions (25°C / 60%RH and 40°C / 75%RH), with observations at weeks 0, 4, 8, 12, and 24. The physicochemical stability of the nanoparticle compositions was tested by observing changes in appearance, pH, viscosity, osmolality, phase separation, particle size, polydispersity index, zeta potential, determination, and purity.

[0444] The conditions used for stability testing are:

[0445] Long-term (25℃ ± 2℃ / 60% RH ± 5% RH) for 3 months and 6 months

[0446] Accelerated (40℃ ± 2℃ / 75% RH ± 5% RH) for 1 month, 2 months, 3 months and 6 months.

[0447] Analysis of perfluorocarbons in nanoparticle compositions

[0448] Analysis of perfluorocarbons in the nanoparticle composition was performed using a gas chromatograph with the following specifications, on a gas chromatograph (manufacturer: Shimadzu, model: Nexis 2030) coupled with a flame ionization detector (FID).

[0449] The results are summarized in Table 2.

[0450] Table 2

[0451] Composition NF1 NF2 NF3 NF4 NF5 NF6 NF7 NF8 NF9 NF10 Observations ++++ + +++ + +++ + ++++ + + + Composition NF11 NF12 NF13 NF14 NF15 NF16 NF17 NF18 NF19 NF20 Observations + + + + + +++ + + + ++ Composition NF21 NF22 NF23 NF24 NF25 NF26 NF27 NF28 NF29 NF30 Observations ++ ++ + + + + + + + +++ Composition NF31 NF32 NF52 NF53 NF54 NF55 NF56 NF57 NF60 NF61 (100 gm) Observations +++ +++ + + + +++ ++++ +++ +++ ++++ Composition NF63 (60g) NF64 (67.27g) NF65 (~135 gm) NF67 (110 gm) NF68 (~260 gm) NF69 (50gm) NF70 (50gm) NF71 (50gm) NF72 (50gm) NF73(150 gm) Observations ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ Composition NF74 (50gm) NF75 (50gm) NF76 (100gm) NF77(100 gm) Observations ++++ ++++ ++++ ++++

[0452] + Phase separation occurs on day 1; ++ Phase separation occurs between day 1 and day 4; +++ Phase separation does not occur until day 4; ++++ Phase separation does not occur until day 10.

[0453] As shown in Table 2, nanoparticle compositions containing cetrimonium bromide (CTAB) (such as NF1 to NF8), nanoparticle compositions containing Tween 80 (such as NF9 to NF19), nanoparticle compositions containing soy lecithin (such as NF14 to NF16, NF19 to NF23, NF31, and NF56), and nanoparticle compositions containing soy phosphatidylcholine (HSPC) (such as NF28 to NF30, NF54, and NF55) did not show significantly improved physical stability. Some phase separation was observed in these nanoparticle compositions. Surprisingly, nanoparticle compositions containing DSPC and DPPC but not CTAB, Tween 80, soy lecithin, or HSPC tended to have better stability and longer pre-phase separation days. Surprisingly, only DSPC / DPPC could form physically stable nanoparticle compositions with perfluorocarbons in the absence of additional surfactants or lipids.

[0454] By comparing NF61 (containing a phosphate buffer) with NF65 through NF79 (all containing citrate buffers), it was unexpectedly found that citrate buffers were significantly less toxic than phosphate buffers and often yielded more stable nanoparticle compositions. Typical cellular concentrations of citrate are quite high, therefore animals generally exhibit high tolerance to citrate buffers.

[0455] Table 3 summarizes other characterization results for representative nanoparticle compositions, including pH, viscosity, dissolved oxygen, particle size, PDI, and Zeta potential.

[0456] Table 3

[0457] NF1 NF2 NF7 NF11 NF30 NF31 NF32 NF55 NF56 pH 5.86 4.72 5.96 7.63 6.58 3.78 7.63 6.58 Osmolality 23 36 13 23 36 Viscosity 1767 cP 1144 cP 40 20 13 40 20 Dissolved Oxygen 8.7 ppm Particle Size (nm) 3256 2108 1219 3256 2108 PDI 1 1 0.678 1.0 1.0 Zeta Potential (mV) 0.598 0.0233 0.677 0.598 0.0233

[0458] Table 3 (continued)

[0459] Composition NF57 NF60 NF61 (100gm) NF62 (60gm) NF63(60g) NF64(67.27g) NF65 (~135gm) NF66 (~260gm) NF67 (110gm) pH 3.78 6.14 6.73 6.29 6.29 6.05 5.94 5.97 5.92 Osmolality 13 365 22 22 160 237 238 261 Viscosity 13 522.5 304.5 304.5 17.0 16.0 17.0 17.8 Dissolved Oxygen NA NA NA NA 8.638 8.299 7.643 Particle Size (nm) 1219 2096 1291 1318 943 Particle Size (nm) + 5x Citrate 309 296 361 320 341 PDI 0.678 0.577 Zeta Potential (mV) 0.677 0.123

[0460] Table 3 (continued)

[0461] NF68 (~260gm) NF69 (50gm) NF70(50 gm) NF71 (50gm) NF72 (50gm) NF73(150 gm) NF74(50 gm) NF78 NF79 pH 5.94 5.90 5.49 6.07 6.07 5.99 6.03 6.05 6.29 Osmolality 297 226 332 247 244 314 264 345 285 Viscosity 26.32 6.9 21.7 14.7 7.0 23.4 9.77 9.5 3.0 Dissolved Oxygen 8.46 8.692 8.794 9.789 9.713 8.947 8.952 9.085 9.179 Particle Size (nm) 1898 738 Particle Size (nm) + 5x Citrate 323 295 309 341 269 322 280 257 273

[0462] As shown in Table 3, surprisingly, the particle size of the nanoparticles in the nanoparticle composition can be reduced by increasing the number of passes in the microfluidization process. Further dilution of the prepared nanoparticle composition can further reduce the particle size to approximately 300 nm to approximately 400 nm. Also surprisingly, increasing the amount of water or aqueous buffer in the nanoparticle composition can reduce the viscosity of the nanoparticle composition. This lower viscosity and smaller particle size are generally preferred for inhalable formulations and are suitable for atomization processes. As shown in Table 3, the stable nanoparticle compositions have a pH range of approximately 5 to approximately 7.5. Measured only in aqueous layers, these compositions typically contain approximately 7 ppm to approximately 10 ppm of dissolved oxygen and exhibit a viscosity of approximately 3 cp to approximately 500 cp.

[0463] Figure 2 shows CryoEM images of representative nanoparticles. The images were captured from different grids diluted 1:5 with lung fluid. As shown in Figure 2, the nanoparticles have a particle size of less than 0.5 μm and exhibit a multi-lamellar structure.

[0464] In summary, two different methods were used to prepare nanoparticle compositions. These required the use of microfluidics, probe acoustic generators, probe homogenizers, thin-film hydration techniques, and various types and concentrations of emulsifiers. Variations affecting emulsion stability (such as particle size and dispersibility) are key characteristics of the fabricated nanoparticle compositions. Various techniques for preparing stable nanoparticle composition systems containing / without surfactants were independently investigated in this study. However, the use of non-lipid surfactants has limitations, potentially contaminating the product and adversely affecting its biocompatibility. To address this issue, surfactant-free nanoemulsification methods have been developed. Nanoparticle compositions can be formed in the absence of surfactants using high-shear-energy-based microfluidics.

[0465] Morphological examination of the developed nanoparticle compositions was performed using Cryo TEM (a technique that captures simultaneous structural and microstructural transitions through high-resolution imaging). Cryo TEM images of NF-66 show that the droplets are spherical and uniform in size (Figure 3B). These nanoparticle compositions were found to be stable for storage at 25°C / 60%RH for up to 6 months.

[0466] The nanoparticle composition NF-62 exhibits high viscosity in the presence of water as the continuous phase. As seen in the Cryo-TEM image (Figure 3A), the Ostwald ripening effect in this preparation causes smaller droplets to shrink or disappear, while larger droplets further enlarge, leading to phase separation on day 28. NF-66 (Figure 3B), prepared with more PFC and citrate buffer, shows even smaller sizes (compared to Figure 3A). DLS data show a slight increase in the hydrodynamic size of NF-66 to 2.2 μm, with a PDI of 0.6. After dilution with citrate buffer at a 1:5 ratio, the average particle size decreased to 320 nm, and the PDI decreased to 0.170.

[0467] Example 16. Efficacy of the exemplary composition in treating acute respiratory distress syndrome (ARDS)

[0468] Inducing ARDS with lipopolysaccharide (LPS)

[0469] Under sedation, mice were intratracheally administered LPS (BMS-345541) at a dose of 1 mg / kg body weight. Briefly, mice were anesthetized with isoflurane, placed in a supine position, and their upper incisors were secured with a suture bundle. Endotracheal intubation was then performed using a gel-loaded pipette tip (plastic / polypropylene). 25 μL of sterile saline solution was introduced into the pipette tip to check for proper placement in the trachea. Slow up-and-down movement of the solution confirmed proper placement. The saline solution was carefully removed using a pipette.

[0470] LPS 1 mg / mL stock solution was diluted with sterile saline and administered via intratracheal instillation according to mouse body weight. Mice weighing 25 g received a mixture of 25 μL LPS and 75 μL sterile saline solution premixed in a test tube. This solution was then carefully introduced into a pipette tip inserted into the trachea. A 200 μL pipette was then used to gently push the solution into the trachea using positive pressure. All mice received a total volume of 100 μL of the LPS and saline mixture, ensuring the same total volume for all animals. Twelve mice were intratracheally instilled with LPS and allowed to recover from anesthesia, and were monitored for adverse signs for 24 hours. One mouse died after the LPS instillation, and the remaining 11 mice were divided into two groups as follows:

[0471] i) Six LPS-infused mice (i.e., diseased mice) received 100 μL of the nanoparticle composition NF-83.

[0472] ii) Five LPS-infused mice (i.e., diseased mice) received sterile saline solution – 100 μL

[0473] Mice received three treatment cycles, 48 ​​hours apart. Forty-eight hours after the last treatment, the mice were euthanized, and bilateral lung tissue was harvested. The lung tissue was evenly distributed for immunohistochemical analysis and Western blotting. For immunohistochemical analysis, the tissue was fixed with 4% paraformaldehyde and cryoprotected with 30% sucrose. For Western blotting analysis, the lung tissue was frozen on dry ice and stored at -80°C.

[0474] result

[0475] The results are shown in Figures 5A (treated with the NF-83 nanoparticle composition) and 5B (treated with sterile saline solution). In Figures 5A and 5B, the bright areas are cross-sections of the alveoli. As shown in Figure 5A, mice treated with the exemplary nanoparticle composition provided herein exhibited more intact alveoli and less epithelial cell damage, while as shown in Figure 5B, untreated mice exhibited disordered and / or collapsed alveoli and epithelial damage. These results demonstrate the role of the exemplary nanoparticle composition in delaying or reversing lung injury and / or lung tissue destruction and in the potential treatment of ARDS.

[0476] Treating mice with ARDS caused by bacterial infection

[0477] Based on promising early results, this treatment regimen was repeated in another group of mice with bacterial infection-induced ARDS. The treatment group received the exemplary nanoparticle composition NF-83 on days 3 and 5, as shown in the LPS group. Based on lung histology, treatment with the exemplary nanoparticle composition NF-83 significantly reduced alveolar structural degradation in both infected and treated mice compared to untreated infected mice. The results are shown in Figure 6.

[0478] Biomarker expression

[0479] As shown in Figure 9A, the exemplary nanoparticle composition NF-83 also increased CFTR1 expression in LPS-treated mice. While CFTR is primarily associated with cystic fibrosis (CF), a genetic condition affecting the lungs and other organs, recent studies have shown that CFTR may also play a role in the pathophysiology of ARDS. Studies have demonstrated that increased CFTR expression in the lungs can have a protective effect against ARDS. These results demonstrate the ability of NF-83 to target the CFTR signaling pathway and its potential as a therapeutic strategy for ARDS.

[0480] Furthermore, as shown in Figure 9B, the exemplary nanoparticle composition NF-83 reduced TRPV1 expression in LPS-treated mice. TRPV1 (transient receptor potential vanillic acid 1) is a member of the transient receptor potential (TRP) family of ion channels, which is expressed in multiple tissues, including the lungs. Studies have shown that inhibition of TRPV1 reduces lung inflammation and damage. TRPV1 also plays a role in the pathophysiology of acute respiratory distress syndrome (ARDS), a life-threatening condition characterized by severe pneumonia and impaired oxygenation. Studies have found that TRPV1 is expressed in lung tissue, and its activation may contribute to lung inflammation and damage. TRPV1 is also associated with the development of pulmonary edema, a hallmark of ARDS characterized by fluid accumulation in the lungs.

[0481] In animal models of ARDS, blocking TRPV1 has been shown to reduce pneumonia and improve oxygenation, suggesting that TRPV1 antagonists may have the potential to serve as therapeutic targets for ARDS. NF-83, which reduces TRPV1 in animals with ARDS, has also demonstrated promising therapeutic potential for ARDS.

[0482] Furthermore, NF-83 treatment was observed to suppress hypoxia-related inflammatory signals. Elevated COX-PTGS2 is a marker of hypoxia-related inflammation. Studies of lung tissue from control, infected, and treated mice showed that NF-83 treatment significantly inhibited COX-PTGS2 expression, indicating its oxygenation function (Figures 9C and 9D).

[0483] Example 17. Dynamics of nanoparticles driven by the tidal breathing pattern of the lungs.

[0484] To evaluate the shuttle function of NF-83 in oxygen delivery and carbon dioxide removal, we used the StimuLung chip lung model, where oxygen flow and CO2 absorption were measured using a fluorescent dye, simulating the exposure of nanoparticles in the nanoparticle composition to air and to deoxygenated blood. The chip features a transparent and flexible substrate housing microchannels simulating lung sacs and capillaries. These microchannels are separated by a porous membrane simulating the alveolar-capillary interface. To represent the application of the NF-83 composition, 5 mL of sample was dispersed with 1 mg of Texas Red DHPE triethylammonium salt, centrifuged, and sonicated in a water bath for 20 min. 6 μL of this solution was diluted with 1 mL of PBS and injected into the bottom channel of the StimuLung chip. To simulate oxygen flow and CO2 absorption, the microchannels of the sac were perfused with a controlled gas flow, representing O2 flow, while the capillary channels were perfused with carbon dioxide to simulate venous blood flow. The NF-83 composition was diffused through a porous membrane into capillary channels, and monitored in real time using optical sensors and a multichannel ECHO fluorescence microscope. Computer vision and particle tracking algorithms were used to track the trajectories of individual NF-83 nanoparticles through the microchannels to elucidate the diffusion of oxygen and CO2 in the system and analyze the flow and distribution of the nanoparticle bulk.

[0485] In this experiment, as shown in Figure 7A, a "boomerang" pattern was observed, in which the motion of oxygen-saturated nanoparticles that had absorbed carbon dioxide was reversed and moved toward the air interface.

[0486] Results from computer vision-based particle tracking methods collectively demonstrate that introducing airflow at the surface of the nanoparticle composition effectively guides oscillatory behavior, utilizing nanoparticles to agitate fluids.

[0487] In addition to analyzing the behavior of particles within aggregates, the trajectories of individual nanoparticles were analyzed to describe their behavior. As shown in Figure 7B (nanoparticle trajectory 0 s to 10 s) and Figure 7C (nanoparticle trajectory 0 s to 30 s), the nanoparticles approach the carbon dioxide interface and then begin to move towards the air interface. This phenomenon and broadly similar patterns were observed at different times on different particles within the same medium and space. Figure 8 shows sample images of the nanoparticles.

[0488] Example 18. Diffusion of an exemplary nanoparticle composition into the alveolar cavity

[0489] Five percent by weight of the calculated total lipid content of Texas Red-X labeled lipids were mixed overnight on a shaker with the exemplary nanoparticle composition NF-83. 100 μL of this solution was injected intratracheally into (n=3) male mice (C57BL / 6J, Jacson Labs). Several hours later, the exemplary nanoparticle composition NF-83 was instilled into the lungs, and the animals (three mice) were euthanized and sectioned. A control was administered 100 μL of the label-free composition with citrate buffer. The distribution of the composition was then imaged. As shown in Figure 11, the exemplary nanoparticle composition NF-83 was able to uniformly reach the alveolar spaces throughout the lung. This demonstrates the potential of the exemplary nanoparticle composition NF-83 in the treatment of lung-related diseases.

[0490] Example 19. Quenching of FITC (Fluorescent Indole)

[0491] FITC is a common fluorescent reagent in biological research, possessing excellent fluorescence and water solubility. It is quenched when it interacts with oxygen.

[0492] The FITC solution sample was treated with the exemplary nanoparticle composition NF-83, and the FITC sample was then quenched (Figure 12). This demonstrates the oxygen-carrying characteristics of the exemplary nanoparticle composition NF-83.

[0493] Example 20. Changes in collagen conformation

[0494] An exemplary nanoparticle composition, NF-83, was prepared and its ability to modulate collagen conformation was tested. Circular dichroism absorbance results are shown in Figure 10. As shown in Figure 10, NF-83 induces conformational changes in collagen, making it more permeable and allowing liquids to pass through extracellular structures and fibrotic tissues. HA refers to hyaluronic acid.

[0495] List of implementation plans

[0496] Implementation Scheme 1. A nanoemulsion composition for oxygen delivery comprising nanoparticles, wherein the nanoparticles comprise a perfluorocarbon component and a lipid component, wherein the perfluorocarbon component comprises perfluoronaphthene (PFD) and perfluorotripropylamine (PFTPA), and optionally wherein the nanoemulsion composition comprises no more than 22% (w / w) of any polymeric surfactant in the total nanoemulsion composition.

[0497] Implementation Scheme 2. The nanoemulsion composition as described in Implementation Scheme 1, wherein the perfluorocarbon component is substantially composed of PFD and PFTPA.

[0498] Implementation Scheme 3. The nanoemulsion composition as described in Implementation Scheme 1 or Implementation Scheme 2, wherein the perfluorocarbon component is a mixture of perfluoronaphthene (PFD) and perfluorotripropylamine (PFTPA).

[0499] Implementation Scheme 4. The nanoemulsion composition as described in Implementation Scheme 3, wherein the weight ratio of PFD to PFTPA is about 2:1 to about 3:1.

[0500] Implementation Scheme 5. The nanoemulsion composition as described in Implementation Scheme 3 or Implementation Scheme 4, wherein the weight ratio of PFD to PFTPA is about 2.4:1.

[0501] Implementation Scheme 6. The nanoemulsion composition of any one of Implementation Schemes 1 to 5, wherein the lipid component comprises distearate phosphatidylcholine (DSPC) and dipalmitoylphosphatidylcholine (DPPC).

[0502] Implementation Scheme 7. The nanoemulsion composition of any one of Implementation Schemes 1 to 6, wherein the lipid component is a mixture of DSPC and DPPC in a weight ratio of about 1:1 to about 4:1.

[0503] Implementation Scheme 8. The nanoemulsion composition of any one of Implementation Schemes 1 to 7, wherein the lipid component is a mixture of DSPC and DPPC in a weight ratio of about 2.8:1.

[0504] Implementation Scheme 9. The nanoemulsion composition of any one of Implementation Schemes 1 to 8, wherein the lipid component comprises no more than 2% (w / w) of hydrogenated soybean phosphatidylcholine (HSPC) of the total nanoemulsion composition.

[0505] Implementation Scheme 10. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 9, further comprising a buffer.

[0506] Implementation Scheme 11. The nanoemulsion composition as described in Implementation Scheme 10, wherein the buffer solution is a 50 mM citrate.

[0507] Implementation Scheme 12. The nanoemulsion composition as described in Implementation Scheme 10 or Implementation Scheme 11, wherein the buffer is about 40% to about 60% (w / w) of the total nanoemulsion composition.

[0508] Implementation Scheme 13. The nanoemulsion composition of any one of Implementation Schemes 10 to 12, wherein the buffer is about 48% (w / w) of the total nanoemulsion composition.

[0509] Implementation Scheme 14. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 13, wherein the nanoemulsion composition comprises no more than 2% (w / w) of phosphate buffer in the total nanoemulsion composition.

[0510] Implementation Scheme 15. The nanoemulsion composition of any one of Implementation Schemes 1 to 14, wherein the perfluorocarbon component comprises about 40% to about 60% (w / w) of the total nanoemulsion composition.

[0511] Implementation Scheme 16. The nanoemulsion composition of any one of Implementation Schemes 1 to 15, wherein the perfluorocarbon component is about 50% (w / w) of the total nanoemulsion composition.

[0512] Implementation Scheme 17. The nanoemulsion composition of any one of Implementation Schemes 1 to 16, wherein the lipid component is about 1.5% to about 3.5% (w / w) of the total nanoemulsion composition.

[0513] Implementation Scheme 18. The nanoemulsion composition of any one of Implementation Schemes 1 to 17, wherein the lipid component is about 2.3% (w / w) of the total nanoemulsion composition.

[0514] Implementation Scheme 19. The nanoemulsion composition of any one of Implementation Schemes 1 to 18, wherein the weight ratio of the perfluorocarbon component to the lipid component is from about 10:1 to about 30:1.

[0515] Implementation Scheme 20. The nanoemulsion composition of any one of Implementation Schemes 1 to 19, wherein the weight ratio of the perfluorocarbon component to the lipid component is about 23:1.

[0516] Implementation Scheme 21. The nanoemulsion composition of any one of Implementation Schemes 1 to 20, wherein the weight ratio of the perfluorocarbon component to the buffer solution is from about 0.8:1 to about 1.5:1.

[0517] Implementation Scheme 22. The nanoemulsion composition of any one of Implementation Schemes 1 to 21, wherein the weight ratio of the perfluorocarbon component to the buffer solution is about 1.1:1.

[0518] Implementation Scheme 23. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 22, having a pH of about 3.5 to about 8.0.

[0519] Implementation Scheme 24. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 23, having a pH of about 6.0 to about 7.5.

[0520] Implementation Scheme 25. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 24, having a viscosity of not more than 100 cp as determined by a viscometer.

[0521] Implementation Scheme 26. The nanoemulsion composition as described in any one of Schemes 1 to 25, having a viscosity of not more than 20 cp as determined by a viscometer.

[0522] Implementation Scheme 27. The nanoemulsion composition of any one of Implementation Schemes 1 to 26, wherein the average particle size of the nanoparticles is not greater than about 3 μm.

[0523] Implementation Scheme 28. An emulsion composition as described in any one of Implementation Schemes 1 to 27, wherein the average particle size of the nanoparticles is about 100 nm to about 500 nm.

[0524] Implementation Scheme 29. The nanoemulsion composition of any one of Implementation Schemes 1 to 28, wherein the polydispersity index (PDI) of the nanoparticles is about 0.5 to about 1.

[0525] Implementation Scheme 30. The nanoemulsion composition of any one of Implementation Schemes 1 to 29, wherein the nanoparticles have an average particle size of not more than 500 nm when diluted with 5×(v / v) citrate buffer.

[0526] Implementation Scheme 31. The nanoemulsion composition as described in Implementation Scheme 30, wherein the nanoparticles have a PDI of about 0.5 to about 1 when diluted with 5 × (v / v) citrate buffer.

[0527] Implementation Scheme 32. The nanoemulsion composition of any one of Implementation Schemes 27 to 31, wherein the average particle size of the nanoparticles is determined by dynamic light scattering.

[0528] Embodiment 33. The nanoemulsion composition of any one of Embodiments 1 to 31, wherein the nanoparticles have a zeta potential of about 0.02 mV to about 0.7 mV.

[0529] Implementation Scheme 34. The nanoemulsion composition of any one of Implementation Schemes 1 to 33, wherein the nanoparticles have a multi-lamellar structure comprising at least two lamellar layers.

[0530] Implementation Scheme 35. The nanoemulsion composition as described in Implementation Scheme 34, wherein each sheet layer comprises perfluorocarbon sandwiched between two lipid layers.

[0531] Implementation Scheme 36. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 35, which is enriched with dissolved oxygen.

[0532] Implementation Scheme 37. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 36, which is enriched with at least about 5 ppm of dissolved oxygen.

[0533] Implementation Scheme 38. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 37, which is enriched with about 7 ppm to about 10 ppm of dissolved oxygen.

[0534] Implementation Scheme 39. The nanoemulsion composition of any one of Implementation Schemes 36 to 38, wherein the concentration of dissolved oxygen is determined by an oxygen probe.

[0535] Implementation Scheme 40. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 39, which is capable of binding with CO2.

[0536] Implementation Scheme 41. The emulsion composition as described in Implementation Scheme 40, wherein the nanoemulsion composition is capable of carrying CO2 and being exhaled.

[0537] Implementation Scheme 42. The nanoemulsion composition as described in any one of Schemes 1 to 41, which is stable at room temperature for at least 4 days without phase separation or chemical oxidation.

[0538] Implementation Scheme 43. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 42, which is stable at room temperature for at least 10 days without phase separation or chemical oxidation.

[0539] Implementation Scheme 44. The nanoemulsion composition of any one of Implementation Schemes 1 to 43, comprising no more than about 2% (w / w) of cetrimonium bromide (CTAB) of the total nanoemulsion composition.

[0540] Implementation Scheme 45. The nanoemulsion composition as described in any one of Schemes 1 to 44, comprising not more than 2% (w / w) of Tween 80 of the total nanoemulsion composition.

[0541] Implementation Scheme 46. The nanoemulsion composition of any one of Implementation Schemes 1 to 45, comprising not more than about 2% (w / w) of soybean lecithin in the total nanoemulsion composition.

[0542] Implementation Scheme 47. The nanoemulsion composition of any one of Implementation Schemes 1 to 46, comprising not more than about 2% (w / w) of poloxamer in the total nanoemulsion composition.

[0543] Implementation Scheme 48. The nanoemulsion composition of any one of Implementation Schemes 1 to 46, comprising not more than about 1% (w / w) of poloxamer in the total nanoemulsion composition.

[0544] Implementation Scheme 49. The nanoemulsion composition of any one of Implementation Schemes 1 to 48, comprising no more than about 2% (w / w) of any other surfactant or lipid component other than DSPC and DPPC of the total nanoemulsion composition.

[0545] Implementation Scheme 50. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 49 is suitable for inhalation, topical application or enema application.

[0546] Implementation Scheme 51. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 50, comprising:

[0547] (i) PFD, which is about 25% to about 40% by weight of the total nanoemulsion composition (e.g., about 35.5%);

[0548] (ii) PFTPA, which is about 10% to 20% by weight of the total nanoemulsion composition (e.g., about 14.5%);

[0549] (iii) DPPC, which is about 0.1% to 1% by weight of the total nanoemulsion composition (e.g., about 0.6%);

[0550] (iv) DSPC, comprising, by weight, about 1.5% to 2.5% (e.g., about 1.7%) of the total nanoemulsion composition; and

[0551] (v) A citrate buffer (50 mM) at pH 6.0, comprising about 35% to about 55% (e.g., about 47.7%) of the total nanoemulsion composition by weight.

[0552] Implementation Scheme 52. The nanoemulsion composition of any one of Implementation Schemes 1 to 51, wherein the average particle size is about 200 nm to about 500 nm when diluted with 5× citrate buffer.

[0553] Implementation Scheme 53. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 51, wherein the average particle size is about 200 nm to about 300 nm when diluted with 5× citrate buffer.

[0554] Implementation Scheme 54. A nanoemulsion composition prepared by mixing about 35.5% (w / w) of PFD, about 14.5% (w / w) of PFTPA, about 0.6% (w / w) of DPPC, about 1.7% (w / w) of DSPC and about 47.7% (w / w) of citrate buffer (50 mM) at pH 6.0.

[0555] Implementation Scheme 55. The nanoemulsion composition as described in Implementation Scheme 54, wherein the mixing is performed in a microfluidic apparatus.

[0556] Implementation Scheme 56. The nanoemulsion composition as described in Implementation Scheme 55, wherein the mixing is performed in a microfluidic apparatus for 2 to 6 cycles at at least about 1500 PSI.

[0557] Implementation Scheme 57. The nanoemulsion composition as described in any one of Implementation Schemes 1 to 56, which is suitable for atomization and inhalation.

[0558] Implementation Scheme 58. The nanoemulsion composition as described in Implementation Scheme 57, wherein the nanoemulsion composition is enriched with O2 before the nanoparticles reach the alveolar epithelial cells, and is enriched with CO2 after contact with the alveolar epithelial cells.

[0559] Implementation Scheme 59. The nanoemulsion composition as described in Implementation Scheme 57 or Implementation Scheme 58, wherein the nanoemulsion composition is exhalable.

[0560] Implementation Scheme 60. A wound healing composition comprising a nanoemulsion composition as described in any one of Implementation Schemes 1 to 56, silver dioxide, and collagen.

[0561] Implementation Scheme 61. The composition of Implementation Scheme 60, wherein the collagen is hydrolyzed bovine collagen type 1 or porous bovine collagen type 1.

[0562] Implementation Scheme 62. The composition of Implementation Scheme 60, wherein the collagen is processed to have a collagen structure similar to fetal skin.

[0563] Implementation Scheme 63. The composition of any one of Implementation Schemes 60 to 52, wherein the nanoemulsion composition comprises about 55% (w / w) of the total wound healing composition.

[0564] Implementation Scheme 64. The composition of any one of Implementation Schemes 60 to 63, wherein the silver dioxide is about 1% (w / w) of the total wound healing composition.

[0565] Implementation Scheme 65. The composition of any one of Implementation Schemes 60 to 64, wherein the collagen is from about 15% (w / w) to about 44% (w / w) of the total wound healing composition.

[0566] Implementation Scheme 66. An enema gel comprising a nanoemulsion composition as described in any one of Implementation Schemes 1 to 56, poloxamer, and a PBS solution.

[0567] Implementation Scheme 67. The gel as described in Implementation Scheme 66, wherein the nanoemulsion composition comprises about 40% (w / w) of the total enema gel.

[0568] Implementation Scheme 68. The gel as described in Implementation Scheme 66 or Implementation Scheme 67, wherein the poloxamer is P407 poloxamer or P188 poloxamer.

[0569] Implementation Scheme 69. The gel of any one of Implementation Schemes 66 to 68, wherein the poloxamer is P407 poloxamer, which accounts for about 17% to about 25% by weight of the total enema gel.

[0570] Implementation Scheme 70. The gel as described in any one of Implementation Schemes 66 to 69, wherein the poloxamer is P407 poloxamer, which accounts for about 25% by weight of the total enema gel.

[0571] Implementation Scheme 71. The gel as described in Implementation Scheme 70, wherein the PBS solution is approximately 35% (w / w) of the total enema gel.

[0572] Implementation Scheme 72. The gel as described in any one of Implementation Schemes 66 to 68, wherein the poloxamer is P188 poloxamer, and its weight percentage is about 20% to about 50% of the total enema gel.

[0573] Implementation Scheme 73. The gel as described in any one of Implementation Schemes 66 to 68 or Implementation Scheme 72, wherein the poloxamer is P188 poloxamer, which accounts for about 50% by weight of the total enema gel.

[0574] Implementation Scheme 74. The gel as described in Implementation Scheme 73, wherein the PBS solution is approximately 10% (w / w) of the total enema gel.

[0575] Implementation Scheme 75. A cream composition comprising a nanoemulsion composition as described in any one of Implementation Schemes 1 to 56 and a pharmaceutically acceptable excipient.

[0576] Implementation Scheme 76. The cream composition of Implementation Scheme 75, wherein the pharmaceutically acceptable excipient comprises petroleum jelly.

[0577] Embodiment 77. The cream composition as described in Embodiment 76, wherein the nanoemulsion composition is provided at about 40% by weight of the total cream composition.

[0578] Implementation Scheme 78. A cream composition as described in Implementation Scheme 76 or Implementation Scheme 77, wherein the petroleum gel comprises about 60% (w / w) of the total cream composition.

[0579] Implementation Scheme 79. The cream composition of Implementation Scheme 75, wherein the pharmaceutically acceptable excipients include colloidal oat flakes and purified coconut oil.

[0580] Implementation Scheme 80. The cream composition of Implementation Scheme 79, wherein the nanoemulsion composition comprises about 40% (w / w) to about 60% (w / w) of the total cream composition.

[0581] Implementation Scheme 81. The cream composition as described in Implementation Scheme 79 or Implementation Scheme 80, wherein the colloidal oat flakes comprise about 20% (w / w) to about 30% (w / w) of the total cream composition.

[0582] Implementation Scheme 82. The cream composition of any one of Implementation Schemes 79 to 81, wherein the purified coconut oil comprises about 20% (w / w) to about 30% (w / w) of the total cream composition.

[0583] Implementation Scheme 83. The cream composition of Implementation Scheme 75, wherein the pharmaceutically acceptable excipients include colloidal Chaga mushroom extract and Fomitopsis pinicola extract.

[0584] Implementation Scheme 84. The cream composition as described in Implementation Scheme 83, wherein the nanoemulsion composition comprises about 40% (w / w) to about 60% (w / w) of the total cream composition.

[0585] Implementation Scheme 85. A cream composition as described in Implementation Scheme 83 or Implementation Scheme 84, wherein the colloidal birch extract comprises about 20% (w / w) to about 30% (w / w) of the total cream composition.

[0586] Implementation Scheme 86. A cream composition as described in any one of Implementation Schemes 83 to 85, wherein the *Pterocarpus styracifolius* extract comprises about 20% (w / w) to about 30% (w / w) of the total cream composition.

[0587] Implementation Scheme 87. A method of delivering oxygen to the tissues of a subject in need, comprising administering to the subject a nanoemulsion composition as described in any one of Implementation Schemes 1 to 59, a wound healing composition as described in any one of Implementation Schemes 60 to 65, an enema gel as described in any one of Implementation Schemes 66 to 74, or a cream composition as described in any one of Implementation Schemes 75 to 86.

[0588] Implementation Scheme 88. The method as described in Implementation Scheme 87, wherein the tissue is located in the skin.

[0589] Implementation Scheme 89. The method as described in Implementation Scheme 87, wherein the tissue is in the colon or gastrointestinal tract.

[0590] Implementation Scheme 90. The method as described in Implementation Scheme 89, wherein the tissue is in the lung.

[0591] Implementation Scheme 91. The method of any one of Implementation Schemes 87 to 91, wherein the method treats hypoxemia.

[0592] Implementation Scheme 92. A method of treating a local condition in a subject in need, comprising administering to the subject a nanoemulsion composition as described in any one of Implementation Schemes 1 to 59, a wound healing composition as described in any one of Implementation Schemes 60 to 65, an enema gel as described in any one of Implementation Schemes 66 to 74, or a cream composition as described in any one of Implementation Schemes 75 to 86.

[0593] Implementation Scheme 93. The method as described in Implementation Scheme 92, wherein the local condition is a wound, open injury, burn, colonic or gastrointestinal condition, or lung condition.

[0594] Implementation Scheme 94. The method as described in Implementation Scheme 93, wherein the lung condition is COVID, chronic obstructive pulmonary disease (COPD), asthma, or lung injury.

[0595] List of implementation plans

[0596] Implementation Scheme A1. A nanoemulsion composition for oxygen delivery comprising nanoparticles, wherein the nanoparticles comprise a perfluorocarbon component and a lipid component, wherein the perfluorocarbon component comprises perfluoronaphthene (PFD) and perfluorotripropylamine (PFTPA), and optionally wherein the nanoemulsion composition comprises no more than about 2% (w / w) of a polymeric surfactant in the total nanoemulsion composition.

[0597] Implementation Scheme A2. The nanoemulsion composition as described in Implementation Scheme A1, wherein the perfluorocarbon component is optionally a mixture of perfluoronaphthene (PFD) and perfluorotripropylamine (PFTPA) in a weight ratio of about 2:1 to about 3:1.

[0598] Implementation Scheme A3. The nanoemulsion composition as described in Implementation Scheme A1 or Implementation Scheme A2, wherein the lipid component is optionally a mixture of DSPC and DPPC in a weight ratio of about 1:1 to about 4:1.

[0599] Implementation Scheme A4. The nanoemulsion composition of any one of Implementation Schemes A1 to A3, wherein the lipid component comprises no more than about 2% (w / w) of hydrogenated soybean phosphatidylcholine (HSPC) of the total nanoemulsion composition.

[0600] Implementation Scheme A5. The nanoemulsion composition as described in any one of Implementation Schemes A1 to A4 further comprises a buffer.

[0601] Implementation Scheme A6. The nanoemulsion composition as described in Implementation Scheme A5, wherein the buffer is a citrate of about 20 mM to about 100 mM, and optionally the buffer is about 40% to about 60% (w / w) of the total nanoemulsion composition.

[0602] Implementation Scheme A7. The nanoemulsion composition of any one of Implementation Schemes A1 to A6, wherein the nanoemulsion composition comprises no more than about 2% (w / w) of phosphate buffer in the total nanoemulsion composition.

[0603] Implementation Scheme A8. The nanoemulsion composition of any one of Implementation Schemes A1 to A7, wherein the perfluorocarbon component comprises about 40% to about 60% (w / w) of the total nanoemulsion composition.

[0604] Implementation Scheme A9. The nanoemulsion composition of any one of Implementation Schemes A1 to A8, wherein the lipid component is about 1.5% to about 3.5% (w / w) of the total nanoemulsion composition.

[0605] Implementation Scheme A10. The nanoemulsion composition as described in any one of Implementation Schemes A1 to A9, having a pH of about 3.5 to about 8.0.

[0606] Implementation Scheme A11. The nanoemulsion composition as described in any one of Implementation Schemes A1 to A10, having a viscosity of not more than about 100 cp.

[0607] Implementation Scheme A12. The nanoemulsion composition of any one of Implementation Schemes A1 to A11, wherein the average particle size of the nanoparticles is not greater than about 3 μm.

[0608] Implementation Scheme A13. The nanoemulsion composition of any one of Implementation Schemes A1 to A12, wherein the nanoparticles have an average particle size of not more than about 500 nm when diluted with 5 × (v / v) citrate buffer.

[0609] Implementation Scheme A14. The nanoemulsion composition of any one of Implementation Schemes A1 to A13, wherein the nanoparticles have a multi-lamellar structure comprising at least two lamellar layers, wherein each lamellar layer comprises perfluorocarbon sandwiched between two lipid layers.

[0610] Implementation Scheme A15. The nanoemulsion composition as described in any one of Implementation Schemes A1 to A14, which is enriched with at least about 5 ppm of dissolved oxygen.

[0611] Implementation Scheme A16. The nanoemulsion composition of any one of Implementation Schemes A1 to A15 is stable at room temperature for at least 4 days without phase separation.

[0612] Implementation Scheme A17. The nanoemulsion composition as described in any one of Implementation Schemes A1 to A16, comprising no more than about 2% (w / w) of cetrimonium bromide (CTAB) of the total nanoemulsion composition.

[0613] Implementation Scheme A18. The nanoemulsion composition as described in any one of Implementation Schemes A1 to A17, comprising not more than about 2% (w / w) of Tween 80 in the total nanoemulsion composition.

[0614] Implementation Scheme A19. The nanoemulsion composition as described in any one of Implementation Schemes A1 to A18, comprising not more than about 2% (w / w) of soybean lecithin in the total nanoemulsion composition.

[0615] Implementation Scheme A20. The nanoemulsion composition as described in any one of Implementation Schemes A1 to A19, comprising not more than about 2% (w / w) of poloxamer in the total nanoemulsion composition.

[0616] Implementation Scheme A21. The nanoemulsion composition as described in any one of Implementation Schemes A1 to A20, comprising not more than about 2% (w / w) of a surfactant or lipid other than DSPC and DPPC.

[0617] Implementation Scheme A22. The nanoemulsion composition as described in any one of Implementation Schemes A1 to A21 is suitable for inhalation, topical application or enema application.

[0618] Implementation Scheme A23. The nanoemulsion composition as described in any one of Implementation Schemes A1 to A22, comprising:

[0619] (i) PFD, which is about 35.5% by weight of the total nanoemulsion composition;

[0620] (ii) PFTPA, which accounts for about 14.5% by weight of the total nanoemulsion composition;

[0621] (iii) DPPC, which accounts for about 0.6% by weight of the total nanoemulsion composition;

[0622] (iv) DSPC, comprising approximately 1.7% by weight of the total nanoemulsion composition; and

[0623] (v) A citrate buffer (50 mM) at pH 6.0, comprising approximately 47.7% by weight of the total nanoemulsion composition.

[0624] Implementation Scheme A24. A wound healing composition comprising a nanoemulsion composition as described in any one of Implementation Schemes A1 to A23, silver dioxide, and collagen.

[0625] Implementation Scheme A25. The composition as described in Implementation Scheme A24, wherein the collagen is optionally about 44% (w / w) of hydrolyzed type I bovine collagen in the total wound healing composition, or optionally about 15% (w / w) of porous type I bovine collagen in the total wound healing composition.

[0626] Implementation Scheme A26. The composition as described in Implementation Scheme A24 or Implementation Scheme A25, wherein the nanoemulsion composition comprises about 30% to about 90% (w / w) of the total wound healing composition.

[0627] Implementation Scheme A27. The composition of any one of Implementation Schemes A20 to A26, wherein the silver dioxide is about 1% (w / w) of the total wound healing composition.

[0628] Implementation Scheme A28. An enema gel comprising a nanoemulsion composition as described in any one of Implementation Schemes A1 to A23, poloxamer, and a PBS solution.

[0629] Implementation Scheme A29. The gel as described in Implementation Scheme A28, wherein the nanoemulsion composition comprises about 40% (w / w) of the total enema gel.

[0630] Implementation Scheme A30. The gel as described in Implementation Scheme A28 or Implementation Scheme A29, wherein the poloxamer is P407 poloxamer, optionally in a weight percentage of about 17% to about 25% of the total enema gel, or P188 poloxamer, optionally in a weight percentage of about 20% to about 50% of the total enema gel.

[0631] Implementation Scheme A31. The gel as described in any one of Implementation Schemes A28 to A30, wherein the PBS solution comprises about 10% (w / w) to about 35% (w / w) of the total enema gel.

[0632] Implementation Scheme A32. A cream composition comprising a nanoemulsion composition as described in any one of Implementation Schemes A1 to A23 and a pharmaceutically acceptable excipient.

[0633] Implementation Scheme A33. The cream composition as described in Implementation Scheme A32, wherein the pharmaceutically acceptable excipient comprises, optionally, 60% (w / w) of petroleum gel in weight percentage of the total cream composition, and the nanoemulsion composition is provided in weight percentage of about 40% of the total cream composition.

[0634] Implementation Scheme A34. The cream composition as described in Implementation Scheme A32, wherein the pharmaceutically acceptable excipient comprises colloidal oat flakes, optionally in a weight percentage of about 20% (w / w) to about 30% (w / w) of the total cream composition, and purified coconut oil, optionally in a weight percentage of about 20% (w / w) to about 30% (w / w) of the total cream composition.

[0635] Implementation Scheme A35. The cream composition as described in Implementation Scheme A34, wherein the nanoemulsion composition comprises about 40% (w / w) to about 60% (w / w) of the total cream composition.

[0636] Implementation Scheme A36. The cream composition of Implementation Scheme A32, wherein the pharmaceutically acceptable excipient comprises, optionally, about 20% (w / w) to about 30% (w / w) of colloidal Chaga mushroom extract and, optionally, about 20% (w / w) to about 30% (w / w) of Fomitopsis pinicola extract of the total cream composition.

[0637] Implementation Scheme A37. The cream composition as described in Implementation Scheme A36, wherein the nanoemulsion composition comprises about 40% (w / w) to about 60% (w / w) of the total cream composition.

[0638] Implementation Scheme A38. A method of delivering oxygen to the tissues of a subject in need, comprising administering to the subject a nanoemulsion composition as described in any one of Implementation Schemes A1 to A23, a wound healing composition as described in any one of Implementation Schemes A24 to A27, an enema gel as described in any one of Implementation Schemes A28 to A31, or a cream composition as described in any one of Implementation Schemes A32 to A37.

[0639] Implementation scheme A39. The method as described in implementation scheme A38, wherein the tissue is in the skin, in the colon or gastrointestinal tract, or in the lungs.

[0640] Implementation Scheme A40. A method of treating a condition in a subject in need, comprising topically applying to the subject a nanoemulsion composition as described in any one of Implementation Schemes A1 to A23, a wound healing composition as described in any one of Implementation Schemes A24 to A27, an enema gel as described in any one of Implementation Schemes A28 to A31, or a cream composition as described in any one of Implementation Schemes A32A to A37.

[0641] Implementation scheme A41. The method as described in implementation scheme A40, wherein the condition is a wound, open injury, burn, colon or gastrointestinal condition or lung condition.

[0642] Implementation scheme A42. The method as described in implementation scheme A41, wherein the lung condition is COVID, chronic obstructive pulmonary disease (COPD), asthma, or lung injury.

Claims

1. A nanoparticle composition for oxygen delivery comprising nanoparticles, wherein the nanoparticles comprise a perfluorocarbon component and a lipid component, wherein the perfluorocarbon component comprises perfluorodecalin (PFD) and perfluorotripropylamine (PFTPA), and optionally wherein the nanoparticle composition comprises no more than about 2% (w / w) of a polymeric surfactant.

2. The nanoparticle composition of claim 1, wherein the perfluorocarbon component comprises perfluorodecalin (PFD) and perfluorotripropylamine (PFTPA) in a weight ratio of optionally about 2: 1 to about 3:

1.

3. The nanoparticle composition of claim 1 or 2, wherein the lipid component comprises DSPC and DPPC in a weight ratio of optionally about 1 : 1 to about 4:

1.

4. The nanoparticle composition of any one of claims 1 to 3, wherein the weight percent of hydrogenated soy phosphatidylcholine (HSPC) in the total nanoparticle composition is no more than about 2% (w / w).

5. The nanoparticle composition of any one of claims 1 to 4, further comprising a buffer component.

6. The nanoparticle composition of claim 5, wherein the buffer component comprises sodium citrate hydrate and citric acid in a weight ratio of about 5: 1 to about 20: 1, optionally about 12:

1.

7. The nanoparticle composition of claim 6, wherein the weight ratio of the perfluorocarbon component to the buffer component in the nanoparticle composition is about 3.5: 1 to about 4.5: 1, optionally 3.8:

1.

8. The nanoparticle composition of any one of claims 1 to 7, wherein the perfluorocarbon component is about 40% to about 60% (w / w) of the total nanoparticle composition.

9. The nanoparticle composition of any one of claims 1 to 8, wherein the lipid component is about 1.5% to about 3.5% (w / w) of the total nanoparticle composition.

10. The nanoparticle composition of any one of claims 1 to 9, having a pH of about 3.5 to about 8.

0.

11. The nanoparticle composition of any one of claims 1 to 10, having a viscosity of no more than about 100 cp.

12. The nanoparticle composition of any one of claims 1 to 11, wherein the average particle size of the nanoparticles is no more than about 3 μιη.

13. The nanoparticle composition of any one of claims 1 to 12, wherein the nanoparticles have an average particle size of no more than about 500 nm when diluted with 5x (v / v) citrate buffer.

14. The nanoparticle composition of any one of claims 1 to 13, wherein the nanoparticles have a multi-lamellar structure comprising at least two lamellar layers, wherein each lamellar layer comprises perfluorocarbon sandwiched between two lipid layers.

15. The nanoparticle composition of any one of claims 1 to 14, enriched with at least about 5 ppm of dissolved oxygen.

16. The nanoparticle composition of any one of claims 1 to 15, wherein the nanoparticles do not phase separate at room temperature for at least 4 days.

17. The nanoparticle composition of any one of claims 1 to 16, comprising no more than about 2% (w / w) cetrimonium bromide (CTAB).

18. The nanoparticle composition of any one of claims 1 to 17, comprising no more than about 2% (w / w) Tween 80.

19. The nanoparticle composition of any one of claims 1 to 18, comprising no more than about 2% (w / w) soy lecithin.

20. The nanoparticle composition of any one of claims 1 to 19, comprising no more than about 2% (w / w) poloxamer.

21. The nanoparticle composition of any one of claims 1 to 20, comprising no more than about 2% (w / w) surfactant or lipid other than DSPC and DPPC.

22. The nanoparticle composition of any one of claims 1 to 21, wherein the nanoparticles are suitable for inhalation, topical administration, or enema administration.

23. The nanoparticle composition of any one of claims 1 to 22, comprising: (i) PFD at about 35.5% by weight of the total nanoparticle composition; (ii) PFTPA at about 14.5% by weight of the total nanoparticle composition; (iii) DPPC at about 0.6% by weight of the total nanoparticle composition; (iv) DSPC at about 1.7% by weight of the total nanoparticle composition; (v) citric acid at about 0.1% by weight of the total nanoparticle composition; (vi) sodium citrate dihydrate at about 1.2% by weight of the total nanoparticle composition; and (viii) water at about 46.4% by weight of the total nanoparticle composition.

24. A wound healing composition comprising the nanoparticle composition of any one of claims 1 to 23, further comprising silver dioxide and collagen.

25. The wound healing composition of claim 24, wherein the collagen comprises hydrolyzed bovine collagen type 1 at about 44% (w / w) by weight of the total wound healing composition, or porous bovine collagen type 1 at about 15% (w / w) by weight of the total wound healing composition, optionally.

26. The wound healing composition of claim 24 or 25, wherein the nanoparticle composition is about 30% to about 90% (w / w) of the total wound healing composition.

27. The wound healing composition of any one of claims 24 to 26, wherein the silver dioxide is about 1% (w / w) of the total wound healing composition.

28. An enema gel prepared by mixing the nanoparticle composition of any one of claims 1 to 23, poloxamer, and a PBS solution.

29. The enema gel of claim 28, wherein the nanoparticle composition is about 40% (w / w) of the total enema gel.

30. The enema gel of claim 28 or 29, wherein the poloxamer comprises P407 poloxamer optionally at about 17% to about 25% by weight of the total enema gel, or P188 poloxamer optionally at about 20% to about 50% by weight of the total enema gel.

31. The gel of any one of claims 28 to 30, wherein the PBS solution is about 10% (w / w) to about 35% (w / w) of the total enema gel.

32. A cream composition comprising the nanoparticle composition of any one of claims 1 to 23 and a pharmaceutically acceptable excipient.

33. The cream composition of claim 32, wherein the pharmaceutically acceptable excipient comprises petroleum jelly optionally at 60% (w / w) of the total cream composition, and the nanoparticle composition is provided at about 40% by weight of the total cream composition.

34. The cream composition of claim 32, wherein the pharmaceutically acceptable excipient comprises colloidal oatmeal optionally at about 20% (w / w) to about 30% (w / w) of the total cream composition and purified coconut oil optionally at about 20% (w / w) to about 30% (w / w) of the total cream composition.

35. The cream composition of claim 34, wherein the nanoparticle composition is about 40% (w / w) to about 60% (w / w) of the total cream composition.

36. The cream composition of claim 32, wherein the pharmaceutically acceptable excipient comprises colloidal birch extract optionally at about 20% (w / w) to about 30% (w / w) of the total cream composition and Fomes officinalis extract optionally at about 20% (w / w) to about 30% (w / w) of the total cream composition.

37. The cream composition of claim 36, wherein the nanoparticle composition is about 40% (w / w) to about 60% (w / w) of the total cream composition.

38. A method of delivering oxygen to a tissue of a subject in need thereof, comprising administering to the subject the nanoemulsion composition of any one of claims 1 to 23, the wound healing composition of any one of claims 24 to 27, the enema gel of any one of claims 28 to 31, or the cream composition of any one of claims 32 to 37.

39. The method of claim 38, wherein the tissue is skin, colon or gastrointestinal tract, or lung.

40. A method of treating a condition associated with a tissue experiencing reduced normal oxygen levels in a subject comprising topically administering a nanoparticle composition of any one of claims 1 to 23, a wound healing composition of any one of claims 24 to 27, an enema gel of any one of claims 28 to 31, or a cream composition of any one of claims 32 to 37, near the tissue experiencing reduced normal oxygen levels.

41. The method of claim 40, wherein the tissue is in the lung, in the colon, in the gastrointestinal tract, or on the skin.

42. The method of claim 40 or 41, wherein the condition is a wound, an open injury, a burn, a colonic or gastrointestinal condition, or a pulmonary condition.

43. The method of claim 42, wherein the pulmonary condition is COVID, chronic obstructive pulmonary disease (COPD), asthma, or lung injury.

44. The method of claim 43, wherein the pulmonary condition is acute respiratory distress (ARDS).

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