Novel process for production of aqueous mesoporous particle compositions comprising lipophilic compounds

Abstract

The present invention describes a process for preparing an aqueous mesoporous particle composition comprising a lipophilic compound, the process comprising the steps of: a. Providing an emulsifier or emulsifier blend in powder form; b. Mixing one or more oils at a temperature above 40 DEG C at which all oils have become liquid, where the melting temperatures of the oils differ and the mixture comprises at least a sufficient amount of medium chain triglycerides, such that the composition formed in step g has a partially liquid oil phase at a temperature of about 4 DEG C; c. Adding a hydrophobic compound or amphiphilic compound in any hydrophobic solvent to the oil mixture; d. Optionally cooling the mixture to room temperature; e. Adding the emulsifier powder and water to the oil mixture and emulsifying the mixture with optional stirring and heating to 30 DEG C to 40 DEG C; f, performing ultrasonic treatment on the emulsified mixture, and optionally performing mixing or fluidization treatment until the average particle size of the mixture is kept stable; g. Cooling the ultrasonically treated mixture to allow for sufficient time to crystallize; and h. Optionally, performing a second ultrasonic treatment while maintaining the mixture at a low temperature, as well as a composition produced by the above process.

Description

Background Technology

[0001] Mesoporous particles comprise particles with an average diameter of 80 nm to 300 nm and are sometimes referred to as nanoparticles (NPs) and nanocarriers (NCs) in the scientific literature. However, for the purposes of this invention, nanoparticles refer to particles with an average diameter of 100 nm or smaller, while mesoporous particles are particles larger than this common nanoscale. Mesoporous particle formulations of highly lipophilic drugs can deliver compounds that were previously unable to be administered at therapeutic levels using conventional formulations. Complex NC constructs, such as liposomes, nanocapsules, polymeric NPs, micelles, and polymeric vesicles, can enhance the observed therapeutic effects of drug compounds by improving solubility, improving pharmacokinetics, or altering biodistribution. Metallic, organic, inorganic, and polymeric structures, including dendritic macromolecules, micelles, and liposomes, are frequently considered when designing targeted drug delivery systems. In particular, drugs with poor solubility and weak absorption are loaded into these mesoporous particles. However, the effectiveness of these mesoporous structures as drug delivery carriers varies depending on size, shape, and other inherent biophysical / chemical properties. For these reasons, there remains a need for a new, stable mesoporous particle composition containing a lipophilic compound that can be stored for a long period and is readily applicable to the production of pharmaceutical compositions containing the lipophilic compound.

[0002] Many drugs are lipophilic and therefore require different carrier systems than hydrophilic drugs to reach their targets. A non-exhaustive list of such compounds includes well-known drugs such as imipramine, pendimethalin, vortioxetine, lurasidone, posaconazole, diazepam (and various other benzodiazepines such as midazolam and oxazepam), propranolol, trazodone, phenytoin, various statins such as atorvastatin, simvastatin, and lovastatin, bifonazole, ciprofloxacin, clarithromycin, tigecycline, and clindamycin. In addition to pharmaceutical compounds, many other compounds, such as plant-derived auxins, also possess hydrophobic or amphiphilic properties.

[0003] In practice, (oral) compositions containing lipophilic substances are typically provided in solution form in an oily solvent, where the compound dissolves to achieve a fairly high concentration. Most known compositions are oil-based, i.e., an oily solution in which the compound is dissolved, or an oil-in-water dispersion in which the compound is in the oil phase. For oral administration, the oily solvent needs to be food-grade and acceptable for oral administration. A composition is defined as oily or oil-based when more than half of its volume is oil, and in the case of a dispersion, the oil phase should be the continuous phase. However, ingesting oil is cumbersome, and due to limited intake, compositions known in the art are high concentrations, for example, from 5 w / w% to 60 w / w%. However, such high-concentration compositions are difficult to administer correctly, and undesirable side effects are frequently observed. Furthermore, the bioavailability of lipophilic compounds in oily formulations is low, meaning that a large portion of the ingested active compound is not utilized.

[0004] In more recent times, numerous methods have been published for providing water-based compositions containing lipophilic substances. This invention provides an aqueous mesoporous particulate composition advantageously suited for containing such lipophilic substances, particularly plant extracts, thereby overcoming many of the disadvantages of currently available aqueous formulations of lipophilic materials.

[0005] Mesoporous particles represent a significant step forward in the delivery of lipophilic drugs to the body. They help drugs function better by making them easier to melt, delivering them to the correct locations in the body, and altering how they are dispersed there. However, some problems remain to be solved. A major challenge is finding ways to manufacture mesoporous particles that are stable over long periods and easy to apply, especially for drugs that do not readily absorb water. Therefore, in this invention, water-soluble mesoporous carriers in which the oil phase remains partially liquid have been developed. Furthermore, such mesoporous particles can be used not only in pharmaceuticals but also in cosmetics and even for non-human applications (such as plant nutrition and protection).

[0006] Currently, most methods for delivering lipophilic substances use oily solutions. This is far from ideal. It is not easily ingested, and the body does not absorb many of the drugs. The drug takes a long time to take effect (approximately 3.5 hours), and the dosage (and thus the effects) can be unpredictable. Moreover, these methods often involve high concentrations of the drug, which can make administration difficult and lead to unwanted side effects. They also undergo a process in the body called first-pass metabolism, which can enhance the drug's interaction with other substances in the body, further complicating matters. Summary of the Invention

[0007] This invention provides a novel method for delivering lipophilic components, i.e., hydrophobic or amphiphilic components, such as lipophilic drugs, using an aqueous composition. It is rapid, customizable, and consistent. Users of the composition typically experience effects within seconds, providing immediate and stable efficacy. It avoids many of the problems inherent in oil-based methods, such as the unpredictable absorption of oils, resulting in more reliable effects. Furthermore, it reduces first-pass metabolism, thus minimizing interactions with other substances in the body.

[0008] Even better, this method requires a (drug) delivery system that can be designed to release the encapsulated compound in a specific manner using triggering factors present within the delivery target or those that can be applied externally. This means that specific tissues can be targeted, thereby making the drug more effective and reducing side effects. This invention solves many of the problems in current drug delivery methods and can change the way water-averse drugs are administered.

[0009] This invention relates to a method for preparing an aqueous mesoporous particle composition (a lipid carrier with a mesoporous structure) containing a lipophilic compound, the method comprising the following steps: a. To provide emulsifiers or emulsion blends in powder form; b. Mixing one or more oils at a temperature above 40 °C where all the oils have become liquid, wherein the oils have different melting temperatures and the mixture contains at least a sufficient amount of medium-chain triglycerides, so that the composition formed in step g has a partially liquid oil phase at a temperature of about 4 °C; c. Add hydrophobic compounds or amphiphilic compounds from any hydrophobic solvent to the oil mixture; d. Optionally, allow the mixture to cool to room temperature; e. Add the emulsifier powder and water to the oil mixture, and emulsify the mixture under optional stirring, and heat to 30°C to 40°C; f. The emulsified mixture is subjected to ultrasonic treatment, and optionally mixed or fluidized, until the average particle size of the mixture remains stable; g. Cool the ultrasonically treated mixture to allow sufficient time for crystallization; and h. Optionally, a second ultrasonic treatment may be performed while keeping the mixture at a low temperature.

[0010] Preferably, in such methods, the emulsifier is an emulsifier blend; preferably, the emulsifier is a non-toxic emulsifier; more preferably, the blend comprises a glycosyl emulsifier, such as sucrose esters and / or cyclodextrin. Particularly preferred are emulsifier blends comprising sucrose esters, cyclodextrin, and lecithin, wherein the lecithin is preferably sunflower lecithin; more preferably, the amount of lecithin is such that the concentration of lecithin in the final ultrasonically treated mixture from step g is less than 5%, preferably less than 2%, more preferably less than 1%. In this embodiment, it is further preferred that the amount of the glycosyl emulsifier is at least two times, preferably at least four times, the amount of lecithin. Also preferred is that the ratio of sucrose esters, cyclodextrin, and lecithin is 2:2:1.

[0011] In another embodiment, the method preferably includes non-toxic oils or fats.

[0012] It is also preferred that the oil mixture contains at least one oil with a melting point above 50°C, preferably above 60°C. In another preferred embodiment, the oil mixture contains an oil with a melting point between room temperature and body temperature.

[0013] To achieve the above conditions, the oil mixture preferably comprises stearic acid, coconut oil, and medium-chain triglycerides. Then, preferably, when mixed with a lipophilic compound, the oil mixture comprises a component with a stearic acid: coconut oil: medium-chain triglycerides: solvent containing the lipophilic mixture in a ratio of 1:2:3:5.

[0014] In another preferred embodiment, a non-toxic antioxidant is added to the oil mixture. Preferably, the antioxidant is an antioxidant blend. More preferably, the total amount of the antioxidant or antioxidant blend does not exceed 10% of the oil mixture, and more preferably, it does not exceed 5% of the oil mixture. More specifically, the antioxidant blend includes linseed oil, hemp seed oil, tocopherol, and / or rosemary extract. Preferably, it includes linseed oil, hemp seed oil, tocopherol, and rosemary extract in a ratio of 2:2:2:1.

[0015] In the method of the present invention, it is further preferred that the water used to prepare the composition is food-grade water.

[0016] In another preferred embodiment, the average particle size of the mesoporous particles in the ultrasonically treated mixture will be from 10 nm to 600 nm, preferably from 50 nm to 150 nm, and more preferably from 80 nm to 130 nm, most preferably about 110 nm.

[0017] It is also preferred to use the method as detailed herein, wherein glycerol is added to the ultrasonically treated mixture, more preferably wherein the concentration of glycerol is greater than 20%, and more preferably greater than 25%.

[0018] The lipophilic compound used in the method of the present invention is preferably a plant-based extract from oil.

[0019] In another embodiment, the invention relates to the method described above, wherein an additional step of performing the ultrasonically treated mixture obtained in dilution step g is carried out to obtain a diluted composition. Preferably, the mixture is diluted with water; more preferably, the mixture is diluted in such a way that the diluent contains between 0.001% and 5%, preferably between 0.005% and 1%, more preferably between 0.01% and 0.5%, and even more preferably between 0.02% and 0.1% of a lipophilic compound. Preferably, the water used for dilution is food-grade. Further preferably, a stabilizer is added to the composition. Preferably, the stabilizer is a food-grade stabilizer; more preferably, the stabilizer is a gum; more preferably, the stabilizer comprises guar gum and / or xanthan gum; more preferably, the concentration of guar gum and / or xanthan gum in the diluted composition is between 0.01% and 0.05%, more preferably about 0.02%. In another preferred embodiment, a preservative is added to the composition. Preferably, the preservative is a food-grade preservative. More preferably, the preservative is selected from the group consisting of: ascorbic acid, sodium ascorbate, isoascorbic acid, sodium isoascorbate, citric acid, sorbic acid, calcium sorbate, benzoic acid, potassium benzoate, acetic acid, isoascorbic acid, sodium isoascorbate, lauroyl arginine ethyl ester, and other preservatives. spoon cap fake flower ears The long-chain glycolipid MUCL 53181, methylparaben, nisin, sulfite, dimethyl dicarbonate, ascorbyl palmitate, and blends thereof, more preferably, wherein the preservative comprises ascorbic acid, citric acid, or sorbic acid, or mixtures thereof, preferably, wherein ascorbic acid, if present, is present at a concentration between 0.01% and 0.1%, preferably about 0.05%, and wherein citric acid, if present, is present at a concentration between 0.005% and 0.05%, preferably about 0.01%, and wherein sorbic acid, if present, is present at a concentration between 0.05% and 0.5%, preferably about 0.1%. Further, it is preferred to add a flavoring compound, preferably a food-grade flavoring compound.

[0020] In another preferred embodiment, panthenol is added to the oil mixture, preferably in an amount between 0.5% and 5% of the oil mixture, more preferably between 1% and 3%.

[0021] The present invention also relates to aqueous mesoporous particle compositions provided by the methods described above. Furthermore, the present invention also relates to pharmaceutical compositions comprising such aqueous mesoporous particle compositions. Attached Figure Description

[0022] Figure 1 A comparison is shown between solid mesoporous particles and mesoporous particles containing an oil phase according to the present invention, wherein a portion of the oil phase is in a liquid state.

[0023] Figure 2 Corrected peak areas of HPLC measurements of the fast-release mesoporous particle composition and the slow-release mesoporous particle composition prepared according to Example 1 are shown. The data indicates that auxin release increases when the sample is treated at 37 °C compared to lower temperatures. Additionally, after 12 and 45 minutes of incubation, the auxin release from the slow-release mesoporous particles is lower than that from the fast-release mesoporous particles. It was observed that the fast-release mesoporous particle composition likely reaches its peak after 12 minutes of incubation at 37 °C, as no further increase was observed.

[0024] Figure 3: Auxin release leads to GFP degradation in BOX632 nematodes. Quantification of the following items: (a) BOX632 ( mib111 eGFP::AID:: in the gut of nematodes bbln-1 (a) mean fluorescence intensity level and corrected total fluorescence intensity level; and (b) the ratio of GFP in the gut to the pharynx after 120 min of mesoporous auxin exposure. MQ represents the non-auxin control group. The Mann-Whitney U test was performed using false discovery rate (FDR) correction with multiple comparisons, where the FDR-corrected... p A value < 0.05 is considered significant. ns = FDR corrected p Values ​​> 0.05 are considered not significant; += FDR corrected. p-value A value less than 0.1 is considered to indicate a trend; * = FDR corrected. p-value <0.05; **=FDR corrected p-value <0.01.

[0025] Figure 4: Auxin release leads to GFP degradation in BOX817 nematodes. Quantification of the following items: (a) BOX817 ( mib171 (a) Mean fluorescence intensity and corrected total fluorescence intensity of eGFP::AID::bbln-1 in the nematode gut; and (b) the ratio of GFP in the gut to the pharynx after 60 min of mesoporous auxin exposure. MQ represents the non-auxin control group. False discovery rate (FDR) was used, utilizing two samples. t The test performs multiple comparison correction, among which the FDR-corrected p A value < 0.05 is considered significant. ns = FDR corrected pValues ​​> 0.05 are considered not significant; += FDR corrected. p-value A value less than 0.1 is considered to indicate a trend; * = FDR corrected. p-value <0.05; **=FDR corrected p-value <0.01. Detailed Implementation

[0026] In this invention, the mesoporous particle composition is prepared by emulsifying an oil composition containing a lipophilic compound of interest with water by adding an emulsifier, and then sonicating the emulsion to produce an aqueous mesoporous particle solution.

[0027] The first step in preparing the aqueous mesoporous particle composition of this invention is to provide an emulsifier or preferably a blend of emulsifiers. The objective of these emulsifiers is to provide a system capable of producing mesoporous particles, and for this purpose, the emulsifier should provide sufficient stability. Furthermore, since the composition is likely to be ultimately administered orally, it should also possess sufficient safety properties and provide an acceptable taste. A preferred emulsifier is lecithin, which is itself a blend of glycerophospholipids such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, and phosphatidic acid. Lecithin can be derived from various sources such as soybean, rapeseed, cottonseed, or sunflower. Lecithin is a food-grade product with GRAS certification and is also recognized as a food additive E322 in Europe. The use of lecithin is advantageous because it allows for pH-based regulation of the release of the active compound. At high pH values ​​(pH > 9), lecithin degradation does not occur, and therefore no release of the active compound occurs, while at lower pH values ​​(such as the pH of living organisms), release may occur (see also Haidar, I. et al., International Journal of Pharmaceutical Sciences, 528 (1-2) 524-535, 2017). However, due to its unpleasant odor, it is desirable that the amount of lecithin in the final composition be relatively low. Therefore, it should be noted that the concentration of lecithin is less than 5% of the final sonicated mixture obtained as described in the method according to this claim, preferably less than 2%, more preferably less than 1%. In addition to this low amount, it is considered advantageous to include other emulsifiers capable of masking the unpleasant odor of lecithin. For this purpose, glycosyl emulsifiers are preferred because they provide sweetness. Any non-toxic and sweet-tasting glycosyl emulsifier can be used, such as sucrose esters, cyclodextrins, sucralose esters, and sophorolipids. To achieve optimal taste masking, the concentration of these glycosyl emulsifiers is at least twice, preferably at least four times, the concentration of lecithin. The most preferred combination of emulsifiers is a blend of sunflower lecithin, sucrose esters (e.g., Ryoto™ glycoester P-1670 available from Mitsubishi Chemical Corporation), and β-cyclodextrin, in a most preferred ratio of 1:2:2. This blend also exhibits lower toxicity than conventional emulsifiers or emulsifier blends, and generates fewer harmful free radicals and degradation products. Furthermore, the presence of cyclodextrin implies a mesoporous particle size of approximately 110 nm, determined by the internal bond angle of the cyclodextrin, thus promoting this particle size. The incorporation of β-cyclodextrin into particles smaller than 110 nm is achieved through torsional energy supplied by ultrasonication (provided in a later step), a thermodynamically unstable arrangement. These particles are expected to spontaneously reorganize themselves over time to form a larger, more stable, and more energy-efficient conformation of approximately 110 nm.

[0028] The particle size is also determined based on the ratio of oil to emulsifier. Figure 1 As can be seen, the size of the mesoporous particles is also determined by the surrounding emulsifier, which acts as a surfactant. This means that the balance between the oil component and the emulsifier also affects the size of the mesoporous particles that can be obtained. It has been observed that a weight ratio of oil to emulsifier between 3.0 and 5.0 yields good results, but preferably this ratio is between 3.2 and 4.0. More preferably, a ratio of about 3.5 results in an average particle size of about 110 nm.

[0029] Another advantage of being able to vary the average particle size of a composition by selecting different emulsifier blends and / or different oil / emulsifier ratios is that mesoporous particles may be sensitive to degradation caused by UV radiation with the same wavelength as their average particle size. Therefore, to promote the degradation of mesoporous particles with an average particle size of 110 nm, irradiation with UV light at a wavelength of 110 nm can be used. This is particularly useful when the composition is used in vitro in humans or animals, for example, in plants or on plants.

[0030] In the second step of the method, a mixture of oils is provided. Similar to the emulsifier blend, the oil mixture should also consist only of non-toxic, food-grade oils. Furthermore, to allow for adjustment of the viscosity and stability of the mesoporous particle solution, oils with different melting temperatures should be used. To obtain good stability of the mesoporous particles, at least one oil with a melting temperature exceeding 50 °C, preferably exceeding 60 °C, such as myristic acid, palmitic acid, stearic acid, or arachidic acid, should be used. Stearic acid (E570), with a melting point close to 70 °C, is preferably used. Stearic acid is one of the most common saturated fatty acids found in nature and in food supplies, and it is commonly used in (non-alcoholic) beverages. Stearic acid is preferred because it advantageously stabilizes the mesoporous particles that will form in this process.

[0031] Further preferred oils in the oil mixture are those with melting points between room temperature and body temperature. For example, such oils can be selected from coconut oil, cocoa butter, palm kernel oil, peanut oil, and babassu oil. Coconut oil is preferred because it is inexpensive and commercially available. Finally, the oil mixture should also contain components that provide a low melting point, ensuring that the oil phase in the final mesoporous carrier emulsion still contains oil that is liquid at approximately 4 °C. The advantage of the oil phase in the mesoporous particles being at least partially liquid is that the lipophilic compounds contained in these mesoporous particles are more readily absorbed and taken into the body. The characteristics of such mesoporous particles are as follows... Figure 1As shown, the oil phase of the particles includes both liquid and solid oils. For this component, an oily substance with a very low melting point should be used, such as olive oil, rapeseed oil, sunflower oil, soybean oil, castor oil, tung oil, cottonseed oil, or medium-chain triglycerides (MCTs). Medium-chain triglycerides (MCTs) are triglycerides of two or three fatty acids with 6 to 12 carbon atoms at the fatty tail, i.e., medium-chain fatty acids (MCFAs). Preferably, medium-chain triglycerides are used because they are fully saturated, meaning they are unlikely to react during sonication or mixing. Furthermore, MCTs are stable over a wide temperature range under all processing conditions. Furthermore, they are an inexpensive oil source and can be safely and rapidly metabolized by the human body into known safe metabolites with extremely favorable safety properties. Moreover, they produce small particle sizes that are easy to sonicate, and high-purity MCTs are readily available. In another preferred embodiment, C8 MCT (caprylic / capric MCT) is used. The oil mixture is prepared by adding all components at a temperature in which all oils / fats are in a liquid state (and below the boiling point of any of the existing components in the existing composition).

[0032] Using food-grade oil components is preferred because it also means triggering the release of mesoporous particles in organisms with enzymes (lipases) that can degrade the oil components of the mesoporous particles.

[0033] A lipophilic compound of interest is added to the oil mixture. This lipophilic compound may or may not be present in the hydrophobic solvent. If such a hydrophobic solvent is used, care should be taken to ensure that the solvent is non-toxic and acceptable for food applications, at least at concentrations usable in the final product. Preferably, the lipophilic compound in the solvent is a plant extract from the oil. The lipophilic compound can be added to the oil mixture as is (i.e., in the hydrophobic solvent), but it can also be added in a dry or semi-dry form if feasible.

[0034] It has been found that mixtures containing stearic acid, coconut oil, and medium-chain triglycerides have yielded excellent results in the production of stable mesoporous particles, wherein the oil phase within the mesoporous particles is at least partially liquid at a temperature of 4 °C. The ratio between oils and fats with different melting points can control the melting behavior of the mesoporous particles, thereby releasing the lipophilic compounds associated with these mesoporous particles upon entry into the body. Increasing the amount of oils with low melting points results in a predominantly liquid oil phase within the mesoporous particles, providing a faster release; while increasing the amount of oils with high melting points results in a predominantly solid oil phase within the mesoporous particles, providing a delayed release of the lipophilic compounds. Experimental evidence provided in Examples 1 and 2 below illustrates this. The component ratios providing a very stable mesoporous particle composition with the desired (rapid) release characteristics can be obtained by mixing stearic acid, coconut oil, medium-chain triglycerides, and a solvent containing the lipophilic compounds in a ratio of approximately 1:2:3:5. However, other ratios may be equally applicable depending on the properties and melting points of the individual components. Technicians will know how to modify the parameters involved in preparing the oil mixture to achieve the desired release characteristics. Particularly advantageous is ensuring that the (solid) mesoporous particles formed after the emulsification and sonication steps melt at body temperature, making the mesoporous particles unstable and decomposing due to enzymatic digestion of the oil and emulsifier components. Furthermore, the use of lecithin in mesoporous emulsions may cause the mesoporous particles to become unstable at low pH levels. Therefore, care should be taken not to use too much lecithin in the emulsifier mixture if the desired outcome is a composition of aqueous mesoporous particles that can safely pass through the stomach after ingestion.

[0035] Optionally, one or more antioxidants may be added to the oil mixture. However, to maintain the antioxidant activity of such compounds, they should only be added when the oil mixture has cooled to approximately RT. Antioxidants prevent free radical-induced damage to cellular and biological targets by preventing the formation of free radicals, scavenging free radicals, or promoting the breakdown of free radicals. Furthermore, antioxidants prevent oxidation reactions that lead to unpleasant odors and tastes in fats and oils, thereby reducing the nutritional quality of food. Therefore, antioxidants play an important role in the long-term storage of compositions containing oils and fats, and also exert beneficial effects in vivo. Fortunately, there are enough lipophilic compounds that can be added to oil mixtures to act as antioxidants (see, for example, Papas AM. Oil-Soluble Antioxidants in Foods. Toxicology and Industrial Health. 1993;9(1-2):123-149; Fan L and Michael Eskim NA. The Use of Antioxidants in the Preservation of Edible Oils, in: Handbook of Antioxidants for Food Preservation, Woodhead Publishing Series in Food Science, Technology and Nutrition, 2015, 373-388). Many plant oils such as olive oil, rapeseed oil, flaxseed oil, peanut oil, sunflower oil, carrot seed oil, palm oil, corn oil, hemp seed oil, or cottonseed oil can be used, but other plant-derived components such as vitamin E (tocopherol), extracts from rosemary, sage, thyme, etc., can also be used. The primary purpose of adding these antioxidants to the oil mixture is to protect the active lipophilic components during sonication without posing a toxic threat to the user of the composition. Although all of the mentioned antioxidants can be added in such amounts, either as a single component or as a blend, to achieve the desired protection, we have found that a mixture of flaxseed oil, hemp seed oil, tocopherol, and rosemary extract provides sufficient antioxidant protection in the method of the present invention. Tocopherol also enhances the tissue absorption of lipophilic compounds.

[0036] Another component that can be added to the oil mixture is panthenol, a provitamin B5. It is a humectant and moisturizing agent commonly found in shampoos and skincare products. In this invention, it enhances the binding of mesoporous particles to water, i.e., it increases the zeta potential of the mesoporous particle solution. This also improves the tissue absorption rate of the lipophilic compound of interest. Other additives may be pyridoxal phosphate or pyridoxine hydrochloride (vitamin B6) or melatonin.

[0037] At a slightly elevated temperature (approximately 30°C to 40°C), mix the oil mixture with the emulsifier (blend) and water. For every 1 liter of oil mixture, a total of 25 grams of emulsifier (blend) and 500 mL of water can be used. The water is preferably food-grade water.

[0038] The addition of these three components results in an emulsion with discontinuous oil droplets containing a large amount of lipophilic components dispersed in a continuous aqueous medium. The mixture is preferably homogenized to obtain an emulsion in which the oil droplets are uniformly dispersed in the continuous phase. Such homogenization can be performed using any type of mixing equipment, such as a high-speed mixer, homogenizer, submersible mixer, overhead mixer, magnetic stirrer, or even (for small batches) a kitchen mixer or egg beater. Homogenization can also be achieved by fluidization. Following this optional homogenization step, an ultrasonic treatment process is initiated. Through this process, the oil droplets in the emulsion are broken down into smaller droplets, eventually forming mesoporous particles. The result is an aqueous medium in which mesoporous-sized oil droplets loaded with lipophilic components can be obtained; that is, the droplets are a mixture of solid and liquid compositions. As shown above, the properties of the oil, as well as the amount and ratio, largely determine the distribution of solid and liquid oils in the mesoporous particles, thereby determining the release characteristics of the mesoporous particles. The sonication process can be performed using any commercially available ultrasonic instrument and should continue until the average particle size of the mesoporous particles no longer decreases, i.e., until the average particle size of the mesoporous particles stabilizes. Care should be taken not to overheat the sonicated mixture. The sonication process itself generates heat, which may jeopardize the formation and stability of the resulting mesoporous droplets. Cooling can be achieved by externally cooling the container in which the sonication process takes place, but a better approach is to cool the resulting mesoporous particle solution immediately at the moment the sonication process is (near) completion. This can be achieved by placing the solution on ice, which can be done during the sonication process; however, it can also be achieved by adding glycerol to a volume of up to 25% of the mesoporous particle solution. Cooling has the added benefit of causing the particles to crystallize, thereby extending the product's shelf life. If crystallization is not permitted, so-called amorphous mesoporous particles are formed (Type 3 mesoporous particles, see Sharma, A. and Baldi, A. Journal of Drug Development, 2018, (7): 1) and Khan, S. et al., Adv. Pharm. Bull. 2023, 13(3): 446-460). However, since crystallization can lead to drug extrusion, which can ultimately result in agglomeration of the mesoporous particles, a second sonication process may be advantageous to recapture the defects formed by the extruded compound of interest. The sonication should then be performed while the mixture is being cooled.

[0039] Depending on the ultrasonic treatment equipment used and the components used in the oil mixture and emulsifier blend, the average particle size of the mesoporous particles will be from 10 nm to 600 nm, preferably from 50 nm to 150 nm, and more preferably from 80 nm to 130 nm, most preferably about 110 nm. The average particle size can be calculated according to ISO 9276-2 (14th edition, September 4, 2019) or according to D50, such as that determined by adjustable resistance pulse sensing (TRPS) using an Izon-Exoid™ device. Other methods for measuring droplet size in mesoporous emulsions can be used, such as dynamic light scattering, mesoporous particle tracking analysis, transmission electron microscopy, scanning electron microscopy, or laser diffraction. Monitoring particle size during ultrasonic treatment is preferably achieved by performing TRPS on samples collected during ultrasonic treatment.

[0040] After sonication, but before the addition of glycerol, the mixture is preferably filtered to remove larger particles and microorganisms, such as bacteria. For this type of filtration, a filter with a cutoff value of, for example, 200 nm is used. Several filter types can be used, such as polyethersulfone (PES) filters, polyvinylidene fluoride (PVDF) filters, polytetrafluoroethylene (PTFE) filters, mixed cellulose ester (MCE) filters, polypropylene (PP) filters, or nylon filters. All such filters can be pre-sterilized or can be sterilized by the user, and these filters are readily commercially available.

[0041] The presence of mesoporous-sized oil droplets (nanostructured lipid carriers, NLCs) enables controlled release of the lipophilic component: depending on the temperature of the aqueous solution exposure, a portion of the lipids will be solid and a portion will be liquid. This achieves controlled release because, as the mesoporous particles crystallize, they displace the lipophilic component from the core into the surrounding medium. This is also what distinguishes the currently claimed system from earlier drug delivery mesoporous particle systems (see, for example, Khan et al.). See above These utilize solid matrices (solid lipid nanoparticles, SLNs) with low encapsulation efficiency and poor drug release kinetics, or liquid matrices (mesoporous emulsions) with poor storage stability, because the mesoporous particles simply aggregate and fuse together, ceasing to be mesoporous. Partial crystallization, depending on the melting temperature of the oil used in the oil mixture, provides stability to the anti-flocculation mesoporous particles without sacrificing their ability to load large amounts of hydrophilic or amphiphilic compounds and maintain encapsulation efficiency.

[0042] When storing products obtained according to the above process, the products should be packaged in aseptic packaging, which can be made of any inert material, such as glass or vacuum packaging materials commonly used for airtight packaging of food products. When packaged in this way, the product has a very long shelf life.

[0043] The dispersion obtained according to the method claimed above is a highly stable solution / dispersion of mesoporous lipid particles loaded with a lipophilic compound of interest, also referred to herein as mesoporous particles. It has a shelf life of up to several years with no significant changes in composition. Furthermore, it is a high-concentration source of the lipophilic compound of interest.

[0044] If necessary, the product can be diluted to reduce the amount of active ingredient to obtain a suitable dosage form. Dilution is typically done by adding (food-grade) water. When additional dilution is performed, stabilizers, colorants, preservatives, and / or flavoring agents may be added. For stabilizers, food-grade stabilizers are preferred. Preferably, the stabilizer is a gum, such as guar gum (E412), gum arabic (E414), xanthan gum (E415), alginate (E400), carrageenan (E407), gum arabic, tragacanth gum (E413), carrageenan gum (E416), locust bean gum (E410), dammar gum, glucomannan (E425), tara gum (E417), gellan gum, or β-glucan. We found that when the combination of guar gum and xanthan gum is used in a diluted composition at a concentration between 0.01% and 0.05%, more preferably about 0.02%, the combination of guar gum and xanthan gum effectively acts as a stabilizer.

[0045] Preservatives can be selected from the following groups: ascorbic acid, sodium ascorbate, isoascorbic acid, sodium isoascorbate, citric acid, sorbic acid, calcium sorbate, benzoic acid, potassium benzoate, acetic acid, isoascorbic acid, sodium ascorbate, lauroyl arginine ethyl ester, and others. Fake flower ears with spoon cap MUCL 53181 comprises long-chain glycolipids, methylparaben, nisin, sulfite, dimethyl dicarbonate, ascorbyl palmitate, and blends thereof. In this invention, it has been found that the combination of ascorbic acid, citric acid, and sorbic acid provides the desired results when ascorbic acid, if present, is present at a concentration between 0.01% and 0.1%, preferably about 0.05%; when citric acid, if present, is present at a concentration between 0.005% and 0.05%, preferably about 0.01%; and when sorbic acid, if present, is present at a concentration between 0.05% and 0.5%, preferably about 0.1%.

[0046] Although the composition prepared according to the method described above already has an acceptable taste, if not a pleasant one, another flavoring compound can be added if desired. Any flavoring compound suitable for food, including beverages, such as flavorings, can be added. These are readily available in any flavor profile, and a skilled technician will know how to apply them and the concentration required to provide a pleasant flavor.

[0047] The obtained product was diluted 1:8 to obtain a product containing approximately 8 mg to 16 mg of the compound in 25 mL (used as a dosage form), with a compound concentration of approximately 0.04% to 0.08% (meaning an undiluted product concentration of approximately 0.3% to 0.6%). However, in the current system, it appears that concentrations of the lipophilic compound of interest up to 5% have been achieved, depending on the nature of the lipophilic complex, the concentration of the lipophilic compound in the solvent added to the oil mixture, the amount of oil mixture, and the amount of water used in the emulsification reaction, etc.

[0048] Through the specific compositions exemplified herein, a superior system for preparing products exhibiting stability, flavor, and safety properties compared to currently available similar products on the market has been discovered. The selected co-emulsifiers (sunflower lecithin, β-cyclodextrin, and sucrose esters) interact synergistically to enhance the stability of mesoporous particles, thereby providing a robust formulation capable of maintaining product quality under a variety of storage conditions. This combination also enhances the flavor characteristics of the beverage, ensuring a pleasant consumer experience. Furthermore, the blend exhibits lower toxicity compared to conventional emulsifiers, thus preventing the formation of harmful free radicals and degradation products during intensive processing. As exemplified herein, the thermal process involving the formation of a hot emulsion phase followed by cooling before homogenization achieves a more efficient process with minimized energy requirements and side reactions, particularly by employing a blend of natural antioxidants to protect the active ingredients, thereby mitigating potential oxidative damage to consumers. Furthermore, several molecules (tocopherol, vitamin B5 precursor) have been incorporated to improve tissue absorption and promote efficient drug release kinetics.

[0049] By adjusting the ratio of solid to liquid lipids, formulations enable precise manipulation of drug release kinetics, providing a customizable delivery experience. The unique temperature-responsive release mechanism within the mesoporous particles ensures a stable product at room temperature, achieving controlled release upon ingestion. Furthermore, the formulation's versatility allows for the loading of a wide range of hydrophobic drugs onto ultra-stable mesoporous particles, expanding potential applications. Optimized mesoporous particle size in the formulation supports efficient tissue penetration and helps overcome drug resistance mechanisms. The unique combination of emulsifiers, lipid carriers, and natural antioxidants in the formulation not only provides a safe and stable delivery system but also enhances bioavailability and release kinetics. Tested formulations show improved compound bioavailability, up to 10 times higher than conventional bioavailability, and demonstrate effects within minutes rather than hours. Moreover, the release kinetics of compounds from mesoporous particles can be engineered to meet the needs of diverse consumers, ranging from rapid to delayed release. The technology also supports targeted delivery of therapeutic agents specifically within tumors, providing a valuable tool for personalized medicine. The mesoporous particle design supports the potential sequential release of multiple therapeutic agents, facilitating coordinated treatment approaches. This technology can be extended to controlled-release drug delivery systems, thereby improving patient compliance and treatment outcomes.

[0050] Furthermore, due to their semi-solid, partially crystalline nature, mesoporous particles possess transdermal bioavailability in areas of relatively thin skin with sufficient blood flow. This allows the particles to maintain their structure while diffusing through various tissue layers in the skin, thereby expanding the delivery pathway of therapeutic agents. The technology can be applied to enhance the efficacy of cosmetic formulations, potentially achieving better skin penetration and longer-lasting effects. Moreover, the mesoporous particle design allows for the encapsulation and preservation of volatile or sensitive substances, thereby extending shelf life and maintaining compound efficacy. Formulations can load and deliver lipophilic, amphiphilic, or charged bioactive compounds, thus expanding potential applications. For example, the technology can be used to improve the oral delivery of drugs with low bioavailability due to first-pass metabolism. The incorporation of bioessential compounds may potentially improve the stability and bioavailability of probiotics or other beneficial gut microbiota, thus allowing for co-administration with these compounds. Formulations may also potentially protect and enhance the delivery of probiotics, thereby supporting their survival in the harsh gastric environment.

[0051] This technology can also be used to produce stable solutions of nanoparticles, i.e., particles with an average diameter of 10 nm to 100 nm. Such small particles can be achieved by omitting the use of β-cyclodextrin from current formulations and adding larger amounts of lecithin and sucrose esters. Depending on the total amount of emulsifier added, nanoparticles between 10 nm and 100 nm can be produced (more emulsifier will produce even smaller particles). However, the smaller the nanoparticles, the lower their loading capacity for hydrophilic compounds due to their larger (hydrophilic) surface area and smaller internal volume. However, due to the increased surface area, these particles will be able to load more amphiphilic and hydrophilic compounds. Smaller particle size helps reduce metabolic clearance (lower renal clearance and less endocytosis), translating into improved and faster bioavailability in vivo. Moreover, due to their smaller size, nanoparticles can move more quickly in vivo, thereby improving their ability to penetrate deep tissues such as solid tumors, where chemotherapy has historically been unavailable. Leaking tumor vessels are also a key factor in the enhanced permeability and retention (EPR) effect, which can promote the passive accumulation of nanoparticles in tumor tissue (Zhu, D. et al., Journal of Nanobiotechnology, 19, 435, 2021). Furthermore, deeper penetration may also be effective for antibiotic therapy of biomembranes.

[0052] Nanoparticles can also be used to overcome drug resistance, which is particularly helpful for chemotherapy resistance and antibiotic resistance, which greatly hinder the efficacy of drug treatments (see Wang, H. et al., Nature Communications, 12 312, 2021). Many different mechanisms enable or promote multidrug resistance, but drug efflux pumps, located on the cell membrane, are considered a key mechanism for effluxing anticancer drugs, for example, from cancer cells. Since drug efflux pumps cannot expel nanoparticles from (cancer) cells, nanoparticles can be used to deliver anticancer drugs to multidrug-resistant cancer cells, thereby overcoming chemotherapy resistance. Similarly, nanoparticles can be used to deliver antibiotics to target bacterial cells. Advantageously, in addition to effective compounds such as chemotherapeutic agents or antibiotics, efflux pump inhibitors (such as ritonavir, pendimethalin, erythromycin, cyclosporine, ketoconazole, tamoxifen, quinine, or HM30181A) are also incidentally included in the nanoparticles.

[0053] Nanoparticles may also easily cross the blood-brain barrier, while mesoporous particles are blocked due to tight junctions, which effectively filter particles larger than 100 nm.

[0054] Although the mesoporous particles of this invention can penetrate the skin, such applications can be even better achieved with nanoparticles that encounter less resistance when passing through the skin. This means that for smaller particles, the application site is less critical.

[0055] Both nanoparticle and mesoporous particle designs may further enable cell-specific targeting of therapeutic drugs by incorporating specific ligands or antibodies onto the surface of nanoparticles. Nanoparticle or mesoporous particle designs may also potentially enable the delivery of gene or RNA therapies, thus expanding potential applications into the emerging field of gene therapy. Such formulations may further potentially enhance vaccine delivery and efficacy by protecting antigens and providing adjuvant effects.

[0056] The potential applications of this technology extend to animal health, thereby potentially improving the delivery and absorption of veterinary therapeutic drugs, as well as herbal therapies and nutrition.

[0057] Example 1 Preparation of aqueous mesoporous particle auxin composition An emulsifier mixture was prepared by mixing 1.25 g of β-cyclodextrin (Landor Trading Comp.), 1.36 g of sucrose ester (Ryoto™ P-1670 from Mitsubishi Chemical Corporation), and 0.87 g of sunflower lecithin (buXtrade). The mixture was then diluted to 102 mL with purified water at 25 °C.

[0058] Additionally, the oily mixture was prepared by melting 1 g of stearic acid, 5.11 g of auxin (indole-3-acetic acid, Sigma-Aldrich), 2 g of coconut oil (Ekoplaza), 3 g of C8 MCT (Lus Health Ingredients), 0.2 g of hemp seed oil (Holland and Barrett), 0.2 g of flaxseed (Holland and Barrett), 0.1 g of natural tocopherol concentrate (soapqueen.nl), and 0.1 g of rosemary extract in descending order of melting point using a heated stirrer. The temperature during this process did not exceed 75 °C, and the entire process lasted approximately 30 minutes.

[0059] The oil phase solution and emulsifier solution were then mixed using a mixer until the mixture was clearly homogeneous. Subsequently, the mixture was further treated in a 1L beaker using an ultrasonic treatment power of 600W (US solid-state ultrasonic instrument), with a cycle of 10 seconds on, 2 seconds off, for a total of 7 minutes and 30 seconds.

[0060] Cool the mixture to room temperature (approximately 20 °C) using an ice-water bath, then sonicate it for an additional 2 minutes under the same conditions. The additional minute of sonication removes any remaining clumps.

[0061] The final product was filtered using a 2 μm polypropylene filter (a woven, non-adhesive filter from VWF) to remove any larger particles. These samples were designated as mesoporous fast samples. To achieve a slower release, “mesoporous slow” samples were prepared in the same manner, but now with 5.11 g of auxin added to an oil mixture of 4 g stearic acid, 2 g coconut oil, and 2 g medium-chain triglycerides.

[0062] Example 2 To test the release of the auxin (fast and slow groups) mesoporous particles prepared in Example 1, the samples were incubated in water baths at 20 °C or 37 °C. Prior to temperature treatment, 7.5 mL volumes of sample were first aliquoted into 15 mL centrifuge tubes placed on ice water. To ensure complete temperature rise, the pH of the test samples was measured after 5 and 10 minutes. This indicated that 10 minutes was sufficient to reach 37 °C. The samples were incubated for 12 or 45 minutes and then cooled directly on ice water to prevent further release. The samples were then transferred to Amicon. ® Ultracentrifuge tubes (30 kDa MWCO, catalog number: UFC503024, Millipore) were centrifuged at 2500 rcf and 4 °C for 20 min. After centrifugation, the fraction passing through the filter (hereinafter referred to as filtrate) was collected in an Eppendorf tube and immediately stored at -20 °C for HPLC measurements. The product remaining in the filter was replenished with 20% ethanol, the volume of which was equal to the volume of filtrate passing through the column. For this purpose, 7.5 mL of 100% hexane was added and the mixture was shaken vigorously. The sample was incubated on a shaker at 20 °C for 30 min. The hexane phase (upper layer) was then transferred to a new tube. 7.5 mL of hexane was added to the bottom phase, and 7.5 mL of water was added to the hexane phase, and both tubes were incubated on a shaker at 20 °C for 30 min. The two hexane phases were combined into a new tube and stored at -20 °C. The filtrate sample was thawed and thoroughly mixed before use. Aliquot 200 µL of sample into 5 mL volumetric flasks and dilute to 5 mL with the mobile phase, then mix thoroughly. Draw 1 mL of sample into a 2 mL syringe and filter into a vial. Repeat the above steps to produce a second sample vial. Inject three 8 µL replicates of the sample and measure using an Agilent 1100 HPLC with a UV-DAD detector at a flow rate of 0.3 mL / min.

[0063] ImageJ software was used for the following measurements: a) the number of pixels on the Y-axis corresponding to 100 mAU; b) the number of pixels on the X-axis corresponding to 0.1 min. For the sample, the peak height and half-peak width were measured in pixels. The peak height was calculated in mAU by dividing the number of pixels at the height peak by the number of pixels at 100 mAU and then multiplying by 100. The peak width was calculated by dividing the number of pixels at half-peak width by the number of pixels at 0.1 min. The peak area was calculated by multiplying the peak height by the half-peak width, in mAU.

[0064] The corrected peak area was calculated by subtracting the peak area of ​​the sample held at 4 °C under the corresponding conditions from the peak area of ​​the sample. In summary, it appears that fast-moving mesoporous particles seem to release their components better. Release occurs at 20 °C but is enhanced at 37 °C. Slow-moving mesoporous particles appear to release their compounds at a slower or lower rate, at least at 20 °C (see [link to relevant documentation]). Figure 2 ).

[0065] Example 3 Study on the release of mesoporous particles by the auxin-induced degradation determinant system To determine the effect of the release compounds from mesoporous particles on... Caenorhabditis elegans To address the impact of [unspecified factors], this invention developed a detection method for quantitatively determining the release of compounds from mesoporous particles. This detection method was performed using transgenic nematode strains expressing various marker proteins and other components. These transgenic strains allowed us to hypothesize the degradation of readout proteins upon exposure to the plant hormone auxin (Reference 1). We were able to test the ability of mesoporous protective compounds (auxins) to become bioactive by analyzing the degradation signals of the readout proteins. We could compare the mesoporous protective compounds with unprotected (natural) compounds.

[0066] In short, the system consists of three components. First, the protein of interest is labeled with a fluorescent protein and an auxin-induced degradation determinant (AID) derived from indole-3-acetic acid (IAA) protein. Second, F-box transport inhibitor response protein 1 (TIR1) is expressed under a tissue-specific promoter, causing protein degradation to occur only in selected tissues. The final component of the system is the plant hormone auxin. In the presence of auxin, TIR1 binds to the AID degradation determinant, leading to ubiquitination and rapid degradation of the target protein. This can be detected by the loss of GFP signaling in the selected tissue. While auxin is typically provided in NGM plates for nematode growth, we adapted the protocol to allow us to expose nematodes at the desired time. Nematodes can be exposed to both the normal plant hormone auxin and auxin encapsulated in mesoporous particles.

[0067] In this study, we selected the endogenous marker eGFP::AID:: bbln-1 As a readout protein and degradation target, BBLN-1 is a small coiled-coil protein that acts as a regulator of luminal morphology and... Xiuli Yin nematode It is strongly expressed in many tissues, particularly the gut (Reference 2). We investigated two different transgenes of this gene that were identical in GFP readout levels but differed in the AID sequence, leading to different degradation dynamics. Therefore, we expect... mib111 Alleles in comparison mib171 Alleles degrade at a slower rate (Reference 1).

[0068] 1. Materials and Methods 1.1. Strains and Cultures Caenorhabditis elegans The bacteria were cultured on NGM plates inoculated with OP50 bacteria and typically maintained at 20 °C. All strains used in this study were genotype BOX632. mibIs48 [P ELT-2 ::TIR-1::tagBFP2- Lox511 ::tbb-2-3'UTR, IV]) ; bbln-1(mib111 [eGFP::AID:: bbln-1 X and BOX817 mibIs48 [P ELT-2 ::TIR-1::tagBFP2- Lox511 ::tbb-2-3'UTR, IV]) ; bbln-1(mib171 [eGFP::AID:: bbln-1 ])X.

[0069] 1.2. Nematode growth To synchronize nematodes, oviparous adults from several plates were washed and bleached according to standard protocol. The nematodes were incubated overnight for 18 to 22 hours in food-free medium. The hatched L1 synchronized nematodes were then cultured on standard nematode growth medium (NGM) plates with OP50 bacteria at 20 °C for 30 hours. Prior to incubation with mesoporous auxin, the nematodes were washed with M9+ Tween (0.05%) and added to glass vials as described in paragraph 1.3.

[0070] 1.3. Treatment with mesoporous growth hormone To prevent stress in the nematodes due to heat treatment, the (rapid group) mesoporin + OP50 bacteria prepared in Example 1 (each 10 mL of OP50 contains 5 µL of 3.8% mesoporin, resulting in a final mesoporin concentration of approximately 50 µM) was pre-incubated for 30 minutes in glass vials at 20 °C or 37 °C. After a brief cooling at room temperature, 200 nematodes were added to each vial and incubated at 20 °C for 60 minutes (BOX817) or 120 minutes (BOX632).

[0071] 1.4. Microscopic examination For analysis and quantification, nematodes were collected in centrifuge tubes and gently centrifuged. 3.7 µL of the nematode pellet was added to 1 µL of 50 mM sodium azide on a 3% agarose slide. Images were acquired using an LDA-Plan 20X / 0.30 Ph1 lens on a Zeiss Axioplan 2 microscope at an exposure time of 3000 ms.

[0072] 1.5. Quantitative Images were analyzed using ImageJ software by selecting regions of interest corresponding to the pharynx, intestine, or background (Figure S1). Area, mean, standard deviation, minimum, maximum, integrated density, and raw integrated optical density were collected from ImageJ. The average green fluorescent protein (GFP) intensity was calculated by subtracting the average fluorescence of the background region outside the nematode from the average fluorescence of the tissue. The corrected intensity was calculated by subtracting the total background fluorescence intensity of a region of the same area from the tissue's integrated density.

[0073] The intensity ratio is calculated by dividing the average intensity or corrected total intensity of the intestine by the corresponding intensity of the pharynx.

[0074] 2. Results 2.1. Auxin In the body Release from mesoporous particles To study mesoporous auxin In the body Following the release of auxin, we exposed synchronous nematodes in the L3 developmental stage to mesoporins and analyzed the readout protein signals over time. After exposing strain BOX632 to auxin, we observed no degradation or change in readout proteins at 30 or 60 minutes. After 120 minutes, we observed statistically significant degradation in the auxin-treated samples compared to the control group. This was independent of whether they were heat-treated and exposed to 37 °C. Figures 3A to 3BThis timescale matches the previously described degradation of GFP in this allele when exposed to conventional auxin, where 120 minutes of auxin exposure led to severe GFP depletion (Reference 1). Our results indicate that auxin is released from mesoporous particles over time, even without treatment at 37 °C. We hypothesize that this is due to environmental changes in the presence of mesoporous particles. Specifically, differences in bacterial food sources or pH or the presence of enzymes in nematodes may lead to the breakdown of mesoporous particles or the release of mesoporous particles from the particles. We did not observe a statistically significant decrease in pharyngeal GFP levels, suggesting that GFP loss is gut-specific and tissue-specific due to the auxin-induced degradation system (Figure S2). In conclusion, we conclude that mesoporous auxin particles can release their contents, leading to… in vivo eGFP::AID:: bbln-1 Degradation of the readout signal.

[0075] 2.2. At 37 °C, the release of mesoporous particles is enhanced. When strain BOX817 was exposed to auxin, we did not observe any degradation of GFP in the gut after 30 minutes. In contrast, we observed a statistically significant decrease in the mean GFP intensity in the gut of nematodes treated at 37 °C after 60 minutes of exposure to auxin compared to the untreated control group. Figures 4A to 4B We did not observe a significant difference in GFP intensity between the control group and the 20 °C treatment group. Furthermore, statistical comparisons between nematodes treated with mesoporous auxin at 20 °C and 37 °C showed a decreasing trend in GFP fluorescence intensity in the 37 °C group. These results indicate that increased temperature (37 °C) leads to enhanced auxin release from mesoporous particles. After 90 minutes, we observed GFP degradation in the presence of mesoporous auxin, independent of temperature. This reinforces our previous findings that auxin can be released from mesoporous particles at 20 °C. In conclusion, our data confirm that auxin release from mesoporous particles leads to GFP degradation in the gut, and that particle release is enhanced at 37 °C.

[0076] 1)Sepers, JJ, Verstappen, NHM, Vo, AA, Ragle, JM, Ruijtenberg, S., Ward, JD, and Boxem, M., 2022b. The mIAA7 degradation determinant enhances auxin-mediated degradation in *C. elegans*. Degradation (The mIAA7 degron improves auxin‐mediated degradation in Caenorhabditis elegans). G3 gene | Genome | Genetics 12:jkac222 https: / / doi.org / 10.1093 / g3journal / jkac222.2 ) 2)Remmelzwaal, S., Geisler, F., Stucchi, R., van der Horst, S., Pasolli, M., Kroll, JR, Jarosinska, OD, Akhmanova, A., Richardson, CA, Altelaar, M., Leube, RE, Ramalho, JJ, and Boxem, M., 2021. BBLN-1 is crucial for intermediate filament tissue and apical membrane morphology. (BBLN‐1 is essential for intermediate filament organization and apical Membrane morphology. Current Biology 31:2334-2346.e9 https: / / doi.org / 10.1016 / j.cub.2021.03.069

Claims

1. An aqueous mesoporous particle composition comprising a hydrophobic compound or an amphiphilic compound, wherein the composition comprises: Emulsifier blends, and An oil mixture comprising the target compound and at least two oils. The oils have different melting temperatures and contain at least sufficient amounts of medium-chain triglycerides to have a partially liquid oil phase at a temperature of about 4 °C, and the oils have been mixed at a temperature above 40 °C; The composition is obtained by emulsifying the oil mixture and water in the presence of an emulsifier blend, subjecting the emulsified mixture to ultrasonic treatment, and optionally mixing or fluidizing the mixture until the average particle size of the mixture remains stable.

2. The aqueous mesoporous particle composition according to claim 1, wherein the mesoporous particles have an average particle size of 10 nm to 600 nm, determined according to ISO 9276-2 and expressed as D50, preferably wherein the emulsifier blend comprises lecithin, sucrose ester and cyclodextrin, more preferably wherein the weight ratio between sucrose ester:cyclodextrin:lecithin is 2:2:

1.

3. The aqueous mesoporous particle composition according to claim 1 or 2, wherein the emulsifier blend comprises a glycosyl emulsifier in an amount of at least twice the amount of lecithin.

4. The aqueous mesoporous particle composition according to any one of claims 1-3, wherein the concentration of lecithin in the final composition is less than 1% by weight.

5. The aqueous mesoporous particle composition according to any one of claims 1-4, wherein the oil mixture comprises at least one oil with a melting point higher than 50 °C, preferably higher than 60 °C, preferably, the oil mixture comprises at least one oil with a melting point higher than 60 °C and at least one oil with a melting point between room temperature and body temperature, more preferably, the oil mixture comprises stearic acid, coconut oil and medium-chain triglycerides, more preferably, the weight ratio of stearic acid: coconut oil: medium-chain triglycerides is 1:2:

3.

6. The aqueous mesoporous particle composition according to any one of claims 1-5, wherein the oil mixture further comprises a non-toxic antioxidant selected from linseed oil, hemp seed oil, tocopherol and rosemary extract, preferably, the antioxidant is a blend comprising linseed oil, hemp seed oil, tocopherol and rosemary extract in a weight ratio of 2:2:2:

1.

7. A method for preparing an aqueous mesoporous particle composition, wherein the mesoporous particles have an average particle size of 10 nm to 600 nm, determined according to ISO 9276-2 and expressed as D50, the composition comprising a lipophilic compound, the method comprising the following steps: a. To provide emulsifiers or emulsion blends in powder form; b. Mixing two or more oils at a temperature above 40 °C where all the oils have become liquid, wherein the oils have different melting temperatures, wherein the oil mixture contains at least one oil with a melting point above 50 °C, and the mixture contains at least a sufficient amount of medium-chain triglycerides, such that the composition formed in step g has a partially liquid oil phase at a temperature of about 4 °C; c. Add hydrophobic compounds or amphiphilic compounds from any hydrophobic solvent to the oil mixture; d. Optionally, allow the mixture to cool to room temperature; e. Add the emulsifier powder and water to the oil mixture, and emulsify the mixture under optional stirring, and heat to 30°C to 40°C; f. The emulsified mixture is subjected to ultrasonic treatment, and optionally mixed or fluidized, until the average particle size of the mixture remains stable; g. Cool the ultrasonically treated mixture to allow sufficient time for crystallization; and h. Optionally, a second ultrasonic treatment may be performed while keeping the mixture at a low temperature.

8. The method according to claim 7, wherein the emulsifier is an emulsifier blend, preferably wherein the emulsifier is a non-toxic emulsifier, more preferably wherein the blend comprises lecithin and a glycosyl emulsifier, such as sucrose esters and / or cyclodextrin; more preferably, wherein the emulsifier is a blend comprising sucrose esters, cyclodextrin and lecithin, preferably comprising sunflower lecithin; more preferably, wherein the amount of lecithin is such that in the mixture from the final sonication treatment of step g, the concentration of lecithin is less than 5% by weight, preferably less than 2%, more preferably less than 1%; more preferably, wherein the weight of the glycosyl emulsifier is at least twice the weight of the lecithin, preferably at least four times; more preferably, wherein the weight ratio between sucrose esters, cyclodextrin and lecithin is 2:2:

1.

9. The method according to claim 7 or 8, wherein the oil mixture comprises at least one oil with a melting point above 60 °C; preferably, wherein the oil mixture further comprises an oil with a melting point between room temperature and body temperature; more preferably, wherein the oil mixture comprises stearic acid, coconut oil, and medium-chain triglycerides; more preferably, wherein when mixed with the lipophilic compound, the oil mixture comprises a component in a weight ratio of stearic acid: coconut oil: medium-chain triglycerides: solvent containing the lipophilic mixture of 1:2:3:

5.

10. The method according to any one of claims 7-9, wherein a non-toxic antioxidant is added to the oil mixture, preferably wherein the antioxidant is an antioxidant blend, more preferably wherein the total amount of the antioxidant or antioxidant blend does not exceed 10% of the amount of the oil mixture, preferably not exceeding 5% of the amount of the oil mixture; more preferably, wherein the antioxidant blend comprises linseed oil, hemp seed oil, tocopherol and / or rosemary extract; preferably wherein the antioxidant blend comprises linseed oil, hemp seed oil, tocopherol and rosemary extract in a ratio of 2:2:2:

1.

11. The method according to any one of claims 7-10, wherein the weight ratio of oil to emulsifier is 3.0 to 5.0, more preferably 3.2 to 4.0, and even more preferably about 3.

5.

12. The method according to any one of claims 7-11, wherein an additional step of performing the ultrasonically treated mixture obtained in dilution step g is performed to obtain a diluted composition, preferably, wherein the mixture is diluted with water, preferably food-grade water; more preferably, wherein the mixture is diluted in such a way that the diluted solution contains between 0.001% and 5%, preferably between 0.005% and 1%, more preferably between 0.01% and 0.5%, more preferably between 0.02% and 0.1% of the lipophilic compound.

13. The method according to any one of claims 7-12, wherein a stabilizer is further added to the composition, preferably wherein the stabilizer is a food-grade stabilizer, more preferably wherein the stabilizer is a gum, more preferably wherein the stabilizer comprises guar gum and / or xanthan gum, more preferably wherein the concentration of guar gum and / or xanthan gum in the diluted composition is between 0.01% and 0.05%, more preferably about 0.02%.

14. The method according to any one of claims 7-13, wherein a preservative is further added to the composition, preferably wherein the preservative is a food-grade preservative, more preferably the preservative is selected from the group consisting of: ascorbic acid, sodium ascorbate, isoascorbic acid, sodium isoascorbate, citric acid, sorbic acid, calcium sorbate, benzoic acid, potassium benzoate, acetic acid, isoascorbic acid, sodium ascorbate, lauroyl arginine ethyl ester, and other preservatives. Fake flower ears with spoon cap The preservatives of MUCL 53181 include long-chain glycolipids, methylparaben, nisin, sulfite, dimethyl dicarbonate, ascorbyl palmitate, and blends thereof; more preferably, the preservatives comprise ascorbic acid, citric acid, or sorbic acid, or blends thereof, preferably the ascorbic acid, if present, is present at a concentration between 0.01% and 0.1%, preferably about 0.05%, and the citric acid, if present, is present at a concentration between 0.005% and 0.05%, preferably about 0.01%, and the sorbic acid, if present, is present at a concentration between 0.05% and 0.5%, preferably about 0.1%.

15. The method according to any one of claims 7-14, wherein a flavoring compound, preferably a food-grade flavoring compound, is further added.

16. The method according to any one of claims 7-15, wherein panthenol is added to the oil mixture, preferably wherein the amount of panthenol added is between 0.5% and 5% of the oil mixture, more preferably between 1% and 3% of the oil mixture.

17. The aqueous mesoporous particle composition according to claim 1 or the method according to claim 7, wherein the lipophilic compound is an oily plant extract.

18. A pharmaceutical composition comprising an aqueous mesoporous particle composition according to any one of claims 1-6 or an aqueous mesoporous particle composition provided by the method according to any one of claims 7-17.

Images