microparticles

Abstract

To provide self-assembled microparticles useful in a wide range of physical and chemical processes where interaction with a substrate is required.SOLUTION: The invention provides a self-assembled microparticle having an acid with two or more acid groups and an organic base. The microparticle may be obtained by contacting a bis-acid and an organic base in a hydrophilic solvent, where the acid is insoluble or sparingly soluble in the hydrophilic solvent and the organic base is soluble in the hydrophilic solvent. Such microparticles have antimicrobial activity and may be used in a wide range of consumer product applications, cell culture and medical products, such as wound dressings.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to microparticles, particularly self-assembled microparticles, methods for producing such microparticles, methods for producing macroporous materials formed by collision of such particles, and uses of the resulting particles and porous structures. These microparticles and porous materials are particularly useful in a wide range of physical and chemical processes requiring interaction with a substrate, such as solid-phase synthesis, solid-phase extraction, solid-phase reagents, seed immobilization (e.g., proteins and nucleic acids), cell culture, wound care (e.g., chronic and acute wounds in and out of the body), burn treatment, medical diagnosis, regenerative medicine, vision correction, controlled release of chemicals (e.g., pharmaceuticals and pesticides), catalytic reactions, and chromatography. [Background technology]

[0002] Solid support materials useful in solid-phase synthesis processes are well known. For example, a wide range of physical and chemical processes, such as the synthesis of organic molecules, especially peptides and oligonucleotides, species immobilization, catalyst support, ion exchange, species extraction from materials, diagnostics, and chromatography, utilize solid support materials.

[0003] Typically, multi-step synthesis of organic molecules involves numerous isolation steps to separate the intermediates produced at each stage before proceeding to the next step. These processes are often time-consuming, expensive, and can be inefficient in terms of yield. Intermediates often require purification to remove excess reagents and reaction by-products, as well as treatments such as precipitation, filtration, and two-phase solvent extraction; solid-phase extraction, crystallization, and chromatography may also be used.

[0004] Solid-phase synthesis often offers several advantages over solution-phase synthesis. For example, isolation procedures used in solution-phase synthesis can be avoided to some extent by reversibly binding the target molecule to a solid support. Excess reagents and some by-products can be removed by filtration and washing of the solid support. The target molecule can be recovered in substantially quantitative yield through several processes that are usually particularly difficult in solution-phase synthesis. Furthermore, the time required to perform operations on a solid support is usually much shorter than the time required to perform the corresponding steps in solution-phase synthesis.

[0005] Immobilization of species in a certain range of processes is also known. For example, polymer supports are commonly used to immobilize catalysts for use in traditional organic chemistry, such as chemical and biocatalytic reactions. Organic chemical reactions can be carried out using immobilized enzymes, or, in the case of chiral resolution, immobilized penicillin amidase has been used, for example, to resolve secondary alcohols (E. Baldaro et al., Tet. Asym. 4, 1031 (1993)), and also immobilization Penicillin G amidase is also used to hydrolyze benzylpenicillin in the production of amoxicillin (Carleysmith, SW and Lilly, MD, Biotechnol. Bioeng., 21, 1057-73, 1979).

[0006] Solid supports are also used to immobilize biomacromolecules for medical and diagnostic applications. This includes the immobilization of proteins, monoclonal, and polyclonal antibodies. Cell culture is typically performed on solid supports with specific surface features and morphologies. Immobilized enzymes can be used as sensors to generate signals. One example is the detection of glucose by a glucose oxidase / peroxidase conjugate system, where the presence of glucose generates hydrogen peroxide, which then becomes a substrate for peroxidase to oxidize a wide range of substrates, giving a colored, fluorescent, or luminescent signal.

[0007] Using various phosphors whose fluorescence is sensitive to specific cations or anions, the concentration of specific ions such as hydrogen ions can be indicated for pH measurement. Polymer particles and porous materials are often used in chromatography, where the solid support is called the stationary phase. In some modes of chromatography, the cost of the stationary phase can be limited. In other modes, the effectiveness of this method may be reduced due to the physical properties of the stationary phase. For example, the soft polymers often used for affinity, ion exchange, and gel permeation chromatography cannot be used at high flow rates due to the easily deformable nature of the particles. The hard macroporous polymers used for many other modes of chromatography are often mechanically brittle and may therefore have a short service life.

[0008] The application of solid supports or stationary phases in chromatographic separations, such as in the complex advanced separations used in the pharmaceutical and biotechnology industries and in the large-scale processes used in the mining industry, is very widespread. Some of the most valuable pharmaceuticals in the pharmaceutical industry are purified by preparative chromatography, and improved chromatographic separations would be technically beneficial and economically advantageous. In the mining and precious metal recovery industries, much of the world's palladium, an important component in a wide range of industrial applications and processes such as catalytic converters and in the manufacture of high-value products, is thought to be purified using immobilized crown ethers (Traczyk, F.P.; Bruening, R.L.; Izatt, N.E., "The Application of Molecular Recognition Technology (MRT) for Removal and Recovery of Metal Ions from Aqueous Solutions"; Fortschritte in der Hydrometallurgie; 1988, Vortrage beim 34. Metallugischen Seminar des Fachausschusses fuer Metallugische Aus-und Weiterbildung der GDMB; 18-20 November 1998; Goslar).

[0009] The use of polymeric particles and macroporous materials in solid-phase extraction and the manufacture of solid-phase reagents is also known in the chemical, pharmaceutical, and biotechnology industries. Known solid supports generally comprise polymeric particles of specific dimensions and physical properties adapted to the intended use. For ease of handling, these polymeric particles are often spherical and have a defined particle size distribution. The spherical nature of the particles improves the flow and filtration characteristics of the polymer. Although the use of solid supports has operational advantages, there are drawbacks to the solid-phase approach. For example, commercially available supports commonly used for the solid-phase synthesis of peptides and oligonucleotides can be expensive, e.g., due to complex manufacturing processes. Microporous polymeric particles and macroporous polymers are commonly used. Microporous polymers have a relatively low level of crosslinking, whereby the polymer particles solvate and swell in a suitable solvent. Macroporous polymers have a high level of crosslinking within the polymer matrix and contain large pores. These polymeric particles are generally hard, have good flow characteristics, and are suitable for use in packed columns.

[0010] Significant clinical challenges have been identified in the treatment and care of wounds. Many known wound care treatments utilize animal-derived collagen, such as bovine, horse, pig, and human collagen. The use of these materials may present a range of ethical, moral, and religious challenges that may hinder their widespread use. Animal-derived collagen may also present technical and commercial problems, as the extraction and processing of these products can be complex, time-consuming, and expensive manufacturing processes. When specific source materials are required for animal-derived products, for example, some commercially available products rely on the use of horse tendons, which may present supply and availability difficulties. The manufacture of animal-derived products may generate biological waste, and specific handling may be required to reduce the risk of biological contamination. [Prior art documents] [Non-patent literature]

[0011] [Non-Patent Document 1] E. Baldaro et al., Tet. Asym. 4, 1031 (1993). [Non-Patent Document 2] Carleysmith, SW and Lilly, MD, Biotechnol. Bioeng., 21, 1057-73, 1979). [Non-Patent Document 3] Traczyk, FP; Bruening, RL; Izatt, NE, "The Application of Molecular Recognition Technology (MRT) for Removal and Recovery of Metal Ions from Aqueous Solutions"; Fortschritte in der Hydrometallurgie; 1988, Vortrage beim 34. 18-20 November 1988; Goslar. [Overview of the project] [Problems that the invention aims to solve]

[0012] There is a need for materials suitable for wound care applications that do not contain animal-derived components, are biocompatible, biodegradable, non-cytotoxic, and preferably antibacterial.

[0013] Known polymer particles used in many of these application areas have several drawbacks. Polymer particles can usually be produced by dispersion or emulsion polymerization processes, in which a solution of monomers is dispersed in an immiscible solvent (continuous phase) before polymerization is initiated. The resulting polymer particles are usually then filtered, washed, and classified to isolate the desired particle size distribution. However, this process can be complex and costly and may be limited by the need to use organic solvents. As used herein, the term "polymer" includes inorganic polymers, such as silica, and organic polymers, such as polyamides.

[0014] These processes have several disadvantages, such as the loss of monomers into the continuous phase, and the generation of a certain range of particle sizes and the undesirable generation of fine particles during polymerization necessitate cumbersome particle size classification, for example, by sieving or air classification.

[0015] In addition to the undesirable costs of losses during manufacturing and preparation, several drawbacks can arise due to the physical properties of known polymer particles. Microporous polymer particles are generally soft and are generally unsuitable for use in chromatographic applications at high flow rates in packed column beds. Furthermore, soft particles can be undesirably compressed during filtration, for example, causing fouling, which can often lead to compression penetration into the sinter or mesh used at the bottom of the column. Hard macroporous and macroretic particles are more suitable for high flow rates in packed column beds. However, due to their hard nature, these particles can be brittle and break under physical stress.

[0016] These problems are exacerbated by packing the polymer particles into the column from bottom to top, which subjects the polymer particles to undesirable and excessive stress. Spherulites typically consist of multiple layers of surfactants in an oil-in-water composition, and the use of solvents and complex temperature control may be required to form them. Particle size can vary widely. [Means for solving the problem]

[0017] Herein, the inventors argue that these and other problems relating to known polymer particles, macroporous materials, and known spherulites are resolved by forming self-assembled microparticles containing fatty acids and bases having two or more carboxylic acid groups, which give a narrow particle size distribution, or by contacting them with self-assembled microparticles. We found that this can be improved by providing a particulate support containing a macroporous material.

[0018] In a first embodiment, the present invention provides a particulate support comprising self-assembled microparticles. Preferably, these microparticles contain an acid having two or more acidic groups and an organic base soluble in a hydrophilic solvent. [Brief explanation of the drawing]

[0019] [Figure 1] Figure 1 is an electron microscope image of the object obtained in Example 1. [Figure 2] Figure 2 is a scanning electron microscope image of the microspherical bodies obtained in Example 5. [Figure 3] Figure 3 is an electron microscope image of the microspherical bodies obtained in Example 8 before crosslinking. [Figure 4] Figure 4 is an electron microscope image of the crosslinked particles obtained in Example 8. [Figure 5] Figure 5 shows the SEM image of the microspherical bodies obtained in Example 12. [Figure 6] Figure 6 shows the metabolic activity assay (CCK-8) of cultured osteoblasts obtained in Example 14. [Figure 7] Figure 7 is a micrograph of cultured osteoblasts obtained in Example 14. [Figure 8] Figure 8 shows the PDAC-brassic acid fine particles formed in Example 15. [Figure 9] Figure 9 shows the test results of the mixed graft in Example 16. [Figure 10] Figure 10 shows the test results of the mixed graft in Example 16. [Figure 11] Figure 11 shows the test results of the mixed graft in Example 16. [Figure 12] Figure 12 shows the test results of the mixed graft in Example 16. [Modes for carrying out the invention]

[0020] Preferably, the acid comprises bis-acid, preferably bis-fatty acid, and preferably contains two or more carboxylic acid groups, but other acid groups can be used. Preferably, the bis-acid is insoluble or sparingly soluble in hydrophilic solvents. Preferably, the acid, preferably bis-fatty acid, can be solubilized by contacting it with an organic base soluble in a hydrophilic solvent.

[0021] The solvent is preferably hydrophilic, and more preferably an aqueous solution, such as an aqueous phase, particularly an oil-in-water emulsion in water. Advantageously, an aqueous solvent, preferably water, allows for the use of fine particles in applications where environmental considerations are important. For example, these fine particles can be incorporated into aqueous products that may be suitable for personal or consumption, medical applications, and, for example, use as biocides. Known biocides may contain environmentally undesirable solvents or components, such as isopropanol and silicones (which require careful use and disposal and subsequent cleaning when used, for example, in cleaning or antimicrobial applications). The aqueous compositions provided by the present invention reduce or avoid the drawbacks associated with products containing organic solvents or silicones.

[0022] In a preferred embodiment, the bis-fatty acid includes a biscarboxyl fatty acid in which the terminal carboxylic acids are linked by a region that is less hydrophilic than the terminal carboxylic acids, preferably hydrophobic. The less hydrophilic region may contain a substituted skeleton, and / or the skeleton may contain a heteroatom, for example, poly-ε-lysine. Preferably, the region linking the carboxylic acids is hydrophobic, preferably a hydrocarbyl group. In a particularly preferred embodiment, the hydrophobic group is an aliphatic hydrocarbyl group. Preferably, the bis-acid has the general formula: HOOC-(CH2) n The compound contains -COOH (wherein n is large enough that bisic acid is sparingly soluble or insoluble in water). Preferably, n is at least 5, more preferably at least 6, and particularly at least 7. Preferably, n is 40 or less, preferably 36 or less, more preferably 25 or less, and particularly 20 or less. Preferably, n is between 7 and 18.

[0023] In a preferred embodiment, the organic acid is C7-C 18 Contains biscarboxyl fatty acids. In another preferred embodiment, the organic acid is C7-C 13 Biscarboxyl fatty acids are used as EDTA, nitrilotriacetic acid, and monocarboxylic acids, preferably C6-C6.18 It is included together with a further acid selected from carboxylic acids such as caproic acid, palmitic acid, and octanoic acid.

[0024] For example, by selecting more than one acid having different n values for the plurality of acids, the size of the fine particles can be adjusted. A longer hydrophobic moiety connecting the acid groups preferably gives larger fine particles. For example, when n is 8 (sebacic acid), particles with a size of 2.6 microns can be obtained, and when n is 11 (brassylic acid), particles with a size of 3.0 microns can be obtained.

[0025] The biscarboxylic fatty acid may also be unsaturated, such as traumatic acid, or substituted, or both unsaturated and substituted. Preferably, the substitution is not one that makes the bis acid soluble in an aqueous solution. When the bis fatty acid is contacted with a solvent-soluble organic base, fine particles are spontaneously formed.

[0026] The bis fatty acid has the general formula: (HO)2OP-(CH2) n -PO(OH)2 bisphosphonic acid, or an unsaturated bisphosphonic acid; general formula: HOOC-(CH2) n -PO(OH)2 monocarboxyl monophosphonic acid, or an unsaturated form of such a bis acid; general formula: (HO)O2S-(CH2) n -SO2(OH) disulfonic acid, or an unsaturated form of such a bis acid; general formula: HOOC-(CH2) n -SO2(OH) monocarboxyl monosulfonic acid, or an unsaturated form of such a bis acid; general formula: (HO)2B-(CH2) n -B(OH)2 bisboronic acid, or an unsaturated bisboronic acid, or a substituted bisboronic acid; general formula: HOOC-(CH2) nThe acid may contain a monocarboxyl monoboronic acid of -B(OH)2, or an unsaturated form of such bisac acid; or a substituted form of such bisac acid. In these acids, n is large enough that the bisac acid becomes sparingly soluble or insoluble in water. Preferably, n is at least 5, more preferably at least 6, and particularly at least 7. Preferably, n is 40 or less, more preferably 36 or less, more preferably 25 or less, and particularly 20 or less. Preferably, n is between 7 and 18.

[0027] Preferably, an organic base is bonded to the bisac acid moiety so that the combination of the two components includes two separate hydrophilic or ionic head regions connected by hydrophobic regions. While we do not wish to be bound by theory, it is thought that the hydrophobic and hydrophilic regions of adjacent bisac acid and organic bases align to form micelles, resulting in the self-assembly of the microparticles of the present invention. Preferably, the microparticles include a multilamellar structure in which further molecules containing bisac acid and organic bases align with the hydrophilic heads of other bisac acid / organic bases to form a multilamellar structure.

[0028] The organic base can be selected from a range of bases that, together with bis-acid, form self-assembled microparticles. Preferably, the organic base includes amines, preferably aliphatic amines or aromatic amines having basic properties or other nitrogen-containing bases. Examples of suitable organic bases include alkylated amines and polyamines, for example, one or two C 1-4 Examples of amines having an -N-alkyl group include methylated amines. Preferred amines include N-methylmorpholine, 4-methylmorpholine (NMM), N,N-dimethylaminoethanol (DMAE), 4-dimethylaminopyridine (DMAP), imidazole or 1-methylimidazole, poly(diallyldimethylammonium chloride) (PDAC), didecyldimethylammonium chloride (DDAC), and dodecyldipropyl One example is lyamine (DDPT).

[0029] In a preferred embodiment, the acid is preferably one or more brassic acid, sebacic acid, and azelaic acid combined with a base selected from methylmorpholine (NMM), N,N-dimethylaminoethanol (DMAE), 4-dimethylaminopyridine (DMAP), imidazole, 1-methylimidazole, poly(diallyldimethylammonium chloride) (PDAC), didecyldimethylammonium chloride (DDAC), and dodecyldipropylenetriamine (DDPT).

[0030] Preferred examples include microparticles containing poly-ε-lysine in combination with one or more of the following: brassic acid and PDAC, brassic acid and DDAC, brassic acid and DDPT, sebacic acid and NMM, sebacic acid, brassic acid, and azelaic acid.

[0031] The inventors have found that the microparticles according to the present invention, which contain an amine having antibacterial properties, are particularly suitable for use in antibacterial compositions and biocides. The level of antibacterial activity of the base can be increased in the form of the self-assembled microparticles according to the present invention compared to conventional formulations.

[0032] In a further embodiment, the present invention provides an antimicrobial composition comprising self-assembled microparticles comprising bisac acid and an antimicrobial base. The present invention also provides the use of self-assembled microparticles comprising bisac acid and an antimicrobial base having a higher level of antimicrobial activity than the antimicrobial base in the form of non-self-assembled microparticles.

[0033] Preferably, by providing an antimicrobial base to the self-assembled microparticles, the antimicrobial activity is increased, resulting in a reduction of at least 10² bacterial load, preferably at least 10⁴ bacterial load, and more preferably at least 10⁵ bacterial load.

[0034] The acid and base are preferably mixed in relative amounts such that the molar ratio of acidic groups in the acid to basic groups in the base is approximately stoichiometric in amount, which allows for the formation of self-assembled particles. The molar amount of acidic groups relative to basic groups may be less or more than the stoichiometric amount, provided that self-assembled particles are formed. If the ratio of acidic groups to basic groups is excessively low or excessively high, the excess components will disrupt the structure of the acid and base, and self-assembled particles will not be formed. The ratio of acidic groups to basic groups that enables the formation of self-assembled particles varies depending on the specific acid and base.

[0035] Those skilled in the art can determine whether self-assembled particles have formed by observing them under a microscope at a magnification sufficient for visual observation, for example, 40x. By varying the relative amounts of acid and base, the minimum and maximum ratios of the multiple components that form the microparticles can be determined. Acids with longer chains can yield more stable microparticles than those containing acids with shorter chains (having the same base and the same molar ratio). Greater stability allows for the use of lower levels of acid, and it is still possible to form microparticles even with a lower ratio of acidic groups to basic groups.

[0036] Preferably, the ratio of acidic groups in the acid to basic groups in the base is 0.6 to 1.4:1, more preferably 0.7 to 1.3:1, more preferably 0.8 to 1.2:1, and most preferably 0.9 to 1.1:1. Sebacic acid and brassic acid are examples of preferred acids. Preferably, fine particles containing sebacic acid with a base have a sebacic acid-to-base ratio of 0.85 to 1.15:1. Fine particles containing brassic acid with a base have a brassic acid-to-base ratio of 0.8 to 1.2:1. In a preferred embodiment, the acid and base are present at levels that give a 1:1 molar ratio of acidic groups to basic groups.

[0037] In a second embodiment, the present invention provides a macroporous material formed by contacting self-assembled microparticles under conditions that result in the formation of a macroporous material. The macroporous material is preferably formed by crosslinking the microparticles.

[0038] The organic base may be reactive in such a way that it can crosslink the self-assembled microparticles to form a macroporous material. The organic base does not need to be reactive, in which case it can preferably be replaced by another reactive species that can then crosslink to form a macroporous material. Solvent-soluble organic bases can be replaced by adding a reactive species such as (but not limited to) an amine containing an organic component. The amine preferably enables crosslinking of the microparticles by forming amide bonds. In a preferred embodiment, the amine containing an organic component is a polymer amine such as (but not limited to) peptides, proteins, polyallylamines, polyethyleneimines, and other polyamines.

[0039] Suitable examples of amines and polyamines include ethylenediamine, poly-ε-lysine, polyallylamine, polyethyleneimine, aminopropyltrialkoxysilane, 3-(2-aminoethylamino)propyltrimethoxysilane, and N-(3-(trimethoxysilyl)-propyl)diethylenetriamine.

[0040] In forming fine particles or macroporous materials, the aforementioned bis-acid can be mixed in any proportion. Furthermore, reactive amines can also be mixed in. Preferably, the fine particles or macroporous material contains functional components adjusted according to the intended use. For example, metal chelating properties are imparted by adding ethylenediaminetetraacetic acid.

[0041] In other embodiments, polyethyleneimine can be used as a support structure in binding or in the synthesis or manipulation of nucleic acids such as DNA and RNA, for example, in sequencing.

[0042] Alkoxysilanes can be used, which can form silica shells in the lamellar layer of fine particles. In another embodiment, the active site of a specific enzyme can be introduced into a peptide within a particle to enable controlled release of the activator. For example, the cleavage site of a wound-based metallinoprotease can be introduced into a wound care-based material to enable controlled release of an antimicrobial agent. This can make it possible.

[0043] In other applications, the microparticles of the present invention can be used to form macrostructures for osteocyte growth. Preferably, the microparticles are bound to hydroxyapatite, preferably in a crystalline form, to attract osteocytes and promote osteocyte culture.

[0044] Many commercially available wound dressings now incorporate animal-derived collagen, which has been shown to improve wound healing. The microparticles and macroporous materials of the present invention are useful as a substitute for animal-derived collagen in wound dressings. The macroporous materials of the present invention have been shown to mimic the biological and physical properties of collagen in tissue repair. However, the components of these materials are not plagued by the growing ethical and religious concerns associated with the use of animal-derived materials in human healthcare.

[0045] The self-assembled microparticles or macroporous materials according to the present invention may also include functional materials supported by polymers. Examples of suitable functional materials include catalysts, initiators for peptide synthesis or oligonucleotide synthesis, pharmacologically active substances, agrochemical active substances, and polymers. Examples include children, enzymes, nucleic acid sequences, and proteins.

[0046] The present invention is particularly useful for supporting precious metal catalysts, such as palladium catalysts. Palladium is a particularly advantageous example. In a further embodiment, the present invention provides a method for producing self-assembled fine particles or macroporous materials in an aqueous medium, comprising contacting a two-acid having two or more acid groups with an organic base in an aqueous medium, preferably in water.

[0047] Preferably, polymerization and crosslinking are initiated by processes known to those skilled in the art. For example, self-assembled microparticles or macroporous materials produced in water using an amine-containing component can be crosslinked using a water-soluble carbodiimide.

[0048] The self-assembled microparticles or macroporous materials of the present invention can be used in any chemical or physical process using a solid support. These self-assembled microparticles or macroporous materials can be used in applications involving electrically conductive and luminescent polymers. A particulate support containing a luminescent polymer can be arranged on a display panel.

[0049] These self-assembled microparticles or macroporous materials are particularly useful in the solid-phase synthesis of organic species, especially polymers. In a preferred embodiment, these self-assembled microparticles or macroporous materials can be used in the synthesis of peptides, oligonucleotides, or oligosaccharides.

[0050] In solid-phase synthesis, processes using solid polymer particles, such as in peptide synthesis, typically involve suspending the particles in a suitable solvent above a porous filter plate and gently agitating the particles to avoid mechanical damage. In particle manufacturing processes, fine particles are often generated, causing clogging of the filter plate and necessitating slow filtration or filter replacement or cleaning. Furthermore, agitation of solid particles can cause fragmentation, leading to the generation of fine particles that exacerbate filter clogging problems. In the pharmaceutical and related industries, current Good Manufacturing Practices (cGMP) strict quality standards require the replacement of the filter plate after each product batch to avoid contamination of subsequent batches by substances removed from the filter plate.

[0051] Preferably, the self-assembled microparticles or macroporous material according to the present invention are substantially monodisperse; that is, the material consists entirely of particles of substantially the same size. Monodisperse microparticles or macroporous material advantageously simplify solid-phase synthesis.

[0052] The present invention further provides the use of self-assembled microparticles or macroporous materials according to the present invention as a solid phase in a chromatography process. Conventionally, chromatography columns are generally packed by preparing a slurry of particles or a stationary phase in a suitable solvent and transferring it into a column where a lower column filter plate is present. Non-uniform sedimentation of the bed due to a broad particle size distribution within the chromatography column can result in a non-uniform and uniformly fractured stationary phase bed, potentially leading to poor and irreproducible separation. The column often needs to be emptied and repacked several times to achieve the desired performance. This is cumbersome and can result in downtime, which is particularly disadvantageous in process-scale operation.

[0053] The substantial monodispersity of the self-assembled microparticles in a preferred embodiment of the present invention makes it possible to produce a slurry and transfer the slurry to a column to form a more uniform bed. Alternatively, a macroporous material formed by impacting self-assembled microparticles can be used. Norris-type chromatography columns can be formed.

[0054] In other embodiments, the interstitial spaces between particles within the monolith can be filled with different components, such as cell culture nutrients. In this example, cells can be cultured on the surface of the self-assembling macroporous material. In this example, the self-assembling macroporous material is often referred to as a scaffold for three-dimensional cell culture. Thus, the materials described herein have applications in regenerative medicine, 3D cell culture, and wound care.

[0055] The self-assembled microparticles and macroporous materials of the present invention are also useful for solid-phase extraction, such as ion extraction and ion exchange, for extracting seeds from a liquid in contact with a support, whether in batch form or as a flow on a support. Solid-phase extraction is typically performed in a column or system having filter plates for separating the solid phase from a mixture under extraction. Problems observed with respect to solid-phase synthesis and chromatography as referred to herein may similarly be observed with respect to solid-phase extraction. The self-assembled microparticles and macroporous materials of the present invention provide similar advantages to those given in chromatography and solid-phase synthesis.

[0056] The self-assembled microparticles and macroporous materials of the present invention can be used to immobilize species such as antibodies, oligonucleotides, enzymes, or phosphors, and can be arranged in an array so that each support analyzes different components of a solution. Self-assembled microparticles and macroporous materials having ligands covalently bound to their surface can be used as "wells." Specific binding of target ligands, such as antigens or complementary DNA or RNA sequences, can then be detected using established methods.

[0057] The self-assembled microparticles and macroporous materials of the present invention can also be used to immobilize biocatalysts. Biocatalysts are often used in columns or systems having filter plates for separating a solid phase from a mixture under extraction conditions. The problems observed with respect to solid-phase synthesis and chromatography as described herein may similarly be observed with respect to solid-phase extraction.

[0058] The self-assembled microparticles and macroporous materials of the present invention are particularly useful in the immobilization of species such as solid-phase reagents, metal and other catalysts, biocatalysts, enzymes, proteins, antibodies including polyclonal and monoclonal antibodies, whole cells, and polymers. The present invention is particularly useful in the support of enzymes, such as lipase CalB, which is commonly used in detergents and personal care products.

[0059] The present invention is also particularly useful in the immobilization of affinity ligands such as protein A. In further applications, the particulate support of the present invention can also be used in chemical catalytic reactions by immobilizing, for example, transition metal catalysts and ligands.

[0060] In further applications, the present invention can be used in cell culture. Large-scale culture of animal cell lines is essential for the manufacture of viral vaccines, biopharmaceuticals, and many biotechnological products. Biological products produced by recombinant DNA technology in animal cell culture include enzymes, synthetic hormones, immunobiological preparations (monoclonal antibodies, interleukins, and lymphokines), and anticancer agents. Many simpler proteins can be produced using rDNA in bacterial culture; however, more complex glycosylated (carbohydrate-denatured) proteins must now be produced in animal cells. An important example of such complex proteins is the hormone erythropoietin. Because the cost of growing mammalian cell cultures is high, companies are continuously trying to improve the technology. Yes, they are.

[0061] Cells can be grown in suspension or as adherent cultures. However, adherent cells require a surface, which can be coated with extracellular matrix components to increase adherence and provide other signals necessary for growth and differentiation. Generally, cells derived from solid tissues are adherent. Organ cultures involve growing cells in a three-dimensional environment, as opposed to two-dimensional culture dishes. While these 3D culture systems are more biochemically and physiologically similar to in vivo tissues, they are technically difficult to maintain due to many factors (e.g., diffusion).

[0062] In a further embodiment, the present invention provides the use of self-assembling microparticles and macroporous materials according to the present invention for culturing cells on the surface of a support. Preferably, stem cells can be cultured on the self-assembling microparticles and macroporous materials of the present invention to reduce uncontrolled differentiation and control desired differentiation. The handling properties of the self-assembling microparticles and macroporous materials and the high usability of the surface area of ​​the support are advantageous in this application.

[0063] The present invention is particularly useful in medical diagnostic tests such as immunoassays. Accordingly, the present invention further provides compounds comprising self-assembled microparticles and / or macroporous materials according to the present invention, which are held in a support by a polymer to selectively react with or bind to a compound to be detected, as well as medical diagnostic agents for detecting the presence of functional substances such as enzymes, for example, horseradish peroxidase.

[0064] The self-assembled microparticles and / or macroporous materials of the present invention can be used in separation processes, such as magnetic separation, flow cytometry, and drug delivery, as well as in a wide range of fields, including detergents, pesticides, and personal care.

[0065] Many medical diagnostic agents rely on solid supports to immobilize various diagnostic ligands. The self-assembled microparticles and / or macroporous materials of the present invention can be used in medical diagnostic methods that physically separate a solid phase through a liquid phase.

[0066] In further applications, self-assembled microparticles and / or macroporous materials can be used as absorbents. In this application, this is particularly advantageous when the support contains an inert absorbent material to which the self-assembled microparticles and / or macroporous materials are bound. Self-assembled microparticles and / or macroporous materials can be used to absorb household spills, such as tea, coffee, and wine, or in large-scale applications, such as absorbing oil from spills. Absorbent supports can be used to absorb and physically remove spills, or, in the case of oil spills in water bodies, to effectively trap the oil and retain it in the residue for recovery and disposal.

[0067] The self-assembled microparticles and / or macroporous materials of the present invention can be used as carriers for supporting compounds, such as pharmaceutical or agrochemical compounds or compositions, that are released over time. This use provides a means for adjusting the compound administration plan according to the amount of compound loaded in the support. In the case of pharmaceuticals, this can be advantageous in assisting the precise delivery of the active substance by continuous sustained release rather than requiring the patient to take, for example, periodic high doses in chemotherapy. In the case of commercially available drugs, microparticles can be used to deliver antinasal congestion agents, disinfectants, and anti-inflammatory adjuvants. In some cases, microparticles are used to deliver natural oils such as peppermint oil and lavender oil as anti-snoring adjuvants.

[0068] In addition to aspiration, these self-assembling microparticles can be used in a suitable form for intravenous drug delivery or vaccine delivery. It can be used for this purpose. The fine particles or macroporous materials of the present invention can be incorporated into compositions for a wide range of applications, such as personal care products, including topical and oral compositions; home care products; and medical products, including wound treatment products.

[0069] Examples of personal care products include hand soaps, hand scrubs, creams, deodorants, shampoos, and conditioners. Examples of home care products include antimicrobial products, surface spray cleaners, and cleaning agents. Topical compositions may include functional materials suitable for topical administration. [Examples]

[0070] The present invention is demonstrated by the following non-limiting embodiments. Example 1: Production of self-assembled microparticles: Brassic acid (1.54g, 6.31 mmol) and 4-dimethylaminopyridine (DMAP, 1.54g, 12.62 mmol) are dissolved in water (10cm). 3 The sample was dissolved in (a solution) and placed on a microscope. A nearly monodisperse spherical object with a diameter of approximately 3 μm was observed (Figure 1).

[0071] Example 2: Production of self-assembled microparticles: Brassic acid (1.54g, 6.31 mmol) and dimethylaminoethanol (DMAE, 1.12g, 12.62 mmol) are dissolved in water (10cm). 3 The sample was dissolved in (a solution) and placed on a microscope. A nearly monodisperse spherical object with a diameter of approximately 3 μm was observed.

[0072] Example 3: Production of self-assembled microparticles: Brassic acid (1.54 g, 6.31 mmol) and 4-methylmorpholine (NMM, 1.275 g, 12.62 mmol) are dissolved in water (10 cm³). 3 The sample was dissolved in (a solution) and placed on a microscope. A nearly monodisperse spherical object with a diameter of approximately 3 μm was observed.

[0073] Example 4: Production of self-assembled microparticles: Furthermore, the above dicarboxylic acid dissolution experiments were conducted using a certain range of acids and a certain range of water-soluble organic bases. Some of the combinations tested are shown below. These combinations had an acid group / basic group molar ratio of 0.9 to 1.1:1. All of these combinations formed spherical objects as described in Example 1.

[0074] Pimelic acid + NMM; Suberic acid + NMM; Azelaic acid + NMM; Sebacic acid + NMM; Sebacic acid + DMAP; Sebacic acid + DMAE; Sebacic acid + imidazole; Dodecanediol + NMM; Dodecanediate + DMAP; Dodecane dioxide + DMAE; C 36 Dimer acid + NMM.

[0075] Example 5: Production of crosslinked self-assembled microparticles: Brassic acid (1.54g, 6.31 mmol) and 4-dimethylaminopyridine (DMAP, 1.54g, 12.62 mmol) are dissolved in water (10cm). 3 The sample was dissolved in water (10cm³) and placed on a microscope. A nearly monodisperse spherical object with a diameter of approximately 3 μm was observed (Figure 1). Poly-ε-lysine (PeK) (2 g, 12.04 mmol NH₂) was dissolved in water (10cm³). 3 ) inside The solution was dissolved and added to the above solution of brassic acid / DMAP microspheroids. The mixture was filtered through a 0.45 μm membrane, and the sample was placed on a microscope. Microspheroids with a diameter of approximately 3 μm were still present. This solution was diluted with water for 100 cm³. 3 Diluted in water (10cm³). N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDCl) (4.6g, 2.4 mmol) and HONSu (1.38g, 1.2 mmol) 3 The material was dissolved in ( ) and added to the above solution. The crosslinking reaction was left overnight, and the resulting particles were washed by tangential flow filtration (TFF) and recovered by freeze-drying (yield 2.35 g). Figure 2 shows a scanning electron microscope image of the obtained microspherical bodies.

[0076] Example 6: Production of cross-linked self-assembled microparticles containing protoporphyrin IX heme B: Brassic acid (0.734g, 3.3 mmol) and 4-dimethylaminopyridine (DMAP, 0.734g, 6.6 mmol) are dissolved in water (10cm³). 3 The sample was dissolved in water (10cm³) and placed on a microscope. A nearly monodisperse spherical object with a diameter of approximately 3 μm was observed (Figure 1). Poly-ε-lysine (PeK) (1 g, 6.02 mmol of NH₂) was dissolved in water (10cm³). 3 The solution was dissolved in ( ) and added to the above solution of brassic acid / DMAP microspheroids. The mixture was filtered through a 0.45 μm membrane and the sample was placed on a microscope. Microspheroids with a diameter of approximately 3 μm were still present. This solution was then subjected to a saturated solution of heme B (50 cm³). 3 Diluted with water (5cm³). N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDCl) (2.3g, 1.2 mmol) and HONSu (0.7g, 0.6 mmol) were mixed with water (5cm³). 3 It was dissolved in ( ) and added to the above solution. The crosslinking reaction was left overnight, and the resulting particles were washed by tangential flow filtration (TFF) and recovered by freeze-drying (yield 0.93 g).

[0077] Example 7: Production of crosslinked self-assembled microparticles: Sebacic acid (0.619g, 6.12 mmol) and NMM (0.62g, 6.12 mmol) are dissolved in water (10cm³). 3 The sample was dissolved in (a solution) and placed on a microscope. A nearly monodisperse spherical object with a diameter of approximately 2.5 μm was observed.

[0078] Poly-ε-lysine (PeK) (1g, 5.83 mmol of NH2) in water (10cm) 3 The solution was dissolved in ( ) and added to the above solution of sebacic acid / NMM microspheroids. The mixture was filtered through a 0.45 μm membrane and the sample was placed on a microscope. Microspheroids with a diameter of approximately 2.5 μm were still present. This solution was diluted with water for 50 cm. 3 Diluted in water (10cm³). EDCl (2.24g, 11.7 mmol) and HONSu (2.0g, 17.4 mmol) were mixed with water (10cm³). 3 It was dissolved in ( ) and added to the above solution. The crosslinking reaction was left overnight, the resulting particles were washed with TFF and recovered by freeze-drying.

[0079] Example 8: Production of crosslinked self-assembled microparticles: Sebacic acid (5.06g, 25 mmol) and imidazole (3.4g, 50 mmol) are mixed with water (50cm³). 3 The sample was dissolved in (a solution) and placed on a microscope. A nearly monodisperse spherical object with a diameter of approximately 2.5 μm was observed.

[0080] Poly-ε-lysine (PeK) (8.576g, 50 mmol NH2) in water (50cm³) 3 The solution was dissolved in ) and added to the above solution of sebaciate / imidazole microspheroids. The mixture was filtered through a 0.45 μm membrane and the sample was placed on a microscope. Microspheroids with a diameter of approximately 2.5 μm were still present (Figure 3). This solution was diluted with water for 500 cm³. 3 Diluted to EDC. Add 1 (4.8g, 25 mmol) to water (20cm 3 The material was dissolved in ( ) and added to the above solution. The crosslinking reaction was left for 1 hour, then 25 mmol of EDCl was added and left overnight. The resulting particles were washed with water by decantation and recovered by freeze-drying (Figure 4).

[0081] Example 9: Production of crosslinked self-assembled microparticles: Sebacic acid (5g, 24.7 mmol) and (3-aminopropyl)trimethoxysilane (8.42g, 46.9 mmol) are dissolved in water (50cm³). 3 The sample was dissolved in (a solution) and placed on a microscope. A nearly monodisperse spherical object with a diameter of approximately 2.5 μm was observed.

[0082] The mixture was left to stand overnight and then acidified with concentrated hydrochloric acid. The addition of hydrochloric acid led to the formation of silica within the particles, creating a sebacic acid / silica complex. Example 10: Production of crosslinked self-assembled microparticles: Sebacic acid (5g, 24.7 mmol) and N-[3-(trimethoxysilyl)propyl]ethylenediamine (5.77g, 51.9 mmol amine) are dissolved in water (50cm³). 3The sample was dissolved in (a solution) and placed on a microscope. A nearly monodisperse spherical object with a diameter of approximately 2.5 μm was observed.

[0083] Dilute this solution with water to 500 cm 3 Diluted in water (100cm³). EDCl (20g, 104 mmol) was added to water (100cm³). 3 It was dissolved in ( ) and added to the above solution. After letting the mixture stand overnight, it was acidified with concentrated hydrochloric acid. By adding hydrochloric acid, silica was formed within the particles, and a sebacic acid / silica complex was created.

[0084] Example 11: Production of cross-linked self-assembled microparticles: Sebacic acid (5g, 24.7 mmol) and N1-(3-trimethoxysilylpropyl)diethylenetriamine (4.37g, 46.9 mmol amine) are dissolved in water (50cm³). 3 The sample was dissolved in (a solution) and placed on a microscope. A nearly monodisperse spherical object with a diameter of approximately 2.5 μm was observed.

[0085] Dilute this solution with water to 500 cm 3 Diluted in water (100cm³). EDCl (20g, 104 mmol) was added to water (100cm³). 3 It was dissolved in ( ) and added to the above solution. After letting the mixture stand overnight, it was acidified with concentrated hydrochloric acid. By adding hydrochloric acid, silica was formed within the particles, and a sebacic acid / silica complex was created.

[0086] Example 12: Production of a self-assembled macroporous crosslinked sheet: Sebacic acid (0.619g, 6.12 mmol) and NMM (0.62g, 6.12 mmol) are dissolved in water (10cm³). 3 The sample was dissolved in (a solution) and placed on a microscope. A nearly monodisperse spherical object with a diameter of approximately 2.5 μm was observed.

[0087] Poly-ε-lysine (PeK) (1g, 5.83 mmol of NH2) in water (10cm) 3The solution was dissolved in ( ) and added to the above solution of sebacic acid / NMM microspheroids. The mixture was filtered through a 0.45 μm membrane and the sample was placed on a microscope. Microspheroids with a diameter of approximately 2.5 μm were still present. EDCl (2.24 g, 11.7 mmol) and HONSu (2.0 g, 17.4 mmol) were added to water (10 cm³). 3 It was dissolved in ( ) and added to the above solution. The crosslinking reaction was left overnight, the resulting sheet was washed with water, and dried by freeze-drying. The SEM shown in Figure 5 clearly shows that a fused microspherical structure of macroporous polymer was formed.

[0088] Example 13: Production of self-assembled macroporous crosslinked sheets: (12-Phosphonododecyl)phosphonic acid (330 mg, 1 mmol) and NMM (404 mg, 4 mmol) were dissolved in water. The samples were placed on a microscope and the presence of microspherical particles in a substantially monodisperse state was confirmed. PeK (343 mg, 2 mmol NH2) was dissolved in water (10 cm³). 3 It was dissolved in (10 cm³) and added to the bisphosphonic acid solution prepared above. At this stage, microspherical bodies were still present. 3 Dissolved EDCl (1.15 g, 6 mmol) was added to the mixture, and the mixture was immediately poured into a tray. At this stage, microspherical particles were still present. After about 2 hours, a sheet was formed, which was thoroughly washed with water. The final sheet had a rubbery texture.

[0089] Example 14: Culture of osteocytes (Example of 3D cell culture): In this example, the self-assembled macroporous sheet produced in Example 12 was manufactured. Further products were produced by combining the sheet from Example 12 with hydroxyapatite nanoparticles available from SigmaAldrich (catalog No. 702153) at a concentration of approximately 10% by weight relative to this scaffolding material. The samples were prepared and their cytocompatibility was tested.

[0090] The 9-day growth curve of osteoblasts showed no significant difference in cell viability during the early stages of the culture period compared to a controlled study of tissue culture in plastic culture dishes. There was no significant difference in cell viability between osteoblasts cultured on either carboxyl or hydroxyapatite coated scaffolds. Cell-cell interactions with the 3D scaffold were more evident at 40x magnification, and actin filament staining highlighted the attachment points where cells were fixed to the 3D scaffold, as shown in Figure 7.

[0091] Figure 7 shows osteoblasts cultured on carboxyl-functionalized and hydroxyapatite-coated 3D scaffolding materials, with (a+b) cells cultured for 48 hours at 10x magnification, (c+d) cells cultured for 48 hours at 40x magnification, (e+f) cells cultured for 7 days at 10x magnification, and (g+h) cells cultured for 7 days at 20x magnification.

[0092] The osteoblasts survived and continued to interact with the scaffold material for the remainder of the culture period. These results indicate that the 3D scaffold material promotes osteoblast growth, whether or not it has a hydroxyapatite coating.

[0093] Osteoblast proliferation: Osteoblasts are placed on a 24-well insert at a rate of 1 × 10⁶ cells. 5 Cells were seeded at a density and cultured under standard tissue culture conditions of 37°C and 5% CO2. Dulbecco's modified Eagle medium (DMEM) (high glucose + 2 mM glutamine) containing 10% FCS supplemented with fungizone and penicillin / streptomycin was used. Cellular F-actin was stained with FITC-labeled (FITC-conjugated) phalloidin, followed by counterstaining with the nuclear stain Hoechst 33342. Images were taken at 24 hours, 48 ​​hours, and 7 days using a Nikon Eclipse Ti-E phase-contrast microscope (Nikon, Tokyo, Japan) (Figure 6). Figure 6 shows carboxyl and hydroxyapha. This shows the metabolic activity assay (CCK-8) of osteoblasts cultured for 9 days on a tightly coated 3D scaffold material.

[0094] Cell proliferation assay: Cell proliferation was monitored at various time points during a 9-day culture period using the CCK-8 assay kit (Dojindo Laboratories, Kumamoto Prefecture, Japan). A standard curve for determining cell count was created according to the manufacturer's guidelines. The cell count was 5 × 10⁶. 4 After sowing at an initial density onto scaffolding material, the material was incubated under standard culture conditions. At each time point, CCK-8 solution (50 mm) was used. 3 ) in the culture medium (500 mm) in each well. 3 ) Next, the cells were incubated under standard culture conditions for 2 hours. Aliquots of the solution from each well (3 × 100 mm) were taken. 3 The sample was pipetteed into labeled wells in a 96-well plate. A suitable control sample was also used. FLUOstar Optima plate reader Using (BMG Labtech, Ortenberg, Germany), background at 600nm Along with the reading, the absorbance at 485 nm was read and the results were recorded.

[0095] Example 15: Biocide formulation: Biocides for personal care, cosmetics, home care, and general disinfection currently have limited contact time with the surface to be treated due to abrasion. For example, surface sprays used for disinfection in hospitals have limited active use. They have a limited lifespan and therefore exhibit reduced activity against hospital-acquired infections such as MRSA, Pseudomonas aeruginosa, and C. difficile. Furthermore, some surface sprays contain organic solvents such as isopropanol or non-biodegradable components such as silicone oil to reduce the removal of the biocide by abrasion.

[0096] Cationic and amphoteric biocides, such as quaternary ammonium compounds, act against pathogens by solubilizing cell membranes, leading to cell lysis and death. Many commercially available biocides exist for disinfection, including cationic compounds such as chlorhexidine, benzalkonium chloride, crimbazole, didecyldimethylammonium chloride, and dodecyldipropylenetriamine. Furthermore, some biocides are polymeric cationic compounds, such as poly(diallyldimethylammonium chloride). These compounds can be readily incorporated into spherical microparticles using the techniques described herein, which may reduce abrasion removal on surfaces, skin, and hair, and allow for controlled release of the biocide. Moreover, biocides containing multiple cationic compounds within the same microparticle are possible, allowing for formulations that can be tailored to specific applications with clearly defined sources of infection, providing targeted formulations.

[0097] The samples produced were as follows: Poly(diallyldimethylammonium chloride) (PDAC) Spheri Somes: PDAC (1.615g, 10 mmol) in water (50cm 3 It was dissolved in ) and NaOH (0.4 g, 10 mmol) was added. Brassic acid (1.22 g, 5 mmol) was added to this solution and allowed to dissolve overnight. This appeared to be a clear solution, but under microscopic observation it was a suspension of approximately 3 μm fine particles, confirming that it was a new formulation of PDAC, and the results shown in Figure 8 demonstrate the formation of PDAC-brassic acid fine particles.

[0098] Didecyldimethylammonium chloride (DDAC): DDAC (9.04cm) 3 A 40% w / w solution (10 mmol) is diluted with water in 50 cm³. 3The solution was diluted, and NaOH (0.4 g, 10 mmol) was added. Brassic acid (1.22 g, 5 mmol) was added to this solution and allowed to dissolve overnight. This appeared to be a cloudy solution, but microscopic observation revealed it to be a suspension of approximately 3 μm fine particles, confirming it to be a new formulation of DDAC.

[0099] Dodecyldipropylenetriamine (DDPT): DDPT (9.97cm) 3 A 30% w / w solution (10 mmol) is diluted with water in 50 cm³. 3 The solution was diluted, and brassic acid (3.66 g, 15 mmol) was added and dissolved overnight. Although it appeared as a clear solution, microscopic observation revealed it to be a suspension of approximately 3 μm particles, confirming it to be a new formulation of DDPT.

[0100] Example 16: Antibacterial wound dressing: The hydrophilicity of porous polymers formed by impacting biscarboxylate fatty acid microparticles is advantageous in absorbable wound dressings. By binding biscarboxylate fatty acids to poly-ε-lysine and crosslinking them to form such a porous matrix, the inherent antibacterial activity of the wound dressing components can be retained and, if necessary, increased. In the cationic form, when poly-ε-lysine is present in excess of fatty acids, this material has been shown to retain nutrient-preserving properties, providing a novel antibacterial wound dressing. The porosity of this material, combined with its cationic properties that can disrupt microbial biofilms, enables improved skin repair as a 3D scaffolding material.

[0101] The antibiofilm capacity of cationic wound dressings was evaluated using a mixed-species CDC reactor model. The product from Example 13 was used in these experiments. As shown below, two types of mixed biofilms were prepared and tested on PBS and controlled anionic bandages.

[0102] Multiple types of biofilms 1: Staphylococcus aureus - NCTC8325; Pseudomonas aeruginosa-NCIMB10434; Acinetobacter baumannii - ATCC19606; Staphylococcus epidermidis.

[0103] Multiple types of biofilms 2: Staphylococcus aureus - NCTC8325; MRSA; VRE-Fecalis-NCTC12201; Candida albicans - ATCC-MYA-2876-SC5313; Page 3 of *Escherichia coli* - NCTC-12923, 6DOT202(03).

[0104] Preparation of mixed graft 1: Using a sterile cotton swab, collect 24-hour cultures of Staphylococcus aureus, Pseudomonas aeruginosa, Acinetobacter baumannii, and Staphylococcus epidermidis from a suitable agar plate and measure 20 cm². 3 The mixture was suspended in tryptone soy broth (TSB). This mixed species suspension was diluted in TSB to 107 ± 5 × 10⁶. 6 cfu·mL -1 The total concentration of the substance was provided and used as a graft for the CDC reactor. To promote biofilm growth, the CDC reactor was incubated at 37°C for 72 hours with shaking at 50 rpm.

[0105] Preparation of mixed graft 2: Using a sterile cotton swab, 24-hour cultures of Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, Baycomycin-resistant Enterococcus, Candida albicans, and Escherichia coli were collected from a suitable agar plate and placed in a 20 cm³ container. 3 The mixture was suspended in TSB. This mixed species suspension was diluted in TSB to 107 ± 5 × 10⁷. 6 cfu·mL -1 The total concentration of the substance was provided and used as a graft for the CDC reactor. To promote biofilm growth, the CDC reactor was incubated at 37°C for 72 hours with shaking at 50 rpm.

[0106] Biofilm treatment: After incubation, the specimens were removed from the CDC reactor and washed three times in sterile phosphate-buffered saline (PBS) to remove plankton-like cells. Next, the washed specimens were processed by sandwiching them between two discs of wound dressing material. Before testing, 400 mm of material was placed on each disc. 3 The bandage was activated by adding PBS + 1% TSB to the sample. 3 The samples were immersed in PBS + 1% TSB. All samples were tested three times. After a 24-hour treatment time, the samples were cut to 1 cm to collect the viable microorganisms attached to them. 3 The samples were placed in PBS and sonicated for 15 minutes. The recovered microorganisms were quantified using serial dilutions and smear plates.

[0107] Mixed graft 1: After treatment with the control bandage (A), the bacterial recovery rate was equivalent to that of the control sample treated with PBS alone, as shown in Figure 9. No viable organisms were recovered from the sample treated with the cationic bandage (B). This represents a decrease greater than 10^5 compared to the PBS-treated control sample. The surviving organisms after treatment were mainly Pseudomonas aeruginosa (Figure 10).

[0108] Mixed graft 2: Treatment with the control bandage (A) resulted in a 10¹²⁷ reduction in the number of viable bacteria compared to the PBS-treated control sample. No viable organisms were recovered from the sample treated with the cationic bandage (B). This represents a greater reduction than 10⁷⁷ compared to the PBS-treated control sample (Figure 11). The viable organisms were a mixed species (Figure 12). The present invention includes the following embodiments. [1] Self-assembled microparticles containing an acid having two or more acidic groups and an organic base. [2] [1] Fine particles having a particle size of 0.5 to 10 microns, preferably 1 to 5 microns. [3] [1] or [2], wherein the molar ratio of acidic groups in the acid to basic groups in the base is 0.6 to 1.4:1. Fine particles as described in any of [4][1] to [3], wherein the molar ratio of acidic groups to basic groups is 0.7 to 1.3:1. Fine particles as described in any of [5][1] to [4], which are suitable for use as a particulate support containing self-assembled fine particles having two or more acid groups and an organic base, and which can be obtained by a method comprising contacting the bis acid with the organic base in a hydrophilic solvent, wherein the acid is insoluble or sparingly soluble in the hydrophilic solvent and the organic base is soluble in the hydrophilic solvent. The fine particles described in [6][5], wherein the solvent comprises an aqueous solution. The fine particles described in [7][5], wherein the solvent comprises an oil-in-water emulsion in the aqueous phase. Fine particles as described in any of [8][1] to [7], wherein the acid contains bis-acid. Fine particles as described in any of [9][1] to [8], wherein the acid contains a bis-fatty acid. Fine particles according to any of

[10] [1] to [9], wherein the acid contains a biscarboxyl fatty acid in which a plurality of terminal carboxylic acids are linked by hydrophobic regions. Fine particles according to any of

[11] [1] to

[10] , wherein the acid group is separated by a saturated or unsaturated aliphatic chain; or a substituted saturated or unsaturated aliphatic chain.

[12] [1] to

[11] are fine particles, wherein the acid is of the general formula: HOOC-(CH 2 ) n Fine particles containing a compound of -COOH (wherein n is large enough that bisic acid is sparingly soluble or insoluble in water). The fine particles described in

[13]

[12] , wherein n is at least 5 and 40 or less. Fine particles as described in any of

[14] [1] to

[13] , wherein the acid comprises brassic acid, sebacic acid, and / or azelaic acid. Fine particles according to any of

[15] [1] to

[14] , wherein the organic base comprises an aliphatic amine or aromatic amine having basic properties or other nitrogen-containing base properties. Fine particles according to any of

[16] [1] to

[15] , wherein the organic base comprises one or more alkylated amines and alkylated polyamines. Fine particles according to any one of

[17] [1] to

[16] , wherein the organic base comprises one or more of N-methylmorpholine, N,N-dimethylaminoethanol, 4-dimethylaminopyridine, imidazole, 1-methylimidazole, poly(diallyldimethylammonium chloride) (PDAC), didecyldimethylammonium chloride (DDAC), dodecyldipropylenetriamine (DDPT), and poly-ε-lysine. A fine particle as described in any of

[18] [1] to

[17] , wherein the fine particle includes a multilamellar structure. Self-organizing microspheroids as described in any of

[19] [1] to

[18] . Fine particles according to any of

[20] [1] to

[19] , wherein the bisic acid reacts with the organic base to form a crosslinked species. A fine particle according to any of

[21] [1] to

[20] , wherein the organic base is replaced by another reactive base, which then reacts to form a crosslinked species. A macroporous material in which fine particles described in any of

[22] [1] to

[21] are brought into contact to form crosslinks between the fine particles, thereby forming a three-dimensional body. Self-assembled microparticles as described in any of

[23] [1] to

[21] and macroporous material as described in

[22] , wherein the functional material absorbed or covalently bonded to the microparticles and / or macroporous material is selected from catalysts, initiators for peptide synthesis, initiators for oligonucleotide synthesis, initiators for solid-phase organic synthesis, pharmacologically active substances, pesticide active substances, proteins, enzymes, or other biomacromolecules. A medical diagnostic agent comprising a functional material bonded to or held by a support, comprising self-assembled microparticles as described in any of

[24] [1] to

[21] or a macroporous material as described in

[22] . A medical diagnostic agent according to

[25]

[24] , comprising an enzyme on which the functional material is supported by the polymer.

[26] A monolith containing the macroporous material described in

[23] , housed within a column.

[27] Use of self-assembled microparticles according to any of [1] to

[21] or macroporous materials according to

[22] in chemical, biological, or physical processes. Use of self-assembled microparticles and / or macroporous materials as described in

[28]

[27] , in a process selected from: solid-phase synthesis of species selected from peptides, oligonucleotides, and oligosaccharides; solid-phase extraction; solid-phase organic chemistry; immobilization of species selected from solid-phase reagents, metal and other catalysts, biocatalysts, enzymes, proteins, antibodies including polyclonal and monoclonal antibodies, whole cells, and polymers; cell culture; preparation of stationary phases for chromatographic separation; or use as an absorbent.

[29] Use of self-organizing microparticles according to any of [1] to

[21] or macroporous material according to

[22] in the care or treatment of any internal or external wound of the body.

[30] Use of self-assembled microparticles and / or macroporous materials according to any of [1] to

[26] or the macroporous material according to

[22] as a support or three-dimensional scaffold material for cell culture, regenerative medicine, or tissue repair.

[31] A method for producing self-assembled fine particles or macroporous material according to any one of [1] to

[21] in an aqueous medium, comprising contacting a diacid having two or more acid groups with an organic base in an aqueous medium, preferably in water. An antimicrobial composition comprising self-organizing microparticles as described in any of

[32] [1] to

[21] .

Claims

1. Self-assembled microparticles containing an acid having two or more acidic groups and an organic base, The molar ratio of acidic groups in the acid to basic groups in the base is 0.7 to 1.3:

1. The acid comprises brassic acid, sebacic acid, and / or azelaic acid. Self-assembled microparticles comprising at least one organic base selected from the group consisting of alkylated amines and alkylated polyamines.

2. Fine particles according to claim 1, wherein the fine particles have a particle size of 0.5 to 10 microns.

3. Fine particles according to claim 1 or 2, wherein the functional material absorbed or covalently bonded to the self-assembled fine particles is selected from catalysts, metal chelating agents, initiators for peptide synthesis, initiators for oligonucleotide synthesis, initiators for solid-phase organic synthesis, pharmacologically active substances, pesticide active substances, proteins, enzymes, or other biomacromolecules.

4. Fine particles according to any one of claims 1 to 3, which are used in therapies for the treatment of the human or animal body.

5. The fine particles according to claim 4, wherein the use includes topical application of the fine particles to the body of a human or animal.

6. A macroporous material comprising fine particles according to any one of claims 1 to 5, wherein the fine particles are crosslinked with each other to form a three-dimensional body.

7. A macroporous material according to claim 6, wherein the organic base of the fine particles is replaced with another reactive base, which then reacts to form a crosslinking species, thereby providing the macroporous material.

8. A medical diagnostic agent comprising self-assembled microparticles according to any one of claims 1 to 5 and a functional material bonded to or held by a support.

9. Use of self-assembled microparticles according to any one of claims 1 to 5 in a chemical, biological, or physical process.

10. Use of self-assembled microparticles according to claim 9, for use in processes selected from: solid-phase synthesis of species selected from peptides, oligonucleotides, and oligosaccharides; solid-phase extraction; solid-phase organic chemistry; immobilization of species selected from solid-phase reagents, metal and other catalysts, biocatalysts, enzymes, proteins, antibodies including polyclonal and monoclonal antibodies, whole cells, and polymers; cell culture; preparation of stationary phases for chromatographic separation; or use as an absorbent.

11. Use of self-assembled microparticles according to any one of claims 1 to 5 as a support or three-dimensional scaffold material for cell culture, regenerative medicine, or tissue repair.

12. A topical composition for use in personal care products or home care products, comprising self-assembled microparticles according to any one of claims 1 to 5.

13. i) Pharmacologically active substances or pesticide active substances; and ii) Self-assembled fine particles according to any one of claims 1 to 5 A composition containing the following:

14. An article or wound treatment product comprising self-organizing microparticles according to any one of claims 1 to 5.

15. i) Fine particles according to any one of claims 1 to 5; and ii) Hydroxyapatite A composition used for the growth of osteocytes, including the components mentioned above.

16. A method for producing self-assembled fine particles according to any one of claims 1 to 5, The method comprises contacting an acid having two or more acidic groups with an organic base in a hydrophilic solvent, wherein the acid is insoluble or sparingly soluble in the hydrophilic solvent, and the organic base is soluble in the hydrophilic solvent. i) The acid comprises brassic acid, sebacic acid, and / or azelaic acid, ii) The organic base comprises at least one selected from the group consisting of alkylated amines and alkylated polyamines, iii) A method wherein the molar ratio of acidic groups in the acid to basic groups in the base is 0.7 to 1.3:1.

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