Amorphous nanomolecular aggregates composed of organic substances, inorganic substances, or salts thereof, and methods for producing the same.

Abstract

The present invention relates to an amorphous nanomolecular aggregate composed of an organic / inorganic substance or a salt thereof, and more specifically, to an amorphous nanomolecular aggregate which is produced by applying shear stress to an organic / inorganic substance or a salt thereof and has excellent solubility and permeability through lipid membranes.

Description

[Technical Field]

[0001] The present invention relates to amorphous nanomolecular aggregates composed of organic / inorganic substances or salts thereof, and more particularly to amorphous nanomolecular aggregates produced by applying shear stress to organic / inorganic substances or salts thereof, which exhibit excellent solubility and permeability to lipid membranes. [Background technology]

[0002] Generally, it is known that in order for a drug, whether in the form of a substance, an inorganic substance, or a salt thereof, to effectively deliver a pharmacologically active ingredient (so-called active ingredient) to its target location in the human body, the following two conditions must be met:

[0003] First, the soluble capacity of the active ingredient in water must be ensured. Since all fluids in the human body are water-based solutions or dispersions, sufficient solubility in water and a sufficiently dispersed phase in water must be ensured for the drug to be delivered to move within the body.

[0004] Furthermore, permeability of the active ingredient through hydrophobic membranes must be ensured. Since cells in the human body are surrounded by hydrophobic membranes such as phospholipid membranes, in order to pass through them, the surface properties of drug molecules or drug structures must be hydrophobic, or they must be extremely small enough to penetrate the cell membrane.

[0005] Conventional methods for such drugs to penetrate cell membranes in a molecular state have often involved encapsulation methods that utilize a third substance, a surfactant / polymer structure, to contain the drug in the form of an emulsion or suspension within microspheres.

[0006] However, such drug encapsulation methods have limitations, including the difficulty of the encapsulation process, which can result in low encapsulation yields. Furthermore, there are problems such as the removal of surfactants and polymer components surrounding the drug in order to release it, or the need for the drug to pass through these components. Additionally, since the surfaces of cells and tissues that require permeability are mostly composed of lipophilic phospholipid components, there is a problem that drug absorption is inefficient when the drug is hydrophilic.

[0007] To address the problems with such drug encapsulation methods, many researchers have utilized a third substance and leveraged physicochemical bonding, as mentioned earlier, rather than relying on the structure of the molecule itself.

[0008] To disperse or dissolve drug molecules in water, methods can be employed such as giving the drug molecules polarity to induce polar interactions with water molecules, or by covalently or ionically bonding (complexing) molecules that are very compatible with water, such as PEG, to part of the molecular structure.

[0009] However, this method has drawbacks: it requires the cumbersome process of altering the drug's molecular structure, leading to increased costs due to the manufacturing process; and because the altered molecular structure differs structurally from the original, it reverts to its original molecular state after absorption in the body, thus failing to exhibit the intended drug effect. Consequently, there are limitations in maintaining the drug's original efficacy because the drug molecule cannot maintain multiple aggregated states without chemical structural changes. Furthermore, conventional drug structures reaching several hundred nanometers in size were difficult to penetrate skin gaps with a size of 80 nm.

[0010] Therefore, various methods for producing such drugs have been studied. In particular, methods for producing drugs on a nanometer scale include top-down techniques such as high-pressure homogenization, milling, and piston-gap homogenizers, and bottom-up techniques such as precipitation and self-assembly.

[0011] The top-down method (downward process) is a technology that reduces particle size through methods such as grinding to produce micrometer-sized particles, but it has disadvantages such as increased costs, the risk of contamination, and product damage due to repeated grinding. On the other hand, the bottom-up method (upward process), in contrast to the top-down method, is a technology that grows structures to nanometer size at the atomic and molecular level, and it has been presented as a solution to overcome the limitations of the conventional top-down method, which reduces the size to nanometer size in a bulk state. However, at the current level of technology, it is difficult to directly obtain the economic advantages of atomic and molecular level technology. Furthermore, although the conventional method of crystal growth has the advantage of low cost and a simple manufacturing process, it can only produce crystalline products, and there is a problem that excessive crystal growth and their aggregation require the addition of other compounds such as surfactants. [Prior art documents] [Patent Documents]

[0012] [Patent Document 1] Korean Published Patent No. 2011-0053775 (May 24, 2011), Nanopowder Dispersion Apparatus Using Focused Ultrasound and Dispersion Method Using the Same. [Overview of the project] [Problems that the invention aims to solve]

[0013] Therefore, in order to solve the aforementioned problems, the inventors of the present invention confirmed that when drug molecules, which are pharmacologically active ingredients, are dissolved in a solvent and then brought very close to each other, the polar groups within the molecules behave as if they were a single entity.

[0014] Furthermore, we confirmed that when polar groups interact with each other due to the proximity of molecules, the drug molecules as a whole become hydrophobic. This reduces the size of the structure to which such drug molecules are bound, lowering the surface tension and thus increasing the degree of dispersion.

[0015] Through this process, we confirmed that it is possible to manufacture small particles at the molecular level using a bottom-up method rather than a top-down method, while producing amorphous, nano-sized drug structures rather than crystalline forms.

[0016] Through this process, we confirmed that it is possible to create structures in which molecules with novel properties are bonded together by various methods that bring molecules closer together, such as manufacturing molecular structures by utilizing the voids in powders or bringing molecules closer together using flexible rolls, thus completing the present invention.

[0017] Therefore, the object of the present invention is to provide a molecular aggregate of a pharmacologically active ingredient in which the surface of the molecular aggregate is hydrophobic and its permeability to a phospholipid membrane is increased, by producing a molecular aggregate with a new structure that utilizes the polar interactions or hydrogen bonds of the molecules, without changing the chemical structure of the molecules of the pharmacologically active ingredient.

[0018] Therefore, the present invention is not manufactured by the conventional top-down method but by a new method. Despite having the advantage of reaching the highest saturation solubility possessed by conventional amorphous drug nanoparticles, it improves the problems with respect to the shelf-life maintenance period and the problem due to the very fast dissolution rate of the amorphous nanoparticle's particle size, and is expected to bring innovation to the field of nanomedicine, that is, ultimately to the field of nanotechnology. Conclusively, the present invention will become a new innovative technology in the field of amorphous nanomedicine and nanotechnology with a size of 50 nm or less.

Means for Solving the Problems

[0019] To achieve the above object, the present invention provides an amorphous nano molecular aggregate in which an organic or inorganic substance is physically bonded, and when the molecular aggregate is formed with a composition containing water, the molecular aggregate has an aggregated structure in the composition, and the average particle size of the molecular aggregate is 50 nm or less.

[0020] The present invention also provides an amorphous nano molecular aggregate in which an organic or inorganic salt is physically bonded, and when the molecular aggregate is formed with a composition containing water, the molecular aggregate has an aggregated structure in the composition, and the average particle size of the molecular aggregate is 50 nm or less.

[0021] The present invention also provides a method for producing a nano molecular aggregate, in which after a composition containing an organic substance, an inorganic substance or a salt thereof; and water; is put into a device capable of applying a shear stress,

[0022] a shear stress is applied to the composition to produce a molecular aggregate in which an organic substance, an inorganic substance or a salt thereof is physically bonded.

Effects of the Invention

[0023] The present invention does not have the annoyance of having to change the molecular structure of organic substances, inorganic substances, or salts thereof such as pharmacologically active ingredients, and by producing a molecular aggregate with a new structure that utilizes polar interactions or hydrogen bonds, the surface of the molecular aggregate can be made hydrophobic, increasing the permeability to phospholipid membranes. Moreover, since the chemical properties themselves are the same as those of the original molecule, the effect of the originally intended drug can be shown as it is, and the manufacturing method is simple and has the advantage of being able to save manufacturing costs.

[0024] Due to such advantages, the molecular aggregate of the present invention can be used in various pharmaceutical compositions.

Brief Description of the Drawings

[0025] [Figure 1] It is a schematic diagram of an apparatus for producing a molecular aggregate of a pharmacologically active ingredient according to an embodiment of the present invention. [Figure 2] It is a schematic diagram of an apparatus for producing a molecular aggregate of a pharmacologically active ingredient according to another embodiment of the present invention. [Figure 3] It is a graph showing the result of measuring NOESY, which is a two-dimensional NOE (Nuclear Overhauser Effect) spectrum, for a pharmacologically active ingredient according to an embodiment of the present invention. [Figure 4] It is a graph showing the result of measuring NOESY, which is a two-dimensional NOE spectrum, for a molecular aggregate of a pharmacologically active ingredient according to an embodiment of the present invention. [Figure 5] It is a figure showing a comparison of NMR spectra according to an embodiment of the present invention. [Figure 6] It is a figure showing a comparison of FT-IR spectra according to an embodiment of the present invention. [Figure 7a] It is a figure showing a comparison of HPLC measurement results according to an embodiment of the present invention. [Figure 7b] It is a figure showing a comparison of HPLC measurement results according to an embodiment of the present invention. [Figure 7c] This figure shows a comparison of HPLC measurement results according to one embodiment of the present invention. [Figure 8] This figure shows a comparison of the NMR spectrum according to another embodiment of the present invention. [Figure 9] This figure shows a comparison of the FT-IR spectrum according to another embodiment of the present invention. [Figure 10] This figure shows a comparison of HPLC measurement results according to another embodiment of the present invention. [Figure 11] This figure shows a comparison of the FT-IR spectrum according to another embodiment of the present invention. [Figure 12] This figure shows a comparison of the FT-IR spectrum according to another embodiment of the present invention. [Figure 13] This is a TEM image of a molecular aggregate according to another embodiment of the present invention. [Figure 14] This graph shows the XRD measurement results of a molecular aggregate according to another embodiment of the present invention. [Figure 15] This graph shows the XRD measurement results of a molecular aggregate according to another embodiment of the present invention. [Figure 16] This graph shows the DSC measurement results of a molecular aggregate according to another embodiment of the present invention. [Figure 17] This is a TEM image of a molecular aggregate according to another embodiment of the present invention. [Figure 18] This is a TEM image of a molecular aggregate according to another embodiment of the present invention. [Figure 19] This is a TEM image of a molecular aggregate according to another embodiment of the present invention. [Figure 20] This is a 3D hologram image showing the permeability of a phospholipid membrane of a molecular aggregate according to another embodiment of the present invention. [Figure 21] This is a schematic diagram of a Wushing chamber used to measure the corneal penetration performance of a molecular aggregate according to another embodiment of the present invention. [Modes for carrying out the invention]

[0026] In this invention, we present molecular aggregates of organic substances, inorganic substances, or salts thereof that have an amorphous, nano-sized drug structure rather than a crystalline form, while being manufactured using a bottom-up method rather than a top-down method at the molecular level. Furthermore, without altering the chemical structure of organic substances, inorganic substances, or salts thereof, we produce molecular aggregates with novel structures that utilize polar interactions or hydrogen bonding, thereby making the surface of the molecular aggregates hydrophobic and increasing their permeability to phospholipid membranes. The following provides a more detailed explanation.

[0027] Apparatus for producing molecular aggregates of organic substances, inorganic substances, or salts thereof. The apparatus for producing molecular aggregates of organic substances, inorganic substances, or salts thereof as referred to herein is characterized by introducing a solution containing organic substances, inorganic substances, or salts thereof into the apparatus, and then applying shear stress to the solution containing organic substances, inorganic substances, or salts thereof to produce molecular aggregates of organic substances, inorganic substances, or salts thereof.

[0028] Methods for producing drugs at the nanometer scale included top-down techniques such as high-pressure homogenization, milling, and piston-gap homogenizers, as well as bottom-up techniques such as precipitation and self-assembly.

[0029] The top-down manufacturing method is a technique that reduces particle size through methods such as grinding to produce micrometer-sized particles, but it has disadvantages such as increased costs, the risk of contamination, and product damage due to repeated grinding.

[0030] On the other hand, bottom-up manufacturing techniques such as precipitation have the advantage of being low-cost and having a simple manufacturing process through crystal growth, but they have the problem that they can only produce crystalline products, and excessive crystal growth and their aggregation require the addition of other compounds such as surfactants.

[0031] Furthermore, in order to effectively deliver pharmacologically active ingredients, which are in the form of organic, inorganic, or salts thereof, to targets within the human body, the solubility in water and permeability to hydrophobic membranes of these organic, inorganic, or salt substances must be ensured. Conventional methods for such drugs to permeate cell membranes in molecular form have often involved giving the drug polarity to induce polar interactions with water molecules, or forming a complex with water-friendly molecules such as PEG via covalent or ionic bonds in part of the molecular structure.

[0032] However, this method has the drawback of requiring the alteration of the drug's molecular structure, and because the altered molecular structure is structurally different from the original molecule, it has the limitation that after being absorbed into the body, it may revert to its original molecular state, thus failing to exhibit the intended drug effect.

[0033] To solve the aforementioned problems, the inventors have confirmed that when drug molecules, in the form of organic, inorganic, or salts thereof, are dissolved in a solvent and then brought very close together, the polar groups within the molecules interact to form molecular aggregates, which then behave like a single entity.

[0034] Through this process, we confirmed that when polar groups interact with each other due to the proximity of molecules, the overall structure becomes hydrophobic. This reduces the size of the structure to which such drug molecules are bound, lowering the surface tension and thus increasing the degree of dispersion.

[0035] We have completed the present invention by confirming that molecular aggregates with novel properties can be created by bringing molecules closer together through various methods that reduce the distance between molecules, such as manufacturing molecular structures using voids in powders and bringing molecules closer together using flexible rolls.

[0036] In other words, the present invention is for producing small particles at the molecular level using a bottom-up method rather than a top-down method, while producing amorphous, nano-sized drug structures rather than crystalline ones.

[0037] First, the present invention provides an apparatus for producing molecular aggregates of organic substances, inorganic substances, or salts thereof by introducing a solution containing organic substances, inorganic substances, or salts thereof into the apparatus and then applying shear stress to the solution containing organic substances, inorganic substances, or salts thereof. The apparatus for producing molecular aggregates of organic substances, inorganic substances, or salts thereof can be used without special limitations as long as it is capable of producing the molecular aggregates of organic substances, inorganic substances, or salts thereof of the present invention by applying shear stress to a solution containing organic substances, inorganic substances, or salts thereof. Preferably, an apparatus that uses a roll mill process or a ball mill process to apply shear stress can be used.

[0038] When preparing a solution containing the aforementioned organic or inorganic substances or salts thereof, it may also be prepared using an oil phase. In this case, the solvent used for preparation in the oil phase may be one or more of the following: oils derived from grain extracts such as castor oil, MCT oil, soybean oil, and peanut oil, or oils derived from herbal extracts that exhibit pharmacological effects, such as ginseng, camellia, green tea, and angelica tree.

[0039] Furthermore, in the present invention, the organic substances, inorganic substances, or salts thereof can be used as pharmacologically active ingredients, and these can be used without any special limitations as long as they are pharmaceutically useful substances or substances that have medical effects. Examples include cyclosporine A, paclitaxel, docetaxel, declusin, meloxicam, itraconazole, celecoxib, capecitabine, travoprost, isoflavones, diclofenac sodium, tyrosine kinase inhibitors such as sunitinib, pazopanib, axitinib, regorafenib, trametinib, ginsenoside Rg1, tacrolimus, alendronate, latanoprost, bimatoprost, atorvastatin calcium, rosuvastatin calcium, entecavir, amphotericin B, omega-3, and deodorant. Various cholic acids such as xycholic acid and ursodeoxycholic acid, and their sodium or potassium salts; steroids such as prednisolone, which are substituted with fluorine or hydrogen; aromatic oils such as eucalyptus oil, lavender oil, lemon oil, sandalwood oil, rosemary oil, chamomile oil, cinnamon oil, and orange oil; alpha-bisabolol, vitamin A (retinol), vitamin E, tocopheryl acetate, vitamin D, vitamin F, or derivatives thereof; and combinations thereof may be used.

[0040] As one specific example of a device that applies shear stress in this manner, Figure 1 shows a schematic diagram illustrating a device for producing molecular aggregates of organic substances, inorganic substances, or salts thereof according to one embodiment of the present invention.

[0041] As shown in Figure 1, an apparatus for producing molecular aggregates of organic substances, inorganic substances, or salts thereof according to one embodiment of the present invention may be equipped with a plurality of rolls so as to be able to apply shear stress to a solution containing organic substances, inorganic substances, or salts thereof. In this case, the plurality of rolls may be two opposing rolls, or one or more additional rolls may be included.

[0042] Specifically, a solution containing the aforementioned organic matter, inorganic matter, or salts thereof (shown as PTX Sol. in Figure 1 as an example) can be introduced between two opposing rolls in the apparatus (roll A and roll B in Figure 1). By rotating the two opposing rolls, shear stress is applied to the organic matter, inorganic matter, or salts contained in the solution, thereby producing molecular aggregates of the organic matter, inorganic matter, or salts thereof according to the present invention.

[0043] In an apparatus for producing molecular aggregates of organic substances, inorganic substances, or salts thereof according to one embodiment of the present invention, the distance between the two opposing rolls may be set to 0.5 to 1000 μm. The distance between the two opposing rolls may be set to, for example, 0.5 μm or more, 1 μm or more, 2 μm or more, 3 μm or more, 4 μm or more, 5 μm or more, 10 μm or more, 1000 μm or less, 900 μm or less, 800 μm or less, 700 μm or less, 600 μm or less, 500 μm or less, 400 μm or less, 300 μm or less, 200 μm or less, 100 μm or less, and preferably to 10 to 100 μm. If the distance between the two opposing rolls is less than 0.5 μm, the discharge amount between the rolls is very small, which is a problem for production speed, and if it is greater than 1000 μm, the shear stress and compressive stress are very small, which is a problem for particle formation.

[0044] As mentioned above, when the solution is passed between the rollers, excessive force is applied between molecules, resulting in a bottom-up particle structure where particles aggregate between molecules, rather than a top-down grinding process.

[0045] Furthermore, in order to more efficiently transfer shear stress to organic substances, inorganic substances, or salts thereof contained in the solution, the apparatus for producing molecular aggregates according to one embodiment of the present invention can be configured such that the two opposing rolls rotate at different speeds. In this case, one of the two opposing rolls can be configured to rotate at a speed of 50 to 250 rpm, and the other at a speed of 200 to 500 rpm. Alternatively, the two opposing rolls can be rotated in a ratio of 1:1.5 to 1:5.

[0046] Furthermore, in an apparatus for producing molecular aggregates according to one embodiment of the present invention, the rotation directions of the two opposing rolls may be set to co-current directions, where they have the same rotation direction, or to counter-current directions, where their rotation directions are different.

[0047] Furthermore, the apparatus for producing molecular aggregates according to one embodiment of the present invention can repeatedly apply shear stress and compressive stress to the contents that are dispensed once, several times.

[0048] Furthermore, the present invention provides an apparatus for producing molecular aggregates by introducing a solution containing organic matter, inorganic matter, or salts thereof into the apparatus in a first direction, introducing another solution in a second direction opposite to the first direction, and then applying shear stress. The apparatus for producing molecular aggregates of the present invention can also be used without special limitations as long as it can produce molecular aggregates of the present invention by applying shear stress to a solution containing organic matter, inorganic matter, or salts thereof, but preferably it can be used in which a roll mill process or a ball mill process is used to apply shear stress.

[0049] In the present invention, the pharmacologically active ingredient can be the same as that mentioned above.

[0050] In the present invention, the water-soluble compound can be one or more selected from the group consisting of citric acid, carbonic acid, lactic acid, acetic acid, phosphoric acid, ascorbic acid, malic acid, tartaric acid, glutaric acid, succinic acid, maleic acid, fumaric acid, malonic acid, HCl, H2SO4, NaH2PO4, NaHCO3, KHCO3, Na2CO3, K2CO3, Na3PO4, K3PO4, NaH2PO4, NH4OH, sodium acetate (NaOAc), KOH, NaOH, and Ca(OH)2.

[0051] As a specific example of a device that applies shear stress in this way, Figure 2 shows a schematic diagram illustrating a device for producing molecular aggregates according to one embodiment of the present invention.

[0052] As shown in Figure 2, the apparatus for producing molecular aggregates according to one embodiment of the present invention may be equipped with multiple rolls so as to be able to apply shear stress to a solution containing organic matter, inorganic matter, or salts thereof. In this case, the multiple rolls may be two opposing rolls (roll A, roll B), or one or more additional rolls (roll C) may be included as shown in Figure 2.

[0053] Specifically, a solution containing the organic, inorganic, or salt thereof (shown as PTX Sol. in Figure 2 as an example) can be introduced to the first roll side, and a solution containing the water-soluble compound (shown as Sucrose Sol. in Figure 2 as an example) can be introduced to the second roll side facing the first roll.

[0054] In this manner, when two opposing rolls rotate over solutions containing organic substances, inorganic substances, or salts thereof, and a solution containing a water-soluble compound, which are fed into the two rolls in opposite directions, shear stress is applied to the organic substances, inorganic substances, or salts thereof and the water-soluble compound contained in these solutions, thereby enabling the production of molecular aggregates of organic substances, inorganic substances, or salts thereof according to the present invention.

[0055] In an apparatus for producing molecular aggregates according to one embodiment of the present invention, the distance between the two opposing rolls may be set to 0.5 to 1000 μm. The distance between the two opposing rolls may be set to, for example, 0.5 μm or more, 1 μm or more, 2 μm or more, 3 μm or more, 4 μm or more, 5 μm or more, 1000 μm or less, 900 μm or less, 800 μm or less, 700 μm or less, 600 μm or less, 500 μm or less, and preferably to 5 to 500 μm.

[0056] If the distance between the two opposing rolls is less than 0.5 μm, the discharge volume between the rolls is very small, resulting in production speed problems. If it is greater than 1000 μm, the shear stress and compressive stress are very small, resulting in problems with particle formation.

[0057] Furthermore, in order to more efficiently transfer shear stress to organic matter, inorganic matter, or salts thereof contained in the solution, the two opposing rolls can be made to rotate at different speeds. In this case, one of the two opposing rolls can be used with a rotation speed of 50 to 150 rpm and the other with a rotation speed of 200 to 500 rpm. Alternatively, the two opposing rolls can be rotated in a ratio of 1:1.5 to 1:5.

[0058] Furthermore, in an apparatus for producing molecular aggregates according to one embodiment of the present invention, the rotation directions of the two opposing rolls may be set to co-current directions having the same rotation direction, or they may be set to counter-current directions having different rotation directions.

[0059] Furthermore, the apparatus for producing molecular aggregates according to one embodiment of the present invention can repeatedly apply shear stress and compressive stress to the contents that are dispensed once, several times.

[0060] In the present invention, the apparatus for producing the molecular aggregate may be further equipped with a third roll that applies shear stress again to the solution passing between the first and second rolls. The third roll can apply shear stress in relation to the second roll, and the distance between the opposing second and third rolls may be set to 0.5 to 1000 μm. The distance between the two opposing rolls may be set to, for example, 0.5 μm or more, 1 μm or more, 2 μm or more, 3 μm or more, 4 μm or more, 5 μm or more, 1000 μm or less, 900 μm or less, 800 μm or less, 700 μm or less, 600 μm or less, 500 μm or less, and preferably to 5 to 500 μm. If the distance between the second and third rolls is less than 0.5 μm, the discharge volume between the rolls is very small, which is a problem for production speed, and if it is greater than 1000 μm, the shear stress and compressive stress are very small, which is a problem for easy particle formation.

[0061] Furthermore, in order to transmit shear stress more efficiently, the second and third rolls can be made to rotate at different speeds. In this case, one of the second and third rolls can be rotated at a speed of 200 to 500 rpm, and the other at a speed of 600 to 1200 rpm. Alternatively, the rotation speeds of the second and third rolls can be made to be in a ratio of 1:1.5 to 1:5.

[0062] In the present invention, the apparatus for producing the molecular aggregate is not limited to the method described above, and can be freely modified as long as it is an apparatus capable of applying shear stress to organic matter, inorganic matter, or salts thereof.

[0063] Molecular aggregates of organic substances, inorganic substances, or salts thereof The inventors have confirmed that when drug molecules, in the form of organic, inorganic, or salts thereof, are dissolved in a solvent and then brought very close together, the polar groups within the molecules interact to form molecular aggregates, which then behave like a single entity.

[0064] Through this process, we confirmed that when polar groups interact with each other due to the proximity of molecules, the overall structure becomes hydrophobic. This reduces the size of the molecular aggregate to which such drug molecules are bound, lowering the surface tension and thus increasing the degree of dispersion.

[0065] We have completed the present invention by confirming that molecular assemblies with novel properties can be created by bringing molecules closer together through various methods that reduce the distance between molecules, such as manufacturing molecular assemblies by utilizing the voids in powders and bringing molecules closer together using flexible rolls.

[0066] Therefore, the nanomolecular aggregates produced by the apparatus referred to herein have the following characteristics:

[0067] First, the nanomolecular aggregates according to the present invention are produced by applying shear stress to a solution containing organic, inorganic, or salts thereof, which are precursors to the molecular aggregates, thereby bringing the intermolecular distances of the organic, inorganic, or salts thereof very close together. This process results in nanoparticle-sized aggregates that possess amorphous characteristics. In other words, by applying shear stress between molecules of the same structure, these molecules are physically bonded together to form nano-sized amorphous molecular aggregates.

[0068] Furthermore, the molecular aggregate and the chromatographic measurements of the organic, inorganic, or salt thereof precursors of the molecular aggregate are the same, while the spectroscopy measurements of the molecular aggregate and the pharmacologically active component precursors of the molecular aggregate are different.

[0069] Common methods for determining the structure of a substance include various instrumental analyses such as elemental analysis, FT-IR, NMR, UV spectroscopy, X-ray, and DSC (differential scanning calorimeter). Among these, NMR and FT-IR are the most effective analytical methods for revealing the chemical structure, that is, the atoms that make up a molecule and their bonds. Typically, X-ray and DSC are often used to investigate secondary structures, such as the crystalline structure formed by molecules. In addition, scanning electron microscopes and transmission electron microscopes are sometimes used to analyze the fine structure.

[0070] Among the methods mentioned, NMR allows us to know the electronic environment of atomic nuclei, which is determined by the electronic structure of a molecule. The results of this invention relate to a method for producing a certain compound and its physical structure, and NMR provides very useful information as a way to check for chemical mutations during the formation of the physical structure. If there is no creation or annihilation of chemical bonds, the compound and its physical structure will basically have similar NMR spectra. Of course, in order to know how close the compounds are to form molecular aggregates, we roughly measured the intermolecular distances of compounds that are usually located at a distance of 0.5 nm using NOE (Nuclear Overhauster Effect) and two-dimensional NOESY spectra.

[0071] Furthermore, according to one embodiment of the nanomolecular aggregate according to the present invention, the molecular aggregate may have an intermolecular distance of 10 Å or less. The intermolecular distance means measuring the average distance of these molecules with respect to the molecules that make up the molecular aggregate, and can be measured, for example, using NOE in NMR. NOE (Nuclear Overhauster Effect) is a phenomenon in which the intensity of hydrogen atoms at a close distance increases when one hydrogen atom is excited using a pulse, and this is inversely proportional to the sixth power of the internuclear distance of the hydrogen atoms. Therefore, even if the hydrogen atoms are within a molecule, if they are far apart, they do not contribute to the increase in peak intensity due to NOE. However, when molecules are very close together, NOE is detected when the distance between intermolecular hydrogen atoms becomes closer than the distance between hydrogen atoms within the molecule, which means that the distance between molecules is very small. It is generally known that NOE is observed when the distance is closer than 1 nm.

[0072] In this invention as well, the intermolecular distance between the NaDC (sodium deoxycholate) compound and the molecular aggregate obtained in this invention is very small, as observed through two-dimensional NOESY NMR, and the angstrom (10) is very small. -10 We confirmed that the data was collected on a scale of approximately m.

[0073] Figures 3 and 4 show the NOESY, a two-dimensional NOE spectrum, for the NaDC molecule and its molecular aggregate, respectively. The areas marked with * and ** in Figures 3 and 4 represent the same location. In Figure 3, no off-diagonal peaks appear in the regions marked with * and ** for the NaDC molecule itself, whereas in Figure 4, off-diagonal peaks appear in the regions marked with * and ** for the NaDC molecular aggregate. This indicates that in pure NaDC, the distance between the two nuclei was far, but when the molecular aggregate is formed, the distance between the two nuclei decreases. Through this, it can be seen that when a molecular aggregate is formed, the distance between molecules becomes very small. Generally, the distance required for two hydrogen atoms to exhibit NOE is known to be approximately 6 Å (angstroms), so it can be seen that the intermolecular distance forming the molecular aggregate of the present invention is also within the range of approximately 6 Å (angstroms).

[0074] In summary, the reason why peaks that were too far away to appear in the molecules themselves appear in the nanomolecular aggregates provided by this invention is due to a change in the physical positions of the organic, inorganic, or salt molecules bound to the nanomolecular aggregate. In other words, it means that the distance between molecules within the molecular aggregate is very small.

[0075] As mentioned above, the position of the peak in the NMR spectrum represents the frequency of rotational motion of the atomic nucleus within a molecule composed of atomic bonds, depending on the magnitude of the magnetic field applied to it. The magnitude of the magnetic field applied to the atomic nucleus varies depending on the properties of surrounding active groups that push or pull electrons. Consequently, as the strength of the magnetic field applied to the nucleus increases, the peak shifts to a higher frequency. Conversely, if there are many electrons around the nucleus and the strength of the magnetic field felt by the nucleus decreases, the rotational motion frequency of the nucleus decreases in the lower direction. When molecular aggregates are physically formed, the density of the electron cloud of the atomic nucleus changes depending on the distance between strongly interacting molecules. Another method is that nuclei near a phenyl ring (C6H5-), where a ring current is formed under a magnetic field, will experience a local change in the magnitude of the magnetic field due to the ring current, which changes the rotational speed of the nucleus. Furthermore, even in cases where the electron density is high due to a double bond, such as C=O, the position of the peak can shift to high-field (up field) or low-field (down field) magnetic fields because the strength of the magnetic field changes depending on the relative position of the surrounding nuclei to the C=O.

[0076] In the case of molecular aggregates produced by the present invention, they can be physically bound together by intermolecular interactions and consist substantially of organic or inorganic materials, without the need for separate binders or additives.

[0077] Judging from these points, when comparing the NMR values ​​measured for the molecular aggregate produced by the present invention with those of the organic or inorganic precursor of the nanomolecular aggregate, the position of some peaks in the NMR spectrum changes. This indicates that there is no change in the chemical structure between the nanomolecular aggregate and the organic or inorganic precursor, but rather a change in the physical structure. Specifically, when comparing the NMR values ​​measured for the nanomolecular aggregate and the organic or inorganic precursor of the nanomolecular aggregate, if the peak shift is 0.005 ppm or more based on 1H NMR, they can be judged to be different.

[0078] On the other hand, in the case of FT-IR, the density of electrons constituting the molecular bonds is unevenly distributed, and it is known that changes in the peak often appear when the dipole moment changes as the molecule moves. It is known that the peaks in the spectrum are excited by various molecular motions such as stretching and bending. Polarity in molecular bonds occurs when they are formed by heteroatoms, and changes in the peak often appear. For example, ether bonds (CO), which are primary bonds, and carbonyl bonds (C=O), which are secondary bonds, between carbon and oxygen show stretching bands when exposed to infrared lasers. Similarly, in the case of carbon and hydrogen, stretching bands also appear because they are bonds between different molecules. In the case of stretching, it can be explained by a model in which two masses are connected by a spring, and when the energy that excites the spring's motion is supplied by infrared light, a peak appears in the spectrum.

[0079] Even when multiple carbon atoms are linked together, such as in benzene where six atoms are bonded, various bending peaks appear, including in-plane and out-of-plane vibrations. In particular, in the case of bending motion, various types of molecular motion can be observed using FT-IR spectroscopy. For example, rocking (movement from side to side), scissorsing (movement like scissors), wagging (swaying back and forth in a plane), and twisting / torsion (repeated twisting motion) can occur.

[0080] The structure of highly polar molecules can undergo interactions such as hydrogen bonding, polar interactions, or pi-pi stacking, and these interactions cause changes in the position and intensity of peaks in the FT-IR spectrum. Therefore, even the same molecular bond can exhibit changes in peak position and intensity depending on the surrounding environment. In other words, if the FT-IR spectra are the same, it can be determined that they belong to the same molecule. Especially at 400 cm⁻¹ -1 ~700 cm -1The area between these points is called the fingerprint zone, and if the peaks are the same within this zone, they are considered to be the same compound.

[0081] The present invention aims to form compounds into molecular aggregates, which are physical aggregates. When compounds are formed into molecular aggregates in this way, the intermolecular distances within the aggregate are very close, within approximately 10 Å or 5 Å. When intramolecular bonds are excited by infrared light, the compound bonds within the molecular aggregate undergo molecular motion such as stretching and bending. At this time, it has been found that in the molecular aggregates of the present invention, various changes occur, such as the appearance or disappearance of peaks that were not present in the original compound spectrum, changes in the position of peaks, and decreases or increases in peak intensity, due to compounds present at a very close distance to the original compound.

[0082] In other words, if one or more peaks are generated or disappear based on the results measured by the FT-IR, it can be determined that they are different.

[0083] When comparing the FT-IR measurements of the nanomolecular aggregate produced by the present invention with those of an organic or inorganic precursor of the nanomolecular aggregate, one or more peaks appear or disappear in the FT-IR spectrum. This indicates that there is no change in the chemical structure between the nanomolecular aggregate and its organic or inorganic precursor, but a change in the physical structure occurs. Furthermore, when comparing the FT-IR measurements of the nanomolecular aggregate and its organic or inorganic precursor, the position of one or more peaks changes at 5 cm. -1 If the above changes occur, it can be determined that the result is different. In other words, the conversion of a compound into a molecular aggregate is due to a physical action, and no new chemical bonds are formed or existing chemical bonds are destroyed. Therefore, such a change in the spectrum can be well explained as the fact that a new molecular aggregate with a novel structure, unlike anything in the past, has been created in this invention.

[0084] However, the nanomolecular aggregates produced by the present invention and the organic or inorganic precursors of the nanomolecular aggregates have the same chemical structure as previously mentioned. This can be seen by performing chromatographic analysis on the nanomolecular aggregates and the organic or inorganic precursors of the nanomolecular aggregates and finding that the measured values ​​are the same. The chromatographic measurements may also be results obtained by HPLC (high-performance liquid chromatography), and it is observed that the organic or inorganic products provided by the present invention and these molecular aggregates exhibit peaks at approximately the same position and have retention times of approximately 10% (i.e., within ±5%). Generally, when the same column, mobile phase, and stationary phase are used, peaks that appear within 10% of the time can be determined to be the same substance.

[0085] Thus, because the nanomolecular aggregate and the organic or inorganic precursor of the nanomolecular aggregate have different physical structures, the peaks of the two target substances in the NMR spectrum are generally similar, or some peaks can be altered. On FT-IR, various forms of changes may be observed, such as the generation and disappearance of peaks, and changes in peak intensity. Furthermore, because the nanomolecular aggregate and the organic or inorganic precursor of the nanomolecular aggregate have the same chemical structure, the chromatographic measurements may be identical.

[0086] In the present invention, there are no particular limitations on the NMR measurement of the nanomolecular aggregate and the organic or inorganic precursor of the nanomolecular aggregate, but as an example, it can be measured using a Bruker 400 MHz Avance.

[0087] In the present invention, there are no particular limitations on the FT-IR measurement of the nanomolecular aggregate and the organic or inorganic precursor of the nanomolecular aggregate, but as an example, it can be measured using a Bruker Alpha 2 ATR.

[0088] Next, the drug molecule-derived or other molecular aggregate according to the present invention is a precursor of the nanomolecular aggregate, characterized in that shear stress is applied to a solution containing an organic or inorganic salt to produce a nanomolecular aggregate having a structure in which the organic or inorganic salt is physically bonded.

[0089] The molecular aggregate produced in this manner is a molecular aggregate to which a pharmacologically active ingredient and a water-soluble compound are bound, characterized in that the chromatographic measurement values ​​of the organic or inorganic salt that is a precursor of the nanomolecular aggregate and the molecular aggregate to which the organic or inorganic salt is bound are the same, and the spectroscopic measurement values ​​of the organic or inorganic salt that is a precursor of the nanomolecular aggregate and the molecular aggregate to which the organic or inorganic salt is bound are different.

[0090] In the aforementioned nanomolecular aggregate, the chromatographic and spectroscopic measurements are the same as those mentioned earlier.

[0091] Similarly, without the need for separate binders or additives, molecules can be physically bound together by intermolecular interactions and consist essentially of organic or inorganic salts.

[0092] In the present invention, the salt of the organic or inorganic substance can be one used as a pharmacologically active ingredient, and the salts of the organic or inorganic substances mentioned above can be used.

[0093] As mentioned earlier, the organic or inorganic salt that is a precursor of the nanomolecular aggregate and the molecular aggregate formed by the physical bonding of the organic or inorganic salt have different physical structures. Therefore, in the NMR spectrum, the peaks of the two target substances are generally similar, although some can be altered. On FT-IR, various forms of changes may be observed, such as the generation and disappearance of peaks and changes in peak intensity.

[0094] For example, when comparing the NMR values ​​measured for an organic or inorganic salt that is a precursor of the nanomolecular aggregate with those of a molecular aggregate to which the organic or inorganic salt is bound, if the position of the peak changes, it can be determined that they are different. Specifically, when comparing the NMR values ​​measured for an organic or inorganic salt that is a precursor of the nanomolecular aggregate with those of a molecular aggregate to which the organic or inorganic salt is bound, if the peak shift is 0.005 ppm or more on a 1H NMR basis, it can be determined that they are different.

[0095] Furthermore, when comparing the FT-IR measurements of an organic or inorganic salt that is a precursor of the nanomolecular aggregate with those of a molecular aggregate to which the organic or inorganic salt is bonded, if one or more peaks appear or disappear, they can be determined to be different. Specifically, when comparing the FT-IR measurements of an organic or inorganic salt that is a precursor of the nanomolecular aggregate with those of a molecular aggregate to which the organic or inorganic salt is bonded, if one or more peaks appear or disappear, they can be determined to be different. -1 If the above changes occur, they can be considered different.

[0096] Furthermore, since the organic or inorganic salt that is a precursor of the nanomolecular aggregate and the molecular aggregate to which the organic or inorganic salt is bonded have the same chemical structure, the chromatographic measurements may be the same. Specifically, if the HPLC measurement results of the organic or inorganic salt that is a precursor of the nanomolecular aggregate and the molecular aggregate to which the organic or inorganic salt is bonded have a retention time of 10% or less, they can be considered to be the same.

[0097] Furthermore, the nanomolecular aggregates according to the present invention may have very small particle sizes, with an average particle size of 50 nm or less, preferably 30 nm or less, more preferably 20 nm or less, very preferably 15 nm or less, and most preferably 10 nm or less or 5 nm or less. The average particle size can be measured through diffraction experiments, preferably using small-angle neutron scattering (SANS). Alternatively, images can be measured using a transmission electron microscope. If the average particle size of the nanomolecular aggregate exceeds 50 nm, there is a problem of reduced dispersibility, transparency, and transmittance. There is no particular lower limit to the average particle size of the nanomolecular aggregate, but those of approximately 1 nm or more can be used. [Examples]

[0098] The manufacturing method of the present invention will be described in more detail below through examples of the present invention. It goes without saying that the present invention is not limited to these examples.

[0099] Equipment used [Table 1]

[0100] manufacturing [Example 1] A 0.2% aqueous solution of NaDC (Sodium Deoxycholate, manufactured by Lipa Biotech, Xi'an, China) was prepared by dissolving 19.8 mg of NaDC in 10 mL of water. As shown in Figure 1, a roll mill consisting of two rolls, roll A and roll B, was prepared, and the prepared NaDC solution was introduced between roll A and roll B at an input rate of 50 ml / min. The rotation speed of roll A was adjusted to 100 rpm, the rotation speed of roll B to 300 rpm, and the distance between roll A and roll B was set to 10 μm.

[0101] The obtained aqueous solution was frozen at -50°C, and then water was removed by running it through a freeze-dryer at 0.1 bar, -70°C, and pressure for 48 hours. The obtained sample was a white powder.

[0102] [Comparative Example 1] Excluding the case where an aqueous NaDC solution was prepared and the rolling process for producing the molecular aggregate was not carried out, NaDC powder was obtained through the same process as in Example 1.

[0103] [Example 2] 500 mg of sunitinib malate (SUNITINIB MALATE, TEVA) was dissolved in 100 mL of ethanol to prepare a sunitinib solution with a concentration of approximately 0.5%. Molecular aggregates were produced by introducing the solution into a roll apparatus as shown in Figure 1. At this time, the introduction rate of each solution was 50 ml / min. The rotation speed of roll A was adjusted to 100 rpm and the rotation speed of roll B to 300 rpm, and the distance between roll A and roll B was set to 10 μm.

[0104] Afterward, the aqueous solution recovered through a roll mill was frozen at -50°C, and then the water was removed by running it through a freeze-dryer at a temperature and pressure of 0.1 bar and -70°C for 48 hours to obtain a "sunitinib / maleate molecular aggregate" in powder form.

[0105] [Comparative Example 2] Excluding the case where an aqueous solution of sunitinib maleate was prepared and the rolling process for producing the molecular aggregate was not carried out, sunitinib maleate powder was obtained through the same process as in Example 2.

[0106] [Example 3] The sample was prepared in the same manner as in Example 2, with the exception that the distance between roll A and roll B was set to 90 μm.

[0107] [Example 4] The sample was prepared in the same manner as in Example 2, with the exception of using niclosamide instead of sunitinib-malate (SUNITINIB MALATE, TEVA) and setting the distance between roll A and roll B to 90 μm.

[0108] [Comparative Example 3] The sample was prepared in the same manner as in Example 2, with the exception that the distance between roll A and roll B was set to 110 μm.

[0109] [Comparative Example 4] The sample was prepared in the same manner as in Example 4, with the exception that the distance between roll A and roll B was set to 110 μm.

[0110] [Example 5] 19.8 mg of DCF-DA (2',7'-Dichlorofluorescein diacetate, Sigma-Aldrich) was dissolved in 10 mL of water to prepare an aqueous NaDC solution of approximately 0.2% concentration. As shown in Figure 1, a roll mill consisting of two rolls, roll A and roll B, was prepared, and the prepared DCF-DA solution was introduced between roll A and roll B at an input rate of 50 ml / min. The rotation speed of roll A was adjusted to 100 rpm, and the rotation speed of roll B was adjusted to 300 rpm, and the distance between roll A and roll B was set to 10 μm.

[0111] The obtained aqueous solution was frozen at -50°C, and then water was removed by running it through a freeze-dryer at 0.1 bar, -70°C, and pressure for 48 hours. The obtained sample was a yellowish fluorescent powder.

[0112] [Comparative Example 5] Excluding the case where an aqueous solution was prepared with DCF-DA and the rolling process for producing the molecular aggregate was not carried out, the DCF-DA powder was obtained through the same process as in Example 5.

[0113] [Example 6] Except for using cyclosporine A instead of NaDC, the molecular aggregate was prepared in the same manner as in Example 1.

[0114] [Comparative Example 6] Excluding the case where an aqueous solution was prepared with cyclosporine A and the rolling process for producing the molecular aggregate was not carried out, the API was obtained using the same process as in Example 6.

[0115] Experimental Example 1: Comparison of the chemical / physical structures of Example 1 and Comparative Example 1 The NaDC molecular aggregates in powder form prepared in Example 1 and the NaDC itself from Comparative Example 1 had their spectra measured using NMR (Nuclear Magnetic Resonance; Bruker 400MHz Avance) and FT-IR (Fourier-transform infrared spectroscopy; Bruker Alpha 2 ATR). The spectral results obtained by NMR are shown in Figure 5, and the spectral results obtained by FT-IR are shown in Figure 6.

[0116] As shown in Figure 5, two spectra of NaDC can be observed. The upper spectrum of Figure 5 shows the H-NMR spectrum of NaDC (pure compound, Comparative Example 1), the pharmacologically active ingredient that is a precursor of the molecular aggregate of the present invention, while the lower spectrum of Figure 5 shows the H-NMR spectrum of the molecular aggregate produced by the manufacturing method of the present invention. Comparing the upper and lower spectra of Figure 5, the upper spectrum shows very sharp peaks and good resolution, while the lower spectrum shows blunt peaks and overlapping. This difference is because the molecular weight at which the structure of the molecular aggregate produced by the manufacturing method of the present invention is formed and substantial molecular motion occurs is different, resulting in a shorter T2 (spin-spin relaxation time).

[0117] The FT-IR spectra of the two substances are shown in Fig. 6. Comparing the spectrum of pure NaDC (Comparative Example 1) shown in blue with the spectrum of the molecular aggregate of Example 1 produced by the production method of the present invention shown in red, characteristically new peaks can be found. 600 cm -1 , 705 cm -1 , 820 cm -1 , 1150 cm -1 The four peaks appearing at indicate that a molecular aggregate was formed and that a new molecular motion mode occurred in addition to the conventional stretching and bending motions.

[0118] Through the spectral results of Figs. 5 and 6, the result values measured by spectroscopic analysis such as NMR or FT-IR for the nano molecular aggregate produced by the production method of the present invention and the pharmacologically active ingredient which is the precursor of the nano molecular aggregate are different from each other, and it was judged that such a difference in the result values was due to the different physical structures of the two substances. Specifically, when comparing the result values measured by NMR, if the position of the peak changes, it is judged as different, and when comparing the result values measured by FT-IR, if one or more peaks are generated or disappear, it can be judged as different. At this time, it was found that the result values are different from each other because the physical structures of the two substances are different.

[0119] Moreover, HPLC (e2695 of Waters) measurement was carried out for the molecular aggregate of NaDC in powder form produced in Example 1 and NaDC of Comparative Example 1, and the results are shown in Fig. 7(a). Like the main peak indicated by the red ellipse in Fig. 7(a), the API (Comparative Example) before producing the structure was subjected to shear stress, and the chromatographic characteristics of the newly produced molecular aggregate are the same, and through this, it can be confirmed that they are the same substance having the same peak. Therefore, as shown in Fig. 7(a), it was found that the HPLC measurement results, the molecular aggregate of NaDC in powder form produced in Example 1 and NaDC of Comparative Example 1 have the same result values.

[0120] Similarly, when a molecular aggregate was prepared using niclosamide prepared in Example 4 of the present invention as the API, shear stress was applied to the API before the preparation of the molecular aggregate. To confirm that the chromatographic properties of the newly prepared molecular aggregate were the same as those of the API before preparation, measurements were performed using HPLC, as in the previous experimental example. As a result, it was found that the same peaks were observed, as shown in the graph in Figure 7(b).

[0121] Furthermore, in the case of the Sunitinib / Maleate molecular aggregate produced in Example 2 of this application and the Sunitinib and Maleate salt produced in Comparative Example 2, shear stress was applied to the API before production of the molecular aggregate, and in order to confirm that the chromatographic properties of the newly produced molecular aggregate were the same, measurements were performed using HPLC in the same manner as in the previous experimental example. As a result, it was found that the same peaks were observed, as shown in the graph in Figure 7(c).

[0122] Therefore, based on the results shown in Figure 7, the nanomolecular aggregate produced by the manufacturing method of the present invention and the pharmacologically active component which is a precursor of the nanomolecular aggregate were found to have the same values ​​when measured by analysis such as chromatography (spectroscopy) including HPLC. It was determined that this result is due to the fact that the chemical structures of both substances are the same.

[0123] Furthermore, looking at the two spectra shown in Figure 5, while changes were observed in terms of sharp peak resolution, the peak positions were almost the same. This means that when comparing pure NaDC with the molecular aggregate newly produced using the manufacturing method of the present invention, no new chemical bonds are formed or chemical bonds are removed, and the chemical structure is also the same.

[0124] Through this, it can be seen that the molecular aggregates produced by the manufacturing method of the present invention have the same chemical structure as the pharmacologically active substance that is the precursor of the molecular aggregate, but possess new properties due to their different molecular physical structures.

[0125] Experimental Example 2: Comparison of the chemical / physical structures of Example 2 and Comparative Example 2 The spectra of the Sunitinib / Maleate molecular aggregate prepared in Example 2 and the Sunitinib and Maleate salt prepared in Comparative Example 2 were measured using NMR and FT-IR. The spectral results obtained by NMR are shown in Figure 8, and the spectral results obtained by FT-IR are shown in Figure 9.

[0126] First, Figure 8 shows the H-NMR spectrum of the Sunitinib / Maleate salt and the H-NMR spectrum of the molecular aggregate prepared according to the present invention. In Figure 8, the spectrum shown at the top is the H-NMR spectrum of the salt mixture of Sunitinib and Maleate, and the spectrum shown at the bottom is the H-NMR spectrum of the molecular aggregate formed by the physical bonding of the two compounds produced through the present invention. Regions where changes occurred are indicated by * and **. When a molecular aggregate is formed, the distance between molecules becomes very close, and these closely spaced molecules cause changes in the nuclear environment of each other, which is evidence that they are close enough that changes in the NMR peaks can be observed.

[0127] Next, Figure 9 shows the FT-IR spectra of the Sunitinib / Maleate salt and the molecular aggregate prepared according to the present invention. As observed in Figure 9, the changes are much more dramatic than the changes in the NMR spectrum. This indicates that, as explained above, the expansion and contraction modes of the molecules are constrained by the formation of molecular aggregates, which not only changes the excitation energy of these motion modes but also alters the generation and disappearance of peaks, as well as their position and intensity. 1300~1700 cm -1The peaks that appear at 900 cm² indicate the stretching and contracting motion of the C=O group in the molecular structure. As can be seen from the chemical structure shown in Figure 9, the mixture of the two molecules contains four distinct carbonyl groups, which are located at different positions. However, in the molecular aggregate prepared using this mixture, these four sharp peaks become broader and separate, and the intensity of the peaks also changes significantly. This means that multiple carbonyl groups are creating various environments through their interactions with each other. -1 , 500 cm -1 A new peak can be observed in the vicinity, at 3000-3500 cm. -1 Observing the stretching and contracting peaks of OH and NH, which are known to appear in this region, we can observe that the peaks generally become thicker, and their positions change so much that it is difficult to define them. This indicates that as two molecules approach each other, the hydrogen bonds are strengthened, and various peaks appear depending on the strength of these bonds, resulting in an increase in the intensity of the thicker peaks compared to a molecular mixture.

[0128] Through the spectral results in Figures 8 and 9, it was determined that the results measured by spectroscopy, such as NMR or FT-IR, for the nanomolecular aggregate produced by the manufacturing method of the present invention and the pharmacologically active component which is a precursor of the nanomolecular aggregate differed from each other. This difference in results was determined to be due to the difference in the physical structures of the two substances. Specifically, when comparing the results measured by NMR, if the position of the peaks changes, it can be determined that the results measured by spectroscopy are different from each other. When comparing the results measured by FT-IR, if one or more peaks are generated or disappear, it can be determined that the results measured by spectroscopy are different from each other. In this case, it was found that the difference in results occurs because the physical structures of the two substances are different.

[0129] Furthermore, based on the results shown in Figure 10, the measured values ​​of the nanomolecular aggregate produced by the manufacturing method of the present invention and the pharmacologically active component which is a precursor of the nanomolecular aggregate were the same when analyzed by chromatography such as HPLC. It was determined that this result is due to the fact that the chemical structures of both substances are the same.

[0130] Experimental Example 3: Comparison of the chemical / physical structures of Example 3 and Comparative Example 3 The spectra of the powder produced in Example 3 and the powder produced in Comparative Example 3 were measured using FT-IR (Fourier-transform infrared spectroscopy; Bruker Alpha 2 ATR), and these were compared with the spectral results of Comparative Example 1, which was the original precursor not produced by the manufacturing method of the present invention, and which had already been measured.

[0131] First, Figure 11 shows a comparison of the FT-IR spectral results of Example 3, Comparative Example 3, and Comparative Example 1, and Figure 12 shows a comparison of the FT-IR spectral results of Example 4, Comparative Example 4, and Comparative Example 1.

[0132] As shown in Figure 11, Comparative Example 3 and Comparative Example 1 showed nearly similar FT-IR spectral results, indicating that when the distance between the two roll mills was adjusted to exceed 100 μm, the molecular aggregate according to the present invention could not be produced. Conversely, in Example 3, adjusting the distance between the two roll mills to 100 μm or less resulted in the production of a new structure with a different physical structure, i.e., the molecular aggregate according to the present invention.

[0133] Next, as shown in Figure 12, Comparative Example 4 and Comparative Example 1 also showed nearly similar FT-IR spectral results, indicating that, regardless of the type of pharmacologically active ingredient, the molecular aggregate according to the present invention is not produced when the distance between the two roll mills is adjusted to exceed 100 μm. Conversely, in Example 4, it was found that adjusting the distance between the two roll mills to 100 μm or less produced a new structure with a different physical structure, i.e., the molecular aggregate according to the present invention.

[0134] Therefore, it was found that, regardless of the type of pharmacologically active ingredient, if the distance between the two roll mills is adjusted to exceed 100 μm, the molecular aggregate according to the present invention is not produced, but if it is adjusted to 100 μm or less, the molecular aggregate according to the present invention is produced.

[0135] Furthermore, the molecular aggregates of niclosamaide prepared in Example 4 were imaged using TEM (FEI Tecnai G2 Spirit Twin) and are shown in Figure 13. The eccentricity measured using the transmission electron microscope image had an average value of approximately 0.85, and its distribution was between 0.48 and 1.0. The minimum and maximum sizes were approximately 2.5 nm and 7.7 nm, respectively, and the average size was approximately 4.8 nm.

[0136] Experimental Example 4: Stability test and particle size confirmation of molecular aggregates from Example 2 The powder-form "Sunitinib / Maleate molecular aggregate" produced in Example 2 was prepared in a 0.05% aqueous solution, and the changes in pH, particle size, content %, and concentration were measured over 6 months under the following conditions, as shown in Table 2 below.

[0137] Method for measuring pH A 0.5 mL sample was taken and analyzed once. Using a 1 mL pipette, 0.5 mL of the sample was added so that it completely covered the pH meter sensor, and the pH was measured.

[0138] Method for measuring particle size (by zetasizer) Take 1 mL of the sample. Carefully place the sample into a cuvette cell, taking care to avoid creating air bubbles. Repeat the analysis a total of 10 times with the same sample. (Basic settings were used for 1 measurement and 10 repeated analyses.) From the processed data, the size distribution by volume portion was used, and the size and PDI values ​​were processed using the average of 10 values.

[0139] [Table 2]

[0140] Measurement conditions for content (Preparation of standard solution for calibration curve) A mobile phase consisting of 20 mM ammonium acetate and acetonitrile was prepared. The HPLC was run with the mobile phase for approximately 30 minutes. Sunitinib samples were dissolved in Vehicle to prepare 0.005%, 0.01%, 0.03%, 0.05%, and 0.1% standard solutions. Area values ​​were measured using the standard solutions under HPLC conditions. A calibration curve was created with the measured area values ​​on the y-axis and the known concentrations on the x-axis. Once the calibration curve was completed, the equation for the y-axis was determined.

[0141] Concentration by HPLC A 1 mL sample was taken and placed in an LC vial. The HPLC was run in mobile phase for approximately 30 minutes. 10 μl was injected, and the HPLC area was measured. The concentration was calculated using the prepared calibration curve.

[0142] [Table 3]

[0143] As shown in Table 2 above, the particle size of the molecular aggregates produced in Example 2 was an average of 2.2 nm, and the PDI was 1. Furthermore, since the pH, content, and particle size were maintained for 6 months, it was found that stability was maintained.

[0144] Experimental Example 5: Comparison of XRD structures of the example and comparative example. Figure 14 shows a comparison of the XRD measurement results of the molecular aggregate of Example 6 of the present invention, manufactured using cyclosporine A, and cyclosporine A itself in Comparative Example 6. As can be seen from the comparison of the graphs, when the peaks that appear due to the holder (SUS material) that grips the specimen (peaks that appear similarly on the right side of the graph) are excluded, it can be seen that, unlike the API (cyclosporine A) in Comparative Example 6 before the process, which exists in a crystal form (sharp peak on the left), the molecular aggregate of Example 6 of the present invention after the process changed to an amorphous form (amorphous peak on the left) due to a change in its physical structure.

[0145] In the case of amorphous crystalline structures like the molecular aggregates of the present invention, the peaks may change due to changes in the size of the reticles inside, but this also only involves a change from one conventional crystal form to another, and does not involve a change to an amorphous form as in the present invention.

[0146] Therefore, as can be seen from Figure 14, the molecular aggregate of the present invention did not change into a solvent form, but rather its physical structure itself changed into an amorphous form.

[0147] Similarly, the XRD measurement results of the molecular aggregate of Example 2 of the present invention, prepared using sunitinib / malate salt as the API, and Comparative Example 2 were compared and are shown in Figure 15.

[0148] Similar to cyclosporine A mentioned earlier, unlike Comparative Example 2, where the API (sunitinib / malate salt) itself exists in a crystalline form before the process, the molecular aggregate of Example 2 of the present invention, after the process, shows a change to an amorphous form due to a change in its physical structure. In other words, it is clear that the molecular aggregate of the present invention did not change to a solvent form, but rather that the physical structure itself changed to an amorphous form.

[0149] Experimental Example 6: Comparison of DSC measurement experiments between the example and comparative example. To clearly demonstrate that the molecular aggregate of this application is not a crystalline (sorbated) product, the applicant of this application conducted the following DSC measurement experiment.

[0150] Specifically, the DSC measurement results of the molecular aggregate of Example 6 of the present invention, which was produced using cyclosporine A as the API, and Comparative Example 6, which is cyclosporine A itself, were compared and are shown in Figure 16. If the molecular aggregate produced through the process of the present invention were a solvent, a peak should appear on the DSC graph indicating that the solvent (the solvent used in the present invention is water or ethanol) is evaporating near its boiling point (approximately 100°C for water and 60°C for ethanol). However, no such peak appears in the DSC measurement results data shown in Figure 16. Therefore, it is clear that the difference in the spectroscopic measurements of the molecular aggregate of the present application and the pharmacologically active component, which is a precursor of the nanomolecular aggregate, is not due to a change caused by sorbation, but rather to a change in physical structure.

[0151] Experimental Example 7: Confirmation of Particle Photographs in the Example To confirm that the molecular aggregate of the present application has a structure in which multiple APIs are physically bonded, the applicant took TEM images of the molecular aggregate of Example 6, which was prepared using cyclosporine A as the API, and the molecular aggregate of Example 2, which was prepared using sunitinib / malate salt. First, a TEM image of the molecular aggregate of Example 6, which was prepared using cyclosporine A as the API, was taken and is shown in Figure 17. Unlike cyclosporine A itself, which is poorly soluble in water and cannot be confirmed by TEM, the molecular structure according to the present invention can be confirmed to be a structure in which multiple APIs are physically bonded, as can be seen in the TEM image in Figure D.

[0152] Similarly, a TEM image of the molecular aggregate of Example 2, prepared using sunitinib / malate salt, was taken and is shown in Figure 18.

[0153] As can be seen in the TEM image in Figure 18, the molecular aggregate according to the present invention can be confirmed to have a structure in which multiple APIs are physically bound together. In contrast, as shown in the TEM image in Figure 19, when Comparative Example 2, which is the sunitinib / malate salt itself, was dissolved in water and photographed with TEM, it was not possible to confirm any structure in which APIs were physically bound together.

[0154] Therefore, it is clear that the structure according to the present invention is a molecular aggregate structure in which multiple APIs are physically bonded together.

[0155] Experimental Example 8: Comparison of Cell Permeability Performance of Example 5 and Comparative Example 5 The cell permeability performance of the DCF-DA molecular aggregate in powder form produced in Example 5 and the DCF-DA itself from Comparative Example 5 was compared. Suspension cells (MV-4-11 human macrophage) were grown to fill a dish to approximately 50%. The DCF-DA from Comparative Example 5 was dissolved in ethanol, and the DCF-DA molecular aggregate from Example 5 was dissolved in distilled water at the same concentration of 0.005% for 30 minutes. After that, the cells were washed with PBS (phosphate buffer solution) to remove extracellular DCF-DA and stop diffusion-induced cell ingestion. After stabilizing the cells with an additional 30 minutes of culture, they were treated with 0.03% hydrogen peroxide (H2O2), and the DCF exhibiting intracellular fluorescence was imaged as a 3D hologram and fluorescence using a 3D microscope (Tomocube HT-2H), which is shown in Figure 20. The experimental results confirmed that the DCF-DA molecular aggregate treatment in Example 5, shown in Figures 20C and 20D, exhibited much stronger fluorescence than the DCF-DA treatment treatment in Comparative Example 5, shown in Figures 20A and 20B. Therefore, it was confirmed that the molecular aggregate produced by the present invention has much higher phospholipid membrane permeability.

[0156] Experimental Example 9: Comparison of Corneal Permeability Performance of Example 2 and Comparative Example 2 Corneal transmission experiments were conducted on the sunitinib-malate molecular aggregate in powder form prepared in Example 2 and on the sunitinib-malate itself from Comparative Example 2.

[0157] The corneal transmission experiment described above was conducted by referring to the content of the already published paper, "Lee et. al, Modulating the Transport Characteristics of Bruch's Membrane With Steroidal Glycosides and its Relevance to Age-Related Macular Degeneration (AMD). Invest. Ophthalmol. Vis. Sci. 2015, 56, 8403., 2015." As shown in Figure 21, the cell for the transmission experiment was constructed by using a usching chamber, which consists of three main parts: a donor chamber for drug administration, a receiver chamber, and a membrane through which the drug permeates. Basically, the drug was introduced into the supply chamber of a Wushing chamber, an 8mm circular sample of the cornea from a pig's eye was taken, other tissues attached to the cornea were removed, and the sample was fixed in place on a membrane. The solution that permeated the membrane was received in the receiving chamber, and at this time, the solution was stored in a constant temperature chamber set to 37°C, close to the temperature at which diffusion occurs, similar to human body temperature. After a certain period of time, the solution concentration in each supply chamber and receiving chamber was measured, and the physical properties of each sample in relation to diffusion were determined. The concentrations for each sample after 18 hours are shown in Table 1 below. The concentration was measured using a UV / VIS spectrum (Thermo, Evolution201 UV / Vis), with the peak height at 420nm used to determine the concentration.

[0158] [Table 4]

[0159] As shown in Table 3 above, when the amount of permeation is expressed as a percentage, it can be seen that the permeability of the molecular aggregate of the pharmacologically active substance is approximately four times better than that of the pharmacologically active substance itself. Therefore, as observed in the cell permeation experiment of DCF-DA in Example 4 above, it was found that the permeability of the molecular aggregate according to the present invention to cells is significantly better than that of the pharmacologically active substance itself.

Claims

1. A molecular aggregate formed by the physical bonding of organic matter, When the molecular aggregate is formed with a composition containing water, the molecular aggregate has an aggregated structure within the composition. The average particle size of the molecular aggregate is 50 nm or less. The molecular aggregate is produced by applying shear stress to a solution containing the organic substance. The organic substance is characterized by being one or more pharmacologically active ingredients selected from the group consisting of NaDC (Sodium Deoxycholate), Sunitinib-malate, DCF-DA (2',7'-Dichlorofluorescein diacetate), or Cyclosporine A. Amorphous nanomolecular aggregates.

2. The HPLC (high-performance liquid chromatography) results for the organic substance itself and the molecular aggregate itself are the same. The nanomolecular aggregate according to claim 1, characterized in that the results measured by NMR or FT-IR methods of the organic substance itself and the molecular aggregate itself are different.

3. The nanomolecular aggregate according to claim 1, characterized in that if the HPLC-measured results of the organic substance itself and the molecular aggregate itself have a retention time of 10% or less, they are determined to be the same.

4. The nanomolecular aggregate according to claim 1, characterized in that when comparing the NMR measured values ​​of the organic substance itself and the molecular aggregate itself, if the position of the peak changes, it is determined that they are different.

5. The nanomolecular aggregate according to claim 4, characterized in that when comparing the NMR measured values ​​of the organic substance itself and the molecular aggregate itself, if the peak shift is 0.005 ppm or more on a 1H NMR standard, they are judged to be different.

6. The nanomolecular aggregate according to claim 1, characterized in that when comparing the FT-IR measured values ​​of the organic substance itself and the molecular aggregate itself, if one or more peaks are generated or disappear, they are determined to be different.

7. When comparing the FT-IR measured values ​​of the organic substance itself and the molecular aggregate itself, if the position of one or more peaks is 5 cm －１ The nanomolecular aggregate according to claim 1, characterized in that if the above changes occur, it is determined to be a different entity.

8. The nanomolecular aggregate according to claim 1, characterized in that the distance between molecules is 10 Å or less.

9. The nanomolecular aggregate according to claim 1, characterized in that the molecular aggregate is made of organic material.

10. A molecular aggregate formed by the physical bonding of salts of organic substances, When the molecular aggregate is formed with a composition containing water, the molecular aggregate has an aggregated structure in the composition. The average particle size of the molecular aggregate is 50 nm or less. The molecular aggregate is produced by applying shear stress to a solution containing a salt of the organic substance. The organic substance is characterized by being one or more pharmacologically active ingredients selected from the group consisting of NaDC (Sodium Deoxycholate), Sunitinib-malate, DCF-DA (2',7'-Dichlorofluorescein diacetate), or Cyclosporine A. Amorphous nanomolecular aggregates.

11. The HPLC (high-performance liquid chromatography) results for the salt of the organic substance itself and the molecular aggregate itself are the same. The nanomolecular aggregate according to claim 10, characterized in that the measured values ​​obtained by NMR or FT-IR methods for the salt of the organic substance itself and the molecular aggregate itself are different.

12. The nanomolecular aggregate according to claim 10, characterized in that if the HPLC-measured results of the salt of the organic substance itself and the molecular aggregate itself have a retention time of 10% or less, they are determined to be the same.

13. The nanomolecular aggregate according to claim 10, characterized in that when comparing the NMR measured values ​​of the salt of the organic substance itself and the molecular aggregate itself, if the position of the peak changes, it is determined that they are different.

14. The nanomolecular aggregate according to claim 13, characterized in that when comparing the NMR measured values ​​of the salt of the organic substance itself and the molecular aggregate itself, if the peak shift is 0.005 ppm or more on a 1H NMR standard, they are judged to be different.

15. The nanomolecular aggregate according to claim 10, characterized in that when comparing the FT-IR measured values ​​of the salt of the organic substance itself and the molecular aggregate itself, if one or more peaks are generated or disappear, they are determined to be different.

16. When comparing the FT-IR measured values ​​of the salt of the organic substance itself and the molecular aggregate itself, if the position of one or more peaks is 5 cm －１ The nanomolecular aggregate according to claim 10, characterized in that if the above changes occur, it is determined to be a different entity.

17. The nanomolecular aggregate according to claim 10, characterized in that the distance between molecules is 10 Å or less.

18. The nanomolecular aggregate according to claim 10, characterized in that the molecular aggregate is made of a salt of an organic substance.

19. After placing a composition containing an organic substance or its salt, and water, into a device capable of applying shear stress, By applying shear stress to the aforementioned composition, a molecular aggregate in which organic matter or a salt thereof is physically bonded is produced. The aforementioned organic substance is one or more pharmacologically active ingredients selected from the group consisting of NaDC (Sodium Deoxycholate), Sunitinib-malate, DCF-DA (2',7'-Dichlorofluorescein diacetate), or Cyclosporine A. The apparatus comprises multiple rolls capable of applying shear stress to a solution containing an organic substance or a salt thereof. After introducing the solution containing the organic substance or a salt thereof between two opposing rolls in the apparatus, A method for producing a nanomolecular aggregate according to claim 1 or 9, characterized by applying shear stress to a solution containing the organic substance or a salt thereof.

20. A method for producing a nanomolecular aggregate according to claim 19, characterized in that the distance between the two opposing rolls is 0.5 to 1000 μm.

21. A method for producing a nanomolecular aggregate according to claim 20, characterized in that the two opposing rolls rotate at different speeds from each other.

22. A method for producing a nanomolecular aggregate according to claim 21, characterized in that the rotational speed of one of the two opposing rolls is 50 to 250 rpm and the rotational speed of the other is 200 to 500 rpm.

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