Pharmaceutical composition, method for improving intestinal bacterial flora, treatment method, and prophylactic method

Abstract

To provide a novel pharmaceutical composition, preferably a pharmaceutical composition capable of improving the intestinal bacterial flora in vivo or increasing a short-chain fatty acid in vivo.SOLUTION: A pharmaceutical composition comprises MXene for improving intestinal bacterial flora in vivo or increasing a short-chain fatty acid in vivo.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present disclosure relates to a pharmaceutical composition, a method for improving the intestinal flora, a treatment method, and a prevention method.

Background Art

[0002] In recent years, the relationship between the intestinal environment and various diseases has been pointed out, and it is expected that the treatment effect of these diseases can be improved by improving the intestinal environment.

[0003] Patent Document 1 describes a pharmaceutical composition for improving the intestinal flora, which contains 1-cyclopropyl-6-fluoro-1,4-dihydro-8-methyl-7-(2-amino-3-cyano-5-pyridyl)-4-oxo-3-quinolinecarboxylic acid or a pharmaceutically acceptable salt thereof as an active ingredient.

[0004] Non-Patent Document 1 describes that by administering genetically engineered Escherichia coli to dextran sulfate sodium-induced colitis (DSS) mice as a model animal for inflammatory bowel disease (IBD), the diversity of the intestinal flora can be promoted, Ruminococcaceae can be proliferated, and as a result, the production of butyric acid can be enhanced.

[0005] Non-Patent Document 2 describes that the diversity of intestinal bacteria can promote the expression of Cldn and reduce the permeability of the blood-brain barrier (BBB).

[0006] Non-Patent Document 3 describes that butyrate can significantly correct hypertensive symptoms in pregnant women.

Prior Art Documents

Patent Documents

[0007]

Patent Document 1

Non-Patent Documents

[0008] [Non-Patent Document 1] Lifu Wang et al., An engineered probiotic secreting Sj16 ameliorates colitis via Ruminococcaceae / butyrate / retinoic acid axis, BIOENGINEERING & TRANSLATIONAL MEDICINE, volume 6, Issue 3 September 2021 [Non-Patent Document 2] V. Braniste et al., The gut microbiota influences blood-brain barrier permeability in mice, 2014, Science Translational Meddicine, 6: 263ra158 [Non-Patent Document 3] Luisa F. Gomez-Arango et al., Increased Systolic and Diastolic Blood Pressure Is Associated With Altered Gut Microbiota Composition and Butyrate Production in Early Pregnancy, Hyperteinsion October 2016 Vol 68, Issue 4 [Summary of the Invention] [Problems to be Solved by the Invention]

[0009] Patent Document 1 describes that the intestinal flora can be improved by administering a predetermined compound. Non-Patent Documents 1 to 3 describe that Ruminococcaceae, which is one of the intestinal bacteria, can be proliferated by directly administering Escherichia coli, and that butyric acid that can be produced thereby can contribute to the suppression of the increase in the permeability of the blood-brain barrier (BBB) and the improvement of hypertensive symptoms. However, there is no description about the action of MXene.

[0010] One of the objects of the present disclosure is to provide a novel pharmaceutical composition, preferably a pharmaceutical composition capable of improving the gut microbiota or increasing short-chain fatty acids in vivo.

Means for Solving the Problems

[0011] The pharmaceutical composition of the present disclosure contains MXene and is used to improve the gut microbiota in vivo.

Effects of the Invention

[0012] The present disclosure can provide a novel pharmaceutical composition, preferably a pharmaceutical composition capable of improving the gut microbiota or increasing short-chain fatty acids in vivo. Further, the present disclosure can provide a treatment method or a prevention method.

[0013] The pharmaceutical composition of the present disclosure contains MXene and can improve the gut microbiota by promoting the growth of intestinal bacteria, or can increase short-chain fatty acids in vivo. Therefore, it is useful for the treatment and / or prevention of various diseases.

[0014] Although it should not be construed as being limited to a specific theory, the MXene contained in the pharmaceutical composition of the present disclosure is considered to have the ability to promote the growth of intestinal bacteria in the intestine, and it is expected that the intestinal bacteria can actually grow. Further, it is expected that the growth of such intestinal bacteria will also promote the growth of short-chain fatty acids that these intestinal bacteria can produce by metabolism. As a result, it is expected that the action of such short-chain fatty acids will promote the repair of the blood vessel barrier. In addition, it is also expected that the blood pressure will decrease by the intake of MXene. And since the above MXene is considered not to be absorbed from the intestinal tract, it is expected to be excreted as it is through the digestive tract together with feces. As described above, originally in a living body, there are two excretion functions, bile excretion and urine excretion, but the metabolic pathway by the pharmaceutical composition of the present disclosure can be said to be a third metabolic pathway, and for example, it is expected to lead to the reduction of the treatment related to dialysis therapy in patients with renal failure. Also, regarding disease-causing substances and the like contained in the contents of the diet, the MXene of the present disclosure is expected to adsorb in the digestive tract and not be absorbed by the intestinal tract.

Brief Description of the Drawings

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Mode for Carrying Out the Invention

[0016] The pharmaceutical composition of the present disclosure contains Mxene. Mxene is typically a layered material having one or more layers. Generally, MXene has the form of particles (which may include powders, flakes, nanosheets, etc.) of such layered materials.

[0017] The above-mentioned Mxene preferably includes two-dimensional particles of a layered material having one or more layers. The layer preferably includes at least one metal selected from Group 3, 4, 5, 6, and 7 metals, and at least one selected from carbon atoms and nitrogen atoms. The above-mentioned Group 3, 4, 5, 6, and 7 metals are preferably at least one selected from the group consisting of Ti, Zr, Hf, V, Nb, Ta, Cr, Mo, Sc, W, and Mn, and more preferably at least one selected from the group consisting of Ti, V, Cr, and Mo.

[0018] The above-mentioned layer preferably has the following compositional formula: M m X n (In the formula, M is at least one Group 3, 4, 5, 6, and 7 metal, X is a carbon atom, a nitrogen atom, or a combination thereof, n is 1 or more and 4 or less, m is greater than n and 5 or less) and includes a layer body represented by the formula.

[0019] The above-mentioned layer preferably further includes a modification or termination T (T is at least one selected from the group consisting of a hydroxyl group, a fluorine atom, a chlorine atom, an oxygen atom, and a hydrogen atom) present on the surface of the above-mentioned layer body.

[0020] The pharmaceutical composition of the present disclosure includes Mxene, preferably two-dimensional particles having a layer body represented by M m X n and T, and is useful for the treatment and / or prevention of various diseases by adsorbing a substance that can cause a disease (disease-causing substance).

[0021] In the present disclosure, the above-mentioned layered material can be understood as a layered compound, and the above-mentioned layer is "M m X n T sIt can also be expressed as “”. s is an arbitrary number, and conventionally, x or z may be used instead of s. Hereinafter, the above-mentioned layered material may also be referred to as MXene, the above-mentioned layer may be referred to as an MXene layer, and the above-mentioned two-dimensional particles may be referred to as MXene two-dimensional particles or MXene particles.

[0022] In the present disclosure, when referring to an element as an “atom”, the oxidation number of the element is not limited to 0 and can be any number within the range of possible oxidation numbers of the element.

[0023] Also, regarding the symbols in the general formulas shown in the present disclosure, unless otherwise specifically mentioned, the definitions of the same symbols are common among the respective general formulas including those symbols.

[0024] In the above formula: M m X n wherein, m can typically be 2, 3, 4, or 5, but is not limited thereto. Also, n can be 1, 2, 3, or 4, but is not limited thereto. In one aspect, m can be 3 and n can be 2.

[0025] In the above formula: M m X n wherein, M is preferably at least one selected from the group consisting of Ti, Zr, Hf, V, Nb, Ta, Cr, Mo, Sc, W, and Mn, and more preferably at least one selected from the group consisting of Ti, V, Cr, and Mo.

[0026] M m X n As M X, the following are known. 1.3 Sc2C, Ti2C, Ti2N, Zr2C, Zr2N, Hf2C, Hf2N, V2C, V2N, Nb2C, Ta2C, Cr2C, Cr2N, Mo2C, Mo 1.3 C, Cr 1.3 C, (Ti, V)2C, (Ti, Nb)2C, W2C, W 1.3 C, Mo2N, Nb 1.3 Y 0.6C (In the above formula, "1.3" and "0.6" respectively mean approximately 1.3 (= 4 / 3) and approximately 0.6 (= 2 / 3).) Ti3C2, Ti3N2, Ti3(CN), Zr3C2, (Ti,V)3C2, (Ti2Nb)C2, (Ti2Ta)C2, (Ti2Mn)C2, Hf3C2, (Hf2V)C2, (Hf2Mn)C2, (V2Ti)C2, (Cr2Ti)C2, (Cr2V)C2, (Cr2Nb)C2, (Cr2Ta)C2, (Mo2Sc)C2, (Mo2Ti)C2, (Mo2Zr)C2, (Mo2Hf)C2, (Mo2V)C2, (Mo2Nb)C2, (Mo2Ta)C2, (W2Ti)C2, (W2Zr)C2, (W2Hf)C2, Ti4N3, V4C3, Nb4C3, Ta4C3, (Ti,Nb)4C3, (Nb,Zr)4C3, (Ti2Nb2)C3, (Ti2Ta2)C3, (V2Ti2)C3, (V2Nb2)C3, (V2Ta2)C3, (Nb2Ta2)C3, (Cr2Ti2)C3, (Cr2V2)C3, (Cr2Nb2)C3, (Cr2Ta2)C3, (Mo2Ti2)C3, (Mo2Zr2)C3, (Mo2Hf2)C3, (Mo2V2)C3, (Mo2Nb2)C3, (Mo2Ta2)C3, (W2Ti2)C3, (W2Zr2)C3, (W2Hf2)C3, (Mo 2.7 V 1.3 )C3 (In the above formula, "2.7" and "1.3" respectively mean approximately 2.7 (= 8 / 3) and approximately 1.3 (= 4 / 3).)

[0027] Typically, in the above formula: M m X n in, M can be Ti or V, X can be a carbon atom or a nitrogen atom, and M can be Ti and X can be a carbon atom. In one embodiment, MXene can be Ti3C2T s (In other words, M is Ti, X is C, n is 2, and m is 3). In this case, the precursor of such MXene (also referred to as "MAX phase") can be Ti3AlC2.

[0028] MXene is the MAX phase of the precursor (in one embodiment, M m AX nrepresented by, where M, m, X and n have the same meanings as above, and A is at least one element of Group 12, 13, 14, 15, or 16) can be produced by removing A atoms contained therein, but the MXene may contain such A atoms. In one aspect, the residual amount of A atoms contained in the MXene can be preferably 10% by mass or less, more preferably 8% by mass or less, and still more preferably 6% by mass or less, relative to the content of A atoms in the precursor.

[0029] In another aspect, the residual amount of A atoms may exceed 10% by mass. For example, those in which A atoms are removed only from a part of the MAX phase are also included in the technical scope of the above MXene. Examples of such MXene include, for example, MXene in which A atoms are removed only near the end in the plane direction of the MAX phase (the direction parallel to the plane of the M m X n layer contained in the MAX phase). In this aspect, the residual amount of A atoms may be, for example, 50% by mass or more, further 80% by mass or more, and particularly 90% by mass or more.

[0030] The lithium content in the above MXene is preferably 0% by mass or more and 0.1% by mass or less, more preferably 0% by mass or more and 0.01% by mass or less, and still more preferably 0% by mass or more and 0.002% by mass or less. When the lithium content is within the above range, the biocompatibility can be improved. The lithium content in the MXene can be measured by inductively coupled plasma atomic emission spectrometry (ICP-AES).

[0031] The above MXene is an aggregate containing particles of one layer of MXene (hereinafter simply referred to as "MXene particles") 10a (single-layer MXene particles) schematically illustrated in Fig. 1(a). Typically, the MXene particles 10a, more specifically, the layer body represented by M m X n represented by M m X nAn MXene layer 7a having a layer body 1a and a modification or termination T3a, 5a present on the surface of the layer body 1a (more specifically, at least one of two surfaces facing each other in each layer). Thus, the MXene layer 7a is also represented as "M m X n T s ", and s is an arbitrary number.

[0032] The above MXene may include one or more layers. Examples of multi-layer MXene particles (multi-layer MXene particles) include, but are not limited to, two-layer MXene particles 10b as schematically shown in Fig. 1(b). 1b, 3b, 5b, 7b in Fig. 1(b) are the same as 1a, 3a, 5a, 7a in Fig. 1(a) described above. Two adjacent MXene layers (for example, 7a and 7b) of the multi-layer MXene particles do not necessarily have to be completely separated and may be in partial contact. The above single-layer MXene particles 10a are those in which the multi-layer MXene particles 10b are individually separated and exist as one layer. The MXene may be a mixture of unseparated multi-layer MXene particles 10b and the above single-layer MXene particles 10a. Typically, at least one of the surfaces of the layer body 1a represented by M m X n can be planar (two-dimensional), and all the surfaces of the layer body 1a can be planar (two-dimensional).

[0033] Although not limiting this embodiment, the thickness of each layer (corresponding to the above MXene layers 7a, 7b) included in the MXene particles is, for example, 0.8 nm or more and 5 nm or less, particularly 0.8 nm or more and 3 nm or less (mainly depending on the number of M atomic layers included in each layer). The thickness of each layer is determined as a number average dimension (for example, a number average of at least 40) based on an atomic force microscope (AFM) photograph or a transmission electron microscope (TEM) photograph.

[0034] For individual laminates of MXene (particularly multilayer MXene particles that may be included), the interlayer distance (or void size, denoted as Δd in Fig. 1(b)) can be, for example, 0.8 nm or more and 10 nm or less, particularly 0.8 nm or more and 5 nm or less, more particularly about 1 nm, and the total number of layers can be 2 or more and 20,000 or less. The interlayer distance in MXene can be measured by determining the interplanar distance (the sum of the interlayer distance and the thickness of each layer) from the position of the peak corresponding to the (002) plane of MXene that exists at 2θ = 10° (deg) or less in the X-ray diffraction measurement of MXene, and subtracting the thickness of each layer from the interplanar distance.

[0035] The above MXene may contain MXene particles with a small number of layers. The above "with a small number of layers" means, for example, that the number of stacked MXene layers is 6 or less. Also, the thickness in the stacking direction of multilayer MXene particles with a small number of layers is preferably 15 nm or less, more preferably 10 nm or less. Hereinafter, this "multilayer MXene particle with a small number of layers" may be referred to as "few-layer MXene particle". Also, single-layer MXene particles and few-layer MXene particles may be collectively referred to as "single-layer and few-layer MXene particles".

[0036] In the above MXene, the proportion of single-layer and few-layer MXene particles with a thickness of 15 nm or less can be 0 volume% or more and 100 volume% or less, further 0 volume% or more and 99 volume% or less, even further 0 volume% or more and 50 volume% or less, particularly 0 volume% or more and 30 volume% or less.

[0037] (Average value of the major axis of the two-dimensional plane of MXene) The major axis of the above MXene is preferably 1 μm or more and 20 μm or less in a plane parallel to each layer (hereinafter also referred to as the "two-dimensional plane"). Hereinafter, the average value of the major axis of the two-dimensional plane may be referred to as the "average flake size".

[0038] The larger the average flake size, the better the orientation of MXene in the material containing MXene. The average value of the major axis of the two-dimensional plane is preferably 1.5 μm or more, more preferably 2.5 μm or more. When delamination treatment of MXene is performed by subjecting MXene to ultrasonic treatment, since most of the MXene is reduced in diameter to about several hundred nanometers in the major axis by the ultrasonic treatment, it is considered that the film formed of the single-layer MXene delaminated by the ultrasonic treatment has low orientation of MXene.

[0039] From the viewpoint of dispersibility in the dispersion medium, the average value of the major axis of the two-dimensional plane is 20 μm or less, preferably 15 μm or less, more preferably 10 μm or less.

[0040] The major axis of the above two-dimensional plane refers to the major axis when each MXene particle is approximated to an elliptical shape in an electron micrograph observed from a direction substantially orthogonal to the plane parallel to each layer of the above MXene, and the average value of the major axis of the above two-dimensional plane refers to the number average of the above major axes of 80 particles or more. As the electron microscope, a scanning electron microscope (SEM) or a transmission electron microscope (TEM) photograph can be used.

[0041] The average value of the major axis of MXene in this embodiment may be measured by dissolving the material containing the MXene in a solvent and dispersing the MXene in the solvent. Alternatively, it may be measured from the SEM image of the above material.

[0042] (Average value of the thickness of MXene) The average value of the thickness of MXene in this embodiment is preferably 1 nm or more and 100 μm or less. The above thickness is preferably 50 μm or less, more preferably 20 μm or less. On the other hand, considering the thickness of single-layer MXene particles, the lower limit of the thickness of MXene can be 1 nm.

[0043] The thickness of the above-mentioned MXene can be understood as the length in the direction substantially perpendicular to the plane parallel to each layer, and the average value of the thickness of the above-mentioned MXene is determined as the number average dimension (for example, at least 40 number averages) based on an atomic force microscope (AFM) photograph or a transmission electron microscope (TEM) photograph.

[0044] The above-mentioned MXene can be produced by the following production methods, but the MXene in the present disclosure is not limited to those produced by the following production methods.

[0045] In one aspect, the production method of the above-mentioned MXene is (a) The following formula: M m AX n (wherein M is at least one metal of Groups 3, 4, 5, 6, 7, X is a carbon atom, a nitrogen atom or a combination thereof, A is at least one element of Groups 12, 13, 14, 15, 16, n is 1 or more and 4 or less, m is greater than n and 5 or less) preparing a precursor represented by, (b) using an etching solution to remove at least a part of A atoms from the above-mentioned precursor by etching to obtain the above-mentioned etched product, (c) washing the above-mentioned etched product to obtain a washed product, including, furthermore, (d) performing an intercalation treatment on the above-mentioned etched product in a dispersion medium using a metal-containing compound to obtain an intercalated product, and, (e) performing a delamination treatment on the above-mentioned intercalated product to obtain a delaminated product may be included.

[0046] In one aspect, the above-mentioned etched product and delaminated product can be used as the above-mentioned MXene, and preferably, the above-mentioned washed product can be used as the above-mentioned MXene.

[0047] The following describes each step.

[0048] · Step (a) First, prepare a predetermined precursor. The predetermined precursor that can be used in this embodiment is a MAX phase, which is a precursor of MXene, The following formula: M m AX n (In the formula, M is at least one metal of Groups 3, 4, 5, 6, and 7, X is a carbon atom, a nitrogen atom, or a combination thereof, A is at least one element of Groups 12, 13, 14, 15, and 16, n is 1 or more and 4 or less, m is greater than n and 5 or less) is represented by.

[0049] The above M, X, n, and m have the same meanings as above. A is at least one element of Groups 12, 13, 14, 15, and 16, usually a Group A element, typically a Group IIIA element and a Group IVA element, and more specifically, may include at least one selected from the group consisting of Al, Ga, In, Tl, Si, Ge, Sn, Pb, P, As, S, and Cd, and is preferably Al or Si.

[0050] The MAX phase has a crystal structure in which a layer composed of A atoms is located between two layers represented by M m X n (each X may have a crystal lattice located within the octahedral array of M). The MAX phase typically has a repeating unit in which a layer of X atoms is arranged one by one between each of the n + 1 layers of M atoms (collectively referred to as the "M m X n layer"), and a layer of A atoms ("A atom layer") is arranged as the next layer after the (n + 1)-th layer of M atoms, but is not limited thereto.

[0051] The above MAX phase can be manufactured by known methods. For example, TiC powder, Ti powder, and Al powder are mixed with a ball mill, and the obtained mixed powder is fired in an Ar atmosphere to obtain a fired body (block-shaped MAX phase). Thereafter, the obtained fired body can be pulverized with an end mill to obtain a powdery MAX phase for the next process.

[0052] · Step (b) In step (b), an etching process is performed to remove at least a part of the A atoms from the precursor (MAX phase) represented by the above M m AX n . As a result, an etched product is obtained in which at least a part of the layer composed of A atoms is removed while the M m X n layer in the precursor is maintained.

[0053] The conditions of the etching process are not particularly limited, and known conditions can be adopted. The etching can be carried out using an etching solution containing F - . Such an etching solution may contain hydrofluoric acid, hydrochloric acid, phosphoric acid, etc. as acids. In one aspect, examples of the etching solution include hydrofluoric acid; a mixed solution of hydrofluoric acid and hydrochloric acid; a mixed solution of lithium fluoride and hydrochloric acid, etc., and any of them may further contain phosphoric acid. As the solvent in the etching solution, water may be used, for example, pure water may be used.

[0054] The etching process may be carried out by mixing the above precursor and the above etching solution and performing it as a slurry.

[0055] In one aspect, in step (b), an intercalation process may be carried out simultaneously. By coexisting a metal-containing compound described later in the etching solution, the etching process and the intercalation process can be carried out simultaneously. In this case, step (e) described later may be further carried out.

[0056] In step (b), when performing the intercalation treatment simultaneously, the content ratio of the metal-containing compound in the total of the precursor, the metal-containing compound, and the etching solution can be, for example, 0.001% by mass or more and 10% by mass or less, further 0.01% by mass or more and 1% by mass or less, particularly 0.1% by mass or more and 1% by mass or less.

[0057] · Step (c) In step (c), the processed material obtained by the etching treatment is washed to obtain a washed processed material. By performing the washing, acids and the like used in the etching treatment can be sufficiently removed.

[0058] The washing can preferably be carried out using water. The amount of water mixed with the etched material and the washing method are not particularly limited. For example, adding water and stirring, centrifuging, etc. can be mentioned. As the stirring method, stirring methods using a hand shake, an automatic shaker, a share mixer, a pot mill, etc. can be mentioned. The degree of stirring such as the stirring speed and the stirring time may be adjusted according to the amount and concentration of the etched material to be treated. The washing with the above water may be carried out once or more, and it is preferable to carry out the washing with water a plurality of times. For example, specifically, the washing with the above water may be carried out by sequentially adding water to the processed material (or the remaining precipitate obtained in the following (iii)) and stirring, step (ii) centrifuging the stirred material, and step (iii) discarding the supernatant after centrifugation, and steps (i) to (iii) may be repeated within a range of 2 times or more, for example, 15 times or less.

[0059] · Step (d) In step (d), using a metal-containing compound containing metal ions, an intercalation treatment is performed on the above-mentioned etched material in a dispersion medium to obtain an intercalation-treated material, and the intercalation treatment is carried out. Thereby, metal ions contained in the metal-containing compound are intercalated between two adjacent M m X n layers, and an intercalation-treated material is obtained.

[0060] The above metal ions may include monovalent metal ions, and examples of such monovalent metal ions include alkali metal ions such as lithium ions, sodium ions, and potassium ions, copper ions, silver ions, gold ions, and the like.

[0061] Examples of the above metal-containing compound include ionic compounds in which the above metal ions are bonded to anions. For example, sulfide salts containing iodides, phosphates, and sulfates of the above metal ions, nitrates, acetates, and carboxylates can be mentioned. As the above metal ions, lithium ions are preferable, as the metal-containing compound, a metal-containing compound containing lithium ions is preferable, an ionic compound of lithium ions is more preferable, and one or more of iodides, phosphates, and sulfide salts of lithium ions are even more preferable. When lithium ions are used as the metal ions, it is considered that the water hydrated to the lithium ions has the most negative dielectric constant, so that it is easy to form a monolayer.

[0062] In the total of the above etching-treated product, metal-containing compound, and dispersion medium, the content of the metal-containing compound can be, for example, 0.001% by mass or more and 10% by mass or less, further 0.01% by mass or more and 1% by mass or less, and particularly 0.1% by mass or more and 1% by mass or less. When the content of the metal-containing compound is within the above range, the dispersibility in the dispersion medium is good.

[0063] The specific method of the intercalation treatment is not particularly limited. For example, the dispersion medium, the etching-treated product, and the metal-containing compound may be mixed and stirred or allowed to stand. For example, stirring at room temperature can be mentioned. Examples of the above stirring method include a method using a stirrer such as a stir bar, a method using a stirring blade, a method using a mixer, and a method using a centrifuge. The stirring time can be set according to the production scale of monolayer and few-layer MXene particles, and can be set, for example, between 12 and 24 hours. The mixing order of the dispersion medium, the etching-treated product, and the metal-containing compound is not particularly limited. However, in one aspect, the metal-containing compound may be mixed after mixing the dispersion medium and the etching-treated product. Typically, the etching solution after the etching treatment can be used as the dispersion medium.

[0064] · Step (e) In step (e), delamination treatment is performed on the intercalation-treated product obtained by performing intercalation treatment to obtain a delamination-treated product. The delamination treatment includes applying a shear stress to the intercalation-treated product to delaminate at least a part between two adjacent M m X n layers. By the delamination treatment, MXene particles can be made into single-layer or few-layer.

[0065] The conditions of the delamination treatment are not particularly limited and can be performed by known methods. For example, as a method of applying a shear stress to the intercalation-treated product, a method of dispersing the intercalation-treated product in a dispersion medium and stirring it can be mentioned. Examples of the stirring method include stirring using ultrasonic treatment, hand shaking, an automatic shaker, etc. The degree of stirring such as the stirring speed and stirring time may be adjusted according to the amount and concentration of the product to be treated. For example, after centrifuging the slurry after the above intercalation and discarding the supernatant, pure water is added to the remaining precipitate, and layer separation is performed by stirring, for example, by hand shaking or an automatic shaker. Removal of the undelaminated material includes a step of centrifuging and discarding the supernatant and then washing the remaining precipitate with water. For example, (i) add pure water to the remaining precipitate after discarding the supernatant and stir, (ii) centrifuge, and (iii) collect the supernatant. The operations of (i) to (iii) are repeated once or more, preferably two or more times and ten times or less to obtain a supernatant containing single-layer or few-layer MXene particles as the delamination-treated product. Alternatively, this supernatant may be centrifuged, the supernatant after centrifugation may be discarded, and a clay containing single-layer or few-layer MXene particles may be obtained as the delamination-treated product.

[0066] In the above method for producing MXene, when an intercalation treatment is carried out, a washing treatment may be further carried out at any stage after the intercalation treatment, preferably at a stage after the delamination treatment. By carrying out such a washing treatment, the metal ions and metal-containing compounds used in the intercalation can be sufficiently removed. Typically, such a washing treatment is carried out after step (e).

[0067] In one aspect, the washing treatment after such an intercalation treatment may be carried out in the same manner as in the above step (c). In another aspect, after the delaminated product is acid-treated, the acid-treated product may be washed in the same manner as in the above step (c). In these cases, the etched product in the above step (c) may be read as the delaminated product or the acid-treated product respectively to carry out the washing treatment.

[0068] The above acid treatment can be carried out by mixing the delaminated product and an acid solution and stirring them. As such an acid, inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, perchloric acid, hydroiodic acid, hydrobromic acid, hydrofluoric acid; organic acids such as acetic acid, citric acid, oxalic acid, benzoic acid, sorbic acid may be appropriately used, and the concentration of the acid in the acid solution may be appropriately adjusted according to the delaminated product. The above stirring can be carried out using a hand shaker, an automatic shaker, a share mixer, a pot mill, etc. The acid treatment may be carried out one or more times, and if necessary, the operation of mixing and stirring with a fresh acid solution (an acid solution not used in the acid treatment) may be carried out within a range of two or more times, for example, 10 times or less.

[0069] The intermediate and the target product in the production method described above may be isolated by commonly used purification methods. Examples of such purification methods include suction filtration; drying such as heat drying, freeze drying, vacuum drying, etc.

[0070] The above-mentioned MXene may have an effect of improving the gut microbiota in vivo. In particular, it may promote the production of gut bacteria in vivo. Such gut bacteria may preferably include gut bacteria that increase short-chain fatty acids (gut bacteria that produce short-chain fatty acids through metabolism). The gut bacteria that increase short-chain fatty acids may preferably include the phylum Firmicutes, class Clostridia, more preferably the family Eggerthellaceae, order Coriobacteriales, class Coriobacteriia, phylum Actinobacteriota, the family Ruminococcaceae, order Oscillospirales, class Clostridia, phylum Firmicutes, the family Lachnospiraceae, order Lachnospirales, class Clostridia, phylum Firmicutes, and more preferably the family Butyricicoccaceae, order Oscillospirales, class Clostridia, phylum Firmicutes.

[0071] Examples of the short-chain fatty acids include fatty acids having 1 to 4 carbon atoms, preferably butyric acid, acetic acid, and propionic acid. The above short-chain fatty acids may have an effect of repairing the vascular barrier. Examples of such vascular barriers include the intestinal barrier and the cerebrovascular barrier.

[0072] In addition, the above-mentioned MXene may have an effect of lowering blood pressure in vivo.

[0073] Although the present disclosure should not be construed as being limited to a specific theory, the effects of short-chain fatty acids include the following (i) to (iii): (i) Repairing the vascular barrier (ii) Enhancing the anti-inflammatory effect (iii) Improving hypertension are considered to be included.

[0074] (i) Regarding this, short-chain fatty acids contribute to maintaining the homeostasis of the blood-brain barrier (especially the cerebrovascular endothelial cells and the blood-brain barrier), can suppress excessive enhancement of permeability, and act protectively on brain tissues. This can be confirmed by the enhancement of Cldn, which is an RNA encoding Claudin, a protein forming the blood-brain barrier (especially the blood-brain barrier). Such vascular barriers include the intestinal barrier and the blood-brain barrier.

[0075] (ii) Regarding this, short-chain fatty acids act as ligands of G-protein coupled receptor (GPR) and have an anti-inflammatory effect via the immune system throughout the body. In the central nervous system, among short-chain fatty acids, especially those with high permeation efficiency through the blood-brain barrier, they have an anti-inflammatory effect due to the inhibitory effect on Histone Deacetylase (HDAC) activated at the damaged site. The effect of butyric acid on such white matter of the brain can be confirmed from the fact that the GPR signaling pathway in the white matter of the brain is significantly enriched by MFT administration in the RNA-sequencing data of hypertensive mice (functional enrichment analysis using Database for Annotation, Visualization and Integrated Discovery (DAVID)).

[0076] (iii) Regarding this, short-chain fatty acids can have an effect of lowering blood pressure. It is a known fact that the white matter of the brain (nerve axons and myelin [complementary]) is damaged by hypertension. Such antihypertensive effect can be confirmed from the point that a protective effect is confirmed on the central component (myelin) of the white matter and the point that the blood pressure decreases in the white matter of hypertensive mice.

[0077] That is, the MXene of the present disclosure can be used for one or more selected from the following (i) to (iii): (i) Repairing the vascular barrier (ii) Enhancing the anti-inflammatory effect (iii) Improving hypertension and can be used for one or more selected therefrom. Improving the above hypertension also includes reducing blood pressure in vivo.

[0078] The above MXene can be used in the treatment or prevention of diseases in which improvement of symptoms can be expected by repairing the vascular gate in vivo. Examples of such diseases include one or more selected from vascular dementia, Parkinson's disease, inflammatory bowel disease, chronic renal failure, irritable bowel syndrome, and ischemic and demyelinating central nervous system diseases.

[0079] The above MXene and pharmaceutical composition can be administered orally.

[0080] The pharmaceutical composition according to this embodiment can be made into various dosage forms according to the usage. Examples of such dosage forms include, for example, powders, granules, fine granules, dry syrups, tablets, capsules, liquids, sublingual agents, etc., and also include injections, ointments, suppositories, patches, etc.

[0081] The pharmaceutical composition according to this embodiment can be configured as a pharmaceutical composition further containing MXene as an active ingredient and a pharmacologically acceptable additive by a known method according to its dosage form. Examples of such additives include excipients, disintegrants, binders, lubricants, diluents, buffers, isotonic agents, preservatives, wetting agents, emulsifiers, dispersants, stabilizers, solubilizing agents, etc. The pharmaceutical composition of the present disclosure can be prepared by appropriately mixing the above MXene and the above additive, or diluting / dissolving the above MXene with an additive.

[0082] The pharmaceutical composition according to this embodiment can be administered systemically or locally, orally or parenterally (nasally, via the lungs, intravenously, enterally, subcutaneously, intramuscularly, transdermally). In one aspect, the pharmaceutical composition according to this embodiment can be administered orally.

[0083] When the pharmaceutical composition of the present disclosure is used for treatment, the dosage of the above-mentioned MXene as the active ingredient is appropriately determined according to the age, sex, weight, disease, degree of treatment, etc. of the patient. For example, in the case of oral administration, the dosage may be appropriately administered once or in several divided doses within a range of generally 100 mg to 10 g / body per day as an effective amount for an adult (assuming a body weight of 60 kg).

[0084] In addition, the pharmaceutical composition containing the above-mentioned MXene can be used for manufacturing a pharmaceutical for treating or preventing diseases.

Example

[0085] The present disclosure will be further specifically described by the following examples, but the present disclosure is not limited thereto.

[0086] Example 1 〔Preparation of MXene〕 In Examples 1 and 2, (1) preparation of the precursor (MAX), (2) etching of the precursor, and (3) washing and drying, which will be described in detail below, were carried out in sequence to prepare MXene two-dimensional particles. (3) Washing and drying were carried out in sequence to prepare MXene two-dimensional particles.

[0087] (1) Preparation of the precursor (MAX) TiC powder, Ti powder, and Al powder (all manufactured by High Purity Chemical Laboratory Co., Ltd.) were put into a ball mill containing zirconia balls at a molar ratio of 2:1:1 and mixed for 24 hours. The obtained mixed powder was fired at 1,350 °C for 2 hours under an Ar atmosphere. The obtained fired body (block) was ground with an end mill to a maximum size of 40 μm or less. Thereby, Ti3AlC2 particles were obtained as the precursor (MAX).

[0088] (2) Etching of the precursor Using the Ti3AlC2 particles (powder) prepared by the above method, etching was carried out under the following etching conditions to obtain a solid-liquid mixture (slurry) containing a solid component derived from Ti3AlC2 powder. (Etching conditions) · Precursor: Ti3AlC2 (passed through a 45 μm sieve) · Etching solution composition: 6 mL of 50% by mass HF, 18 mL of H2O 36 mL of HCl (12 M) · Precursor input amount: 3.0 g · Etching container: 100 mL I - boy · Etching temperature: 35 °C · Etching time: 24 h · Stirrer rotation speed: 400 rpm

[0089] (3) Washing and drying The above - mentioned slurry was divided into two parts and inserted into two 50 - mL centrifuge tubes respectively. After centrifugation was carried out at 3,500 G using a centrifuge, the supernatant was discarded. 40 mL of pure water was added to each centrifuge tube, and the operation of centrifuging again at 3,500 G to separate and remove the supernatant was repeated 11 times. After the final centrifugation, the supernatant was discarded, and Ti3C2T s - water - medium clay was obtained. The obtained clay was freeze - dried into dry powder to obtain dry powder of MXene.

[0090] Animal experiment 1: Administration experiment on hypertensive mice Hypertension causes inflammation in cerebral capillaries and chronic hypoperfusion, resulting in a decline in cognitive function, and this symptom is called vascular dementia. In animal experiment 2, it was confirmed whether the administration of MXene to genetically hypertensive mice could remove inflammatory substances in the body and restore cognitive function. The administration period was 56 days, and the administration dose was 8% of the food intake. After the end of the administration period, blood pressure fluctuations and cognitive behavior analysis (Novel Object Recognition Test) were carried out. In addition, a Ctl group without MXene administration was also prepared as a comparative example.

[0091] (Blood pressure change) The changes in systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the MXene group and the Ctl group were measured. The comparison was carried out by taking the difference based on the blood pressure value on the first day. The results are shown in Figure 3. A decrease in blood pressure was observed in the MXene group from around the 4th week.

[0092] (Behavior analysis) In the novel object recognition test, a black box with a vertical width of 400 mm, a horizontal width of 400 mm, and a height of 400 mm was used. On the first and second days, the mouse was left in the box for 10 minutes for acclimation. On the third day, two identical substances (old substances) were placed in the cage, and the mouse was left for 10 minutes. On the fourth day, one of the objects was changed to a new substance, and the mouse was left for 10 minutes. The time spent exploring both objects was measured, and the ratio of the exploration time of the new substance to the total exploration time (Times of Entries Discrimination Index) was calculated using the following formula: Times of Entries Discrimination Index = [n / (n + f)] - 0.5 [where n represents the number of times the new object was touched, and f represents the number of times the conventional object was touched.] Based on this, it was calculated and used as an index of memory learning ability. The behavior area of the mouse is shown in Figure 4, and the Times of Entries Discrimination Index is shown in Figure 5.

[0093] As shown in Figure 5, in the Novel Object Recognition Test, the Times of Entries Discrimination Index was higher and a positive number in the MXene group. This indicates that the number of times of approaching the new substance with interest is higher than that of the old substance. That is, it suggests that the new substance can be recognized as new and the cognitive function has recovered.

[0094] (Confirmation of gut microbiota) As shown in FIGS. 6(a) to (d), the abundance of the intestinal microbiota in mice was compared between groups by 16S-rRNA analysis. It was confirmed that the families Eggerthellaceae of the order Coriobacteriales, class Coriobacteriia, phylum Actinobacteriota, Ruminococcaceae of the order Oscillospirales, class Clostridia, phylum Firmicutes, Lachnospiraceae of the order Lachnospirales, class Clostridia, phylum Firmicutes, and Butyricicoccaceae of the order Oscillospirales, class Clostridia, phylum Firmicutes were proliferating.

[0095] (Confirmation of the diversity of the intestinal microbiota) As shown in FIGS. 7(a) and (b), the diversity of the intestinal microbiota in mice obtained by 16S-rRNA analysis was compared. The α-diversity was visualized by PCoA (Principal Coordinates Analysis) after analysis based on the Bray-Curtis distance using the Shannon Index for β-diversity. It was confirmed that the diversity of the microbiota changed between the two groups by the administration of MXene.

[0096] (Confirmation of short-chain fatty acids) Metabolome analysis using CE-MS (capillary electrophoresis mass spectrometer) was performed on mouse serum to confirm changes in biological components due to MXene. As shown in FIGS. 8(a) to (c), the proliferation of short-chain fatty acids, butyric acid, acetic acid, and propionic acid, was confirmed in the MXene group.

[0097] (Confirmation of RNA changes) RNA_seq of corpus callosum cells was performed using a next-generation sequencer. As shown in FIG. 9, the proliferation of Cldn5, a gene encoding the protein (Claudin) that forms the Blood Brain Barrier, was confirmed.

[0098] (Confirmation of the degree of brain tissue damage) The corpus callosum staining evaluation of the specimen was performed. By staining myelin that functionalizes nerve axons, the degree of damage can be confirmed. The more myelin remains, the greater the brightness during staining, and it can be judged that the damage is small. For image analysis, in order to correct the variation during imaging, the brightness of the white matter part based on the part called the cortex was compared. As shown in Figure 10, it was confirmed that the brightness was significantly higher in the MXene group and the damage to the white matter was suppressed.

[0099] In response to the above results, in order to measure the amount of myelin, WB measurement of MBP was performed. The method was a general WB method. Proteins were extracted from the corpus callosum tissue, transferred to a membrane after gel electrophoresis, and bands were detected by antibody reaction. The measurement compared the amount of MBP based on the amount of β-actin as the total protein amount among specimens. Quantification by image processing was performed based on the markers floating on the membrane. As shown in Figures 11(a) and (b), an increase in MBP was confirmed in the MXene administration group.

[0100] It was observed that MXene increased the intestinal growth of Actinobacteriota phylum Coriobacteriia class Coriobacteriales order Eggerthellaceae family, Firmicutes phylum Clostridia class Oscillospirales order Ruminococcaceae family, Firmicutes phylum Clostridia class Lachnospirales order Lachnospiraceae family, and Firmicutes phylum Clostridia class Oscillospirales order Butyricicoccaceae family, which are short-chain fatty acid-producing bacteria. It is considered that the increased short-chain fatty acids promoted the decrease in blood pressure and the repair of the BloodBrainBarrier, contributing to the improvement of cognitive function.

[0101] Furthermore, in the examples, it was confirmed that MXene regulates intestinal bacteria and proliferates short-chain fatty acid (SCFAs)-producing bacteria, especially butyric acid-producing bacteria. Here, the actions of SCFAs (especially butyric acid) are considered to be roughly divided into three.

[0102] First, it contributes to maintaining the homeostasis of cerebrovascular endothelial cells and the blood-brain barrier, suppresses excessive enhancement of permeability, and acts protectively on brain tissues. This can be confirmed by the enhancement of Cldn5, which is an RNA encoding Claudin, a protein forming the blood-brain barrier.

[0103] Second, it acts as a ligand for G-protein coupled receptor (GPR) and has an anti-inflammatory effect via the immune system throughout the body. In the central nervous system, among short-chain fatty acids, it has a particularly high permeation efficiency through the blood-brain barrier and has an anti-inflammatory effect due to its inhibitory effect on Histone Deacetylase (HDAC) activated at the damaged site. In fact, in the RNA-sequencing data of hypertensive mice in the above example (functional enrichment analysis using Database for Annotation, Visualization and Integrated Discovery (DAVID)), the GPR signaling pathway in the cerebral white matter was significantly enriched by MFT administration, and it was confirmed that butyric acid acts on the cerebral white matter.

[0104] Third, it has an effect of lowering blood pressure. It is a known fact that the cerebral white matter (nerve axons and myelin [complementary]) is damaged by hypertension. This time, considering from the fact that a protective effect was confirmed on the central component (myelin) of the white matter and the blood pressure decreased in the cerebral white matter of hypertensive mice, it can be considered that it has an antihypertensive effect.

[0105] From the above points, it was confirmed that MXene has a multi-target protective effect on damaged white matter by regulating the gut microbiota and proliferating short-chain fatty acid (SCFAs)-producing bacteria, especially butyric acid-producing bacteria.

[0106] This disclosure includes the following. [1] A pharmaceutical composition containing MXene, for improving the gut microbiota in vivo or increasing short-chain fatty acids in vivo. [2] The Mxene includes two-dimensional particles having one or more layers, wherein the layer includes at least one metal selected from Group 3, 4, 5, 6, and 7 metals and at least one selected from carbon atoms and nitrogen atoms, and the pharmaceutical composition according to [1]. [3] The layer has the following formula: M m X n (wherein M is at least one metal of Group 3, 4, 5, 6, and 7, X is a carbon atom, a nitrogen atom, or a combination thereof, n is 1 or more and 4 or less, m is greater than n and 5 or less) and includes a layer body represented by the formula, and the pharmaceutical composition according to [2]. [4] The layer further includes a modification or termination T (T is at least one selected from the group consisting of a hydroxyl group, a fluorine atom, a chlorine atom, an oxygen atom, and a hydrogen atom) present on the surface of the layer body, and the pharmaceutical composition according to [3]. [5] The Mxene includes Ti3C2, and the pharmaceutical composition according to [1]. [6] The improvement of the gut microbiota includes growing gut bacteria that increase short-chain fatty acids, and the pharmaceutical composition according to any one of [1] to [5]. [7] The gut bacteria that metabolize the short-chain fatty acids include Clostridia class of the phylum Firmicutes, and the pharmaceutical composition according to any one of [1] to [6]. [8] The intestinal bacteria that metabolize the short-chain fatty acids are the pharmaceutical composition according to any one of [1] to [7], including the Eggerthellaceae family of the order Coriobacteriales, class Coriobacteriia, phylum Actinobacteriota; the Ruminococcaceae family of the order Oscillospirales, class Clostridia, phylum Firmicutes; the Lachnospiraceae family of the order Lachnospirales, class Clostridia, phylum Firmicutes; or the Butyricicoccaceae family of the order Oscillospirales, class Clostridia, phylum Firmicutes. [9] The short-chain fatty acids are the pharmaceutical composition according to any one of [1] to [8], including butyric acid.

[10] Furthermore, the following (i) to (iii): (i) Repairing the vascular barrier (ii) Enhancing the anti-inflammatory effect (iii) Improving hypertension The pharmaceutical composition according to any one of [1] to [9], which is used for one or more selected from the above.

[11] The vascular barrier is the intestinal barrier or the blood-brain barrier, and it is the pharmaceutical composition according to any one of [1] to

[10] .

[12] Furthermore, it is the pharmaceutical composition according to any one of [1] to

[11] , which is used for reducing blood pressure.

[13] The pharmaceutical composition according to any one of [1] to

[12] , which is for oral administration.

[14] A method for improving the intestinal microbiota in vivo or increasing short-chain fatty acids in vivo, including administering an effective amount of MXene to a subject.

[15] The MXene is administered orally, and it is the method according to

[14] .

[16] Including administering an effective amount of MXene to a subject, A method for treating or preventing a disease in which improvement of symptoms can be expected by repairing the vascular barrier.

[17] The method according to

[16] , wherein the disease includes one or more selected from vascular dementia, Parkinson's disease, inflammatory bowel disease, chronic renal failure, irritable bowel syndrome, and ischemic / demyelinating central nervous system diseases.

[18] The MXene is for the treatment or prevention method according to

[16] or

[17] , which is administered orally.

Explanation of reference numerals

[0107] 1a, 1b layer body (M m X n layer) 3a, 5a, 3b, 5b modification or termination T 7a, 7b MXene layer 10, 10a, 10b MXene particles (two-dimensional particles of layered material)

Claims

1. A pharmaceutical composition comprising MXene for improving the gut microbiota in vivo or increasing short-chain fatty acids in vivo.

2. The MXene comprises two-dimensional particles having one or more layers, The layer comprises at least one metal selected from Group 3, 4, 5, 6, 7 metals and at least one selected from carbon atoms and nitrogen atoms, The pharmaceutical composition according to claim 1.

3. The layer has the following formula: M ｍ X ｎ (wherein M is at least one Group 3, 4, 5, 6, 7 metal, X is a carbon atom, a nitrogen atom or a combination thereof, n is 1 or more and 4 or less, m is greater than n and 5 or less) The pharmaceutical composition according to claim 2, comprising a layer body represented by

4. The layer further comprises a modification or termination T (T is at least one selected from the group consisting of a hydroxyl group, a fluorine atom, a chlorine atom, an oxygen atom and a hydrogen atom) present on the surface of the layer body, The pharmaceutical composition according to claim 3.

5. The Mxene is Ti ３ C ２ The pharmaceutical composition according to claim 1, comprising

6. The improvement of the gut microbiota includes proliferating gut bacteria that increase short-chain fatty acids, The pharmaceutical composition according to claim 1.

7. The gut bacteria that metabolize the short-chain fatty acids include Clostridia class of the phylum Firmicutes, The pharmaceutical composition according to claim 1.

8. The gut bacteria that metabolize the short-chain fatty acids include Eggerthellaceae family of the order Coriobacteriales of the class Coriobacteriia of the phylum Actinobacteriota, Ruminococcaceae family of the order Oscillospirales of the class Clostridia of the phylum Firmicutes, Lachnospiraceae family of the order Lachnospirales of the class Clostridia of the phylum Firmicutes, or Butyricicoccaceae family of the order Oscillospirales of the class Clostridia of the phylum Firmicutes, The pharmaceutical composition according to claim 1.

9. The short-chain fatty acids include butyric acid, The pharmaceutical composition according to claim 1.

10. Furthermore, the following (i) to (iii): (i) Repairing the vascular barrier (ii) Enhancing the anti-inflammatory effect (iii) Improving hypertension The pharmaceutical composition according to claim 1, used for one or more selected from

11. The vascular barrier is an intestinal barrier or a blood-brain barrier, The pharmaceutical composition according to claim 1.

12. Furthermore, the pharmaceutical composition according to claim 1, which is used for reducing blood pressure.

13. The pharmaceutical composition according to any one of claims 1 to 12, which is for oral administration.

14. A method for improving the gut microbiota in vivo or increasing short-chain fatty acids in vivo, comprising administering an effective amount of MXene to a subject.

15. The method according to claim 14, wherein the MXene is administered orally.

16. Comprising administering an effective amount of MXene to a subject, A method for treating or preventing a disease in which improvement of symptoms can be expected by repair of the blood-brain barrier.

17. The method according to claim 16, wherein the disease includes one or more selected from vascular dementia, Parkinson's disease, inflammatory bowel disease, chronic renal failure, irritable bowel syndrome, and ischemic / demyelinating central nervous system diseases.

18. The method for treatment or prevention according to claim 16, wherein the MXene is administered orally.

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