Zinc polyphosphate and method for producing same

Abstract

The present invention relates to a novel zinc polyphosphate, a method for using the same, and a method for producing the same. The present invention relates to a zinc polyphosphate and, more specifically, to: a zinc polyphosphate characterized by exhibiting a halo peak at 2 [theta] = 4-7 degrees in X-ray diffraction, and having a structure represented by formula (1), a pharmaceutical composition containing the zinc polyphosphate as an active ingredient, and a method for producing the zinc polyphosphate; the method for producing zinc polyphosphate comprises (1) a step for adding zinc ions to a polyphosphoric acid solution under alkaline conditions, preferably pH 8-11, and (2) a step for recovering the solid obtained in step (1) as zinc polyphosphate.

Description

Technical Field:

[0004] The present invention relates to novel zinc polyphosphates and methods for producing the same. Background Art

[0005] In inflammatory bowel diseases (IBD) typified by ulcerative colitis (UC) and Crohn's disease (CD), anti-inflammatory agents are used as standard therapeutic drugs, but there is still no drug that directly induces mucosal healing. Anti-inflammatory agents have been symptomatic therapies in the treatment of IBD. In contrast, recently, the true treatment goal of IBD is said to be "mucosal healing". Based on long-term consumption experience, probiotics typified by lactic acid bacteria are highly safe and are known to have certain intestinal regulating effects, etc., but there are still many unclear points about the mechanism of action of probiotics on the intestinal state. In order to clarify this, the identification and analysis of bioactive molecules produced by probiotics are underway.

[0006] As a molecule that strengthens the intestinal barrier function, the inventors of the present application identified long-chain polyphosphate from Lactobacillus maltaromicus. In addition, it was confirmed that long-chain polyphosphate improves the decline of intestinal barrier function and intestinal damage caused by DSS treatment (Patent Document 1), and achieves mucosal healing in patients with refractory ulcerative colitis (Non-Patent Document 1). In addition, it has been reported that the Ca salt of polyphosphate acts on damaged digestive tract mucosa in inflammatory bowel disease and exhibits a specific platelet aggregation effect on the damaged mucosa (Patent Document 2).

[0007] Polyphosphate is known to have wound healing and anti-inflammatory effects, and it is expected to develop wound covering materials and dental materials using amorphous or nanoparticles composed of calcium polyphosphate (Patent Document 3).

[0008] Polyphosphate is also known as a blood coagulation inducing factor. If polyphosphate is released from platelets in the blood, it activates factor XII protease in the blood and induces a coagulation reaction. It is assumed that natural polyphosphate has various salts such as Ca salt, Mg salt, Na salt, and K salt, and it is also known that there are amorphous nanoparticles (Non-Patent Documents 2 and 3). Donovan et al. produced nanoparticles from polyphosphates with different chain lengths and reported the effects on the size and blood coagulation of polyphosphate (Non-Patent Document 4).

[0009] Prior Art Documents

[0010] Patent Documents

[0011] Patent Document 1: WO 2011 / 125619

[0012] Patent Document 2: WO 2021 / 141066

[0013] Patent Document 3: WO 2016 / 079006

[0014] Non-patent literature

[0015] Non-patent literature 1: Fujita et al., “Long-Chain Polyphosphate Is a Potential Agent for Inducing Mucosal Healing of the Colon in Ulcerative Colitis.” Clin Pharmacol Ther. 2019 Sep.

[0016] Non-patent literature 2: Feng et al., “Biogenic Polyphosphate Nanoparticles from a Marine Cyanobacterium Synechococcus sp. PCC 7002: Production, Characterization, and Anti-Inflammatory Properties In Vitro.” Mar Drugs. 2018 Sep 10; 16(9).

[0017] Non-patent literature 3: Feng et al., “Biogenic Polyphosphate Nanoparticles from Synechococcus sp. PCC 7002 Exhibit Intestinal Protective Potential in Human Intestinal Epithelial Cells In Vitro and Murine Small Intestine Ex Vivo.” J Agric Food Chem. 2018 Aug 1; 66(30):8026-8035.

[0018] Non-patent literature 4: Donovan et al., “Size-controlled synthesis of granular polyphosphate nanoparticles at physiologic salt concentrations for blood clotting.” Biomacromolecules. 2014 Nov 10; 15(11):3976-84. Summary of the invention

[0019] Problems to be solved by the invention

[0020] An object of the present invention is to provide a novel zinc polyphosphate, its use as a pharmaceutical product, etc., and a manufacturing method thereof.

[0021] Means for Solving the Problem

[0022] The inventors of the present application found that zinc polyphosphate obtained under certain conditions shows characteristic X-ray diffraction peaks and has an excellent intestinal barrier enhancing effect.

[0023] Based on the above insights, in the first aspect, the present invention provides the following [1] to

[12] .

[0024] [1] Zinc polyphosphate, characterized in that in X-ray diffraction, a halo peak is shown at 2θ = 4 to 7°.

[0025] [2] The zinc polyphosphate according to [1], having a Zeta potential of -20 mV or less.

[0026] [3] The zinc polyphosphate according to [1], wherein the zinc content is 10 to 60%, preferably 20 to 55%, more preferably 30 to 50%.

[0027] [4] The zinc polyphosphate according to [1], wherein the average chain length of the polyphosphate part of the zinc polyphosphate is 1.1 phosphate units or more and less than 100 phosphate units, preferably 1.1 phosphate units or more and less than 50 phosphate units, more preferably 1.2 to 30 phosphate units, and further preferably 1.2 to 15 phosphate units.

[0028] [5] The zinc polyphosphate according to [1], wherein the average particle size of the zinc polyphosphate is 1 to 1000 nm, more preferably 20 to 500 nm.

[0029] [6] The zinc polyphosphate according to [1], wherein the half-life of the intestinal barrier enhancing activity under the conditions of 60°C or lower and 75% humidity is 2 weeks or more, preferably 4 weeks or more, and further preferably 6 months or more.

[0030] [7] A pharmaceutical composition containing the zinc polyphosphate according to any one of [1] to [6] as an active ingredient.

[0031] [8] A method, which is a manufacturing method of zinc polyphosphate, comprising:

[0032] (1) A step of adding zinc ions to a polyphosphoric acid solution under alkaline conditions, preferably at pH 8 to 11, more preferably at pH 9 to 10.5, and further preferably at pH 9.5 to 10.0; and

[0033] (2) A step of recovering the solid obtained in step (1) as zinc polyphosphate.

[0034] [9]A method for manufacturing zinc polyphosphate, comprising:

[0035] (1) A step of adding zinc ions to a polyphosphoric acid solution under non-alkaline conditions;

[0036] (2) Subsequently, adjusting the aforementioned solution to alkaline conditions, preferably pH 8 to 11, more preferably pH 9 to 10.5, and further preferably pH 9.5 to 10.0, and allowing the reaction to proceed for more than 2 hours, preferably 3 hours or more, more preferably 4 hours or more; and

[0037] (3) A step of recovering the solid obtained in step (2) as zinc polyphosphate.

[0038]

[10] The method according to [8] or [9], wherein the obtained zinc polyphosphate exhibits a halo peak at 2θ = 4 to 7° in X-ray diffraction.

[0039]

[11] The zinc polyphosphate according to [1], which is obtained by the method according to [8] or [9].

[0040]

[12] The zinc polyphosphate according to [1] or

[11] , wherein in X-ray diffraction, signal peaks are also exhibited at positions of 2θ = 30 to 36° and / or 56 to 62°.

[0041] Effects of the Invention

[0042] By the present invention, a novel zinc polyphosphate having high intestinal barrier enhancing activity and a method for manufacturing the same are provided. Description of the Drawings

[0043] [Figure 1] Figure 1 shows the NMR spectra of various zinc polyphosphates. A: Test15-2, B: Test15-6, C: Test15-12, D: Test40-1, E: Test40-6-1, F: Test40-6-2, G: Short-chain Test15-12

[0044] [Figure 2] Figure 2 shows the X-ray diffraction spectra of various zinc polyphosphates. A: Test15-2, B: Test15-6, C: Test15-12, D: Test40-1, E: Test40-6-1, F: Test40-6-2, G: Short-chain Test15-12

[0045] [Figure 3] Figure 3 shows the particle size distribution of zinc polyphosphate. A: Test15-6, B: Test40-6-1, C: Test40-6-2

[0046] Figure 4 Figure 4 Indicating the intestinal barrier enhancing effect of zinc polyphosphate (intestinal extravasation​​3 Amount of H - mannitol). From left to right are untreated (control group), oxidative stress load (NH 2 Cl), oxidative stress load + Test15 - 12 treatment (NH 2 Cl + Test15 - 12)

[0047] Figure 5 Figure 5 Indicates the comparison of the intestinal barrier enhancing effect of Test40 - 6 - 1 and Test40 - 6 - 2 (intestinal extravasation 3 Amount of H - mannitol). From left to right are the control group, oxidative stress load · untreated (NH 2 Cl), oxidative stress load · Test40 - 6 - 1 treatment (NH 2 Cl, Test40 - 6 - 1), oxidative stress load · Test40 - 6 - 2 treatment (NH 2 Cl, Test40 - 6 - 2)

[0048] Figure 6 Figure 6 Indicates the intestinal barrier enhancing effect of zinc polyphosphate Test15 - 6 after storage under the conditions of 40°C 75% RH or 60°C 75% RH (RH: relative humidity) (intestinal extravasation 3 Amount of H - mannitol). From left to right are the control group, oxidative stress load (NH 2 Cl), oxidative stress load · Test15 - 6 treatment (NH 2 Cl, Test40 - 6 - 1), oxidative stress load · Test15 - 6 treatment, storage at 40°C 75% RH for 24 weeks (NH 2 Cl, Test15 - 6, 40°C 75% RH 24W), oxidative stress load · Test15 - 6 treatment, storage at 60°C 75% RH for 24 weeks (NH 2 Cl, Test15 - 6, 60°C 75% 24W)

[0049] Figure 7 Figure 7 Indicates the intestinal barrier enhancement rate of zinc polyphosphate Test15 - 12 and short - chain Test15 - 12. From left to right are untreated (control group), oxidative stress load (NH 2 Cl), oxidative stress load + Test15 - 12 treatment (NH 2 Cl + Test15 - 12), oxidative stress load + short - chain Test15 - 12 treatment (NH 2 Cl + Test15 - 12) Detailed implementation mode

[0050] 1. Zinc polyphosphate​​​​​​

[0051] The present invention relates to zinc polyphosphate, which is characterized in that in X-ray diffraction, a halo peak is shown at 2θ = 4 to 7°. "Zinc polyphosphate" refers to the zinc salt of polyphosphoric acid. In the aforementioned "zinc polyphosphate", it is not necessary for the hydroxyl groups of the phosphoric acid (-[PO(OH)O]-) constituting the polyphosphoric acid to all form zinc salts. As long as the object of the present invention is not violated, a part thereof can be zinc polyphosphate containing other metal salts, such as sodium polyphosphate, calcium polyphosphate, etc. In addition, as long as the object of the present invention is not violated, it can be zinc polyphosphate containing zinc inorganic salts such as zinc hydroxide, zinc oxide, zinc phosphate, various inorganic salts such as calcium phosphate, and inorganic carbon compounds such as calcium carbonate.

[0052] (1) Polyphosphoric acid

[0053] The "polyphosphoric acid" constituting the zinc polyphosphate is a condensed phosphoric acid compound obtained by dehydration condensation of phosphoric acid (H 3 PO 4 ). It can be linear or cyclic, and in addition, it can have branches. The polyphosphoric acid is preferably linear.

[0054] In this specification, the number (n) of "phosphate units" contained in the linear part of the polyphosphoric acid is used to represent the average chain length of the polyphosphoric acid. For example, an average chain length of 10 or more phosphate units (n = 10) or more means that the number of repetitions of the phosphate units contained in the linear part of the polyphosphoric acid is 10 or more.

[0055] There is no particular limitation on the lower limit of the "average chain length" of the polyphosphoric acid constituting the zinc polyphosphate of the present invention. For example, it is 1.1 or more phosphate units, preferably 1.2 or more phosphate units (for example, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0 or more phosphate units). There is also no particular limitation on the upper limit. For example, the average chain length is less than 100 phosphate units, preferably 50 or less phosphate units, more preferably 30 or less phosphate units, and further preferably 15 or less phosphate units. For example, the "average chain length" of the polyphosphoric acid of the present invention is in the range of 1.1 or more and less than 100 phosphate units, preferably in the range of 1.1 or more and less than 50 phosphate units, more preferably in the range of 1.2 to 30 phosphate units, and further preferably in the range of 1.2 to 15 phosphate units. The zinc polyphosphate of the present invention has a high intestinal barrier enhancing ability even when short-chained, and can effectively prevent or improve (cure, relieve) inflammatory bowel disease.

[0056] The zinc polyphosphate of the present invention is preferably poorly soluble in water. In addition, the zinc polyphosphate of the present invention preferably forms nanoparticles, but it can also be in forms other than nanoparticles such as glassy fragments.

[0057] Polyphosphoric acid can be chemically synthesized, or synthesized in vitro using biological molecules such as enzymes, or synthesized using microorganisms that produce polyphosphoric acid. In the case of synthesizing linear polyphosphoric acid, it is preferable to synthesize in vitro using biological molecules such as enzymes or using microorganisms, etc., because linear polyphosphoric acid can be obtained with high efficiency.

[0058] As a method for chemically synthesizing polyphosphoric acid, for example, a method of heating a reaction solution containing sodium phosphate as a raw material to perform dehydration condensation can be cited. As the heating temperature, for example, it can be set to 150 to 350 °C.

[0059] As a method for synthesizing polyphosphoric acid in vitro using biological molecules such as enzymes, for example, the following method can be cited: Using polyphosphate kinase (PPK), which is an enzyme for synthesizing polyphosphoric acid, as "biological molecules such as enzymes", and using ATP as a raw material, polyphosphoric acid is synthesized by the enzymatic action of PPK. It has been reported that many probiotics such as Lactobacillus rhamnosus GG strain and strains belonging to Lactobacillus brevis possess PPK, and the gene sequence of this enzyme is also publicly available in the DB.

[0060] PPK can be any substance that can synthesize polyphosphoric acid using ATP as a substrate, can be obtained from any strain expressing PPK, or can be purchased as a commercially available PPK. As PPK, for example, it can be PPK from Propionibacterium freudenreichii subsp. shermanii.

[0061] The enzymatic reaction based on PPK is reversible, but in the case where there is a large amount of ADP in the reaction solution compared to ATP, in order for the ADP / ATP ratio to reach equilibrium, the decomposition reaction of polyphosphoric acid will become dominant. Therefore, in order to efficiently synthesize polyphosphoric acid, it is preferable to keep the ADP concentration in the reaction solution low. For example, an ATP continuous regeneration reaction system based on creatine kinase and pyruvate kinase can be coupled. Other conditions such as the composition of the reaction solution, reaction temperature, and reaction time can be appropriately set in a manner optimized for PPK activity and according to the synthesis scale, etc.

[0062] As an example, the reaction conditions are shown below in the case of coupling a continuous ATP regeneration reaction system based on pyruvate kinase when using PPK from Propionibacterium freudenreichii subsp. shermanii to synthesize polyphosphate. Mix 680 μL of 2 mol / L Tris-HCl pH 9.0, 0.1 g of phosphoenolpyruvate, 72 mg of adenosine 5'-triphosphate disodium trihydrate, 160 μL of 1 mol / L phosphate buffer pH 6.0, 60 μL of 2 mol / L magnesium chloride, 1 mL of 2 mol / L acetate buffer pH 6.0, and 2.1 mL of purified water in a 10 mL sample tube, and heat at 40 °C for 30 minutes. After heating, add 2.5 μL of 240 U / mL polyphosphate kinase, and further heat at 40 °C for 5 minutes. Then, 2.5 μL of 1690 U / mL pyruvate kinase can be added, keep it at 40 °C, and set the reaction time to 0.5 to 36 hours. The reaction time can be appropriately set according to the molecular weight and yield of the target polyphosphate. For example, in order to obtain high-molecular-weight polyphosphate with a high yield, it is preferable to set the reaction time to about 20 hours.

[0063] As a method of synthesis using microorganisms, etc., for example, a method of culturing a polyphosphate-producing microorganism under appropriate culture conditions to allow the microorganism to produce polyphosphate can be cited. Examples of polyphosphate-producing microorganisms include Lactobacillus rhamnosus GG strain, Lactobacillus brevis SBC8803 strain, strains belonging to the genus Lactobacillus, strains belonging to the genus Bifidobacterium, strains belonging to the genus Enterococcus, strains belonging to the genus Lactococcus, strains belonging to the genus Pediococcus, strains belonging to the genus Leuconostoc, strains belonging to the genus Streptococcus, strains belonging to the genus Bacteroides, strains belonging to the genus Eubacterium, strains belonging to the genus Clostridium, etc.

[0064] By culturing the microorganism in an appropriate medium and at an appropriate culture temperature condition that can maintain the proliferation of the microorganism used, polyphosphate can be synthesized. The synthesized polyphosphate can be recovered from the culture medium after culturing, or the cultured microorganism can be disrupted to recover the synthesized polyphosphate.

[0065] In the purification process of the synthesized polyphosphoric acid, purification methods commonly used in the art such as size exclusion chromatography, ion exchange chromatography, affinity chromatography, high performance liquid chromatography (HPLC), dialysis, salting out, ammonium sulfate precipitation, precipitation, crystallization, etc. can be appropriately combined and used. The purification method to be used can be appropriately determined according to the method used in the polyphosphoric acid synthesis process, the target purity, the target yield, and the like.

[0066] Preferably, the polyphosphoric acid used in the production of the zinc polyphosphate of the present invention is a linear polyphosphoric acid obtained by an enzymatic synthesis method.

[0067] (2) Zinc polyphosphate

[0068] The "zinc content rate" in the zinc polyphosphate of the present invention is preferably 10 to 60%, more preferably 20 to 55%, and still more preferably 30 to 50%. Regarding the "zinc content rate", in the case of using enzymatically synthesized long-chain polyphosphoric acid as a raw material, it is particularly preferably 32.5 to 42.5%, and in the case of using chemically synthesized short-chain polyphosphoric acid as a raw material, it is particularly preferably 40 to 50%. The zinc polyphosphate within this range can achieve a high intestinal barrier enhancing effect. It should be noted that the so-called "zinc content rate" represents the proportion of the weight % of zinc element in the zinc polyphosphate drug substance excluding excipients and the like.

[0069] The zinc polyphosphate may contain divalent metal salts such as calcium salts and magnesium salts of polyphosphoric acid, and monovalent metal salts such as sodium salts and potassium salts of polyphosphoric acid in part. In this case, the metal content rate other than zinc is in the range of 0 to 30%, preferably in the range of 0 to 20%, more preferably in the range of 0 to 10%, and still more preferably in the range of 0 to 7%. It should be noted that the so-called "metal content rate other than zinc" represents the proportion of the weight % of metal elements other than zinc in the zinc polyphosphate drug substance excluding excipients and the like.

[0070] The Zeta potential of the zinc polyphosphate of the present invention is preferably -20 mV or less, and particularly preferably -30 mV or less. The inventors of the present application have found that the change in the Zeta potential of the divalent metal salt of polyphosphoric acid is related to the change in the intestinal barrier enhancement rate of the divalent metal salt of polyphosphoric acid. Therefore, based on the change in the Zeta potential of the zinc polyphosphate during storage, the intestinal barrier enhancing activity of the zinc polyphosphate can be predicted.

[0071] In the present invention, the zinc polyphosphate is preferably poorly soluble in water, so that it can effectively reach the intestinal mucosa as the action site and be taken up by epithelial cells through endocytosis, thereby expecting a high effect. In addition, the water-insoluble zinc polyphosphate salt can simplify the drug substance manufacturing processes such as recovery, washing, and drying, and is therefore also advantageous in terms of manufacturing cost.

[0072] In the present invention, zinc polyphosphate is preferably "nanoparticles". "Nanoparticles" refer to particles with a particle size in the nanometer range. Most nanoparticles are smooth spherical bodies with relatively consistent particle sizes and also have ideal physical properties in terms of formulation.

[0073] Specifically, the "average particle size" of the zinc polyphosphate of the present invention is preferably 1 nm to 1000 nm, more preferably 20 nm to 500 nm, and the particle size in this range can be formed as a result of the aggregation of fine particles. The average particle size can be determined according to the particle size distribution by known techniques.

[0074] The zinc polyphosphate involved in the present invention is characterized by having a high "intestinal barrier enhancing effect" and also being excellent in terms of storage stability. The intestine has an "intestinal barrier" function of defending against the direct uptake of pathogenic bacteria and harmful substances. The "intestinal barrier enhancing effect" or "intestinal barrier enhancing activity" refers to the effect or activity that enhances this intestinal barrier function. As shown in the examples described later, the "intestinal barrier enhancing effect" and "intestinal barrier enhancing activity" can be evaluated by comparing the intestinal permeability with a control group. For example, in ex vivo experiments, the excised intestine is filled with RPMI culture medium (FUJIFILM Wako Pure Chemical Corporation) supplemented with the test substance and cultured, and then 3 H-mannitol (PerkinElmer), monochloramine solution (prepared by mixing distilled water, NH 4 Cl (FUJIFILM Wako Pure Chemical Corporation), NaOCl (FUJIFILM Wako Pure Chemical Corporation), and RPMI culture medium (FUJIFILM Wako Pure Chemical Corporation)) is added to the culture medium to impose an oxidative stress load, and the amount of 3 H-mannitol leaked into the outside of the intestine is measured and compared with the amount of 3 H-mannitol in the case where the test substance is not added, whereby the "intestinal barrier enhancing effect" and "intestinal barrier enhancing activity" can be evaluated. The "intestinal barrier enhancing activity" can also be evaluated by calculating the following intestinal barrier enhancement rate.

[0075] Intestinal barrier enhancement rate (%) = {1 - (cpm 样品 - cpm 对照组 ) / (cpm NH2Cl - cpm 对照组 )} × 100

[0076] cpm 样品 : Amount of 3 H-mannitol under oxidative stress with polyphosphoric acid added

[0077] cpm NH2Cl : Amount of 3 H-mannitol under oxidative stress (without adding polyphosphoric acid)

[0078] cpm 对照组 : Normal 3 Amount of H-mannitol (without addition of polyphosphoric acid and without oxidative stress load)

[0079] For example, the half-life of the zinc polyphosphate of the present invention for enhancing intestinal barrier activity (effect) under the conditions of 40 °C and 75% humidity is 2 weeks or more, preferably 4 weeks or more, and more preferably 6 months or more. Alternatively, the half-life of the zinc polyphosphate of the present invention for enhancing intestinal barrier activity (effect) under the conditions of 60 °C or lower and 75% humidity is 2 weeks or more, preferably 4 weeks or more, and more preferably 6 months or more.

[0080] The zinc polyphosphate of the present invention can also maintain the above-mentioned intestinal barrier enhancing activity under acidic conditions. For example, the half-life of the zinc polyphosphate of the present invention for enhancing intestinal barrier activity in artificial gastric juice is 120 minutes or more, preferably 360 minutes or more.

[0081] (3) X-ray diffraction pattern

[0082] The zinc polyphosphate of the present invention is characterized in that in the X-ray diffraction pattern, a halo peak is present at the position of 2θ = 4 to 7°. The halo peak refers to a broad peak indicating characteristics different from those of typical amorphous substances. The zinc polyphosphate of the present invention preferably shows a halo signal peak at any one of the positions of 2θ = 30 to 36° and 56 to 62°, and preferably shows halo signal peaks at these two positions.

[0083] 2. Pharmaceutical composition containing zinc polyphosphate as an active ingredient

[0084] The present invention also provides a pharmaceutical composition containing the above-mentioned zinc polyphosphate as an active ingredient. As described above, zinc polyphosphate has intestinal barrier enhancing activity and storage stability. Therefore, the pharmaceutical composition of the present invention is useful as a "pharmaceutical composition for preventing or treating inflammatory bowel disease" or a "pharmaceutical composition for enhancing intestinal barrier", and is also expected to be used as a pharmaceutical composition for treating other diseases.

[0085] "Inflammatory bowel disease (IBD: Inflammatory Bowel Disease)" collectively refers to chronic or remission-relapsing intestinal inflammatory diseases, and generally refers to two diseases, ulcerative colitis (UC: Ulcerative Colitis) and Crohn's disease (CD: Crohn's Disease). Ulcerative colitis is a diffuse non-specific inflammation of unknown cause that forms erosions and ulcers in the mucosa of the large intestine, and mostly recurs and remits. Crohn's disease is a general term for chronic inflammatory diseases of unknown cause that cause chronic inflammation and ulcers in the mucosa of the large intestine and small intestine.

[0086] The zinc polyphosphate of the present invention has both high intestinal barrier function-improving activity. Therefore, it effectively treats damaged digestive tract mucosa, achieves induction remission and maintenance remission of inflammatory bowel disease. The present invention also provides such a "pharmaceutical composition for induction remission / maintenance remission of inflammatory bowel disease".

[0087] The pharmaceutical composition of the present invention inhibits the expression of inflammatory cytokines in the intestine, such as one or more than two inflammatory cytokines selected from IL1β, TNF, IL6, and IL12B. The present invention also provides a "pharmaceutical composition for inhibiting the expression of inflammatory cytokines in the intestine" containing the zinc polyphosphate of the present invention as an active ingredient.

[0088] The pharmaceutical composition of the present invention may contain zinc polyphosphate as an active ingredient, as well as a pharmaceutically acceptable carrier and additive. Examples of such a carrier and additive include, but are not limited to, excipients, binders, lubricants, solvents, disintegrants, solubilizers, suspending agents, emulsifiers, isotonic agents, stabilizers, preservatives, antioxidants, flavoring agents, coloring agents, buffers, flowability promoters, etc. Other commonly used carriers and additives can be appropriately used.

[0089] Specifically, examples of excipients include organic excipients such as sugars like lactose, glucose, D-mannitol, starches, and celluloses like crystalline cellulose, and inorganic excipients such as calcium carbonate and kaolin.

[0090] Examples of binders include pregelatinized starch, gelatin, gum arabic, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, crystalline cellulose, D-mannitol, trehalose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, polyvinyl alcohol, etc.

[0091] Examples of lubricants include fatty acid salts such as stearic acid and stearates, talc, silicate classes, etc.

[0092] Examples of solvents include purified water, physiological saline, phosphate buffer solution, etc.

[0093] Examples of disintegrants include low-substituted hydroxypropylcellulose, chemically modified cellulose, starches, etc.

[0094] Examples of solubilizers include polyethylene glycol, propylene glycol, trehalose, benzyl benzoate, ethanol, sodium carbonate, sodium citrate, sodium salicylate, sodium acetate, etc.

[0095] Examples of suspending agents or emulsifiers include sodium lauryl sulfate, gum arabic, gelatin, lecithin, glycerol monostearate, polyvinyl alcohol, polyvinylpyrrolidone, celluloses such as sodium carboxymethylcellulose, polysorbates, polyoxyethylene hydrogenated castor oil, etc.

[0096] Examples of the isotonic agent include sodium chloride, potassium chloride, sugars, glycerol, urea, and the like.

[0097] Examples of the stabilizer include polyethylene glycol, sodium dextran sulfate, other amino acids, and magnesium carbonate which also serves as an acidity regulator.

[0098] Examples of the preservative include parabens, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid, and the like.

[0099] Examples of the antioxidant include ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite as water-soluble antioxidants, ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, α-tocopherol as fat-soluble antioxidants, and citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid as metal chelators.

[0100] Examples of the flavoring and odor-masking agent include sweeteners, fragrances, etc. commonly used in the pharmaceutical field, and examples of the coloring agent include coloring agents commonly used in the pharmaceutical field.

[0101] The pharmaceutical composition of the present invention can be safely administered orally or parenterally (e.g., intraorally, esophagus, stomach, small intestine, large intestine, rectum, etc.) by formulating it into the following pharmaceutical preparations: tablets (including sugar-coated tablets, film-coated tablets, sublingual tablets, intraoral disintegrating tablets), powders, granules, capsules (including soft capsules, microcapsules), liquids, lozenges, syrups, emulsions, suspensions, injections (e.g., subcutaneous injections, intramuscular injections, intraperitoneal injections, etc.), external preparations (e.g., nasal administration preparations, transdermal preparations, ointments), suppositories (e.g., rectal suppositories, vaginal suppositories), foams (enemas), pellets, nasal agents, pulmonary agents (inhalants), etc. Preferably, the pharmaceutical composition of the present invention includes oral administration, suppository administration, and rectal administration, and oral administration is particularly preferred.

[0102] The pharmaceutical composition of the present invention can be a release-controlled preparation such as an immediate-release preparation or a sustained-release preparation. In addition, when formulating an oral preparation, coating can be performed as needed for the purpose of taste masking, enteric solubility, or sustained release. Examples of the coating base used in the coating include a sugar coating base, a water-soluble film coating base, an enteric film coating base, and a sustained-release film coating base.

[0103] The pharmaceutical composition of the present invention can be administered to humans or non-human mammals. The dosage and administration method are not particularly limited and can be appropriately determined according to the state, age, etc. of the administered individual.

[0104] The dosage of the pharmaceutical composition of the present invention is appropriately determined according to its purpose of use, administration route, etc. In the case of administration to humans, for example, for daily administration, as zinc polyphosphate, the dosage can be selected in the range of 0.01 mg / kg to 4 mg / kg, preferably 0.015 mg / kg to 2 mg / kg, more preferably 0.03 mg / kg to 1 mg / kg. Alternatively, for example, it can be administered once to several times a day in the range of 0.6 to 240 mg / day per patient, preferably 0.9 to 120 mg / day, more preferably 1.8 to 60 mg / day.

[0105] The zinc polyphosphate of the present invention is the main body of intestinal barrier enhancing activity and is composed of zinc polyphosphate with excellent storage stability. Therefore, compared with natural polyphosphoric acid, an unstable complex mixture such as sodium polyphosphate, high drug efficacy and excellent storage stability can be expected.

[0106] As long as the object of the present invention is not impaired, the pharmaceutical composition of the present invention can be used in combination with other drugs. Examples of drugs that can be used in combination with the pharmaceutical composition of the present invention include drugs commonly used in the treatment of inflammatory bowel diseases such as ulcerative colitis and Crohn's disease, for example, 5-aminosalicylic acid (5-ASA) preparations such as mesalazine and sulfasalazine; steroid preparations such as prednisolone and methylprednisolone; anti-TNFα preparations such as infliximab and adalimumab; anti-integrin antibody preparations such as vedolizumab; tofacitinib as a JAK inhibitor; thiopurine preparations such as mercaptopurine and azathioprine; immunosuppressive agents such as cyclosporine and tacrolimus, but are not limited to these. The administration time of the drug used in combination with the pharmaceutical composition of the present invention is not limited and can be administered simultaneously or at intervals.

[0107] The zinc polyphosphate of the present invention can be safely ingested orally and can therefore also be used as an ingredient in specific health foods, special-purpose foods, nutritional supplements, health foods, functional foods, foods for patients, etc.

[0108] 3. Method for manufacturing zinc polyphosphate

[0109] The zinc polyphosphate of the present invention can be obtained by reacting polyphosphoric acid with zinc chloride under alkaline conditions. For example, the pH of the polyphosphoric acid solution is adjusted to 7.5 or more, preferably 8 to 11, zinc chloride is added, and the obtained zinc polyphosphate is recovered. Alternatively, under non-alkaline conditions, for example, at a pH of 4 to 7 or less, zinc chloride is added to the polyphosphoric acid solution, and then the pH is adjusted to 7.5 or more, preferably 8 to 11, and the obtained zinc polyphosphate is recovered.

[0110] For example, the zinc polyphosphate of the present invention can be manufactured by the following steps:

[0111] (1) A step of adding zinc ions to a polyphosphoric acid solution under alkaline conditions, preferably at a pH of 8 to 11, more preferably at a pH of 9 to 10.5, and further preferably at a pH of 9.5 to 10.0; and

[0112] (2) A step of recovering the solid obtained in step (1) as zinc polyphosphate.

[0113] Alternatively, the zinc polyphosphate of the present invention can be produced by the following steps:

[0114] (1) A step of adding zinc ions to a polyphosphoric acid solution under non-alkaline conditions;

[0115] (2) Subsequently, adjusting the aforementioned solution to alkaline conditions, preferably at a pH of 8 to 11, more preferably at a pH of 9 to 10.5, and further preferably at a pH of 9.5 to 10.0, and allowing it to react (stirring or standing) for more than 2 hours, preferably 3 hours or more, and more preferably 4 hours or more; and

[0116] (3) A step of recovering the solid obtained in step (2) as zinc polyphosphate.

[0117] The polyphosphate used can be obtained by the method described in “(1) Polyphosphoric acid” of 1. As described above, the obtained polyphosphoric acid is short-chainized compared to the polyphosphoric acid (such as sodium polyphosphate) used as the starting material.

[0118] The zinc polyphosphate of the present invention has an effect of enhancing the intestinal barrier function, inhibits the expression of inflammatory cytokines in the intestine (for example, 1 or more than 2 selected from IL1β, TNF, IL6, and IL12B), and administering to a subject in need can treat or prevent inflammatory bowel disease and the like.

[0119] Examples

[0120] Hereinafter, the present invention will be specifically described by way of examples, but the present invention is not limited to these examples.

[0121] Example 1: Preparation of Zinc Polyphosphate (PPA-Zn)

[0122] 1. Preparation of Sodium Polyphosphate (PPA-Na) (enzymatic method)

[0123] Mix 680 μL of 2 mol / L Tris-HCl pH 9.0, 0.1 g of phosphoenolpyruvate, 72 mg of adenosine 5'-triphosphate disodium trihydrate, 160 μL of 1 mol / L phosphate buffer pH 6.0, 60 μL of 2 mol / L magnesium chloride, 1 mL of 2 mol / L acetate buffer pH 6.0, and 2.1 mL of purified water in a 10 mL sample tube, and heat at 40 °C for 30 minutes. After heating, add 2.5 μL of 240 U / mL polyphosphate kinase, and further heat at 40 °C for 5 minutes. Then, add 2.5 μL of 1690 U / mL pyruvate kinase and react at 40 °C for 20 hours. Add 1.7 mL of 2 mol / L sodium chloride to the reaction solution, stir at 10 °C for 10 minutes, and let stand. Remove the supernatant, add 400 μL of purified water to the precipitate, dissolve it, add 400 μL of 3 mol / L sodium chloride to precipitate the precipitate. Further add 400 μL of purified water to the precipitate, dissolve it, add 400 μL of 3 mol / L sodium chloride to precipitate the precipitate. After removing the supernatant, lyophilize the precipitate to obtain 92 mg of long-chain sodium polyphosphate (PPA-Na) with a chain length of 600 mer or more.

[0124] 2. Preparation of various zinc polyphosphates

[0125] 2.1 Preparation of zinc polyphosphate Test15-12

[0126] Add 50 g of water to 1 g of the sodium polyphosphate (PPA-Na) prepared in 1. to dissolve it. Add 1 mol / L aqueous sodium hydroxide solution, and when adjusting the pH to 10.0, add 5 mL of 1 mol / L zinc chloride solution dropwise over 30 minutes. During the dropwise addition, add aqueous sodium hydroxide solution as appropriate to maintain the pH at 9.8 - 10.1. After the dropwise addition, stir at room temperature for 4 hours, then let stand at a temperature of 10 °C or below for 16 hours, and recover the precipitate as the precipitate by centrifugation (8000×g, 4 °C, 10 minutes). Further wash the precipitate with aqueous ethanol (33 v / v% ethanol), recover the precipitate by centrifugation again, and perform freeze-drying to obtain 836 mg of zinc polyphosphate Test15-12 as a white powder.

[0127] 2.2 Preparation of zinc polyphosphate Test15-6

[0128] To 10 g of the sodium polyphosphate (PPA-Na) prepared in 1., add 500 g of water to dissolve it. Add 1 mol / L sodium hydroxide aqueous solution. When adjusting the pH to 10.0, dropwise add 50 mL of 1 mol / L zinc chloride solution over 30 minutes. During the dropwise addition, add the sodium hydroxide aqueous solution in a timely manner to maintain the pH at 9.8 - 10.1. After the dropwise addition, stir at room temperature for 4 hours, then let it stand at a temperature below 10 °C for 16 hours. Recover the precipitate by centrifugation (12000×g, 4 °C, 10 minutes). Further wash the precipitate with aqueous ethanol (33 v / v% ethanol), recover the precipitate by centrifugation again, and perform freeze-drying to obtain 7.8 g of zinc polyphosphate Test15-6 as a white powder.

[0129] 2.3 Preparation of zinc polyphosphate Test15-2

[0130] Through the same operation as in 2.1, 812 mg of zinc polyphosphate Test15-2 was prepared from 1 g of sodium polyphosphate. However, the centrifugation conditions for precipitate recovery were set to 15000×g, 4 °C, 10 minutes.

[0131] 2.4 Preparation of zinc polyphosphate Test40-1

[0132] To 1 g of the sodium polyphosphate (PPA-Na) prepared in 1., add 50 g of water to dissolve it (pH 5.3). Dropwise add 5 mL of 1 mol / L zinc chloride solution to it over 30 minutes. After the dropwise addition, add 7.3 mL of 1 mol / L sodium hydroxide aqueous solution to adjust the pH to 10.0, and further stir for 4 hours. Let it stand at a temperature below 10 °C for more than 16 hours, and recover the precipitate using a centrifuge. Further wash the recovered precipitate with aqueous ethanol (33 w / w% ethanol), recover the precipitate by centrifugation (15000×g, 4 °C, 10 minutes) again, and perform freeze-drying to obtain 703 mg of zinc polyphosphate Test40-1.

[0133] 2.5 Preparation of zinc polyphosphate Test40-6-1

[0134] To 1 g of the sodium polyphosphate (PPA-Na) prepared in 1., add 50 g of water to dissolve it (pH 5.4). Dropwise add 5 mL of 1 mol / L zinc chloride solution to it over 30 minutes. After the dropwise addition, dropwise add 7.3 mL of 1 mol / L sodium hydroxide aqueous solution to adjust the pH to 9.9. Stir at room temperature for 2 hours, and recover the supernatant and the precipitate by centrifugation (4000×g, 4 °C, 10 minutes) respectively. Wash the precipitate with water and ethanol in sequence, and perform freeze-drying to obtain 403 mg of zinc polyphosphate Test40-6-1.

[0135] 2.6 Preparation of zinc polyphosphate Test40-6-2

[0136] The supernatant recovered above was allowed to stand at a temperature below 10°C for 16 hours, and the precipitate thus formed was recovered by centrifugation (15,000×g, 4°C, 10 minutes) as a precipitate. The precipitate was further washed with water, and the precipitate was recovered again by centrifugation and freeze-dried to obtain 48 mg of zinc polyphosphate Test40-6-2.

[0137] 3. Analysis of zinc polyphosphate

[0138] 3.1 Based on zinc polyphosphate 31 Confirmation of degree of polymerization by 31P NMR measurement

[0139] Dissolve about 10 mg of zinc polyphosphate in about 0.7 mL of about 190 mmol / L citric acid-sodium buffer (D 2 2O) at pH 5.5 as an analytical sample for measurement 31 31P NMR.

[0140] (i) Use the equipment Bruker BioSpin 400 MHz

[0141] (ii) For the analysis and resolution conditions, assign the signal at -5 ppm to the terminal phosphate group, assign the signals at -15 to -25 ppm to the internal phosphate groups, and calculate the total integrated value of the phosphate groups assuming the integrated value of the terminal phosphate group is set as the reference 2. Since zinc polyphosphate is prepared from linear PPA-Na in an aqueous solution, cyclic polyphosphates are scarce, so it is considered that the calculated total integrated value of the phosphate groups is approximate to the degree of polymerization.

[0142] The NMR spectra are shown in Fig. 1 (A - F), and the calculated degrees of polymerization (total integrated values of phosphate groups) are shown in Table 1. The degrees of polymerization of various zinc polyphosphates are calculated to be in the range of shorter chains of 6.6 - 11.9, and are significantly shorter-chained compared to the sodium polyphosphate (600 mer) used.

[0143] 3.2 X-ray diffraction of zinc polyphosphate

[0144] Press 10 - 50 mg of zinc polyphosphate onto a glass plate and set it in a crystal diffraction apparatus for measurement.

[0145] (i) Use the equipment MiniFlexII manufactured by Rigaku Corporation

[0146] (ii) For the analysis conditions, the cathode: Cu, tube voltage: 30 kV, tube current: 15 mA, monochromatization: Ni filter, sampling width: 0.020°, scanning speed: 10°C / min, wavelength: Measurement of diffraction angle range (2θ): 2 to 60°, divergence slit: 1.25°, scattering slit: 8.0 mm, receiving slit: open

[0147] The results of X-ray diffraction are shown in Fig. 2 (A - F). In Test15-2, Test15-6, Test15-12, Test40-1, and Test40-6-2, signal peaks of halos were observed at positions where 2θ = 4 - 7°, 30 - 36°, and 56 - 62°. On the other hand, in Test40-6-1, signal peaks of halos were observed at positions where 2θ = 30 - 36° and 56 - 62°, but no peak was observed at 2θ = 4 - 7°.

[0148] 3.3 Measurement of pH, Zeta potential, and particle size of zinc polyphosphate

[0149] Suspend 5 mg of zinc polyphosphate in 1 mL of purified water. Let the suspension stand still, and measure the pH of the supernatant. After measuring the pH, ultrasonically disperse the suspension and dilute it with purified water to 2 mg / mL. Use this as the sample for analysis, and measure the Zeta potential and particle size.

[0150] (i) Use equipment Malvern, ZEN3600

[0151] (ii) Analysis conditions: Dispersant: water, refractive index of dispersant RI: 1.330, viscosity (cp): 0.8872, dielectric constant of dispersant: 78.5, temperature: 25 °C

[0152] The results of pH and Zeta potential measurements are summarized together with the degree of polymerization in Table 1.

[0153] [Table 1]

[0154] Zeta potential, pH, and NMR results

[0155]

[0156] The results of particle size measurement are shown in Fig. 3. The average particle size of zinc polyphosphate was mostly detected at around several hundred nm, but sometimes peaks and shoulders were also detected at around smaller several tens of nm, showing a bimodal distribution pattern. It is considered that fine particles sometimes aggregate to form large particles.

[0157] Test Example 1: Intestinal barrier enhancing effect of zinc polyphosphate Test15-12

[0158] Ex vivo intestinal loop study

[0159] The intestines of C57BL / 6 mice were excised and cut into four equal lengths. The ends were ligated with silk sutures (suture number: 4-0, Alfresa Corporation), and the intestines were filled with RPMI culture medium (FUJIFILM Wako Pure Chemical Corporation) supplemented with 1 μg / mL of zinc polyphosphate Test15-12. After culturing in the RPMI culture medium at 37 °C for 2 hours, one of the ligated parts was incised, and filled with a monochloramine solution supplemented with 1 μCi / mL of 3 3H-mannitol (PerkinElmer), and ligated again, thereby imposing an oxidative stress load. It should be noted that the monochloramine solution was prepared by mixing 920 μl of distilled water, 1000 μl of 40 mM NH 4 Cl (FUJIFILM Wako Pure Chemical Corporation), 80 μl of 5.0% NaOCl (FUJIFILM Wako Pure Chemical Corporation), and 8000 μl of RPMI culture medium. At 15 minutes and 30 minutes after filling, the amount of 3 3H-mannitol leaked outside the intestine was measured using a liquid scintillator (PerkinElmer, Liquid Scintillation Analyzer Tri-Carb 4910TR). Similarly, the amount of 3 3H-mannitol under oxidative stress load without the addition of polyphosphate, and the amount of 3 3H-mannitol in the state without the addition of polyphosphate and without oxidative stress load (control group) ( Figure 4 ) were measured.

[0160] As Figure 4 shown, by adding zinc polyphosphate Test15-12, the amount of 3 3H-mannitol leaked outside the intestine under oxidative stress load was at the same level as that of the control group, and zinc polyphosphate Test15-12 showed a similarly high intestinal barrier enhancing effect.

[0161] Test Example 2: Intestinal barrier enhancing effects of zinc polyphosphate Test40-6-1 and Test40-6-2

[0162] Everted intestinal loop study

[0163] According to the method described in Test Example 1, the intestinal barrier enhancing effects of zinc polyphosphate Test40-6-1 and Test40-6-2 were evaluated ( Figure 5 ).

[0164] As Figure 5 shown, zinc polyphosphate Test40-6-1 did not exert an intestinal barrier enhancing effect. On the other hand, Test40-6-2 showed a high intestinal barrier enhancing effect.

[0165] Test Example 3: Stability of Zinc Polyphosphate

[0166] Comparison of Stability of Test15-6 under Conditions of Storage at 40°C + Humidity 75% and Storage at 60°C + Humidity 75% (In Vitro)

[0167] 1. Accelerated Stability Test

[0168] Weigh 20 mg each of zinc polyphosphate Test15-6 in an Eppendorf tube and store it in an open state in a small environmental test chamber (ESPEC Corporation, set at 40°C 75% RH) and a constant temperature and humidity chamber (Yamato Scientific Co., Ltd., set at 60°C 75% RH) for 24 weeks.

[0169] 2. In Vitro Intestinal Loop Study (After Storage at 40°C 75% RH and 60°C 75% RH)

[0170] According to the method described in Test Example 1, evaluate the intestinal barrier enhancing effect of Test15-6 after storage at 40°C 75% RH and 60°C 75% RH ( Figure 6 ).

[0171] As Figure 6 shown, zinc polyphosphate Test15-6 maintained its intestinal barrier enhancing effect for more than 24 weeks under these conditions.

[0172] Test Example 4: Elemental Analysis of Various Zinc Polyphosphates

[0173] Weigh 20 mg each of various zinc polyphosphates (Test15-2, Test15-6, Test15-12, Test40-1, Test40-6-1, Test40-6-2) using a precision electronic balance and conduct elemental analysis at Yebisai Co., Ltd. (Sapporo). The moisture content was studied using the drying loss method (105°C, drying for 2 hours). The measurement test solution was obtained as follows: Add nitric acid and hydrogen peroxide to the sample, heat and decompose it using a microwave sample pretreatment device, and make up the volume to 20 ml with distilled water. Regarding the sodium, calcium, and magnesium contents, they were studied using flame atomic absorption spectrometry with an atomic absorption spectrophotometer (PerkinElmer, Analist 200), the zinc content rate was studied using ICP mass spectrometry with an ICP mass spectrometer (Agilent, Agilent 7500cx), and the phosphorus content rate was studied using molybdenum blue spectrophotometry with a spectrophotometer (Shimadzu Corporation, UV-1800) (Table 2).

[0174] [Table 2]

[0175] Elemental Analysis Results

[0176] Sodium (%) Calcium (%) Magnesium (%) Zinc (%) Phosphorus (%) Moisture content (%) PPA-Na 17.80 0.01 1.89 0.001 30.6 5.0 Test15-2 6.27 <0.005 0.737 36.9 14.9 7.9 Test15-6 4.56 <0.005 0.837 39.6 15.4 9.6 Test15-12 5.78 <0.005 0.832 34.6 15.2 3.9 Test 40-1 4.87 <0.005 0.835 37.3 13.8 1.5 Test 40-6-1 2.85 <0.005 0.770 40.4 12.2 3.8 Test 40-6-2 2.55 <0.005 0.677 42.6 13.5 9.4

[0177] Test Example 5: Intestinal barrier enhancement effect of short-chain Test15-12

[0178] Test15-12 prepared in 2.1 was stored in a sealed container in a thermostatic chamber at 40°C for 13 days to obtain short-chain Test15-12. 31 When the degree of polymerization was confirmed by P NMR measurement, the degree of polymerization was shortened to 2.0 ( Figure 1G When the X-ray diffraction of zinc polyphosphate was measured according to the method described in 3.2, the short-chain Test15-12 continued to have a halo peak at the position of 2θ=4 to 7° in the X-ray diffraction spectrum ( Figure 2G ). When the pH and Zeta potential of the shortened Test15-12 were measured according to the method described in 3.3, the Zeta potential was -30.8 mV and the pH was 8.3.

[0179] The intestinal barrier enhancing effect of zinc polyphosphate Test15-12 and short-chain Test15-12 was studied according to the method described in Experimental Example 1. Figure 7 As shown, by adding short-chain Test15-12, intestinal leakage during oxidative stress was reduced. 3 The amount of H-mannitol was suppressed to the same level as the control group, and the short-chain Test15-12 showed a high intestinal barrier strengthening effect equivalent to that of Test15-12 ( Figure 7 ). Based on this result, it was confirmed that zinc polyphosphate has a high intestinal barrier strengthening ability even in the form of a short chain.

[0180] Industrial Applicability

[0181] Zinc polyphosphate showing a halo peak at 2θ=4-7° in X-ray diffraction has an intestinal barrier-enhancing effect and is also excellent in storage stability. It is useful in the treatment and prevention of inflammatory bowel diseases represented by ulcerative colitis and Crohn's disease, and is also expected to be a pharmaceutical composition for the treatment of other diseases.

[0182] All publications, patents and patent applications cited in this specification are incorporated herein by reference.

Claims

1. Zinc polyphosphate, characterized in that, in X-ray diffraction, it shows a halo peak at 2θ = 4 - 7°.

2. The zinc polyphosphate according to claim 1, having a Zeta potential of -20 mV or less.

3. The zinc polyphosphate according to claim 1, wherein, the zinc content is 10 - 60%.

4. The zinc polyphosphate according to claim 1, wherein, the average chain length of the polyphosphate moiety of the zinc polyphosphate is 1.1 phosphate units or more and less than 100 phosphate units.

5. The zinc polyphosphate according to claim 1, wherein, the average particle size of the zinc polyphosphate is 1 - 1000 nm.

6. The zinc polyphosphate according to claim 1, wherein, the half-life of the intestinal barrier enhancing activity is 2 weeks or more under the conditions of 60°C or lower and 75% humidity.

7. A pharmaceutical composition comprising the zinc polyphosphate according to any one of claims 1 - 6 as an active ingredient.

8. A method, which is a method for manufacturing zinc polyphosphate, comprising: (1) A step of adding zinc ions to a polyphosphoric acid solution under alkaline conditions, preferably under conditions of pH 8 - 11; and (2) A step of recovering the solid obtained in step (1) as zinc polyphosphate.

9. A method, which is a method for manufacturing zinc polyphosphate, comprising: (1) A step of adding zinc ions to a polyphosphoric acid solution under non-alkaline conditions; (2) Subsequently, adjusting the solution to alkaline conditions, preferably pH 8 - 11, and reacting for more than 2 hours; and (3) A step of recovering the solid obtained in step (2) as zinc polyphosphate.

10. The method according to claim 8 or 9, wherein, the obtained zinc polyphosphate shows a halo peak at 2θ = 4 - 7° in X-ray diffraction.

11. The zinc polyphosphate according to claim 1, which is obtained by the method according to claim 8 or 9.

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