Composition for preventing or treating endometriosis

Abstract

The present invention provides a composition, in particular, a pharmaceutical composition and a food and drink composition, for preventing or treating endometriosis, the composition containing polyphosphoric acid or a salt thereof. The salt of the polyphosphoric acid may be a metal salt of polyphosphoric acid, may contain, as a constituent metal thereof, at least one among sodium, potassium, magnesium, calcium, and zinc, and is preferably calcium polyphosphate or zinc polyphosphate. According to the present invention, the prevention or treatment of endometriosis based on a novel action mechanism, which is highly safe to a living body, is possible.

Description

Composition for preventing or treating endometriosis

[0001] The present invention relates to a composition for preventing or treating endometriosis, which contains polyphosphoric acid or a salt thereof.

[0002] Endometriosis is a disease in which endometrial tissue (ectopic endometrioid tissue, also known as endometriotic tissue) appears in areas other than the uterine cavity or myometrium, and not only does it adhere to surrounding tissues, causing pelvic pain, menstrual pain, pain during intercourse, pain with bowel movements, etc., but also leads to infertility in approximately half of patients. Endometriosis often develops in women in their 20s and 30s, with the peak incidence reported being between 30 and 34 years old.

[0003] The standard treatments for endometriosis are pain relief with analgesics, hormone therapy to suppress ovarian function and inhibit the growth and activity of endometriotic tissue, and surgical removal. Drug therapy and surgical procedures that do not involve total hysterectomy are limited to symptomatic treatments aimed at controlling the disease, and many patients with drug therapy experience a recurrence of endometriosis after stopping the medication, and approximately half of patients who undergo conservative surgical procedures to preserve fertility also experience a recurrence.

[0004] Furthermore, hormone therapy uses hormones such as combined oral contraceptives, progestins such as dienogest, gonadotropin-releasing hormone (GnRH) agonists, GnRH antagonists, and androgens. However, these hormones have side effects such as thrombosis and weight gain, and also make it difficult to become pregnant while taking them.

[0005] Polyphosphate is known to be utilized as an energy source by microorganisms and to have various physiological activities, such as being involved in the coagulation reaction of human blood. The present inventors have identified long-chain polyphosphate derived from malt-derived lactic acid bacteria as a molecule that strengthens the intestinal barrier function. They have demonstrated that long-chain polyphosphate improves the deterioration of intestinal barrier function and intestinal injury caused by dextran sulfate sodium treatment (Patent Document 1), and further demonstrated that long-chain polyphosphate promotes mucosal healing in patients with refractory ulcerative colitis (Non-Patent Document 1). They have also reported that calcium salts of polyphosphate act on damaged gastrointestinal mucosa in inflammatory bowel disease and exert a platelet aggregation effect specific to the damaged mucosa (Patent Document 2), and that zinc polyphosphate, which exhibits a characteristic X-ray diffraction peak, has an excellent intestinal barrier-enhancing effect (Patent Documents 3 and 4).

[0006] Polyphosphate is known to have wound healing and anti-inflammatory effects, and there are hopes for the development of wound dressings and dental materials using amorphous or nanoparticles composed of calcium polyphosphate salts (Patent Document 5).

[0007] Polyphosphate is also known as a blood coagulation-inducing factor. When polyphosphate is released from blood platelets, it activates factor XII protease in the blood, inducing a coagulation reaction. Natural polyphosphate is thought to exist in various salts, such as calcium salt, magnesium salt, sodium salt, and potassium salt, and amorphous nanoparticles are also known to exist (Non-Patent Documents 2 and 3). Donovan et al. prepared nanoparticles from polyphosphates of different chain lengths and reported on the size of polyphosphates and their effect on blood coagulation (Non-Patent Document 4).

[0008] WO2011 / 125619WO2021 / 141066WO2024 / 090537WO2024 / 090538WO2016 / 079006

[0009] 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.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).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. Donovan et al., “Size-controlled synthesis of granular polyphosphate nanoparticles at physiological salt concentrations for blood clotting.” Biomacromolecules. 2014 Nov 10;15(11):3976-84.

[0010] The present invention provides a new means for preventing or treating endometriosis that is highly safe for the living body.

[0011] The present inventors have found that salts of polyphosphate are useful for the prevention and treatment of endometriosis, particularly for inhibiting the proliferation of endometriotic tissue.

[0012] The present disclosure provides the following: Item 1. A pharmaceutical composition for preventing or treating endometriosis, comprising polyphosphoric acid or a salt thereof. Item 2. The pharmaceutical composition according to Item 1, wherein the polyphosphoric acid or a salt thereof is poorly soluble or insoluble in water. Item 3. The pharmaceutical composition according to Item 1 or 2, wherein the polyphosphoric acid or a salt thereof has an average particle size of 1 to 3,000 nm. Item 4. The pharmaceutical composition according to any one of Items 1 to 3, wherein the polyphosphoric acid or a salt thereof has a zeta potential of 0 mV or less. Item 5. The pharmaceutical composition according to any one of Items 1 to 4, comprising linear polyphosphoric acid or a salt thereof. Item 6. Item 6. The pharmaceutical composition according to any one of Items 1 to 5, wherein the average chain length of the polyphosphate or a salt thereof is at least 1.1 phosphate units, at least 1.2 phosphate units, at least 1.3 phosphate units, at least 1.4 phosphate units, at least 1.5 phosphate units, at least 1.6 phosphate units, at least 1.7 phosphate units, at least 1.8 phosphate units, at least 1.9 phosphate units, at least 2 phosphate units, at least 2.5 phosphate units, at least 3 phosphate units, at least 4 phosphate units, at least 5 phosphate units, at least 6 phosphate units, at least 7 phosphate units, at least 8 phosphate units, at least 9 phosphate units, or at least 10 phosphate units. Item 7. The pharmaceutical composition according to any one of Items 1 to 6, wherein the average chain length of the polyphosphoric acid or a salt thereof is 10,000 phosphate units or less, 1,000 phosphate units or less, 900 phosphate units or less, 800 phosphate units or less, 700 phosphate units or less, 600 phosphate units or less, 500 phosphate units or less, 400 phosphate units or less, 300 phosphate units or less, 200 phosphate units or less, 100 phosphate units or less, 90 phosphate units or less, 80 phosphate units or less, 70 phosphate units or less, 60 phosphate units or less, 50 phosphate units or less, 40 phosphate units or less, 30 phosphate units or less, 20 phosphate units or less, or 15 phosphate units or less. Item 8. The pharmaceutical composition according to any one of Items 1 to 5, wherein the average chain length of the polyphosphoric acid or a salt thereof is 1.1 phosphate units or more, 1.2 phosphate units or more, 1.3 phosphate units or more, or 1.4 phosphate units or more. Item 9. The pharmaceutical composition according to any one of Items 1 to 5 and 8, wherein the average chain length of the polyphosphate or a salt thereof is 10 phosphate units or less, 5 phosphate units or less, 4 phosphate units or less, 3.5 phosphate units or less, 3 phosphate units or less, 2.5 phosphate units or less, 2 phosphate units or less, or 1.5 phosphate units or less.Item 10. The pharmaceutical composition according to any one of Items 1 to 5, wherein the average chain length of the polyphosphoric acid or a salt thereof is at least 1.5 phosphate units, at least 1.6 phosphate units, at least 1.7 phosphate units, at least 1.8 phosphate units, at least 1.9 phosphate units, at least 2 phosphate units, or at least 2.5 phosphate units. Item 11. The pharmaceutical composition according to any one of Items 1 to 5 and 10, wherein the average chain length of the polyphosphoric acid or a salt thereof is at most 15 phosphate units, at most 10 phosphate units, at most 5 phosphate units, at most 4 phosphate units, or at most 3.5 phosphate units. Item 12. The pharmaceutical composition according to any one of Items 1 to 11, which contains a metal salt of polyphosphate, wherein the metal salt of polyphosphate contains at least one of sodium, potassium, magnesium, calcium, and zinc as its constituent metal. Item 13. The pharmaceutical composition according to any one of Items 1 to 12, which contains calcium polyphosphate. Item 14. The pharmaceutical composition according to any one of Items 1 to 12, which contains zinc polyphosphate. Item 15. Item 14. The pharmaceutical composition according to Item 14, wherein the zinc polyphosphate exhibits a broad peak at 2θ=4 to 7° in an X-ray diffraction chart. Item 16. The pharmaceutical composition according to Item 14 or 15, wherein the zinc polyphosphate has a zinc content of 10 to 60%, preferably 20 to 55%, more preferably 30 to 50%. Item 17. The pharmaceutical composition according to any one of Items 1 to 12, which contains sodium polyphosphate. Item 18. The pharmaceutical composition according to any one of Items 1 to 17, for inhibiting the proliferation of endometriotic tissue. Item 19. The pharmaceutical composition according to any one of Items 1 to 18, for use in a subject in whom preservation of fertility is desired. Item 20. The pharmaceutical composition according to any one of Items 1 to 19, which is for oral administration. Item 21. A food or drink composition containing polyphosphoric acid or a salt thereof, for use in a subject in need of prevention or treatment of endometriosis. Item 22. Item 22. The food or drink composition according to Item 21, which is a food for specified health uses, a food for special dietary uses, a nutritional supplement, a supplement, a health food, a food with functional claims, or a food for medical use. Item 23. The food or drink composition according to Item 21 or 22, which contains a culture supernatant or cells of Lactobacillus brevis SBC8803 (accession number: FERM BP-10632).Item 24. The food or beverage composition according to any one of Items 21 to 23, which contains heat-killed cells of Lactobacillus brevis SBC8803 (accession number: FERM BP-10632).

[0013] According to the present invention, it is possible to prevent or treat endometriosis based on a new mechanism of action.

[0014] Figure 1 shows an X-ray diffraction chart of zinc polyphosphate. Figure 2A shows the protocol for an oral administration test to evaluate the therapeutic effects of zinc polyphosphate and Lactobacillus brevis SBC8803 cells using endometriosis model mice. Figure 2B shows the results of the oral administration test (weight change of endometriotic tissue) whose protocol is shown in Figure 2A. Figure 3A shows the protocol for an oral administration test to evaluate the preventive effects of zinc polyphosphate and Lactobacillus brevis SBC8803 cells using endometriosis model mice. Figure 3B shows the results of the oral administration test (weight change of endometriotic tissue) whose protocol is shown in Figure 3A. Figure 4A shows the protocol for an oral administration test to evaluate the therapeutic effects of zinc polyphosphate, calcium polyphosphate, and Lactobacillus brevis SBC8803 cells using endometriosis model mice. Figure 4B shows the results of the oral administration test (weight change of endometriotic tissue) whose protocol is shown in Figure 4A. Figure 5A shows the protocol for an oral administration study of zinc polyphosphate on fertility, and Figure 5B shows the results (pregnancy rate, number of fetuses per implantation) of the oral administration study whose protocol is shown in Figure 5A.

[0015] The following description may be based on representative embodiments or specific examples, but the present invention is not limited to such embodiments or specific examples. In this specification, numerical ranges expressed using "to" or "-" mean ranges that include the numerical values ​​at both ends as upper and lower limits, unless otherwise specified. The upper and lower limits of each numerical range exemplified in this specification can be combined in any way.

[0016] Polyphosphoric Acid and Its Salts In the present disclosure, the term "polyphosphoric acid" refers to a substance containing multiple phosphate units (represented by the formula -[P(=O)(O-)-O]-) as building blocks.

[0017] In the present disclosure, the term "salt of polyphosphate" refers to a substance containing multiple phosphate units and cations as constituent units. In the salt of polyphosphate, the cation is bonded to an oxygen anion. The salt of polyphosphate may contain one or more types of cations. Examples of cations contained in the salt of polyphosphate include metal ions (e.g., monovalent metal ions such as sodium and potassium, and divalent metal ions such as calcium, zinc, and magnesium) and organic ions (e.g., ammonium ions).

[0018] The polyphosphate salt used in the present disclosure may be any pharmaceutically acceptable salt, and is preferably a metal salt containing a metal ion as a cation. The polyphosphate metal salt preferably contains at least one of sodium, potassium, magnesium, calcium, and zinc as its constituent metal, more preferably at least one of sodium, calcium, and zinc, and even more preferably at least one of calcium and zinc.

[0019] The individual phosphate units contained in polyphosphoric acid and salts thereof may be bonded to each other directly or via an atom or functional group other than the phosphate unit, for example, a divalent or higher valent cation (e.g., a divalent metal ion such as calcium or zinc). In this specification, when matters common to both polyphosphoric acid and salts of polyphosphoric acid are explained, they are collectively referred to as "polyphosphoric acid and salts thereof."

[0020] The polyphosphoric acid and salts thereof may be linear or cyclic, and may have one or more branches. Preferably, the polyphosphoric acid and salts thereof are linear.

[0021] Polyphosphates and salts thereof may contain only one type of molecule having the same structure and chain length, or may contain multiple types of molecules having different structures or chain lengths. The chain length of polyphosphates and salts thereof is expressed in terms of the number of consecutive phosphate units directly bonded to each other in the linear portion. For example, a chain length of 10 phosphate units or more (n=10) means that the linear portion of the polyphosphate molecule contains 10 or more consecutive phosphate units.

[0022] When polyphosphoric acid and its salts contain only one type of molecule having the same structure and chain length, the average chain length of the polyphosphoric acid and its salts is the same as the chain length of the molecule. When polyphosphoric acid and its salts contain multiple types of molecules having different structures or chain lengths, the average chain length of the polyphosphoric acid and its salts can be calculated by multiplying the chain length of each molecule by the proportion of that molecule in all molecules, and then adding up the obtained values ​​for all types.

[0023] The average chain length can be determined by known analytical methods such as size exclusion chromatography, P NMR for linear chains, or electrospray ionization mass spectrometry (ESI-MS) for relatively short chains. For analytical methods, see, for example, Christ et al., Anal. Chem. 2020, 92, 4167-4176, DOI: 10.1021 / acs.analchem.9b05144.

[0024] For example, using 31P NMR, the average chain length of polyphosphate and its salts can be determined as follows: From a 31P NMR spectrum obtained using a sample dissolved in an appropriate solvent, such as heavy water, the integral value of the signal corresponding to P in the terminal phosphate group of the polyphosphate molecule (terminal phosphate signal) and the total integral value of all signals corresponding to P in the phosphate groups of the polyphosphate molecule (total phosphate group signals) are obtained, and the ratio of the total integral value of all phosphate group signals, assuming the integral value of the terminal phosphate group signal is 2, is calculated. This ratio corresponds to the average chain length of linear polyphosphate. The solvent used for sample preparation may contain a pH adjuster such as a buffer, as long as it does not contain a phosphate-containing compound.

[0025] The average chain length of polyphosphates and salts thereof used in the present disclosure is not limited, and may be, for example, 1.1 phosphate units or more, 1.2 phosphate units or more, 1.3 phosphate units or more, 1.4 phosphate units or more, 1.5 phosphate units or more, 1.6 phosphate units or more, 1.7 phosphate units or more, 1.8 phosphate units or more, 1.9 phosphate units or more, 2 phosphate units or more, 2.5 phosphate units or more, 3 phosphate units or more, 4 phosphate units or more, 5 phosphate units or more, 6 phosphate units or more, 7 phosphate units or more, 8 phosphate units or more, 9 phosphate units or more, or 10 phosphate units or more, and may be, for example, 10,000 phosphate units or less, 1,000 phosphate units or less, 900 phosphate units or less, or 10,000 phosphate units or less. The phosphate units may be less than 800 phosphate units, less than 700 phosphate units, less than 600 phosphate units, less than 500 phosphate units, less than 400 phosphate units, less than 300 phosphate units, less than 200 phosphate units, less than 100 phosphate units, less than 90 phosphate units, less than 80 phosphate units, less than 70 phosphate units, less than 60 phosphate units, less than 50 phosphate units, less than 40 phosphate units, less than 30 phosphate units, less than 20 phosphate units, less than 15 phosphate units, less than 10 phosphate units, less than 5 phosphate units, less than 4 phosphate units, less than 3.5 phosphate units, less than 3 phosphate units, less than 2.5 phosphate units, less than 2 phosphate units, or less than 1.5 phosphate units.

[0026] The polyphosphoric acid and salts thereof used in the present disclosure may have a mass of greater than 10 kDa. Polyphosphoric acid and salts thereof having a mass of greater than 10 kDa can also be expressed as polyphosphoric acid and salts thereof that do not permeate an ultrafiltration membrane with a molecular weight cutoff of 10 kDa when subjected to ultrafiltration. Here, ultrafiltration refers to the process of adding a sample to a spin column equipped with an ultrafiltration membrane made of PES (polyethersulfone) and centrifuging the sample to filter out molecules in the sample.

[0027] The polyphosphoric acid and its salts used in the present disclosure may be readily soluble, sparingly soluble, or insoluble in water. Sparingly soluble or insoluble polyphosphoric acid salts have the advantages of being easy to recover, wash, dry, and otherwise handle, and of having low hygroscopicity and excellent storage stability.

[0028] The polyphosphoric acid and salts thereof used in the present disclosure are not limited in their molecular arrangement, and may be crystalline, quasicrystalline, or amorphous.

[0029] The polyphosphate and its salts used in the present disclosure may form microparticles (hereinafter also referred to as polyphosphate microparticles). The average particle size of the polyphosphate microparticles may be, for example, 1 nm or more, 10 nm or more, 20 nm or more, 50 nm or more, 100 nm or more, 200 nm or more, or 500 nm or more, and may be, for example, 3000 nm, 2500 nm, 2000 nm, 1500 nm, 1000 nm, 800 nm, 600 nm, or 500 nm or less. The polyphosphate microparticles may be aggregates of fine polyphosphate particles, and the aggregates may have an average particle size within the above numerical range. Polyphosphate microparticles with a particle size on the order of nanometers are also called polyphosphate nanoparticles.

[0030] The average particle size refers to the harmonic mean particle size (diameter) based on scattered light intensity, and can be obtained by dynamic light scattering (DLS) measurement and cumulant analysis. Measurement by dynamic light scattering and cumulant analysis can be performed using a commercially available DLS device or the like according to the methods described in JIS Z 8828:2019, ISO 22412:2017, etc.

[0031] The polyphosphoric acid and salts thereof used in the present disclosure may have a zeta potential of, for example, 0 mV or less, preferably -20 mV or less, and more preferably -30 mV or less. The polyphosphoric acid and salts thereof used in the present disclosure may have a zeta potential of, for example, -100 to 0 mV or less, preferably -80 to -20 mV, and more preferably -60 to -30 mV. The zeta potential can be measured by electrophoretic light scattering (ELS, also known as laser Doppler electrophoresis).

[0032] In one embodiment, the metal salt of polyphosphate is zinc polyphosphate. Zinc polyphosphate is primarily composed of polyphosphoric acid and zinc, with zinc being the most abundant constituent metal. Zinc polyphosphate may be composed solely of polyphosphoric acid and zinc, or may be composed of polyphosphoric acid, zinc, and other metals, such as sodium, calcium, and magnesium. The zinc content in zinc polyphosphate is preferably 10-60%, more preferably 20-55%, and even more preferably 30-50%. Here, the zinc content refers to the mass % of zinc element in zinc polyphosphate. Furthermore, the content of other metals in zinc polyphosphate is in the range of 0-30%, preferably 0-20%, more preferably 0-10%, and even more preferably 0-7%. Here, the content of other metals refers to the mass % of metal elements other than zinc in zinc polyphosphate.

[0033] The zinc polyphosphate may further contain various inorganic salts such as inorganic zinc salts (e.g., zinc hydroxide, zinc oxide, zinc phosphate, etc.) and calcium phosphate, and may also contain inorganic carbon compounds such as calcium carbonate.

[0034] The zinc polyphosphate preferably exhibits a broad peak at 2θ = 4 to 7° in an X-ray diffraction chart. In the present disclosure, a broad peak refers to a peak having a half-width of 1 degree or more in an X-ray diffraction chart, and is clearly distinguishable from a sharp linear peak. In addition to the broad peak at 2θ = 4 to 7°, the zinc polyphosphate preferably exhibits a broad peak at either 2θ = 30 to 36° or 56 to 62°, and more preferably exhibits broad peaks at both of these positions. Furthermore, the zinc polyphosphate preferably further exhibits a plateau-like, gentle signal spanning at least the range of 2θ = 10 to 36° in an X-ray diffraction chart, in addition to the one or more broad peaks described above.

[0035] The zinc polyphosphate is preferably in the form of fine particles. The zinc polyphosphate fine particles may have an average particle size of preferably 1 nm to 1000 nm, more preferably 20 nm to 800 nm, even more preferably 20 nm to 500 nm, and even more preferably 100 nm to 500 nm. The zinc polyphosphate fine particles may have a zeta potential of, for example, -100 to 0 mV or less, preferably -80 to -20 mV, and more preferably -60 to -30 mV.

[0036] The average chain length of the zinc polyphosphate can be 1.1 to 1000 phosphate units, 1.1 to 50 phosphate units, 1.1 to 30 phosphate units, 1.1 to 15 phosphate units, 1.1 to 5 phosphate units, 1.1 to 4 phosphate units, 1.1 to 3.5 phosphate units, 1.1 to 3 phosphate units, 1.1 to 2.5 phosphate units, or 1.1 to 2 phosphate units. The zinc polyphosphate is preferably sparingly soluble in water.

[0037] The zinc polyphosphate may be one disclosed in WO2024 / 090537 A1 or WO2024 / 090538 A1, which has both intestinal barrier function and excellent storage stability.

[0038] In one embodiment, the metal salt of polyphosphate is calcium polyphosphate. Calcium polyphosphate is mainly composed of polyphosphate and calcium, with calcium being the most abundant constituent metal. Calcium polyphosphate may be composed only of polyphosphate and calcium, or may be composed of polyphosphate and calcium and other metals, such as sodium, magnesium, and zinc.

[0039] The calcium polyphosphate may further contain calcium inorganic salts (for example, calcium hydroxide, calcium oxide, calcium phosphate, etc.), and may also contain inorganic carbon compounds such as calcium carbonate.

[0040] Calcium polyphosphate is preferably in the form of fine particles. The average particle diameter of the calcium polyphosphate fine particles may be preferably 10 nm to 3000 nm, more preferably 100 nm to 2500 nm, even more preferably 200 nm to 2000 nm, and even more preferably 500 nm to 1500 nm. The zeta potential of the calcium polyphosphate fine particles may be, for example, -100 to 0 mV or less, preferably -80 to -20 mV, and more preferably -60 to -30 mV.

[0041] The average chain length of calcium polyphosphate can be 1.1 to 1000 phosphate units, 1.1 to 200 phosphate units, 1.1 to 50 phosphate units, 1.1 to 30 phosphate units, 1.1 to 20 phosphate units, or 1.1 to 15 phosphate units. Calcium polyphosphate is preferably poorly soluble in water. Calcium polyphosphate is preferably amorphous.

[0042] The calcium polyphosphate may be that disclosed in WO2021 / 141066 A1, which has both intestinal barrier function and excellent storage stability.

[0043] In one embodiment, the metal salt of polyphosphate is sodium polyphosphate. Sodium polyphosphate is mainly composed of polyphosphate and sodium, with sodium being the most abundant constituent metal. Sodium polyphosphate may be composed only of polyphosphate and sodium, or may be composed of polyphosphate and sodium and other metals, such as calcium, magnesium, and zinc.

[0044] The average chain length of the sodium polyphosphate can be 1.1 to 10,000 phosphate units, 1.1 to 1,000 phosphate units, 3 to 1,000 phosphate units, 10 to 750 phosphate units, or 10 to 500 phosphate units. The sodium polyphosphate is preferably readily soluble in water.

[0045] Polyphosphoric acid and salts thereof can be produced by known chemical synthesis techniques, biochemical techniques, or biological techniques.

[0046] An example of a chemical synthesis method is a method in which a reaction solution containing a phosphate such as sodium phosphate is heated to a dehydration condensation temperature of, for example, 150 to 350°C.

[0047] An example of a biochemical method is synthesis by reacting adenosine triphosphate (ATP) with polyphosphate kinase (PPK). Commercially available PPK may be purchased, or PPK produced by known microorganisms capable of producing polyphosphate, such as Lactobacillus rhamnosus, Lactobacillus brevis, and Propionibacterium freudenreichii subsp. shermanii, may be used.

[0048] Although the enzymatic reaction catalyzed by PPK is reversible, when a large amount of ADP is present in the reaction solution relative to ATP, the decomposition reaction of polyphosphate becomes dominant so that the ADP / ATP ratio reaches equilibrium. Therefore, for efficient polyphosphate synthesis, it is preferable to maintain a low ADP concentration in the reaction solution. For example, a continuous ATP regeneration reaction system using creatine kinase or pyruvate kinase may be coupled. Other conditions, such as the composition of the reaction solution, reaction temperature, and reaction time, can be appropriately set to optimize PPK activity and depending on the synthesis scale.

[0049] As an example, the reaction conditions for coupling polyphosphate synthesis using PPK from Propionibacterium freudenreichii subsp. shermanii with a continuous ATP regeneration reaction using pyruvate kinase are shown below. 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 incubate at 40°C for 30 minutes. After incubation, add 2.5 μL of 240 U / mL polyphosphate kinase and incubate at 40°C for an additional 5 minutes. Subsequently, 2.5 μL of 1690 U / mL pyruvate kinase is added, and the mixture is reacted for 0.5 to 36 hours at 40° C. The reaction time may be appropriately set depending on the molecular weight and yield of the target polyphosphate. For example, to obtain high-molecular-weight polyphosphate in good yield, the reaction time is preferably about 20 hours.

[0050] An example of a biological method is to culture a microorganism capable of producing polyphosphate under appropriate culture conditions to produce polyphosphate. Examples of microorganisms capable of producing polyphosphate include strains belonging to the genera Lactobacillus, Bifidobacterium, Enterococcus, Lactococcus, Pediococcus, Leuconostoc, Streptococcus, Bacteroides, Eubacterium, and Clostridium, such as Lactobacillus rhamnosus GG and Lactobacillus brevis SBC8803. Polyphosphate can be recovered from the culture medium, supernatant, or precipitate after culturing a microorganism capable of producing polyphosphate under conditions suitable for polyphosphate production.

[0051] The salt of polyphosphate can be obtained by allowing a cation to coexist in the reaction system during the production of polyphosphate by the above-mentioned chemical synthesis method, biochemical method, or biological method. Alternatively, the salt of polyphosphate can be obtained by reacting polyphosphate with a chloride salt of the cation under alkaline conditions.

[0052] For example, zinc polyphosphate can be obtained by adjusting the pH of a polyphosphoric acid solution to 7.5 or higher, preferably 8 to 11, to prepare an alkaline solution of polyphosphoric acid, and then adding zinc chloride to the solution and allowing it to react. Alternatively, zinc polyphosphate can be obtained by adding zinc chloride to a polyphosphoric acid solution having a pH of 4 or higher (e.g., pH 4 to 7.5), and then adjusting the solution to an alkaline pH, for example, to 7.5 or higher, preferably 8 to 11, and allowing it to react. For details, see WO2024 / 090537 A1 and WO2024 / 090538 A1.

[0053] For example, calcium polyphosphate can be obtained by adding calcium chloride to an alkaline solution of polyphosphoric acid and reacting it, similar to zinc polyphosphate. For details, see WO2021 / 141066 A1.

[0054] Polyphosphoric acid and salts thereof can be purified by separation and concentration methods such as size exclusion chromatography, ion exchange chromatography, affinity chromatography, high performance liquid chromatography (HPLC), dialysis, salting out, ammonium sulfate precipitation, precipitation, and crystallization.

[0055] The polyphosphate or a salt thereof used in the present disclosure may be produced by Lactobacillus brevis SBC8803 strain. The SBC8803 strain has been deposited at the National Institute of Advanced Industrial Science and Technology (AIST) International Patent Organism Depositary under accession number FERM BP-10632. This microorganism has the ability to accumulate polyphosphate within the bacterial cell and release polyphosphate outside the bacterial cell, and therefore the culture medium and its supernatant, live bacterial cells, and dead bacterial cells after cultivation contain polyphosphate or a salt thereof.

[0056] [Composition] The present disclosure provides compositions, particularly pharmaceutical compositions and food and beverage compositions, containing polyphosphate or a salt thereof. The compositions of the present disclosure contain polyphosphate or a salt thereof, preferably a metal salt of polyphosphate, as an active ingredient and can be used in subjects in need of endometriosis prevention or treatment for the prevention or treatment of endometriosis. In addition, since the compositions of the present disclosure do not affect the fertility of the subject to which they are administered, they can be used in subjects in need of endometriosis prevention or treatment and in whom preservation of fertility is desired for the prevention or treatment of endometriosis while preserving fertility. As used herein, the term "prevention" encompasses all types of medically acceptable preventive interventions aimed at preventing or suppressing the onset or development of a disease. The term "treatment" also encompasses all types of medically acceptable therapeutic interventions aimed at curing a disease or condition, temporarily alleviating it, etc. Therefore, the prevention or treatment of endometriosis encompasses medically acceptable interventions for various purposes, including improving symptoms such as pain and infertility, slowing or halting progression, preventing the onset of endometriosis, and preventing recurrence. In particular, the compositions of the present disclosure can be used to inhibit the proliferation of endometriotic tissue in endometriosis.

[0057] The composition of the present disclosure contains an amount of polyphosphate or a salt thereof effective for preventing or treating endometriosis. The effective amount of polyphosphate or a salt thereof can be determined appropriately depending on the dosage, the age, weight, severity, and other factors of the subject. The composition may contain only polyphosphate, only a salt of polyphosphate, or both polyphosphate and a salt of polyphosphate.

[0058] The composition of the present disclosure may contain, in addition to polyphosphoric acid or a salt thereof, pharmaceutically acceptable additives. Examples of pharmaceutically acceptable additives include excipients, binders, lubricants, solvents, disintegrants, solubilizers, suspending agents, emulsifiers, isotonicity agents, stabilizers, preservatives, antioxidants, flavoring agents, coloring agents, buffers, and flow enhancers. Pharmaceutically acceptable additives are well known to those skilled in the art, and those skilled in the art can appropriately select and use them within the scope of their ordinary practice.

[0059] Specific examples of excipients include organic excipients such as sugars such as lactose, glucose, and D-mannitol, starches, and celluloses such as crystalline cellulose, and inorganic excipients such as calcium carbonate and kaolin.

[0060] Examples of binders include pregelatinized starch, gelatin, gum arabic, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, crystalline cellulose, D-mannitol, trehalose, hydroxypropyl cellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, and polyvinyl alcohol.

[0061] Examples of lubricants include stearic acid, fatty acid salts such as stearates, talc, silicates, and the like.

[0062] Examples of the solvent include purified water, physiological saline, and phosphate buffer solution.

[0063] Disintegrants include low-substituted hydroxypropyl cellulose, chemically modified celluloses and starches.

[0064] Examples of solubilizing agents include polyethylene glycol, propylene glycol, trehalose, benzyl benzoate, ethanol, sodium carbonate, sodium citrate, sodium salicylate, and sodium acetate.

[0065] Examples of suspending agents or emulsifying agents include sodium lauryl sulfate, gum arabic, gelatin, lecithin, glycerin monostearate, polyvinyl alcohol, polyvinylpyrrolidone, celluloses such as sodium carboxymethylcellulose, polysorbates, and polyoxyethylene hydrogenated castor oil.

[0066] Examples of isotonic agents include sodium chloride, potassium chloride, sugars, glycerin, and urea.

[0067] Stabilizers include polyethylene glycol, dextran sodium sulfate, other amino acids, and magnesium carbonate, which also acts as an acidity regulator.

[0068] Examples of preservatives include parahydroxybenzoates, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, and sorbic acid.

[0069] Antioxidants include water-soluble antioxidants such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, and sodium sulfite; fat-soluble antioxidants such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, and α-tocopherol; and metal chelating agents such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, and phosphoric acid.

[0070] Examples of flavoring agents include sweeteners and fragrances that are commonly used in the fields of medicine and food and drink, and examples of coloring agents include coloring agents that are commonly used in the fields of medicine and food and drink.

[0071] The composition of the present disclosure may be in any dosage form suitable for the route of administration, and can be formulated into dosage forms such as tablets (including sugar-coated tablets, film-coated tablets, sublingual tablets, and orally disintegrating tablets), powders, granules, capsules (including soft capsules and microcapsules), liquids, troches, syrups, emulsions, suspensions, injections (e.g., intravenous injections, subcutaneous injections, intramuscular injections, intraperitoneal injections, etc.), topical preparations (e.g., nasal preparations, transdermal preparations, ointments), suppositories (e.g., rectal suppositories, vaginal suppositories), foams (enemas), pellets, nasal preparations, pulmonary preparations (inhalants), and the like.

[0072] The compositions of the present disclosure can be administered orally or parenterally (for example, intravenously, intraperitoneally, vaginally, rectally, etc.), and are preferably administered orally.

[0073] Compositions for oral administration may be in the form of oral preparations, such as tablets, powders, granules, capsules, liquids, troches, syrups, emulsions, suspensions, and the like.

[0074] The composition of the present disclosure may be a controlled-release formulation such as an immediate-release formulation or a sustained-release formulation. Furthermore, when used as an oral formulation, coating may be performed as necessary for the purposes of masking, enteric coating, or sustained release. Examples of coating bases used for coating include sugar-coating bases, water-soluble film-coating bases, enteric film-coating bases, and sustained-release film-coating bases.

[0075] The composition of the present disclosure may contain other drugs for preventing or treating endometriosis, etc. Examples of other drugs for preventing or treating endometriosis include combined oral contraceptives, progestins such as dienogest, gonadotropin-releasing hormone (GnRH) agonists, GnRH antagonists, and androgens.

[0076] The compositions of the present disclosure are administered to a subject in need of prevention or treatment of endometriosis, particularly a subject at risk of or currently suffering from endometriosis. The subject may be a subject for whom fertility preservation is desired. The subject may be a human or a non-human animal, and examples of non-human animals include mammals such as rodents including mice, rats, hamsters, and guinea pigs, primates including chimpanzees and rhesus monkeys, livestock including pigs, cows, goats, horses, and sheep, and pets including dogs and cats. The compositions are preferably administered to humans.

[0077] The dosage of the composition when administered to humans can be selected, for example, from the range of 0.005 mg / kg to 15 mg / kg, preferably 0.01 mg / kg to 15 mg / kg, more preferably 0.05 mg / kg to 15 mg / kg, even more preferably 0.05 mg / kg to 5 mg / kg, and even more preferably 0.05 mg / kg to 2.5 mg / kg, in terms of polyphosphate per administration per day. Alternatively, the composition can be administered, for example, in the range of 0.3 to 900 mg / day, preferably 0.6 to 900 mg / day, more preferably 3 to 900 mg / day, even more preferably 3 to 300 mg / day, and even more preferably 3 to 150 mg / day, in terms of polyphosphate per patient, once a day or in divided doses.

[0078] The dosage of a composition containing zinc polyphosphate when administered to a human can be selected, for example, from the range of 0.005 mg / kg to 5 mg / kg, preferably 0.01 mg / kg to 4 mg / kg, more preferably 0.01 mg / kg to 2.5 mg / kg, even more preferably 0.015 mg / kg to 2 mg / kg, even more preferably 0.03 mg / kg to 1 mg / kg, and even more preferably 0.05 mg / kg to 1 mg / kg, in terms of zinc polyphosphate per administration per patient. Alternatively, the composition can be administered once or several times a day in a range of 0.3 to 300 mg / day, preferably 0.6 to 240 mg / day, more preferably 0.6 to 150 mg / day, even more preferably 0.9 to 120 mg / day, even more preferably 1.8 to 60 mg / day, and even more preferably 3 to 60 mg / day, in terms of zinc polyphosphate per patient.

[0079] Polyphosphoric acid and its salts can be used as active ingredients in medicines. Furthermore, because they can be safely taken orally, they can also be used as active ingredients or participating components in food and beverage compositions, such as foods for specified health uses, foods for special dietary uses, nutritional supplements, dietary supplements, health foods, foods with functional claims, and foods for patients. Thus, the present disclosure provides pharmaceutical compositions and food and beverage compositions for preventing or treating endometriosis, pharmaceutical compositions and food and beverage compositions for preventing endometriosis or ameliorating symptoms, and pharmaceutical compositions and food and beverage compositions for inhibiting the proliferation of endometriotic tissue, each containing polyphosphoric acid or a salt thereof.

[0080] The food and beverage composition preferably contains a culture supernatant or cells of Lactobacillus brevis strain SBC8803 as an active ingredient or participating component. The cells are preferably subjected to a process for at least partially disrupting them so that the polyphosphate or a salt thereof contained therein is released to the outside. Examples of such a process include heat treatment at 100°C or higher for several minutes or more (e.g., autoclaving at a temperature in the range of 110-125°C for 10 minutes or more), drying processes such as freeze-drying or spray-drying, and physical disruption processes such as ultrasonication or French press. The food and beverage composition preferably contains heat-killed cells of Lactobacillus brevis strain SBC8803.

[0081] [Method for preventing or treating endometriosis] The present disclosure further provides a method for preventing or treating endometriosis, and a method for preventing or ameliorating symptoms of endometriosis, which comprises administering the composition containing polyphosphate or a salt thereof to a subject in need of endometriosis prevention or treatment, specifically a subject at risk of developing or currently developing endometriosis. The present invention also provides a method for inhibiting the proliferation of endometriotic tissue, which comprises administering the composition containing polyphosphate or a salt thereof to the subject.

[0082] In addition, the present invention provides the use of the above-mentioned polyphosphoric acid or a salt thereof in the manufacture of a composition for preventing or treating endometriosis; the use of the above-mentioned polyphosphoric acid or a salt thereof in the manufacture of a composition for preventing or ameliorating symptoms of endometriosis; the use of the above-mentioned polyphosphoric acid or a salt thereof in the manufacture of a composition for inhibiting the proliferation of endometriotic tissue; the use of the above-mentioned polyphosphoric acid or a salt thereof for the prevention or treatment of endometriosis; the use of the above-mentioned polyphosphoric acid or a salt thereof for the prevention or amelioration of symptoms of endometriosis; and the use of the above-mentioned polyphosphoric acid or a salt thereof for inhibiting the proliferation of endometriotic tissue.

[0083] The present invention will be described in more detail by the following examples, but the present invention is not limited to these examples.

[0084] Example 1. Preparation of Polyphosphates (A) Preparation of Long-Chain Sodium Polyphosphate: 680 μL of 2 mol / L Tris-HCl (pH 9.0), 0.1 g of phosphoenolpyruvic acid, 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 were mixed in a 10 mL sample tube and heated at 40°C for 30 minutes. After heating, 2.5 μL of 240 U / mL polyphosphate kinase was added and the mixture was further heated at 40°C for 5 minutes. 2.5 μL of 1690 U / mL pyruvate kinase was then added and the mixture was incubated at 40°C for 20 hours. 1.7 mL of 2 mol / L sodium chloride was added to the reaction mixture and stirred at 10°C for 10 minutes. After stirring, the supernatant was removed, and 400 μL of purified water was added to the precipitate to dissolve it. 400 μL of 3 mol / L sodium chloride was then added to precipitate the precipitate. 400 μL of purified water was added to dissolve the precipitate, and 400 μL of 3 mol / L sodium chloride was added to precipitate the precipitate. After removing the supernatant, the precipitate was lyophilized to obtain long-chain sodium polyphosphate (also known as Poly-P Na or PPA-Na). From the chromatogram obtained by HPLC using a gel filtration column, the average chain length of the long-chain sodium polyphosphate was estimated to be 600-800.

[0085] (B) Preparation of Zinc Polyphosphate: 1 g of long-chain sodium polyphosphate (average chain length 600 or greater; Kamui Pharma Co., Ltd.), prepared as in (A) above, was added to 50 g of water to prepare an aqueous solution. 1 mol / L aqueous sodium hydroxide was added to adjust the pH to 10, and 50 mL of 1 mol / L zinc chloride solution was added dropwise over approximately 30 minutes. During the dropwise addition, sodium hydroxide was added periodically to maintain the pH at approximately 10. After the dropwise addition, the mixture was stirred at room temperature for 4 hours, then allowed to stand at a temperature below 10°C for 16 hours. The precipitate was then centrifuged (12,000 × g, 4°C, 10 minutes) to recover the precipitate. The precipitate was washed with aqueous ethanol (33% v / v ethanol) and centrifuged twice. This process was repeated twice, followed by lyophilization to obtain zinc polyphosphate (also known as Poly-P Zn or PPA-Zn) as a white powder.

[0086] (C) Confirmation of the degree of polymerization of zinc polyphosphate by NMR measurement. Approximately 10 mg of zinc polyphosphate was dissolved in approximately 0.7 mL of approximately 190 mmol / L citrate-sodium buffer (DO), pH 5.5, and used as an analytical sample for 31P NMR analysis. (i) Instrument: Bruker BioSpin 400 MHz. (ii) Analysis and Characterization Conditions: The signal at -5 ppm represents the terminal phosphate group, and the signals from -15 to -25 ppm represent internal phosphate groups. The total integral value of the terminal phosphate group was calculated by setting the integral value of the terminal phosphate group to 2. Zinc polyphosphate is prepared from linear sodium polyphosphate in aqueous solution. Cyclized polyphosphate is rare, and most polyphosphates are linear. Therefore, the calculated total integral value of the phosphate group is likely to approximate the degree of polymerization of zinc polyphosphate. Furthermore, since zinc polyphosphate is linear, the degree of polymerization is equal to the average chain length. (iii) Results: The degree of polymerization of the zinc polyphosphate prepared in (B) above was calculated to be 8.9 by NMR measurement. Based on manufacturing method (A), by changing detailed conditions such as the stirring time after the dropwise addition of sodium hydroxide and the collection, washing, and drying of the precipitate, we were able to produce zinc polyphosphate with a degree of polymerization in the range of 5.6 to 12.0.

[0087] (D) Powder X-ray crystal diffraction of zinc polyphosphate (i) Instrument: Rigaku MiniFlex II (ii) Analysis conditions: Anticathode: Cu, tube voltage: 30 kV, tube current: 15 mA, monochromator: Ni filter, sampling width: 0.020°, scanning speed: 10° / min, wavelength: 1.541836 Å, diffraction angle range (2θ): 2–60°, divergence slit: 1.25°, scattering slit: 8.0 mm, receiving slit: open (iii) Results: The zinc polyphosphate prepared in (B) above exhibited broad signal peaks at 2θ = 4–7°, 30–36°, and 56–62° (Figure 1). Zinc polyphosphate with a degree of polymerization ranging from 5.6 to 12.0, prepared by the method in (A) but varying the stirring time after sodium hydroxide addition and the recovery, washing, and drying of the precipitate, also exhibited broad signal peaks at the same positions. Such a signal peak is not observed with conventionally known polyphosphates such as sodium polyphosphate and calcium polyphosphate.

[0088] (E) Elemental Analysis of Polyphosphates. 20 mg of sodium polyphosphate and zinc polyphosphate prepared in (A) and (B) were weighed on a precision electronic balance and subjected to elemental analysis at Nogai Kagaku Co., Ltd. (Sapporo). Water content was determined by loss on drying (105°C, 2 hours). The test solution was prepared by adding nitric acid and hydrogen peroxide to the sample, decomposing it in a microwave sample pretreatment device, and then adjusting the volume to 20 ml with distilled water. Sodium, calcium, and magnesium contents were determined by flame atomic absorption spectrometry using an atomic absorption spectrometer (PerkinElmer, Analist 200). Zinc content was determined by inductively coupled plasma mass spectrometry (Agilent, Agilent 7500cx). Phosphorus content was determined by molybdenum reduction blue absorptiometry using a spectrophotometer (Shimadzu, UV-1800). The elemental contents (mass%) are shown in Table 1.

[0089] (F) Measurement of pH, zeta potential, and particle size of zinc polyphosphate. Five mg of zinc polyphosphate was suspended in 1 mL of purified water. The suspension was allowed to stand, and the pH of the supernatant was measured. After pH measurement, the suspension was ultrasonically dispersed and diluted to 2 mg / mL with purified water. This was used as an analytical sample to measure the zeta potential and particle size. Instrument: Malvern, ZEN3600. Analysis conditions: Dispersant: Water, Dispersant RI: 1.330, Viscosity (cp): 0.8872, Dispersant Dielectric Constant: 78.5, Temperature: 25°C. The pH of zinc polyphosphate was 9.46, and the zeta potential was -33.0 mV. The mean particle size based on intensity was 175.9 (d.nm).

[0090] (G) Preparation of Calcium Polyphosphate. 1.2 g of long-chain sodium polyphosphate (average chain length ≥ 600; Kamui Pharma Co., Ltd.) prepared in the same manner as in (A) above was added to 60 g of water to prepare an aqueous solution. 1 mol / L sodium hydroxide solution was added to adjust the pH to 10. 0.865 g of calcium chloride hydrate was added to 7.5 mL of purified water to prepare a calcium chloride solution. The entire amount was added dropwise to the long-chain sodium polyphosphate solution using a peristaltic pump at a rate of approximately 0.2 g / min. During the addition, sodium hydroxide solution was added periodically to maintain the pH at approximately 10. After the addition, the mixture was stirred at room temperature for 4 hours. The resulting suspension was then transferred to a centrifugal ultrafiltration unit (Amicon Ultra-15, nominal molecular weight cutoff: 3 kDa, sample volume: 15 mL) and concentrated and desalted by centrifugation (3,040 × g, 20 °C, 3 hours). The filter unit containing the concentrated precipitate was redispersed by adding 2 mL of ethanol (99.5%), followed by centrifugation at 3,040 × g and 20°C for 1 hour, after which the ethanol wash was removed. This ethanol washing procedure was repeated twice. After washing, the precipitate was vacuum-dried together with the filter unit, and the resulting solid was sieved through a sieve (mesh opening: 500 μm) to obtain calcium polyphosphate (also known as Poly-P Ca or PPA-Ca) as a white powder.

[0091] (H) Zeta Potential and Particle Size Measurement of Calcium Polyphosphate. The average particle size and zeta potential of the prepared calcium polyphosphate particles were measured using a dynamic light scattering (DLS) particle size analyzer (Zetasizer model Nano-ZS, Ver. 6.01, Malvern Instruments). Calcium polyphosphate was suspended in distilled water at a concentration of approximately 1 mg / mL and filtered through a PVDF membrane filter (Millex-SV Filter Unit, Low Protein Binding Durapore (PVDF) Membrane, pore size: 5.0 μm, Merck Millipore Ltd.). After vortexing, the supernatant dispersion was subjected to both dynamic light scattering and zeta potential measurements. Measurements were performed at 25°C using a disposable collapsible capillary cell (DTS1070). The average particle size of calcium polyphosphate was 1252 nm, and the zeta potential was -36.2 mV.

[0092] Example 2. Therapeutic Effect of Oral Administration of Zinc Polyphosphate or SBC8803 Strain. A 3 mm incision was made on the side of 7-week-old female BALB / cAnNCrl mice (Jackson Laboratories Japan), and both ovaries were removed. The incisions were then sutured with needle-tipped thread (COVIDIEN) to create donor and recipient mice. One week after ovariectomy, the uterus of the donor mouse and 200 μl of whole blood were collected from the inferior vena cava. The collected uterus was divided into two halves and incised longitudinally. The endometrial layer was peeled off with a scalpel and shredded with scissors. A 3 mm incision was made in the abdomen of the recipient mouse to create a transplant port. The shredded uterine fragment was mixed with 100 μl of whole blood and transplanted into the abdominal cavity through the transplant port using a 1000 μl pipette (Eppendorf). After transplantation, the transplant port was sutured with needle-tipped thread (COVIDIEN). In endometriosis model mice into which endometrial fragments were transplanted, endometriosis developed 21 days after the transplantation (Somigliana E et al., Hum Reprod, 14, 2944-2950 (1999)).

[0093] The day of uterine fragment transplantation into endometriosis model mice was counted as day 0. From day 21 to day 30, zinc polyphosphate (Poly-P Zn: 5 μg / 100 μl / head) and Lactobacillus brevis SBC8803 bacterial cell powder (product name "SBL88 TM Lactic acid bacteria (heat-treated): 10mg / 100μl / head; Sapporo Beer), dienogest (DNG: 100μg / 100μl / head; D230, TCI) were orally administered. 0.5w / v% methylcellulose (133-17815, Fujifilm Wako Pure Chemical Industries) was used as the solvent, and the control and untreated groups were orally administered the solvent alone (100μl / head). TM "Lactic acid bacteria (heat-treated)" is a heat-sterilized strain of SBC8803, and is 1 x 10 12 Contains more than 1000 dead bacteria per gram.

[0094] From ovariectomy to dissection, mice were intramuscularly injected with 100 μg / kg of β-estradiol (E2758, Sigma-Aldrich) dissolved in DMSO (046-21981, Fujifilm Wako Pure Chemical Industries, Ltd.) once a week. Thirty-one days after uterine implantation, mice were euthanized by cervical dislocation under isoflurane anesthesia, and endometriotic lesions were excised and weighed. The experimental protocol for this example is shown in Figure 2A.

[0095] The weight of endometriotic lesions was significantly reduced in both the zinc polyphosphate group and the SBC8803 heat-treated cell powder group (Fig. 2B).

[0096] Example 3. Preventive effect of oral administration of zinc polyphosphate or SBC8803 strain Endometriosis model mice were prepared in the same manner as in Example 2. The day of uterine fragment transplantation into the endometriosis model mice was designated as day 0. From day 0 to day 21, zinc polyphosphate (Poly-P Zn: 5 μg / 100 μl / head), Lactobacillus brevis SBC8803 bacterial cell powder (trade name "SBL88TM lactic acid bacteria (heat-treated)": 10 mg / 100 μl / head; Sapporo Beer), and dienogest (DNG: 100 μg / 100 μl / head; D230, TCI) were orally administered once a day. 0.5 w / v% methylcellulose (133-17815, Fujifilm Wako Pure Chemical Industries) was used as the solvent, and the control group and untreated group were orally administered only the solvent (100 μl / head).

[0097] From ovariectomy to dissection, mice were intramuscularly injected with 100 μg / kg of β-estradiol (E2758, Sigma-Aldrich) dissolved in DMSO (046-21981, Fujifilm Wako Pure Chemical Industries, Ltd.) once a week. On day 22 after uterine implantation, mice were euthanized by cervical dislocation under isoflurane anesthesia, and endometriotic lesions were excised and weighed. The experimental protocol for this example is shown in Figure 3A.

[0098] The weight of endometriotic lesions was significantly reduced in both the zinc polyphosphate-administered group and the SBC8803 heat-treated cell powder-administered group (Fig. 3B).

[0099] These results indicate that zinc polyphosphate and SBC8803 bacteria inhibit the growth of endometriotic tissue and exert both therapeutic and preventive effects against endometriosis.

[0100] Example 4 Therapeutic Effect of Oral Administration of Calcium Polyphosphate Endometriosis model mice were prepared in the same manner as in Example 2. The day of uterine fragment transplantation into the endometriosis model mice was set as day 0, and from day 21 to day 30, zinc polyphosphate (Poly-P Zn: 5 μg / 100 μl / head), calcium polyphosphate (Poly-P Ca: 20 μg / 100 μl / head), Lactobacillus brevis SBC8803 bacterial cell powder (trade name "SBL88TM lactic acid bacteria (heat-treated)": 10 mg / 100 μl / head; Sapporo Beer), and dienogest (DNG: 100 μg / 100 μl / head; D230, TCI) were orally administered once a day. The vehicle used was 0.5 w / v% methylcellulose (133-17815, Fujifilm Wako Pure Chemical Industries, Ltd.), and the control group and the untreated group were orally administered with the vehicle alone (100 μl / head).

[0101] From ovariectomy to dissection, mice were intramuscularly injected with 100 μg / kg of β-estradiol (E2758, Sigma-Aldrich) dissolved in DMSO (046-21981, Fujifilm Wako Pure Chemical Industries, Ltd.) once a week. Thirty-one days after uterine implantation, mice were euthanized by cervical dislocation under isoflurane anesthesia, and endometriotic lesions were excised and weighed. The experimental protocol for this example is shown in Figure 4A.

[0102] The weight of endometriotic lesions was significantly reduced in all groups administered with zinc polyphosphate, calcium polyphosphate, and heat-treated SBC8803 bacterial cell powder (Fig. 4B).

[0103] Example 5. Effect of Zinc Polyphosphate on Fertility. Five-week-old female BALB / cCrSlc mice (Japan SLC) were orally administered zinc polyphosphate (Poly-P Zn: 5 μg / 100 μl / head) once daily, starting on day 0. 0.5 w / v% methylcellulose (133-17815, Fujifilm Wako Pure Chemical Industries, Ltd.) was used as the solvent, and the control group received orally administered solvent alone (100 μl / head). A comparison group received a subcutaneous injection of leuprorelin acetate (LA: Tokyo Chemical Industry Co., Ltd., 20 μg / 50 μl / head) once daily. The administration period lasted from day 0 until the day before euthanasia. Mating began on day 14 after the start of administration and ended upon confirmation of a vaginal plug. For mice in which a vaginal plug was not confirmed, mating was terminated on day 28 after the start of administration. On the 15th day after mating, 1% Chicago Sky Blue 6B (Fujifilm Wako Pure Chemical Industries) was injected into the tail vein (100 μl / head), and the mice were euthanized by cervical dislocation under isoflurane anesthesia. The uterus was removed, and the number of implantations, fetuses, and resorbed fetuses was counted. The test protocol for this example is shown in Figure 5A.

[0104] There was no difference in pregnancy rate or number of fetuses per implantation between the zinc polyphosphate group and the control group, confirming that it had no effect on fertility (Figure 5B).

[0105] All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety.

Claims

1. A pharmaceutical composition for preventing or treating endometriosis, comprising polyphosphoric acid or a salt thereof.

2. The pharmaceutical composition according to claim 1, wherein the polyphosphoric acid or a salt thereof is poorly soluble or insoluble in water.

3. The pharmaceutical composition according to claim 1, wherein the average particle size of the polyphosphoric acid or a salt thereof is 1 to 3,000 nm.

4. The pharmaceutical composition according to claim 1, wherein the zeta potential of the polyphosphoric acid or a salt thereof is 0 mV or less.

5. The pharmaceutical composition according to claim 1, which contains linear polyphosphate or a salt thereof.

6. The pharmaceutical composition according to claim 1, wherein the average chain length of the polyphosphate or a salt thereof is not less than 1.1 phosphate units and not more than 10,000 phosphate units.

7. The pharmaceutical composition according to claim 1, which contains a metal salt of polyphosphate, wherein the metal salt of polyphosphate contains at least one of sodium, potassium, magnesium, calcium, and zinc as its constituent metal.

8. The pharmaceutical composition according to claim 1, which contains calcium polyphosphate.

9. The pharmaceutical composition according to claim 1, containing zinc polyphosphate.

10. The pharmaceutical composition according to claim 9, wherein the zinc polyphosphate exhibits a broad peak at 2θ=4 to 7° in an X-ray diffraction chart.

11. The pharmaceutical composition according to claim 9, wherein the zinc content of the zinc polyphosphate is 10 to 60%.

12. A pharmaceutical composition according to any one of claims 1 to 11 for inhibiting the proliferation of endometriotic tissue.

13. A pharmaceutical composition according to any one of claims 1 to 11 for use in a subject for whom preservation of fertility is desired.

14. A pharmaceutical composition according to any one of claims 1 to 11, for oral administration.

15. A food or beverage composition containing polyphosphate or a salt thereof for use in a subject in need of prevention or treatment of endometriosis.

16. The food and beverage composition according to claim 15, which is a supplement.

17. A food or beverage composition according to claim 15 or 16, which contains a culture supernatant or cells of Lactobacillus brevis SBC8803 (accession number: FERM BP-10632).

18. A food or beverage composition according to Item 15 or 16, comprising heat-killed cells of Lactobacillus brevis SBC8803 (accession number: FERM BP-10632).

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