Combined administration method and pharmaceutical composition for treating high serum phosphate related diseases

Abstract

The present invention provides methods and pharmaceutical compositions for reducing serum phosphate levels in mammals and humans comprising the combined use of component (a) a phosphate binder; and component (b) a phosphate transport inhibitor, wherein a therapeutically effective amount of at least one of the component (a) and component (b) administered to the subject is lower than a therapeutically effective amount of the component administered in the absence of the other component.

Description

Technical Field

[0001] This invention relates to the fields of disease treatment and pharmaceuticals. Specifically, this invention relates to methods for lowering serum phosphate and methods and pharmaceutical compositions for treating hyperphosphatemia-related diseases. Background Technology

[0002] Hyperphosphatemia is a common condition in patients with renal insufficiency, especially those on dialysis. It generally results from: excessive phosphate intake, excessive phosphate release due to tissue damage, reduced phosphate excretion in late-stage renal failure, and / or symptoms arising from increased phosphate absorption through the kidneys.

[0003] Hyperphosphatemia is a pathophysiologically elevated level of phosphate in the blood, resulting in an electrolyte imbalance. Elevated serum phosphate levels can lead to the deposition of calcium phosphate in blood vessels and other tissues. Chronic hyperphosphatemia can cause calcification of tissues and blood vessels, which can lead to impaired blood flow, myocardial infarction, and / or stroke. Patients with long-term uncontrolled hyperphosphatemia gradually develop extensive soft tissue calcification due to the deposition of calcium / phosphate products in the skin, joints, tendons, and ligaments. Ocular deposition of calcium / phosphate products also occurs. Hyperphosphatemia can also lead to hypocalcemia, secondary hyperparathyroidism, and progressive metabolic bone disease.

[0004] Patients often require phosphate binders to lower serum phosphate levels. Current technology necessitates the use of large doses of phosphate binders, typically in large numbers of tablets, or, where fewer tablets are needed, much larger and more difficult to administer. Furthermore, patients with renal insufficiency, especially those on dialysis, are only permitted to drink small amounts of water daily, making it difficult to easily take large doses of medication. Patients with renal insufficiency have a reduced ability to excrete excess fluid through urine. Excess fluid can cause hypertension, which can damage the cardiovascular system, particularly the left ventricle, and thus indirectly contribute to increased mortality from cardiovascular disease.

[0005] Phosphate transport inhibitors, which inhibit the transport of phosphate ions (Pi) in the gastrointestinal tract and / or kidneys, have also been used to lower serum phosphate levels. Reported phosphate transport inhibitors include phosphodiesterase inhibitors, P2Y agonists, adenylate cyclase receptor agonists, cholinergic agonists, prostaglandin EP4 receptor agonists, dopamine D1 agonists, melatonin receptor agonists, and 5HT4 agonists.

[0006] Several methods and pharmaceuticals for lowering serum phosphate levels already exist in this field. There is a greater need for even better methods and pharmaceuticals, particularly those for lowering serum phosphate and treating hyperphosphatemia-related diseases using smaller or safer dosages. Summary of the Invention

[0007] The present invention unexpectedly discovered that the combination of a specific phosphate binder and a phosphate transport inhibitor can exert a significant synergistic effect in reducing serum phosphate levels, significantly reducing the therapeutically effective dose of either the phosphate binder or the phosphate transport inhibitor compared to administration of either alone, or significantly increasing the reduction in serum phosphate levels. This provides a method and pharmaceutical composition for reducing serum phosphate levels in mammals and humans and for treating hyperphosphatemia-related diseases.

[0008] Specifically, the present invention provides a method for reducing serum phosphate levels in mammals and humans and for treating hyperphosphatemia-related diseases in subjects, comprising administering to said subjects:

[0009] Component (a) phosphate binder; and

[0010] Component (b) Phosphate transport inhibitor.

[0011] In this article, phosphate binders refer to substances that interact with phosphate ions, including protonated phosphate ions, thereby preventing them from being absorbed from the gastrointestinal tract into the bloodstream and into the body.

[0012] Phosphate binders that can be used in this invention include metal salt-based phosphate binders, such as calcium salts, magnesium salts, aluminum salts, iron salts, lanthanum salts, and bismuth salts. In one aspect of this invention, metal salt-based phosphate binders include, for example, lanthanum carbonate, calcium carbonate, calcium acetate, calcium acetate / magnesium carbonate, ferric citrate, superferric hydroxide, magnesium ferric bicarbonate, aluminum hydroxide, SBR-759, and PA-21. Furthermore, phosphate binders that can be used in this invention also include organic polymers of phosphate-bound salts that function as anion exchangers, such as sevelamer (pharmaceutically acceptable salts of which include sevelamer carbonate), AMG223 (Amgen), and MCI-196 (Colestilan, Mitsubishi). Suitable aluminum salts are all pharmaceutically acceptable salts that satisfy the above requirements, particularly oxides, especially aluminum hydroxide gel (Algedrat), and / or hydroxides. Suitable magnesium salts are all pharmaceutically acceptable salts that satisfy the above requirements, preferably chlorides, sulfates, hydroxides, oxides, carbonates, and especially heavy magnesium carbonate. Preferred metal salt-based phosphate binders are ferric hydroxide, ferric hydroxide, and ferric citrate, as well as iron preparations stabilized or bound to carbohydrates or humic acids, such as basic magnesium ferric carbonate and calcium salts. Among metal salts, calcium salts are particularly good phosphate binders. Furthermore, magnesium salts also have independent phosphate-binding properties, and their presence can reduce the amount of calcium-based phosphate binder required. Therefore, a combination of calcium acetate and magnesium carbonate is a preferred composition.

[0013] Phosphate binders that can be used in this invention also include anion-binding polymers such as those disclosed in EP1834976B1, which bind target anions (e.g., phosphates) and generally have the following characteristics: a) an expansion ratio of less than about 5; b) a gel pore volume distribution measured in physiological media, characterized in that a portion of the pore volume is contacted by non-interacting solutes, has a molecular weight greater than about twice the molecular weight (MW) of the target anion, and less than about 20% of the gel weight; and c) less than about 60% ion binding interference to the target anion when measured in a gastrointestinal mimic relative to a non-interfering buffer. Examples of polymers include amine monomers, such as those selected from allylamine, ethyleneamine, ethyleneimine, 1,3-diaminopropane, and NNN', N'-tetra(3-aminopropyl)1,4-diaminobutane, 1,2,3,4-tetraaminobutane, etc. Anion-binding polymers used in this invention also include anion-binding polymers containing cross-linked polyamines, such as the following cross-linked polyamine polymers having polymeric monomers of the following formula:

[0014]

[0015] N,N,N',N'-Tetra(3-aminopropyl)butane-1,4-diamine

[0016] (N,N,N',N'-tetrakis(3-aminopropyl)butane-1,4-diamine)

[0017] The monomer is cross-linked using a cross-linking agent. In one embodiment, the cross-linking agent is epichlorohydrin and / or 1,3-dichloropropane. In another embodiment, the cross-linking agent is 2-(chloromethyl)oxirane.

[0018] In one embodiment, the anionic polymer containing cross-linked polyamines is bixalomer.

[0019] In this article, phosphate transport inhibitors refer to compounds, especially small molecule compounds, that can inhibit the transport of phosphate ions (Pi) in the gastrointestinal tract and / or kidneys.

[0020] Phosphate transport inhibitors that can be used in this invention include inhibitors of sodium / hydrogen exchanger 3 (NHE3)-mediated sodium and hydrogen ion antitransport, or NHE3 inhibitors, as disclosed in WO2014169094A2. These inhibitors are ligands of NHE3 and, upon binding to NHE3, particularly to the epithelium of the gastrointestinal tract, inhibit sodium and hydrogen ion antitransport. In some embodiments, the NHE3 inhibitor is a compound of formula X:

[0021]

[0022] Wherein, NHE is an NHE-bound small molecule comprising (i) a heteroatom-containing portion and (ii) a cyclic or heterocyclic scaffold or carrier portion directly or indirectly bonded thereto, the heteroatom-containing portion being selected from substituted guanidine portions and substituted heterocyclic portions, which may optionally be fused with the scaffold or carrier portion to form a fused bicyclic structure; L is a bond or linking group connecting the core to the NHE-bound small molecule, and n is an integer of 2 or greater.

[0023] In some embodiments, the NHE3 inhibitor is a compound disclosed in Table E1 of WO2014169094A2.

[0024] In some embodiments, the NHE3 inhibitor is a compound of the following formula or a pharmaceutically acceptable salt thereof.

[0025]

[0026] 1-[2-[2-[[3-[(4S)-6,8-dichloro-2-methyl-3,4-dihydro-1H-isoquinoline-4-yl]phenyl]sulfonylamino]ethoxy]ethoxy]ethyl]-3-[4-[2-[2-[3-[(4S)-6,8-dichloro-2-methyl-3,4-dihydro-1H-isoquinoline-4-yl]

[0027] [Phenyl]sulfonylamino]ethoxy]ethoxy]ethylcarbamoylamino]butyl]urea

[0028] In some embodiments, the NHE3 inhibitor is the hydrochloride salt of the above-mentioned compound, i.e.

[0029]

[0030] 1-[2-[2-[[3-[(4S)-6,8-dichloro-2-methyl-3,4-dihydro-1H-isoquinoline-4-yl]phenyl]sulfonylamino]ethoxy]ethoxy]ethyl]-3-[4-[2-[2-[2-[[3-[(4S)-6,8-dichloro-2-methyl-3,4-dihydro-1H-isoquinoline-4-yl]phenyl]sulfonylamino]ethoxy]ethoxy]ethylcarbamoylamino]butyl]urea dihydrochloride

[0031] In some embodiments, the NHE3 inhibitor is tenapanor or tenapanor dihydrochloride.

[0032] In one aspect of the invention, the components in the combined method and pharmaceutical composition for reducing serum phosphate levels in mammals and humans are: component (a) sevelamer or a pharmaceutically acceptable salt thereof; and component (b) tenapanol or tenapanol hydrochloride.

[0033] In one aspect of the invention, the components in the combined method and pharmaceutical composition for reducing serum phosphate levels in mammals and humans are: component (a) bisalom or a pharmaceutically acceptable salt thereof; and component (b) tenapanol or tenapanol hydrochloride.

[0034] The subjects for whom the methods and drugs (pharmaceutical compositions) described herein are needed include those diagnosed with abnormal (elevated) serum phosphate levels. The subjects to be treated can be mammals. The subjects to be treated can be humans or non-human primates such as monkeys. Mammals can be other animals, such as rats, mice, rabbits, pigs, dogs, etc. The mammals can be domesticated animals, such as cats or dogs.

[0035] In one aspect of the invention, in the method and pharmaceutical composition provided by the invention for reducing serum phosphate levels in mammals and humans, the therapeutically effective amount of at least one of components (a) and (b) applied to the subject is lower than the therapeutically effective amount of said component applied in the absence of the other component. In yet another aspect of the invention, the therapeutically effective amounts of both components (a) and (b) applied to the subject are lower than the therapeutically effective amounts of said component applied in the absence of the other component.

[0036] In another aspect of the invention, in the method and pharmaceutical composition for reducing serum phosphate levels in mammals and humans provided by the invention, at least one of components (a) and (b) applied to the subject, particularly both, are at least 5%, at least 10%, at least 25%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% lower than the therapeutically effective amount of said component applied in the absence of the other component.

[0037] In one aspect of the invention, in the method and pharmaceutical composition provided by the invention for reducing serum phosphate levels in mammals and humans, the therapeutically effective amount of at least one of the components (a) and (b) applied to the subject is 5% to 90%, 10% to 90%, 25% to 90%, or 50% to 90% lower than the therapeutically effective amount of the component applied when the other component is absent.

[0038] For example, in the methods and pharmaceutical compositions for reducing serum phosphate levels in mammals and humans provided by the present invention, the therapeutically effective amount of the phosphate binder is at least 5%, at least 10%, at least 25%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% lower than the therapeutically effective amount of the phosphate binder applied in the absence of the phosphate transport inhibitor. That is, in the pharmaceutical compositions of the present invention, the dose of the phosphate binder is at least 5%, at least 10%, at least 25%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% lower than the normal dose (recommended dose) of the phosphate binder used alone. In one aspect of the present invention, in the methods and pharmaceutical compositions for reducing serum phosphate levels in mammals and humans provided by the present invention, the therapeutically effective amount of the phosphate transport inhibitor is 5% to 90%, 10% to 90%, 25% to 90%, or 50% to 90% lower than the therapeutically effective amount of the phosphate transport inhibitor applied in the absence of the phosphate binder. In the pharmaceutical composition of the present invention, the dose of the phosphate transport inhibitor is 5% to 90%, 10% to 90%, 25% to 90%, or 50% to 90% lower than the normal dose (recommended dose) of using the phosphate transport inhibitor alone.

[0039] In the methods and pharmaceutical compositions for reducing serum phosphate levels in mammals and humans provided by this invention, the phosphate binder and the phosphate transport inhibitor are administered simultaneously. In another aspect of this invention, the phosphate binder and the phosphate transport inhibitor are administered separately.

[0040] In the method and pharmaceutical composition for reducing serum phosphate levels in mammals and humans provided by the present invention, the phosphate binder and the phosphate transport inhibitor are formulated in the same pharmaceutical composition, or the phosphate binder is formulated in a first pharmaceutical composition and the phosphate transport inhibitor is formulated in a second pharmaceutical composition.

[0041] In one aspect of the invention, the method and pharmaceutical composition provided by the invention for reducing serum phosphate levels in mammals and humans comprises: component (a) bisalom or a pharmaceutically acceptable salt thereof; and component (b) tenapanol or tenapanol hydrochloride. Preferably, the therapeutically effective amount of bisalom in the combined pharmaceutical composition of the invention is at least 5%, at least 10%, at least 25%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% lower than the therapeutically effective amount administered in the absence of tenapanol, for example, 5% to 90%, 10% to 90%, 25% to 90%, or 50% to 90%. Preferably, the therapeutically effective amount of tenapanol in the combination pharmaceutical composition of the present invention is at least 5%, at least 10%, at least 25%, at least 50%, at least 60%, at least 70%, at least 80%, or at least 90% lower than the therapeutically effective amount when bisalom is not present, for example, 5% to 90%, 10% to 90%, 25% to 90%, or 50% to 90%.

[0042] The methods and pharmaceutical compositions provided by this invention can reduce serum phosphate levels in mammals and humans. Therefore, this invention also provides methods and pharmaceutical compositions for treating hyperphosphatemia-related diseases. These hyperphosphatemia-related diseases include any disease or other condition caused by abnormally elevated serum phosphate levels due to abnormal phosphate absorption in the gastrointestinal tract and / or kidneys. In one aspect of this invention, the hyperphosphatemia-related disease is hyperphosphatemia. In another aspect of this invention, the hyperphosphatemia-related disease is hypocalcemia, secondary hyperparathyroidism, and progressive metabolic bone disease. In another aspect of this invention, the hyperphosphatemia-related disease is tissue and vascular calcification, cardiovascular diseases such as myocardial infarction and / or stroke. In another aspect of this invention, the hyperphosphatemia-related disease is nephropathy, proteinuria, hyperparathyroidism, etc.

[0043] The active ingredient in the pharmaceutical composition provided by this invention is an agent that lowers serum phosphate levels in mammals and humans. Although the active ingredient in the therapeutic pharmaceutical composition of this invention can be administered in the form of a raw material compound, it is preferred that the active ingredient, optionally in the form of a physiologically acceptable salt, be introduced into the pharmaceutical composition together with one or more adjuvants, excipients, carriers, buffers, diluents and / or other conventional pharmaceutical excipients.

[0044] The pharmaceutical compositions of the present invention can be administered via any convenient route suitable for the desired therapy. Preferred routes of administration include oral administration, particularly in tablet, capsule, lozenge, powder, and liquid form; and parenteral administration, particularly by subcutaneous, subcutaneous, intramuscular, and intravenous injection. The pharmaceutical compositions of the present invention can be prepared by those skilled in the art using standard methods and conventional techniques suitable for the desired formulation. If desired, compositions suitable for sustained release of the active ingredient can be used.

[0045] The pharmaceutical compositions of the present invention may be those suitable for oral, rectal, bronchial, nasal, pulmonary, local (including buccal and sublingual), transdermal, vaginal, or parenteral (including skin, subcutaneous, intramuscular, intraperitoneal, intravenous, intraarterial, intracerebral, and intraocular injection or infusion) administration, or pharmaceutical compositions suitable for administration via a sustained-release system. Examples of suitable sustained-release systems include a semi-permeable matrix of a solid hydrophobic polymer containing the compounds of the present invention, which may be in the form of a shaped article, such as a film or microcapsule.

[0046] Therefore, the active ingredient in the pharmaceutical composition of the present invention can be formulated together with conventional adjuvants, carriers, or diluents into a pharmaceutical composition and its unit dosage form. Such forms include solids, and especially tablets, filled capsules, powders, and pellets, as well as liquids, especially aqueous or non-aqueous solutions, suspensions, emulsions, elixirs, and capsules filling the aforementioned forms, all for oral administration, suppositories for rectal administration, and sterile injectable solutions for parenteral administration. Such pharmaceutical compositions and their unit dosage forms may include conventional ingredients in conventional proportions, with or without additional active compounds or ingredients, and such unit dosage forms may contain any suitable effective amount of the active ingredient corresponding to the desired daily dose range.

[0047] For preparing a pharmaceutical composition from the active ingredient in the pharmaceutical composition of the present invention, a pharmaceutically acceptable carrier may be a solid or a liquid. Solid forms of formulations include powders, tablets, pills, capsules, pouches, suppositories, and dispersible granules. The solid carrier may be one or more substances that can also be used as a diluent, flavoring agent, solubilizer, lubricant, suspending agent, binder, preservative, tablet disintegrant, or encapsulation material.

[0048] Aqueous suspensions suitable for oral administration can be prepared by dispersing finely pulverized active ingredients in water containing viscous substances such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, or other well-known suspending agents.

[0049] This also includes solid formulations intended to be converted into a liquid form for oral administration before immediate use. Such liquid forms include solutions, suspensions, and emulsions. In addition to the active ingredient, such formulations may also contain colorants, flavoring agents, stabilizers, buffers, artificial and natural sweeteners, dispersants, thickeners, solubilizers, etc.

[0050] For topical application to the epidermis, the compounds of this invention can be formulated as ointments, creams, lotions, or transdermal patches. For example, ointments and creams can be formulated with an aqueous or oil-based base plus a suitable thickener and / or gelling agent. Lotions can be formulated with an aqueous or oil-based base and typically also contain one or more emulsifiers, stabilizers, dispersants, suspending agents, thickeners, or colorants.

[0051] Alternatively, the active ingredient in the pharmaceutical composition of the present invention may be provided in dry powder form, such as a powder mixture of the compound in a suitable powder matrix (e.g., lactose, starch, starch derivatives (e.g., hydroxypropyl methylcellulose), and polyvinylpyrrolidone (PVP)). Suitably, the powder carrier will form a gel within the nasal cavity. The powder composition may be presented in unit dosage forms, such as capsules or cartridges (e.g., gelatin capsules or cartridges), or in blister packs from which the powder can be administered via an inhaler.

[0052] When needed, a composition suitable for providing sustained release of active ingredients can be applied.

[0053] Pharmaceutical formulations are preferably unit dosage forms. In this type of form, the formulation is subdivided into unit doses containing appropriate amounts of the active ingredient. A unit dosage form can be a packaged formulation containing discrete amounts of the formulation, such as packaged tablets, capsules, or powder in vials or ampoules. Furthermore, a unit dosage form can be the capsule, tablet, sachets, or lozenges themselves, or can be a packaged form of any of these dosage forms in suitable quantities.

[0054] Tablets or capsules for oral administration and liquids for intravenous administration and continuous infusion are preferred compositions.

[0055] Therapeutic effective dose refers to the amount of active ingredient that relieves symptoms or condition. Therapeutic efficacy and toxicity, such as ED50 and LD50, can be determined using standard pharmacological procedures in cell cultures or laboratory animals. The dose ratio between therapeutic and toxic effects is the therapeutic index, which can be expressed as the LD50 / ED50 ratio.

[0056] The dosage must be carefully adjusted according to the individual's age, weight, and condition, as well as the route of administration, dosage form, and administration regimen, and the expected results. The exact dosage should be determined by the physician.

[0057] The actual dosage depends on the nature and severity of the disease being treated, the exact method of administration and dosage form, and, within the physician's judgment, can be changed by gradually increasing the dosage according to the specific circumstances of the invention to produce the desired therapeutic effect.

[0058] In one embodiment of the invention, the suitable single dose of the phosphate binder (e.g., sevelam or bisalome) in the combination is about 100 mg to 5000 mg, preferably about 250 mg to 3500 mg, for example about 500 to 2500 mg; and the suitable dose for the total daily dose is about 300 mg to 15000 mg, preferably about 1000 mg to 10000 mg, for example 1500 to 7500 mg.

[0059] In one embodiment of the invention, the suitable single dose of the phosphate transport inhibitor (e.g., tenapanol or tenapanol hydrochloride) in the combination is about 1 mg to 50 mg, preferably about 5 mg to 20 mg, for example about 10 to 30 mg; and the suitable dose for the total daily dose is about 2 mg to 100 mg, preferably about 10 mg to 50 mg, for example about 20 to 100 mg.

[0060] In some embodiments of the present invention, due to the synergistic effect of the phosphate binder and transport inhibitor in reducing serum phosphate levels in mammals and humans, the dosage of each component may be appropriately reduced when used in combination compared to when used alone. For example, in the combination provided by the present invention, the dosage of the phosphate binder (e.g., sevelamer or bisalome) is half that of the phosphate binder used alone, for example, about 250 mg to 1250 mg per single dose, three times daily, for a total dose of about 750 mg to 3750 mg per day. As another example, in the combination provided by the present invention, the dosage of the phosphate transport inhibitor (e.g., tenapanol or tenapanol hydrochloride) is half that of the phosphate transport inhibitor used alone, for example, about 5 mg to 15 mg per single dose, twice daily, for a total dose of about 10 mg to 30 mg per day.

[0061] In this invention, "treatment" includes: an ongoing process or outcome of improving, alleviating, reducing, or preventing symptoms associated with hyperphosphatemia; an ongoing process or outcome of improving symptoms associated with hyperphosphatemia; an ongoing process or outcome of normalizing bodily functions in a disease or condition that causes specific impairment of bodily function; or an ongoing process or outcome of improving one or more clinically measurable parameters that cause a disease. In one embodiment, the purpose of treatment is to prevent or slow (alleviate) an undesirable physiological condition, symptom, or disease, or to achieve a beneficial or desired outcome. This outcome may be, for example, medical, physiological, clinical, physical therapy, occupational therapy, directed at a healthcare professional or patient; or parameters understood in the art as "quality of life" or activities of daily living. In this invention, beneficial or desired clinical outcomes include, but are not limited to, alleviating symptoms; reducing / minimizing the severity of the condition, symptom, or disease; stabilizing the state of the condition, symptom, or disease; delaying the onset of the condition, symptom, or disease or slowing its progression; improving or mitigating the condition, symptom, or disease; and alleviating (whether partial or overall), whether detectable or undetectable; or enhancing or improving the condition, symptom, or disease. In one embodiment, treatment includes evoking a clinically effective response without excessive levels of side effects. In one embodiment, treatment also includes prolonging survival compared to the expected survival without treatment. In one embodiment, treatment refers to administering a drug or performing a medical procedure on a patient. In this invention, treatment can be prevention, cure of infirmity or disease, or improvement of a patient's clinical condition, including reducing disease duration or severity, or subjectively improving the patient's quality of life or prolonging the patient's survival. Attached Figure Description

[0062] Figure 1 The grouping for the animal experiments is explained below. Rats were divided into 11 groups and administered the excipient, tenapanol or bisalom alone, tenapanol in combination with different concentrations of bisalom (0%, 0.5%, 1%, and 2%), and tenapanol in combination with different concentrations of sevelamer (0.75%, 1.5%, and 3%).

[0063] Figure 2 A schematic diagram of an animal experiment design scheme.

[0064] Figure 3 This is a diagram showing the results of an animal experiment. Detailed Implementation

[0065] The following will further illustrate the essence and beneficial effects of the present invention with reference to embodiments. These embodiments are only used to illustrate the present invention and are not intended to limit the present invention.

[0066] Example 1: Materials and Methods

[0067] Drugs and reagents

[0068] Tenapanor: Ardelyx, Inc., XPHOZAH (tenapanor) tablets 10mg;

[0069] Bisalom: Astallas Pharmaceuticals, Kitlin カプル250mg,

[0070] Capsules 250mg;

[0071] Sevelamer: Sanofi (China) Investment Co., Ltd., Sevelamer Carbonate 800mg;

[0072] Standard rodent feed with added phosphate: 0.692g of sodium dihydrogen phosphate was added to every 100g of standard experimental rat feed (Speford (Beijing) Biotechnology Co., Ltd.), and the phosphorus content of the feed was 1.101%.

[0073] Animal materials

[0074] One hundred and ten male rats, approximately eight weeks old, were selected. Before grouping, they were housed in isolation cages (two rats per cage) for at least 48 hours to acclimatize. The animals were housed in a controlled environment with a temperature of 20–25°C and a humidity of 35–55%. Throughout the study, the rats had free access to a standard rodent diet supplemented with 0.4% inorganic phosphorus (1:1 sodium-potassium ratio, 1.1% w / w total phosphorus). The Sprague–Dawley rats had free access to deionized water. Rats' body weight was recorded at the start of the study and daily throughout the experiment.

[0075] All animal experiments were approved by the Laboratory Animal Ethics Review Committee of Xuzhou Medical University.

[0076] Experimental Design

[0077] Research plan as follows Figure 1 and Figure 2 As shown.

[0078] in Figure 1 Explanation of animal grouping in the experiment. Figure 2 A schematic diagram of an animal experiment design. (Example) Figure 1 As shown, SD rats were randomly divided into 11 groups. In the experiment (as shown...) Figure 2As shown in the figure, each group received the excipient (0.01% Tween 80) or tenapanol (0.15 mg / kg BID dose) by gavage three times daily, in combination with four diets containing different concentrations of bisalom (0%, 0.5%, 1%, and 2%) and three different concentrations of sevelamer (0.75%, 1.5%, and 3%), for 11 days.

[0079] During the entire treatment period, animals were housed in pairs in home cages for the first 5 days to receive medication. On the 6th day of treatment, they were transferred to individual metabolic cages for the remainder of treatment. 24-hour food intake, water intake, and urine output were measured on the 6th day and the last four days of treatment (days 8–11).

[0080] Detection methods

[0081] The excretion of sodium and phosphorus in urine was quantified using ion chromatography. The 24-hour ion excretion was calculated by multiplying the ion concentration by the 24-hour urine volume. The urinary ion excretion was normalized to the daily dietary ion intake to eliminate the influence of fluctuations in food intake.

[0082] Urine ion analysis by ion chromatography

[0083] The sodium and phosphorus content of urine samples was analyzed on an ion chromatography system coupled with a conductivity detector.

[0084] The chromatographic separation of cations was performed using an IonPacCS12A (Thermo Fisher) 2×250 mm analytical column with isocratic elution using 25 mM methanesulfonic acid.

[0085] Anions were separated by chromatography using an IonPac AS18 (Thermo Fisher) 2×250 mm analytical column with isocratic elution using 35 mM potassium hydroxide.

[0086] Concentrations were extrapolated from the standard curves (prepared in 10 mM hydrochloric acid) for each analyte ion based on retention time and peak area.

[0087] Drug solutions and food preparations

[0088] An appropriate weight of tenapanol was added to 0.1% Tween 80 to prepare a fresh dosing solution containing a concentration of 0.015 mg / ml weekly. The dosing volume for all groups was 5 ml / kg.

[0089] Weigh out standard phosphate-fortified rodent feed and place it in a mixing bowl. Add an appropriate amount of bisalom (0%, 0.5%, 1%, or 2% w / w bisalom) or seviram (0.75%, 1.5%, or 3% w / w seviram) to the powdered feed in the mixing bowl. Place the bowl on a vertical mixer and mix for 10 minutes, then granulate.

[0090] Dosage

[0091] Tenapano was administered orally to animals twice daily (Bid) via a standard tube feeding syringe (38mm, 20G) between 7:00 AM and 2:00 PM.

[0092] Animals were removed from their maintenance or metabolism cages for drug administration and then immediately returned to their source cages. Rats had free access to a diet containing bisalom and sevelamer.

[0093] Statistical analysis

[0094] All continuous data in each group are expressed as mean ± standard deviation. One-way ANOVA was used to compare the experimental groups, followed by Tukey post-hoc test for corrected multiple comparisons and pairwise comparisons between groups.

[0095] Example 2 Experimental Results

[0096] Animal in vivo experimental results as follows Figure 3 As shown.

[0097] Figure 3 The observations and statistical results of 24-hour urinary phosphorus excretion in rats grouped as described in Example 1 during the last four days of administration (i.e., days 8-11 post-administration). The ordinate shows the urinary phosphorus concentration as the average of the 24-hour urinary phosphorus concentrations detected over four days for each group of animals.

[0098] The results showed that, compared with the mediator control, terapranol alone significantly reduced urinary phosphorus excretion; bisalome significantly and dose-dependently reduced urinary phosphorus excretion.

[0099] The results also showed that when sevelamer was combined with tranapranol, sevelamer dose-dependently reduced urinary phosphorus excretion. The reduction was significantly greater in the combination at all sevelamer dose levels than in the tranapranol alone group or in bisalomer monotherapy at different dose levels.

[0100] The results also showed that when bisalom was combined with tranapano, bisalom also reduced urinary phosphorus excretion in a dose-dependent manner.

[0101] Combination therapy at all bisalome dosage levels was significantly superior to monotherapy with tranapano and monotherapy with bisalome at all dosage levels. In comparisons of low, medium, and high sevelamer dosages (0.75%, 1.5%, and 3%), the urinary phosphorus excretion of the low, medium, and high dose (0.5%, 1%, and 2%) bisalome combined with tranapano was significantly greater than that of the corresponding combination therapy.

[0102] in conclusion

[0103] The foregoing description of the present invention should not be construed as limiting it. Unless otherwise indicated, the present invention will be practiced using conventional techniques such as organic chemistry, polymer chemistry, and biotechnology, and it is obvious that the invention can be implemented in other ways besides those specifically described in the foregoing description and examples. Other aspects and modifications within the scope of the invention will be apparent to those skilled in the art. Many changes and variations are possible based on the teachings of the present invention, and therefore fall within the scope of the invention.

[0104] Unless otherwise specified, the temperature unit "degree" in this article refers to Celsius, or °C.

Claims

1. Use of the following components in combination for the manufacture of a medicament for lowering serum phosphate levels in mammals and humans: component (a) a phosphate binder; and component (b) a phosphate transport inhibitor.

2. Use according to claim 1, wherein the therapeutically effective amount of at least one of component (a) and component (b) administered to the subject is lower than the therapeutically effective amount of the component administered in the absence of the other component; Preferably, the therapeutically effective amount of both component (a) and component (b) administered to the subject is lower than the therapeutically effective amount of the component administered in the absence of the other component.

3. Use according to claim 2, wherein the therapeutically effective amount of at least one of component (a) and component (b), particularly both, administered to the subject is at least 10%, preferably at least 25%, more preferably at least 50% lower than the therapeutically effective amount of the component administered in the absence of the other component, for example, wherein the therapeutically effective amount of at least one of component (a) and component (b), preferably both, administered to the subject is from 10% to 90%, preferably from 25% to 90%, more preferably from 50% to 90% lower than the therapeutically effective amount of the component administered in the absence of the other component.

4. Use according to any one of claims 1 to 3, wherein the phosphate binder is a metal salt-based phosphate binder (such as lanthanum carbonate, calcium carbonate, calcium acetate, calcium acetate / magnesium carbonate, ferric citrate, super-iron oxyhydroxide, magnesium iron bicarbonate, aluminium hydroxide, SBR-759 and PA-21), a phosphate-binding organic polymer with anion-exchanger function (such as sevelamer carbonate (including sevelamer carbonate), AMG 223 and MCI-196), an anion-binding polymer (such as colestipol), and an anion-binding polymer containing cross-linked polyamines (such as bismuth subsalicylate).

5. Use according to any one of claims 1 to 3, wherein the phosphate transport inhibitor is an inhibitor of NHE3-mediated sodium and hydrogen ion antiport, for example tenapanor or tenapanor hydrochloride.

6. Use according to any one of claims 1 to 3, the components being: component (a) bismuth subsalicylate or a pharmaceutically acceptable salt thereof; and component (b) tenapanor or tenapanor hydrochloride, or the components being: component (a) sevelamer or a pharmaceutically acceptable salt thereof; and component (b) tenapanor or tenapanor hydrochloride.

7. Use of the following components in combination for the manufacture of a medicament for the treatment of a high serum phosphate-related disease in mammals and humans: component (a) a phosphate binder; and component (b) a phosphate transport inhibitor, wherein the phosphate binder or phosphate transport inhibitor being as defined in claims 1 to 6.

8. Use according to claim 7, wherein the high serum phosphate-related disease is hyperphosphatemia, hypernatremia, water and sodium retention, edema, hypocalcemia, secondary hyperparathyroidism, progressive metabolic bone disease, tissue and vascular calcification, cardiovascular disease such as myocardial infarction and / or stroke, nephropathy, proteinuria, hyperparathyroidism, Preferably, wherein the high serum phosphate related disease is hyperphosphatemia.

9. A pharmaceutical composition for treating a high serum phosphate related disease, comprising the following components: Component (a) a phosphate binder; and Component (b) a phosphate transport inhibitor, wherein said phosphate binder or phosphate transport inhibitor being as defined in claims 1 to 6.

10. The pharmaceutical composition according to claim 9, wherein the high serum phosphate related disease is hyperphosphatemia, hypernatremia, water sodium retention, edema, hypocalcemia, secondary hyperparathyroidism, progressive metabolic bone disease, tissue and vascular calcification, cardiovascular disease such as myocardial infarction and / or stroke, nephropathy, proteinuria, hyperparathyroidism.

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