Ammonium salt polymer, preparation method thereof and application of ammonium salt polymer as bile acid chelating agent

Abstract

The invention discloses an ammonium salt polymer, a preparation method thereof and application of the ammonium salt polymer as a bile acid chelating agent. The ammonium salt polymer is obtained by polymerizing a monomer of a formula (I) and a monomer of a formula (II), m, n, p, q and r are independently selected from 1, 2 or 3, the ammonium salt polymer contains monovalent, divalent or trivalent anions as counter ions, and the ammonium salt polymer can be used for treating or preventing dyslipidemia (such as hyperlipidemia), cholestasis, bile acid diarrhea and other diseases. # imgabs0 #

Description

Ammonium salt polymer, preparation method thereof and use as bile acid sequestrant Technical Field

[0001] The present invention belongs to the field of polymer pharmaceuticals. Specifically, the present invention relates to ammonium salt polymers, methods for their preparation, and their use as bile acid sequestrants. The ammonium salt polymers can be used to treat or prevent diseases such as dyslipidemia (e.g., hyperlipidemia), cholestatic liver diseases, and bile acid diarrhea (BAD).

[0002] Background of the Invention

[0003] Bile acid is an important component of bile. The physiological functions of bile acid mainly include promoting the digestion and absorption of lipids, inhibiting the precipitation of cholesterol in bile (stones), etc. It also participates in glucose metabolism and lipid metabolism. Therefore, bile acid is associated with a variety of diseases, such as dyslipidemia (such as hyperlipidemia), cholestatic liver disease, bile acid diarrhea, etc. 1. Dyslipidemia

[0004] Dyslipidemia refers to abnormal lipoprotein metabolism in the human body. Aside from a few secondary cases caused by systemic diseases, the vast majority are primary cases, resulting from genetic defects or interactions with environmental factors. The incidence of dyslipidemia is high in my country and is on the rise. This is closely related to the significant improvement in living standards and changes in dietary habits among Chinese people.

[0005] Hyperlipidemia, also known as hyperlipoproteinemia, is the most common form of dyslipidemia. It refers to a condition characterized by abnormally elevated levels of any one or all lipids and / or lipoproteins in the blood, including elevated total cholesterol (TC), elevated triglycerides (TG), elevated low-density lipoprotein cholesterol (LDL-c), or decreased high-density lipoprotein cholesterol (HDL-c). Clinically, plasma cholesterol levels exceeding 230 mg / 100 ml and triglycerides exceeding 140 mg / 100 ml are collectively referred to as hyperlipidemia.

[0006] Hyperlipidemia causes insidious, progressive, and systemic damage to the body, bringing with it numerous problems. When blood lipids remain elevated for a long time, they and their breakdown products gradually deposit on blood vessel walls, accompanied by the formation of fibrous tissue. This narrows the blood vessels, reduces their elasticity, and can ultimately lead to blockage. If lipid plaques deposit in the coronary arteries, they can cause ischemic symptoms in the heart and induce coronary heart disease; if they deposit in the brain, they can induce cerebral infarction; if they deposit in the lower limbs, they can cause pain and even necrosis; if they deposit in the blood vessels of the eyes, they can cause blindness; and if they deposit in the liver, they can cause fatty liver disease. Hyperlipidemia is a recognized risk factor for stroke, coronary heart disease, myocardial infarction, and sudden death. Hyperlipidemia can also trigger acute pancreatitis.

[0007] Given the adverse consequences of hyperlipidemia, patients need to be treated with lipid-lowering drugs. The mechanisms of action of these drugs primarily include inhibiting cholesterol biosynthesis, reducing cholesterol levels in the body, increasing HDL, and inhibiting cholesterol absorption. For example, statins (such as lovastatin, simvastatin, atorvastatin, and rosuvastatin) inhibit hydroxymethylglutaryl-CoA reductase, the rate-limiting enzyme in cholesterol biosynthesis, thereby reducing cholesterol biosynthesis in the body. Phenoxyacetic acid lipid-lowering drugs (such as clofibrate and fenofibrate) also interfere with cholesterol synthesis in the body, significantly reducing triglycerides and total cholesterol. Nicotinic acid analogs (such as acipimox) can increase HDL. Bile acid-binding resins (such as cholestyramine and coleseelam hydrochloride) can reduce cholesterol levels in the body.

[0008] Among the above-mentioned lipid-lowering drugs, bile acid-binding resin is a relatively safe oral medication with a good lipid-lowering effect. Bile acid is an important component of bile and plays an important role in fat metabolism. During the human digestive process, cholesterol is the only precursor of bile acid. Bile acid produced by the breakdown of cholesterol is secreted into the intestines, which absorb large amounts of bile acid and then re-enter the liver through the enterohepatic circulation. Bile acid-binding resin is a polymer that cannot be absorbed by the human body. It has a high adsorption capacity for bile acid, thereby excreting it from the body and preventing its reabsorption. The large consumption of bile acid will promote the conversion of more cholesterol into bile acid, thereby achieving the purpose of lowering cholesterol in the body.

[0009] Colesevelam hydrochloride is the latest representative of this class of drugs. Its chemical name is N,N,N-trimethyl-6-(2-propenylamino)-1-hexylamine copolymer hydrochloride, and its chemical structure is as follows:

[0010]

[0011] Colesevelam hydrochloride not only lowers cholesterol levels in the body but also increases the clearance of low-density lipoprotein (LDL) from the blood. Colesevelam hydrochloride, marketed under the trade name WELCHOL, is approved as a bile acid sequestrant for the treatment of primary hyperlipidemia and type 2 diabetes.

[0012] Aging is a major challenge currently facing all countries, accompanied by a continuous increase in the number of patients with hyperlipidemia, so more lipid-lowering drugs are needed.

[0013] 2. Cholestatic liver disease

[0014] Cholestasis refers to a pathological state in which bile formation, secretion, and excretion are impaired due to various factors inside and outside the liver, resulting in the inability of bile to flow normally into the duodenum and enter the blood. Clinically, it can manifest as itching, fatigue, darker urine, and jaundice. In the early stages, there are often no symptoms, only elevated serum ALP and GGT levels. As the disease progresses, hyperbilirubinemia may occur, and severe cases can lead to liver failure or even death.

[0015] Hepatobiliary diseases with cholestasis as their primary manifestation due to various causes are generally referred to as cholestatic liver diseases. The diagnostic criteria for cholestatic liver diseases generally follow the 2009 European Association for the Study of the Liver (EASL) Clinical Practice Guidelines for the Management of Cholestatic Liver Diseases: "ALP exceeding 1.5×ULN and GGT exceeding 3×ULN." However, it should be noted that in some specific cholestatic liver diseases, such as PFIC types 1 and 2 and BRIC, GGT may not be elevated.

[0016] Cholestatic liver diseases can be divided into hereditary and acquired cholestatic liver diseases. Hereditary cholestatic liver diseases include progressive familial intrahepatic cholestasis (PFIC), benign recurrent intrahepatic cholestasis (BIRC), congenital bile acid synthesis disorder, Alagille syndrome (ALGS), neonatal cholestasis caused by citrin protein deficiency, and ARC syndrome. Acquired cholestatic liver diseases are often caused by factors such as infection, drugs, hormones, pregnancy, tumors, and biliary obstruction. They include primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC), autoimmune hepatitis (AIH), drug-induced cholestasis (DRIC), biliary atresia (BA), and intrahepatic cholestasis of pregnancy (ICP).

[0017] Currently, the primary treatment for cholestatic liver disease is ursodeoxycholic acid (UDCA), which has been shown to reduce cholestasis in various cholestatic diseases and is the standard first-line treatment for PBC. However, it is ineffective for most pruritus and is only effective in approximately 50% of PBC patients. Nearly 40% of PBC patients have an inadequate response to UDCA. In addition, 5% to 10% of PBC patients are intolerant to UDCA. For PBC patients who have an inadequate response or intolerance to UDCA, only obeticholic acid (OCA) is available. Obeticholic acid is a farnesoid X receptor (FXR) agonist with side effects such as severe skin itching, fatigue, abdominal pain and discomfort, and rash, which limits its use.

[0018] Maralixibat, an apical sodium-dependent bile acid transporter (ASBT) inhibitor, is approved for the treatment of pruritus in children 1 year of age and older with ALGS, but is associated with adverse reactions such as diarrhea, abdominal pain, vomiting, fat-soluble vitamin deficiencies, abnormal liver tests, gastrointestinal bleeding, and fractures. Odevixibat, an ileal bile acid transporter (IBAT) inhibitor approved for the treatment of progressive familial intrahepatic cholestasis (PFIC), is associated with adverse reactions such as abnormal liver tests, diarrhea, abdominal pain, vomiting, and fat-soluble vitamin deficiencies.

[0019] With the significant increase in the incidence of cholestatic liver disease in recent years, new and effective therapeutic agents are urgently needed. Currently, the peroxisome proliferator-activated receptor (PPAR) agonists elafibranor (GFT505) and seladelpar (MBX-8025) are undergoing clinical trials and are expected to become new clinical treatments for cholestatic liver disease. However, preliminary clinical trial results indicate that these drugs also have relatively serious side effects.

[0020] In contrast, bile acid binding resin is not absorbed by the intestine and is therefore a safer oral drug that can adsorb bile acids in the intestine, reduce bile acid reabsorption, accelerate bile acid excretion, and alleviate cholestasis, thereby showing a therapeutic effect on cholestatic liver disease.

[0021] 3. Bile acid diarrhea

[0022] Bile acid diarrhea (BAD), also known as bile acid malabsorption (BAM), is a chronic diarrhea caused by abnormal bile acid metabolism, which leads to excessive bile acids in the large intestine. This irritates the intestinal mucosa, increasing permeability, intestinal fluid secretion, and intestinal motility. In 1967, Hofmann of the Mayo Clinic first proposed the concepts of "biliary diarrhea" and "biliary enteropathy." Later, people divided bile acid-related diarrhea into three types according to the cause: Type I is bile acid absorption disorder caused by ileal diseases such as Crohn's disease and ileal resection; Type II is idiopathic or primary BAD, the cause of which is unknown. It is worth noting that 25%-50% of irritable bowel syndrome with predominant diarrhea (IBS-D) with diarrhea as the main feature show a certain degree of BAM; Type III is bile acid absorption disorder caused by other gastrointestinal diseases, including cholecystectomy, small intestinal bacterial overgrowth, microscopic colitis, etc.

[0023] Depending on the severity of malabsorption, large amounts of bile acids can enter the colon through the ileum, resulting in several pathogenic effects. Bile acids, particularly hydrophobic bile acids, are cytotoxic due to their detergent properties. Physiological concentrations of colonic bile acids have been shown to stimulate propagating motility waves, accelerate colonic transit, and enhance the urge to defecate. Excessive bile acids also induce chloride secretion from colonocytes, leading to secretory diarrhea, a cardinal feature of BAD.

[0024] Currently recommended treatments for BAD include bile acid sequestrants, FXR agonists, loperamide, and a low-fat diet. The first-generation bile acid sequestrant, cholestyramine, is a common treatment for BAD, with numerous clinical trials demonstrating an efficacy of approximately 70%. However, its unpleasant taste leads to poor medication compliance. The next-generation bile acid sequestrant, colesevelam hydrochloride, launched in 2000, has been used clinically as both an initial and second-line treatment for BAD. However, additional bile acid sequestrants are still needed to provide patients with more treatment options.

[0025] The present invention aims to address the above-mentioned unmet market needs.

[0026] Summary of the Invention

[0027] In a first aspect, the present invention provides an ammonium salt polymer obtained by polymerizing a monomer of formula (I) and a monomer of formula (II),

[0028]

[0029] in:

[0030] m, n, p, q, and r are each independently selected from 1, 2, or 3.

[0031] The ammonium salt polymer contains monovalent, divalent or trivalent anions as counter ions, and the anions can be derived from organic acids or inorganic acids. The anions derived from inorganic acids include, but are not limited to: monovalent anions, which are preferably selected from chloride ions (Cl - ), bicarbonate ion (HCO3 - ), hydrogen sulfate ion (HSO4 - ), acetate ion (C2H3O2 - ); divalent anions, which are preferably selected from carbonate ions (CO3 2- ), sulfate ion (SO4 2- ); a trivalent anion, which is preferably a phosphate ion (PO4 3- ); the anion derived from an organic acid may be derived from malic acid, citric acid, fumaric acid, maleic acid, etc.

[0032] Those skilled in the art will understand that the ammonium cations and anions contained in the polymer are counter ions to each other.

[0033] In one embodiment of the first aspect, the ammonium salt polymer is obtained by polymerizing a monomer of formula (I) and a monomer of formula (II) in a molar ratio of 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, further preferably 10:1 to 10:4, and most preferably 10:2.

[0034] In one embodiment of the first aspect, in the monomer of formula (I), m and n are each independently selected from 1 and 2; preferably, m and n are both 1.

[0035] In one embodiment of the first aspect, in the monomer of formula (II), p, q and r are each independently selected from 1 and 2; preferably, p, q and r are all 1.

[0036] The glass transition temperature of the ammonium salt polymer of the present invention is from about 60°C to about 130°C, preferably from about 70°C to 120°C, more preferably from about 70°C to about 105°C.

[0037] The infrared spectrum of the ammonium salt polymer of the present invention preferably has the following absorption peaks (cm -1 ):2918,2855,2359,2072,1627,1529,1402,1362,1200,1000,909,810. More preferably, the infrared spectrum of the ammonium salt polymer of the present invention is shown in Figure 1.

[0038] The ammonium salt polymer of the present invention 13The C solid nuclear magnetic resonance spectrum preferably has the following peaks (ppm): 28.2, 42.9, 50.5, 163.3. More preferably, the ammonium salt polymer of the present invention 13 The solid-state NMR spectrum of C is shown in Figure 2.

[0039] The ammonium salt polymer of the present invention can adsorb bile acid in vitro and in vivo. For example, the ammonium salt polymer of the present invention has an in vitro bile acid adsorption capacity of 7-10 mmol / g, more preferably 7-8 mmol / g.

[0040] In a second aspect, the present invention provides a method for preparing the ammonium salt polymer of the present invention, which comprises the following steps: (1) acidifying the monomer of formula (I) and the monomer of formula (II) with an acid; and (2) polymerizing the acidified monomer obtained in step 1 to obtain the ammonium salt polymer of the present invention, as shown in General Scheme 1.

[0041] General Solution 1

[0042]

[0043] where X - represents the monovalent, divalent or trivalent anion mentioned above as a counter ion for ammonium.

[0044] The acidification in step 1 can be carried out as follows: a monomer of formula (I) and a monomer of formula (II) in a molar ratio of 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, more preferably 10:1 to 10:4, and most preferably 10:2 are dropwise added to a suitable acid at a temperature of 30° C. or lower, preferably 15° C. or lower. After the dropwise addition is completed, the mixture is stirred for a period of time at a temperature of 5-30° C., preferably 10-20° C., to obtain a mixture of the two acidified monomers.

[0045] The suitable acid used in step 1 can be an inorganic acid or an organic acid, such as hydrochloric acid, sulfuric acid, acetic acid, malic acid, citric acid, fumaric acid, maleic acid, etc.

[0046] The polymerization reaction of step 2 can be carried out as follows:

[0047] 2.1. Dissolve the initiator in water and add it to the acidified monomer mixture obtained in step 1 to obtain an aqueous mixture;

[0048] 2.2. In a separate container, dissolve the emulsifier in an organic solvent to obtain an oil phase mixture, wherein the organic solvent is immiscible with water and has a boiling point higher than the polymerization temperature, and the organic solvent is preferably selected from toluene, n-heptane, cyclohexane, xylene, n-octane, and any mixture thereof;

[0049] 2.3. adding the aqueous phase mixture to the oil phase mixture to carry out a polymerization reaction to produce the desired ammonium salt polymer; preferably, the polymerization reaction is carried out at a temperature of, for example, 40-100 ° C, preferably 50-85 ° C;

[0050] 2.4. The reactants from step 2.3 are subjected to conventional post-treatment to isolate the ammonium salt polymer of the present invention. For example, the post-treatment can be performed by filtering the reactants, drying, and pulverizing to obtain the ammonium salt polymer of the present invention. Preferably, the post-treatment can be performed by filtering the reactants, slurrying the wet cake once with methanol or ethanol, filtering, slurrying the wet cake again with purified water and filtering, repeating this process 2-3 times, drying, and pulverizing to obtain the ammonium salt polymer of the present invention.

[0051] Optionally, step 2 may further comprise:

[0052] 2.5. Convert the wet cake in step 2.4 or the ammonium salt polymer separated in step 2.4 into the ammonium salt polymer of the present invention containing other anions.

[0053] Step 2.5 can be carried out by conventional methods known in the art. For example, when the acid used in step 1 is hydrochloric acid, in the ammonium salt polymer generated in step 2.3, the anion X - is chloride ion. It can be converted into ammonium salt polymer containing other anions by step 2.5. For example, it can be converted into bicarbonate and / or carbonate by the following steps: the wet cake in step 2.4 or the ammonium salt polymer separated in step 2.4 is added to a saturated sodium bicarbonate aqueous solution or a sodium carbonate aqueous solution, stirred for a period of time, filtered, dried, and crushed to obtain an anion X - It is an ammonium salt polymer of bicarbonate and / or carbonate.

[0054] The initiator used in step 2.1 is not particularly limited and can be any suitable initiator. For example, the initiator can be selected from 2,2'-azobisisobutylamidine dihydrochloride (V-50), 2,2'-azabis(2-imidazoline) dihydrochloride (VA-044), ammonium persulfate, potassium persulfate, hydrogen peroxide, and any mixture thereof.

[0055] The emulsifier used in step 2.2 is not particularly limited and can be any substance having emulsifying properties that facilitates mixing of the aqueous phase and the oil phase, such as a surfactant. For example, the emulsifier can be selected from polyoxyethylene sorbitan fatty acid esters (e.g., Tween 40, Tween 80, etc.) or sorbitan fatty acid esters (e.g., Span 40, Span 60, Span 80, etc.).

[0056] In a third aspect, the present invention provides a pharmaceutical composition comprising the ammonium salt polymer of the present invention and optionally a pharmaceutically acceptable carrier.

[0057] The pharmaceutical composition can be used as a bile acid sequestrant, and in particular can be used to treat or prevent diseases associated with excessive bile acid.

[0058] The pharmaceutical composition can be formulated into solid preparations (including but not limited to capsules, tablets, pills, granules, powders) or liquid preparations (including but not limited to suspensions) for oral administration by methods well known in the art.

[0059] For oral preparations, the pharmaceutically acceptable carrier may be:

[0060] a) fillers, such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose;

[0061] b) lubricants, for example silicon dioxide, talc, stearic acid, magnesium or calcium stearate and / or polyethylene glycol;

[0062] c) binders, for example magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidone;

[0063] d) disintegrants, for example starches, agar, alginic acid or its sodium salt, or effervescent mixtures; and / or

[0064] e) coloring and / or flavoring agents;

[0065] f) Suspending agents.

[0066] In a fourth aspect, the present invention provides use of the ammonium salt polymer of the present invention in preparing a medicament for use as a bile acid sequestrant and use of the ammonium salt polymer of the present invention in preparing a medicament for treating or preventing diseases associated with excessive bile acid.

[0067] In a fifth aspect, the present invention provides a method for treating or preventing a disease associated with excess bile acid, the method comprising administering an effective amount of the ammonium salt polymer of the present invention to an individual in need thereof.

[0068] The diseases associated with excessive bile acids described herein are selected from dyslipidemia (e.g., hyperlipidemia), cholestatic liver diseases (e.g., progressive familial intrahepatic cholestasis (PFIC), benign recurrent intrahepatic cholestasis (BIRC), congenital bile acid synthesis disorders, Alagille syndrome (ALGS), neonatal cholestasis caused by Citrin protein deficiency, ARC syndrome, primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC), autoimmune hepatitis (AIH), drug-induced cholestasis (DRIC), biliary atresia (BA), and intrahepatic cholestasis of pregnancy (ICP)), bile acid diarrhea, and diabetes.

[0069] The inventors surprisingly discovered that the ammonium salt polymer of the present invention has excellent bile acid adsorption, significantly better than colesevelam hydrochloride. Moreover, the in vivo and in vitro effects are well consistent.

[0070] The ammonium salt polymer of the present invention is not absorbed into the bloodstream, but adsorbs bile acids in the intestines and is excreted from the body in feces. This effectively reduces intestinal bile acid reabsorption, improves bile stasis inside and outside the liver, inhibits the progression of cholestatic liver diseases such as PSC, and alleviates complications such as itching. Due to its non-systemic exposure, the ammonium salt polymer of the present invention avoids the toxic side effects of small molecules such as odevixibat that enter the bloodstream, exhibiting a good safety profile, long-term use, and good compliance.

[0071] Furthermore, the preparation method of the ammonium salt polymer of the present invention is simple. Specific implementation plan

[0072] Embodiment 1. An ammonium salt polymer obtained by polymerizing a monomer of formula (I) and a monomer of formula (II),

[0073]

[0074] in:

[0075] m, n, p, q, r are each independently selected from 1, 2 or 3,

[0076] The ammonium salt polymer contains monovalent, divalent or trivalent anions as counter ions, and the anions are selected from chloride ions (Cl - ), bicarbonate ion (HCO3 - ), hydrogen sulfate ion (HSO4 - ), acetate ion (C2H3O2 - ), carbonate ions (CO3 2- ), sulfate ion (SO4 2- ), phosphate ions (PO4 3-) and anions derived from malic acid, citric acid, fumaric acid, and maleic acid.

[0077] Embodiment 2. The polymer of embodiment 1, wherein m, n, p, q and r are each independently selected from 1 and 2, preferably wherein m, n, p, q and r are all 1.

[0078] Embodiment 3. The polymer according to embodiment 1 or 2, wherein the ammonium salt polymer is obtained by polymerizing a monomer of formula (I) and a monomer of formula (II) in a molar ratio of 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, more preferably 10:1 to 10:4, and most preferably 10:2.

[0079] Embodiment 4. The polymer of any one of embodiments 1 to 3, wherein the polymer has a glass transition temperature of about 60°C to about 130°C, preferably about 70°C to 120°C, and more preferably about 70°C to about 105°C.

[0080] Embodiment 5. The polymer according to any one of embodiments 1 to 4, wherein the anion contained in the polymer is a bicarbonate ion (HCO3 - ) and / or carbonate ions (CO3 2- ), and wherein m, n, p, q and r are all 1.

[0081] Embodiment 6. The polymer according to any one of embodiments 1 to 5, wherein the polymer has an in vitro bile acid adsorption capacity of 7-10 mmol / g, preferably 7-8 mmol / g.

[0082] Embodiment 7. A method for preparing the polymer of any one of embodiments 1 to 6, comprising the steps of:

[0083] (1) acidifying the monomer of formula (I) and the monomer of formula (II) with an acid, preferably, the molar ratio of the monomer of formula (I) to the monomer of formula (II) is 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, further preferably 10:1 to 10:4, and most preferably 10:2;

[0084] (2) polymerizing the acidified monomer obtained in step (1) to obtain the desired ammonium salt polymer.

[0085] Embodiment 8. The method according to embodiment 7, wherein step (1) is carried out as follows: at a temperature of 30°C or lower, preferably at a temperature of 15°C or lower, a monomer of formula (I) and a monomer of formula (II) in a molar ratio of 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, further preferably 10:1 to 10:4, and most preferably 10:2 are added dropwise to a suitable acid; after completion of the addition, the mixture is stirred at a temperature of 5-30°C, preferably at a temperature of 10-20°C, for a period of time to obtain a mixture of the two acidified monomers.

[0086] Embodiment 9. The method according to embodiment 7 or 8, wherein the acid in step (1) is selected from hydrochloric acid, sulfuric acid, acetic acid, malic acid, citric acid, maleic acid and fumaric acid.

[0087] Embodiment 10. The method according to any one of embodiments 7 to 9, wherein step (2) comprises the following sub-steps:

[0088] (2.1) dissolving an initiator in water and adding the mixture to the acidified monomers obtained in step (1) to obtain an aqueous phase mixture;

[0089] (2.2) in another container, dissolving the emulsifier in an organic solvent to obtain an oil phase mixture, wherein the organic solvent is immiscible with water and has a boiling point higher than the polymerization temperature, and the organic solvent is preferably selected from toluene, n-heptane, cyclohexane, xylene, n-octane, and any mixture thereof;

[0090] (2.3) adding the aqueous phase mixture to the oil phase mixture to carry out a polymerization reaction to generate the desired ammonium salt polymer;

[0091] (2.4) performing conventional post-treatment on the reactants of step (2.3) to isolate and obtain the desired ammonium salt polymer; for example, the conventional post-treatment can be performed as follows: filtering the reactants, drying, and crushing to obtain the ammonium salt polymer of the present invention; preferably, the conventional post-treatment can be performed as follows: filtering the reactants, slurrying the wet cake with methanol or ethanol once, filtering, slurrying the wet cake with purified water and filtering, repeating this process 2-3 times, drying, and crushing to obtain the desired ammonium salt polymer;

[0092] Optionally,

[0093] (2.5) converting the wet cake in step (2.4) or the ammonium salt polymer separated in step (2.4) into an ammonium salt polymer containing other anions.

[0094] Embodiment 11. A method for preparing the ammonium salt polymer of embodiment 5, the method comprising the steps of:

[0095] (1) acidifying a monomer of formula (I) and a monomer of formula (II) in a molar ratio of 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, further preferably 10:1 to 10:4, and most preferably 10:2 with concentrated hydrochloric acid to obtain a mixture of the two acidified monomers;

[0096] (2.1) dissolving an initiator in water and adding the mixture to the acidified monomers obtained in step (1) to obtain an aqueous phase mixture;

[0097] (2.2) in another container, dissolving an emulsifier in an organic solvent to obtain an oil phase mixture, wherein the organic solvent is immiscible with water and has a boiling point higher than the polymerization temperature, and the organic solvent is selected from toluene, n-heptane, cyclohexane, xylene, n-octane, and any mixture thereof;

[0098] (2.3) adding the aqueous phase mixture to the oil phase mixture, and performing a polymerization reaction at a temperature of 40-100° C., preferably 50-85° C., to generate the desired ammonium salt polymer;

[0099] (2.4) Filter the reactant from step (2.3), slurry the wet cake with methanol or ethanol once, filter, and then slurry the wet cake with purified water and filter, repeating 2-3 times to obtain a wet cake;

[0100] (2.5) adding the wet filter cake obtained in step (2.4) to a saturated aqueous sodium bicarbonate solution and / or an aqueous sodium carbonate solution, stirring for a period of time, filtering, drying, and crushing to obtain an ammonium salt polymer containing bicarbonate ions and / or carbonate ions.

[0101] Embodiment 12. The method according to any one of embodiments 7 to 11, wherein the initiator is selected from 2,2'-azobisisobutylamidine dihydrochloride (V-50), 2,2'-azabis(2-imidazoline) dihydrochloride (VA-044), ammonium persulfate, potassium persulfate, hydrogen peroxide and any mixture thereof; and the emulsifier is selected from polyoxyethylene sorbitan fatty acid esters (such as Tween 40, Tween 80, etc.), sorbitan fatty acid esters (such as Span 40, Span 60, Span 80, etc.) and any mixture thereof.

[0102] Embodiment 13. The ammonium salt polymer according to any one of embodiments 1 to 6, for use as a medicament, preferably as a medicament for treating or preventing diseases associated with excessive bile acids, preferably the diseases associated with excessive bile acids are selected from dyslipidemia (e.g., hyperlipidemia), cholestatic liver diseases (e.g., progressive familial intrahepatic cholestasis (PFIC), benign recurrent intrahepatic cholestasis (BIRC), congenital bile acid synthesis disorders, Alagille syndrome (ALGS), neonatal cholestasis caused by Citrin protein deficiency, ARC syndrome, primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC), autoimmune hepatitis (AIH), drug-induced cholestasis (DRIC), biliary atresia (BA) and intrahepatic cholestasis of pregnancy (ICP)), bile acid diarrhea and diabetes.

[0103] Embodiment 14. A pharmaceutical composition comprising the polymer of any one of embodiments 1 to 6, and optionally comprising a pharmaceutically acceptable carrier.

[0104] Embodiment 15. Use of the polymer of any one of Embodiments 1 to 6 for use as a bile acid sequestrant or for the preparation of a medicament for treating or preventing a disease associated with excessive bile acid, wherein the disease is preferably selected from dyslipidemia (e.g., hyperlipidemia), cholestatic liver disease (e.g., progressive familial intrahepatic cholestasis (PFIC), benign recurrent intrahepatic cholestasis (BIRC), congenital bile acid synthesis disorder, Alagille syndrome (ALGS), neonatal cholestasis caused by Citrin protein deficiency, ARC syndrome, primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC), autoimmune hepatitis (AIH), drug-induced cholestasis (DRIC), biliary atresia (BA) and intrahepatic cholestasis of pregnancy (ICP)), bile acid diarrhea and diabetes.

[0105] Embodiment 16. A method for treating or preventing a disease associated with excessive bile acid, the method comprising administering an effective amount of a polymer of any one of embodiments 1 to 6 to a subject in need thereof, wherein the disease is preferably selected from dyslipidemia (e.g., hyperlipidemia), cholestatic liver disease (e.g., progressive familial intrahepatic cholestasis (PFIC), benign recurrent intrahepatic cholestasis (BIRC), congenital bile acid synthesis disorder, Alagille syndrome (ALGS), neonatal cholestasis caused by Citrin protein deficiency, ARC syndrome, primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC), autoimmune hepatitis (AIH), drug-induced cholestasis (DRIC), biliary atresia (BA) and intrahepatic cholestasis of pregnancy (ICP)), bile acid diarrhea and diabetes.

[0106] Embodiment 17. The invention described above.

[0107] definition

[0108] As used herein, the term "subject" refers to an animal. Preferably, the subject is a mammal, such as a primate (e.g., human, monkey, or gorilla), cow, sheep, goat, horse, dog, cat, rabbit, rat, mouse, etc. More preferably, the subject is a human.

[0109] As used herein, the term "treat" refers to alleviating or curing a disease or its symptoms, or preventing the progression of a disease or its symptoms.

[0110] As used herein, the term "prevent" or "prevent" means to delay or prevent the onset of a disease or its symptoms.

[0111] As used herein, the term "effective amount" refers to an amount of the ammonium salt polymer of the present invention that significantly reduces bile acid levels in the body and prevents or ameliorates a disease or its symptoms associated with excessive bile acid levels. The "effective amount" of the ammonium salt polymer of the present invention depends on a variety of factors, including, but not limited to, the type and severity of the disease being treated or prevented, the patient's overall health and response, and the like. Typically, for a 70 kg human, the "effective amount" of the ammonium salt polymer of the present invention is 0.5-24 g / day. However, if necessary, the ammonium salt polymer of the present invention may be administered in amounts outside this dosage range. Clinicians can readily determine the effective amount for a specific patient.

[0112] As used herein, the term "about" means that the numerical value given thereafter can be expanded up and down by 20%, preferably up and down by 10%. For example, "about 100" means 80 to 120, preferably 90 to 110; "about 60°C" means 48°C to 72°C, preferably 54°C to 66°C.

[0113] Example

[0114] The following examples are illustrative of the present invention and are not to be construed as limiting the scope of protection of the present invention in any way. Disclosed data (e.g., amount, temperature, etc.) strive to ensure their accuracy, but there may be some experimental errors and offsets. Unless otherwise stated, temperature is degrees Celsius and pressure is atmospheric pressure or near atmospheric pressure. Unless otherwise stated, all reagents used in the examples are commercially available.

[0115] The abbreviations used in this document have the following meanings:

[0116] DAA Diallylamine

[0117] TAA Triallylamine

[0118] DAA-Cl Diallylammonium Chloride

[0119] TAA-Cl Triallyl ammonium chloride

[0120] DMDAAC Dimethyldiallyl ammonium chloride

[0121] BTHB 1-Bromo-6-(trimethylammonium)hexyl bromide

[0122] VA-044 2,2'-Azabis(2-imidazoline) dihydrochloride

[0123] V-50 2,2'-azobisisobutylamidine dihydrochloride

[0124] Span 60, Sorbitan Monostearate

[0125] RPM revolutions per minute

[0126] NaOH sodium hydroxide

[0127] NaHCO3 sodium bicarbonate

[0128] DDC 3,5-diethoxycarbonyl-1,4-dihydro-2,4,6-trimethylpyridine

[0129] TBA total bile acid

[0130] TBIL total bilirubin

[0131] Concentrated hydrochloric acid with a concentration of 36%-38%

[0132] ANIT α-naphthylisothiocyanate

[0133] GC gas chromatography

[0134] CMC-Na Carboxymethyl Cellulose Sodium

[0135] SDS Sodium dodecyl sulfate

[0136] Example 1

[0137] (Polymer WS012-A was prepared according to General Scheme 1)

[0138] Add 1.76 kg of concentrated hydrochloric acid to the reaction flask, controlling the internal temperature to not exceed 15°C. Add 1.46 kg (15 mol) of DAA and 0.41 kg (3 mol) of TAA dropwise to the reaction flask. After the addition is complete, stir at 10-20°C for approximately 0.5-1 hour. Add 0.146 kg of V50 and 1.46 kg of water to a beaker. Stir to dissolve. Add the solution from the reaction flask to the beaker and stir to mix thoroughly.

[0139] 8.76 kg of n-heptane and 0.133 kg of Span60 were added to the reactor and heated to 40-50 ° C while stirring to dissolve. The mechanical stirring speed was adjusted to 200-250 RPM, the contents of the beaker were added to the reactor, the temperature was raised to 50-55 ° C, and the reaction was stirred at 50-55 ° C for 15-24 hours. After GC detection of the reaction solution showed that the monomers DAA and TAA were basically completely converted, the reaction mixture was filtered. The wet cake was slurried with about 5.84 kg of ethanol for 1-2 hours and filtered. Then, the wet cake was slurried with about 18.0-20.0 kg of water for 1-2 hours and filtered. Repeat the slurrying and filtering operation 2-3 times. The wet cake was added to about 30.0 kg of saturated NaHCO3 aqueous solution, stirred at 25-35 ° C for 18-24 hours and filtered, and this step was repeated 2 times. The wet cake was slurried with about 18.0-20.0 kg of water for 1-2 hours and filtered. After repeating the beating and filtering operation 2-3 times, the wet cake was rinsed with purified water, dried under forced air at 45-55°C, crushed, and sieved to obtain 1.8 kg of polymer WS012-A as a light yellow solid.

[0140] Attempts were made to dissolve the prepared polymer using various conventional solvents such as water, methanol, N,N-dimethylformamide, dimethyl sulfoxide, etc., but it was found that none of them could dissolve the polymer.

[0141] The prepared polymer WS012-A was characterized using a SHIMADZU IRSpirit-T Fourier transform infrared spectrometer according to infrared spectrophotometry (General Rules 0402 of Part Four of the 2020 edition of the Chinese Pharmacopoeia). The infrared spectrum obtained is shown in Figure 1, which has the following characteristic absorption peaks (cm -1 ):2918,2855,2359,2072,1627,1529,1402,1362,1200,1000,909,810.

[0142] The infrared spectrum showed that the product contained structures such as methylene, tertiary ammonium salt, secondary ammonium salt, CN, and carbonate.

[0143] The polymer WS012-A prepared in this example was tested using a Jnm-600ECZ solid-state nuclear magnetic analyzer. The sample was filled into a 3.2 mm ZrO2 rotor. The experimental parameters were set to a magic angle spinning speed of 15 kHz, a cycle delay time of 5 seconds, 1024 scans at 293 K, and the chemical shift was measured using the glycine carbonyl at 176.04 ppm as an external standard. 13 The solid-state NMR spectrum of C is shown in FIG2 , which has the following peaks (ppm): 28.2, 42.9, 50.5, 163.3.

[0144] 13 C solid-state nuclear magnetic resonance spectrum showed that tertiary carbon, methylene carbon, CN and carbonic acid existed in this sample.

[0145] The prepared product WS012-A was measured by differential scanning calorimetry (DSC). Instrument model: METTLER TOLEDO DSC3 differential scanning calorimeter. Analysis method: N2 atmosphere, 50ml / min. Scanning procedure: Heat from 30°C to 180°C at 10°C / min, hold constant for 5 minutes, then cool to 30°C at 10°C / min, hold constant for 5 minutes, then heat to 180°C at 30°C / min, record the second heating curve, and analyze the curve to determine the characteristic temperature of the glass transition. An aluminum sample pan was used. The obtained DSC spectrum is shown in Figure 3. The results show that the glass transition temperature of polymer WS012-A is 99.5°C.

[0146] Example 2

[0147] (Alternative method for preparing polymer WS012-A according to general scheme 1)

[0148] Add 101.54 g of concentrated hydrochloric acid to reaction flask 1. Control the internal temperature to not exceed 15°C. Add 82.59 g (0.85 mol) of DAA and 23.33 g (0.17 mol) of TAA dropwise to reaction flask 1. After the addition is complete, stir at 10-20°C for approximately 0.5-1 hour. Add 5.93 g of VA-044 and 54.0 g of water to a conical flask. Stir to dissolve. Add the aqueous solution to reaction flask 1 and stir to mix thoroughly.

[0149] Add 580mL of toluene and 4.1g of Span60 to reaction flask 2 and heat to 40-50°C under stirring to dissolve. Adjust the mechanical stirring speed to 300-400RPM, add the material in reaction flask 1 to reaction flask 2, heat to 50-55°C, and stir the reaction at 50-55°C for 15-24 hours. After GC detection of the reaction solution shows that DAA and TAA are basically completely converted, the internal temperature of the reaction is raised to 80-85°C, and the reaction is stirred at 80-85°C for 1-2 hours, then cooled and filtered. After beating the wet cake with about 1L of water for 0.5-1 hour, filter. Repeat the beating and filtering operation 3-5 times. Add the wet cake to 500mL of 10% NaOH aqueous solution and stir at room temperature for about 8-12 hours. Then, filter the reaction mixture, and after beating the wet cake with about 1L of water for 0.5-1 hour, filter. The wet cake was added to approximately 1 L of saturated aqueous NaHCO₃ solution and stirred at 20-30°C for 12-18 hours, then filtered. This step was repeated 1-2 times. The wet cake was slurried with approximately 1 L of water for 0.5-1 hour, then filtered. This slurrying and filtering process was repeated 2-3 times, and the wet cake was rinsed with purified water. The filter cake was air-dried at 45-55°C, crushed, and sieved to yield 80 g of polymer WS012-A as a pale yellow solid.

[0150] The product was detected using the SHIMADZU IRSpirit-T Fourier transform infrared spectrometer and Jnm-600ECZ solid-state nuclear magnetic analyzer in Example 1. The results showed that the infrared spectrum and 13 The C solid-state NMR spectrum was identical to that of the polymer of Example 1 (spectrum not shown).

[0151] In addition, the solubility of the product of this example in various solvents was also measured. The results showed that, like the product of Example 1, the polymer was not soluble in any solvent.

[0152] Example 3

[0153] (Alternative method for preparing polymer WS012-A according to general scheme 1)

[0154] Add 101.54 g of concentrated hydrochloric acid to reaction flask 1. Control the internal temperature to not exceed 15°C. Add 82.59 g (0.85 mol) of DAA and 23.33 g (0.17 mol) of TAA dropwise to reaction flask 1. After the addition is complete, stir at 10-20°C for approximately 0.5-1 hour. Add 5.93 g of VA-044 and 54.0 g of water to a conical flask. Stir to dissolve. Add the aqueous solution to reaction flask 1 and stir to mix thoroughly.

[0155] Add 600mL of n-heptane and 8.36g of Span60 to reaction flask 2, and heat to 40-50°C while stirring to dissolve. Adjust the mechanical stirring speed to 300-400RPM, add the material in reaction flask 1 to reaction flask 2, heat to 50-55°C, and stir the reaction at 50-55°C for 15-24 hours. After GC detection of the reaction solution shows that DAA and TAA are basically completely converted, the reaction mixture is filtered. Slurry the wet cake with about 1L of methanol for 0.5-1 hour and filter. Then, slurry the wet cake with about 1L of water for 0.5-1 hour and filter. Repeat the slurrying and filtering operation 2-3 times. Add the wet cake to 500mL of 10% NaOH aqueous solution and stir at room temperature for about 8-12 hours. Then, filter the reaction mixture, and slurry the wet cake with about 1L of water for 0.5-1 hour and filter. The wet cake was added to approximately 1 L of saturated aqueous NaHCO₃ solution and stirred at 20-30°C for 12-18 hours, then filtered. This step was repeated 1-2 times. The wet cake was slurried with approximately 1 L of water for 0.5-1 hour, then filtered. This slurrying and filtering process was repeated 2-3 times, and the wet cake was rinsed with purified water. The filter cake was air-dried at 45-55°C, crushed, and sieved to yield 75 g of polymer WS012-A as a pale yellow solid.

[0156] The product was detected using the SHIMADZU IRSpirit-T Fourier transform infrared spectrometer and Jnm-600ECZ solid-state nuclear magnetic analyzer in Example 1. The results showed that the infrared spectrum and 13 The C solid-state NMR spectrum was identical to that of the polymer of Example 1 (spectrum not shown).

[0157] In addition, the solubility of the product of this example in various solvents was also measured. The results showed that, like the product of Example 1, the polymer was not soluble in any solvent.

[0158] Example 4

[0159] (Alternative method for preparing polymer WS012-A according to general scheme 1)

[0160] Add 101.54 g of concentrated hydrochloric acid to reaction flask 1. Control the internal temperature to not exceed 15°C. Add 82.59 g (0.85 mol) of DAA and 23.33 g (0.17 mol) of TAA dropwise to reaction flask 1. After the addition is complete, stir at 10-20°C for approximately 0.5-1 hour. Add 5.93 g of VA-044 and 54.0 g of water to a conical flask. Stir to dissolve. Add the aqueous solution to reaction flask 1 and stir to mix thoroughly.

[0161] 480mL of cyclohexane and 7.17g of Span60 were added to reaction flask 2 and heated to 40-50°C under stirring to dissolve. The mechanical stirring speed was adjusted to 300-400RPM, the material in reaction flask 1 was added to reaction flask 2, the temperature was raised to 50-55°C, and the reaction was stirred at 50-55°C for 15-24 hours. After GC detection of the reaction solution showed that DAA and TAA were basically completely converted, the reaction mixture was filtered. The wet cake was slurried with about 1L of methanol for 0.5-1 hour and filtered. Then, the wet cake was slurried with about 1L of water for 0.5-1 hour and filtered. Repeat the slurrying and filtering operation 2-3 times. The wet cake was added to 500mL of 10% NaOH aqueous solution and stirred at room temperature for about 8-12 hours. Then, the reaction mixture was filtered, and the wet cake was slurried with about 1L of water for 0.5-1 hour and filtered. The wet cake was added to approximately 1 L of saturated aqueous NaHCO₃ solution and stirred at 20-30°C for 12-18 hours, then filtered. This step was repeated 1-2 times. The wet cake was slurried with approximately 1 L of water for 0.5-1 hour, then filtered. This slurrying and filtering process was repeated 2-3 times, and the wet cake was rinsed with purified water. The filter cake was air-dried at 45-55°C, crushed, and sieved to yield 90 g of polymer WS012-A as a pale yellow solid.

[0162] The product was detected using the SHIMADZU IRSpirit-T Fourier transform infrared spectrometer and Jnm-600ECZ solid-state nuclear magnetic analyzer in Example 1. The results showed that the infrared spectrum and 13 The C solid-state NMR spectrum was identical to that of the polymer of Example 1 (spectrum not shown).

[0163] In addition, the solubility of the product of this example in various solvents was also measured. The results showed that, like the product of Example 1, the polymer was not soluble in any solvent.

[0164] Example 5

[0165] (Preparation of polymer WS012-B)

[0166]

[0167] Add 304.6 g of concentrated hydrochloric acid to reaction flask 1. Control the internal temperature to not exceed 15°C. Then, dropwise add 247.7 g (2.55 mol) of DAA and 69.99 g (0.51 mol) of TAA to reaction flask 1. After the addition is complete, stir at 10-20°C for approximately 0.5-1 hour. Add 17.79 g of VA-044 and 180 g of water to a conical flask. Stir to dissolve. Then, add the aqueous solution to reaction flask 1 and stir to mix thoroughly.

[0168] Add 1740mL of toluene and 12.3g of Span60 to reaction flask 2 and heat to 40-50°C under stirring to dissolve. Adjust the mechanical stirring speed to 300-400RPM, add the material in reaction flask 1 to reaction flask 2, heat to 50-55°C, and stir the reaction at 50-55°C for 15-24 hours. After GC detection of the reaction solution shows that the reaction is complete, the temperature in the reaction flask is raised to 80-85°C, and the reaction is stirred at 80-85°C for 1-2 hours, then cooled and filtered. The wet cake is slurried with about 4L of water for 0.5-1 hour and filtered. Repeat the slurrying and filtering operation 3-5 times. The wet cake is added to 4L of 10% NaOH aqueous solution and stirred at room temperature for about 8-12 hours. Then, the reaction mixture is filtered, and the wet cake is slurried with about 1L of water for 0.5-1 hour and filtered. The wet cake is dried by air blowing at 45-55° C., crushed and sieved to obtain a solid, which is an ammonium salt polymer in which the anion is chloride ion.

[0169] The solid, 55.0g 1-bromo-6-(trimethylammonium)hexyl bromide (0.18mol) and 2.4L methanol are added to a reaction flask, heated to 65-70°C, and stirred at 65-70°C for 24-36 hours. After the reaction is complete, the reaction mixture is filtered. The wet cake is slurried with about 1L of methanol for 0.5-1 hour and filtered. Then, the wet cake is slurried with about 1L of water for 0.5-1 hour and filtered. Repeat the slurrying and filtering operation 2-3 times. The wet cake is added to about 4L of saturated NaHCO3 aqueous solution, stirred at 20-30°C for 12-18 hours and filtered, and this step is repeated 1-2 times. The wet cake is slurried with about 1L of water for 0.5-1 hour and filtered. After repeating the slurrying and filtering operation 2-3 times, the wet product is rinsed with purified water. The filter cake was dried by forced air at 45-55° C., and the dried product was crushed and sieved to obtain 170 g of polymer WS012-B as a light yellow solid.

[0170] The product of this example was detected using the SHIMADZU IRSpirit-T Fourier transform infrared spectrometer in Example 1, and its infrared spectrum is shown in FIG4 .

[0171] The product WS012-B prepared in this example was measured using differential scanning calorimetry (DSC) in Example 1, and the obtained DSC spectrum is shown in Figure 5. The results show that the glass transition temperature of the polymer WS012-B is 58.8°C.

[0172] Example 6

[0173] (Preparation of ammonium salt polymer WS012-C)

[0174]

[0175] To a 3L reaction flask, add 178.9g of concentrated hydrochloric acid, controlling the internal temperature to not exceed 15°C. Add 246.8g (1.8mol) of TAA dropwise to the reaction flask. After the addition is complete, stir at 10-20°C for approximately 0.5-1 hour. Add 15.7g of VA-044 and 45g of water to a conical flask. Stir to dissolve, then add the aqueous solution to the reaction flask. Add 242.5g (0.9mol) of a 60% aqueous solution of dimethyldiallylammonium chloride, and stir to mix thoroughly.

[0176] 1.6L toluene and 11.8g Span60 were added to the reaction flask and heated to 40-50°C under stirring to dissolve. The mechanical stirring speed was adjusted to 200-300RPM, the contents of the reaction flask were added to the reactor, the temperature was raised to 50-55°C, and the reaction was stirred at 50-55°C for 15-24 hours. After GC detection of the reaction solution showed that TAA was basically completely converted, the reaction mixture was filtered. The wet cake was slurried with about 3L methanol for 0.5-1 hour and filtered. Then, the wet cake was slurried with about 10L water for 0.5-1 hour and filtered. The slurrying and filtering operations were repeated 1-2 times. The wet cake was added to about 7.5L saturated NaHCO3 aqueous solution, stirred at 20-30°C for 12-18 hours and filtered. This step was repeated 1-2 times. The wet cake was slurried with about 10L water for 0.5-1 hour and filtered. After repeating the beating and filtering operation 2-3 times, the wet product was rinsed with purified water. The filter cake was dried at 45-55°C under forced air, crushed, and sieved to obtain about 200 g of polymer WS012-C as a light yellow solid.

[0177] The product of this example was detected using the SHIMADZU IRSpirit-T Fourier transform infrared spectrometer in Example 1, and its infrared spectrum is shown in FIG6 .

[0178] The product WS012-C prepared in this example was measured using differential scanning calorimetry (DSC) in Example 1, and the obtained DSC spectrum is shown in Figure 7. The results show that the glass transition temperature of the polymer WS012-C is 177.8°C.

[0179] Examples 7 to 13

[0180] In Examples 7-13, polymers were prepared using different amounts of DAA and / or TAA according to the method described in Example 1, and the products were tested using the test methods listed in Example 1. The amounts of the reactants used in Examples 7-13 and the experimental results are as follows:

[0181]

[0182] The results showed that in Example 2, only DAA was used as a monomer for the reaction, and no solid polymer product was formed; Examples 3-13 all formed polymers in the form of light yellow solids, and the DSC detection method described in Example 1 was used to detect the polymers, showing their glass transition temperatures as shown in the last column of the table above.

[0183] Example 14

[0184] (In vitro bile acid adsorption test)

[0185] 1. Solution preparation:

[0186] Bile acid mixed solution: Each 1L of 0.68% phosphate buffered saline (pH 6.8) contains 6mmol of sodium glycocholate, 6mmol of sodium glycochenodeoxycholate and 2mmol of sodium taurodeoxycholate.

[0187] Linear solutions: Dilute the bile acid mixture to obtain concentrations of 0.06, 0.18, 0.3, 0.48, and 0.6 mmol sodium glycocholate, 0.06, 0.18, 0.3, 0.48, and 0.6 mmol sodium glycochenodeoxycholate, and 0.02, 0.06, 0.1, 0.16, and 0.2 mmol sodium taurodeoxycholate per 1 L of purified water. Use these as linear solutions 1 to 5.

[0188] Test solution: Take 100 mg of the test sample, accurately weigh it, and place it in a 250 ml iodine volumetric flask. Add 100 ml of the bile acid mixed solution and stir it in a water bath (37°C ± 0.5°C) with a magnetic stirrer. Sample it after 5 hours and take 5 ml of the solution. Let it come to room temperature. (After taking it out of the room temperature, let it stand for at least 5 minutes, then tilt the iodine volumetric flask and shake it to remove the water droplets on the top of the iodine volumetric flask. Shake it well while rotating for at least 15 seconds.) Filter it and take 1.0 ml of the filtrate. Place it in a 10 ml volumetric flask, dilute it to the mark with purified water, shake it well and inject it into the sample.

[0189] Chromatographic conditions

[0190]

[0191] 2. Measure the bile acid binding capacity using the standard curve method.

[0192] The bile acid concentration (unit: mmol / L) of the linear solution was used as the abscissa and the peak area was used as the ordinate to make a linear regression equation Y=aX+b.

[0193]

[0194] Where:

[0195] a is the slope of the linear regression equation;

[0196] b is the intercept of the linear regression equation;

[0197] A SPL is the peak area of ​​the test solution;

[0198] C0 is the initial concentration of each bile acid in the test solution, μmol / L;

[0199] V is the volume of bile acid conjugated solution added, 0.1 L;

[0200] S is the dilution volume of the test sample, 10 ml;

[0201] m is the sample weight of the test product;

[0202] W is the moisture value of the test sample, %.

[0203] 3. Experimental Results

[0204]

[0205] The experimental results show that the WS012-A type ammonium salt polymer of the present invention has the highest in vitro total bile acid adsorption capacity, which is better than WS012-B type ammonium salt polymer, WS012-C type ammonium salt polymer, WS012-D type ammonium salt polymer and colesevelam hydrochloride.

[0206] Example 15

[0207] (Effects of WS012-A on DDC-induced primary sclerosing cholangitis (PSC) in mice)

[0208] 1. Experimental methods:

[0209] Thirty-six male C57BL / 6J mice (18-22 g, 6-8 weeks old) were randomly assigned to six experimental groups according to body weight after adaptive feeding: a normal control group, a model control group, a positive control drug group 1, a positive control drug group 2, a WS012-A low-dose group, and a WS012-A high-dose group, with six mice in each group. The normal control group and the model control group were given a vehicle (0.5% CMC-Na aqueous solution), the positive control group 1 was given 0.3 mg / kg of Odevixibat (Hanxiang Biotechnology Co., Ltd.), the positive control group 2 was given 300 mg / kg of colesevelam hydrochloride (Formosa Laboratories, Inc.), and the WS012-A low-dose group and high-dose group were given 300 mg / kg and 1000 mg / kg of WS012-A prepared in Example 2, respectively. Models were established according to known methods (Peter Fickert, et al., The American Journal of Pathology, Vol. 171, No. 2, August 2007, pages 525-536). Specifically, except for the normal control group, mice in all other groups were fed 0.1% DDC for modeling and had free access to food and water for 14 consecutive days. Drug administration began simultaneously with modeling and continued once daily for 14 days. All experimental groups were gavage-administered the corresponding test substance or vehicle at a volume of 20 ml / kg. The day after the last dose, blood was collected under anesthesia for testing of alkaline phosphatase (ALP), total bile acid (TBA), and total bilirubin (TBIL). Feces were collected within 24 hours between days 13 and 14 for testing of TBA content. Fecal pretreatment was as follows: 50 mg of feces was weighed from each case, 2% SDS aqueous solution was added at a mass-to-volume ratio of 1:20, and after sonication for 20 minutes, the supernatant was centrifuged at 1000 rpm for 5 minutes, and the supernatant was collected for testing.

[0210] All parameters are expressed as mean ± SEM. Graph Pad Prism 8 software was used to plot the parameters of the different groups before and after administration. Data were analyzed using SPSS 22.0 statistical software. The Levene test was used to test for homogeneity of variance. When the variances were homogeneous (P ≥ 0.05), the Dunnett's LSD method of one-way analysis of variance (ANOVA) was used to compare group differences. When the variances were unequal (P < 0.05), the Mann-Whitney U test (MW method) of the Kruskal-Wallis H rank sum test (KW method) was used to compare group differences.

[0211] 2. Experimental results:

[0212] Effects of the test substances on blood parameters of the PSC model in mice induced by 0.1% DDC diet (Mean±SEM, n=6)

[0213]

[0214] Compared with the model group, *P<0.05, **P<0.01, ***P<0.001; compared with the colesevelam hydrochloride group, #P<0.05, ##P<0.01, ###P<0.001

[0215] The experimental results of this embodiment are shown in FIG8 .

[0216] Figure 8A shows that compared with the normal group, the serum ALP in the model group was significantly increased (P<0.001); compared with the model group, the ALP in the Odevixibat group was significantly decreased (P<0.001), and the serum ALP in the colesevelam hydrochloride group was slightly increased, which was not statistically significant. The ALP in both the low-dose and high-dose WS012-A groups decreased, showing a certain dose-effect relationship, among which the decrease in ALP in the high-dose WS012-A group was statistically significant (P<0.05).

[0217] Figure 8B shows that compared with the normal group, the serum TBA level in the model group was significantly increased (P<0.001); compared with the model group, the TBA level in the Odevixibat group was significantly decreased (P<0.05), the serum TBA level in the colesevelam hydrochloride group was basically unchanged, and the TBA level in both the low-dose and high-dose WS012-A groups was significantly decreased, showing a certain dose-effect relationship. The decrease in TBA in the high-dose WS012-A group was statistically significant (P<0.05).

[0218] Figure 8C shows that compared with the normal group, the serum TBIL level in the model group was significantly increased (P<0.001). Compared with the model group, the TBIL level in the Odevixibat group was significantly decreased (P<0.05). The serum TBIL level in the colesevelam hydrochloride group was increased, while the TBIL level in both the low-dose and high-dose WS012-A groups was significantly decreased, showing a certain dose-effect relationship. The decrease in TBIL level in the high-dose WS012-A group was statistically significant (P<0.05). Moreover, the TBIL level in both the low-dose and high-dose WS012-A groups was significantly lower than that in the colesevelam hydrochloride group (P<0.01 and P<0.001, respectively).

[0219] Figure 8D shows that compared with the normal group, the excretion of TBA in feces of the model group was significantly decreased (P<0.001); compared with the model group, the excretion of TBA in feces of the Odevixibat group was significantly increased (P<0.001), the excretion of TBA in feces of the colesevelam hydrochloride group was significantly increased (P<0.05), and the excretion of TBA in feces of the WS012-A low-dose group and the high-dose group were significantly increased (P<0.05).

[0220] These results demonstrate that: ① Compared with the normal group, serum ALP, TBA, and TBIL levels in the model group were significantly increased, and fecal TBA excretion was significantly decreased, indicating that DDC feeding can induce significant changes in physiological indicators of cholestasis in mice, indicating successful model establishment. ② Compared with the model group, ALP, TBA, and TBIL levels were significantly decreased, and fecal TBA excretion was significantly increased in the odevixibat group, a first-line drug for the treatment of progressive familial intrahepatic cholestasis (PFIC), indicating that this drug has excellent therapeutic effects on cholestasis. ③ Compared with the model group, serum ALP and TBIL levels were increased in the colesevelam hydrochloride group, which is detrimental to the treatment of cholestasis. Serum TBA levels remained almost unchanged, with no significant effect on these parameters. Fecal TBA excretion was significantly increased, suggesting a beneficial effect on the treatment of cholestasis. Overall, colesevelam hydrochloride has no significant effect on cholestasis and is not suitable for the treatment of cholestasis. ③ Compared with the model group, ALP, TBA, and TBIL levels were significantly reduced in both the low- and high-dose WS012-A groups, demonstrating a dose-response relationship. The high-dose WS012-A group showed statistically significant reductions in ALP, TBA, and TBIL. Fecal TBA excretion was also statistically significantly increased in both the low- and high-dose WS012-A groups. Overall, WS012-A has a therapeutic effect on cholestasis, and this therapeutic effect is dose-dependent. ④ Compared with the colesevelam hydrochloride group, the low-dose WS012-A group had a superior effect on all four parameters.

[0221] 3. Experimental conclusions:

[0222] In a DDC-fed PSC mouse model, oral administration of WS012-A at doses of 300 mg / kg and 1000 mg / kg once daily for 14 days increased fecal TBA excretion, reduced serum ALP, TBA, and TBIL, and alleviated the pathological changes of hepatic cholestasis, demonstrating superior efficacy to colesevelam hydrochloride, a similar high-molecular-weight bile acid binder.

[0223] Example 16

[0224] (Effects of WS012-A on ANIT-induced acute cholestasis in rats)

[0225] 1. Experimental methods:

[0226] 48 male SD rats (180-220 g, 6-8 weeks old) were randomly divided into 5 experimental groups according to body weight after adaptive feeding, namely normal control group, model control group, positive control drug group 1, positive control drug group 2, WS012-A group, and WS012-C group, with 8 rats in each group. The normal control group and the model control group were given a solvent (0.5% CMC-Na aqueous solution), the positive control group 1 was given 0.3 mg / kg of Odevixibat, the positive control group 2 was given 1000 mg / kg of colesevelam hydrochloride, the WS012-A group was given 1000 mg / kg of WS012-A prepared in Example 1, and the WS012-C group was given 1000 mg / kg of WS012-C prepared in Example 6. All experimental groups were given the corresponding test substance or solvent by gavage at a volume of 20 ml / kg, once a day for 7 days. The model was established according to methods known in the art (Yoshiji Ohta, et al, Toxicology 139 (1999) 265–275). Except for the rats in the normal control group, all other rats received a single intraperitoneal injection of 80 mg / kg of ANIT on the fourth day of administration. On the seventh day of administration (72 ± 1 h after injection of the ANIT modeling agent), the animals were anesthetized, and whole blood was collected from the jugular venous plexus. The serum was obtained by centrifugation and assayed for alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and total bile acid (TBA).

[0227] All parameters are expressed as mean ± SEM. Graph Pad Prism 8 software was used to plot the parameters of the different groups before and after administration. Data were analyzed using SPSS 22.0 statistical software. The Levene test was used to test for homogeneity of variance. When the variances were homogeneous (P ≥ 0.05), the Dunnett's LSD method of one-way analysis of variance (ANOVA) was used to compare group differences. When the variances were unequal (P < 0.05), the Mann-Whitney U test (MW method) of the Kruskal-Wallis H rank sum test (KW method) was used to compare group differences.

[0228] 2. Experimental results:

[0229] Blood parameters after administration (Mean±SEM, n=8)

[0230]

[0231] Compared with the model group, *P<0.05, **P<0.01, ***P<0.001; compared with the colesevelam hydrochloride group, #P<0.05

[0232] The experimental results of this embodiment are shown in FIG9 .

[0233] Figure 9A shows that compared with the normal group, the model group had a significant increase in serum ALT (P < 0.001). Compared with the model group, the odevixibat group, the colesevelam hydrochloride group, the WS012-A group, and the WS012-C group all had decreased ALT, with the colesevelam hydrochloride group (P < 0.05), the WS012-A group (P < 0.01), and the WS012-C group (P < 0.01) all showing statistical significance. WS012-A and WS012-C were more effective than colesevelam hydrochloride in lowering serum ALT.

[0234] Figure 9B shows that compared with the normal group, the serum AST in the model group was significantly increased (P<0.001). Compared with the model group, the AST in the Odevixibat group, the colesevelam hydrochloride group, the WS012-A group, and the WS012-C group were all decreased, among which the AST in the colesevelam hydrochloride group was not statistically significant, while that in the WS012-A group and the WS012-C group was statistically significant (P<0.05), and the WS012-A group had the best effect.

[0235] Figure 9C shows that compared with the normal group, the serum ALP level in the model group was significantly increased (P<0.001); compared with the model group, the serum ALP level in the Odevixibat group, the colesevelam hydrochloride group, the WS012-A group, and the WS012-C group was significantly decreased (P<0.01), among which the WS012-A group had the best effect.

[0236] Figure 9D shows that compared with the normal group, the serum TBA level in the model group was significantly increased (P<0.01); compared with the model group, the serum TBA levels in the Odevixibat group, the colesevelam hydrochloride group, the WS012-A group, and the WS012-C group were significantly decreased (P<0.05 or P<0.01), and the serum TBA levels in the WS012-A group and the WS012-C group were lower than that in the colesevelam hydrochloride group.

[0237] The above results indicate that: ① Compared with the normal group, serum ALT, AST, ALP, and TBA in the model group were significantly increased (P < 0.05-0.001), indicating that ANIT feeding led to significant changes in cholestatic physiological indicators and that the model was successfully established. ② Compared with the model group, serum ALT, AST, ALP, and TBA in the odevixibat, colesevelam hydrochloride, and WS012-A groups were all decreased. The decreases in serum ALP and TBA in the odevixibat group were statistically significant (P < 0.01), the decreases in serum ALT, ALP, and TBA in the colesevelam hydrochloride group were statistically significant (P < 0.05-0.01), and the decreases in serum ALT, ALP, and TBA in the WS012-A and WS012-C groups were statistically significant (P < 0.05-0.01).

[0238] 3. Experimental conclusions:

[0239] In a rat acute cholestasis model induced by a single intraperitoneal injection of ANIT (80 mg / kg), WS012-A and WS012-C were administered orally by gavage at a dose of 1000 mg / kg once daily for 7 days. WS012-A significantly reduced serum ALT (P < 0.01), ALP (P < 0.01), and TBA (P < 0.05), and to a lesser extent, AST. WS012-C also significantly reduced ALT (P < 0.01), AST (P < 0.05), ALP (P < 0.01), and TBA (P < 0.01). Overall, both WS012-A and WS012-C have a cholestatic effect, with efficacy superior to that of the similar high-molecular-weight bile acid binder colesevelam hydrochloride and the small-molecule drug odevixibat (an IBAT inhibitor).

[0240] Example 17

[0241] (Effects of WS012-A on high-fat diet-induced hyperlipidemia in rats)

[0242] 1. Experimental methods:

[0243] Forty-eight male golden hamsters (100-150 g, 6-7 weeks old) were acclimated for approximately 7 days. Eight animals were retained as a blank control group and continued to be fed a basal diet. The remaining animals served as the experimental group and were fed a high-fat diet (basal diet + 15% lard + 20% sucrose + 1.2% cholesterol + 0.2% sodium cholate). The animals' mental and dietary status were regularly observed. After 7 weeks of high-fat diet, the animals were fasted overnight. The next day, blood was drawn from the orbital venous plexus to measure plasma TC, TG, and LDL-C levels. The hamsters were then grouped based on the results. Thirty-two hamsters with hyperlipidemia were screened based on their blood biochemical indicators and body weight and divided equally into four groups: a model control group (Model), test substance group 1 (WS012-A prepared in Example 2, 1000 mg / kg), test substance group 2 (WS012-B prepared in Example 5, 1000 mg / kg), and a positive control group (colesevelam hydrochloride, 1000 mg / kg). The corresponding solvent or test substance was administered orally at a dosage volume of 20.0 ml / kg, once a day for 4 weeks. The blank control group was fed with ordinary feed, and the other groups continued to be fed with high-fat feed. During the administration period, body weight was measured once a week to observe changes in body weight. Blood was collected before administration and 1, 2, 3, and 4 weeks after administration to detect total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) in serum; feces were collected once 2 weeks and 4 weeks after administration (within 24 hours) to detect total bile acid (TBA) in the fecal supernatant; after the last feces collection, the animals were euthanized and the liver was taken for observation.

[0244] All parameters are expressed as mean ± SEM. Graph Pad Prism 8 software was used to plot the parameters of the different groups before and after administration. Data were analyzed using SPSS 22.0 statistical software. The Levene test was used to test for homogeneity of variance. When the variances were homogeneous (P ≥ 0.05), the Dunnett's LSD method of one-way analysis of variance (ANOVA) was used to compare group differences. When the variances were unequal (P < 0.05), the Mann-Whitney U test (MW method) of the Kruskal-Wallis H rank sum test (KW method) was used to compare group differences.

[0245] 2. Experimental results:

[0246] The experimental results of this embodiment are shown in FIG10 .

[0247] Figure 10A shows that compared with the normal group, the model group had significantly elevated total cholesterol (TC) levels before and during weeks 1 to 4 of drug administration (P < 0.001). TC levels also showed a slowly increasing trend as the high-fat diet induction time increased. Compared with the model group, TC levels in the positive control group (colesevelam hydrochloride) decreased somewhat after drug administration. This was only significantly reduced in the colesevelam hydrochloride group at week 3 (P < 0.01), with no significant differences observed at other sampling time points. TC levels in the WS012-A group decreased significantly from week 1 to 4 of drug administration (P < 0.05 to 0.001), and in the WS012-B group decreased significantly from week 2 to 4 of drug administration (P < 0.05 to 0.001). Compared with the positive control (colesevelam hydrochloride) group, TC levels in both WS012-A and WS012-B groups decreased significantly at week 4 of drug administration (P < 0.01). The above results showed that WS012-A and WS-12-B were superior to colesevelam hydrochloride in reducing TC, among which WS012-A had the best effect.

[0248] Figure 10B shows that compared with the normal group, the model group had significantly elevated triglyceride (TG) levels before and during weeks 1 to 4 of treatment (P < 0.05). After treatment, no significant difference in TG was observed in the colesevelam hydrochloride group compared with the model group (P > 0.05). Throughout the treatment period, TG levels in the WS012-A group were lower than those in the model group. At weeks 1, 2, and 4 of treatment, TG levels in the WS012-A group were significantly lower than those in the model group (P < 0.05). No significant difference in TG was observed in the WS012-B group compared with the model group during the entire treatment period (P > 0.05). Compared with the positive control (colesevelam hydrochloride) group, TG levels in the WS012-A group were significantly lower at week 4 of treatment (P < 0.01). These results demonstrate that WS012-A has a significant TG-lowering effect and is superior to colesevelam hydrochloride.

[0249] Figure 10C shows that compared with the normal group, the model group had significantly elevated low-density lipoprotein (LDL-C) levels before and during weeks 1 to 4 of treatment (P < 0.001). Compared with the model group, the colesevelam hydrochloride group showed a moderate decrease in LDL-C levels after treatment, with a significant decrease only at week 3 (P < 0.05), with no significant differences at other time points (P > 0.05). LDL-C levels in the WS012-A group were significantly lower at weeks 1 to 4 of treatment (P < 0.05 to 0.001). Throughout the treatment cycle, LDL-C levels in the WS012-B group were lower than those in the model group, but significant decreases were observed only at weeks 1 and 4 of treatment (P < 0.01 to 0.001). Compared with the colesevelam hydrochloride group, WS012-A significantly reduced LDL-C at weeks 1, 2, and 4 (P < 0.05-0.001); WS012-B significantly reduced LDL-C at weeks 1 and 4. These results indicate that both WS012-A and WS012-B significantly lower LDL-C and are superior to colesevelam hydrochloride, with WS012-A showing the greatest effect.

[0250] Figures 10D and 10E show that compared with the normal group, liver weight and liver index in the model group increased significantly after administration (P < 0.001), indicating successful model establishment. Compared with the model group, liver weight and liver index decreased in both WS012-A and WS012-B after administration, with the decrease in liver weight and liver index in the WS012-A group being statistically significant (P < 0.05). Compared with the positive control, colesevelam hydrochloride, liver weight and liver index decreased in both WS012-A and WS012-B after administration, but only the decrease in liver weight (P < 0.05) and liver index (P < 0.01) in the WS012-A group was significant. These results indicate that both WS012-A and WS012-B have the effect of ameliorating high-fat diet-induced hepatomegaly, but only WS012-A is significantly superior to colesevelam.

[0251] Figure 10F shows that the livers of animals in the normal control group were bright red, smooth, and soft. The livers of golden hamsters in the model control group were significantly larger, whitish, and generally swollen, with slightly rounded edges and a brittle texture. Liver tissue in animals in the colesevelam hydrochloride positive control group showed slight differences from the model group, but the differences were minor. The livers were significantly larger, whitish, and generally swollen, with slightly rounded edges and a brittle texture. Liver weight and liver coefficient did not decrease. Compared with the model and normal control groups, the livers of animals in the WS012-A and WS012-B groups were partially bright red with slightly whitish edges, and their overall volume was intermediate between the model and normal control groups, with WS012-A showing the greatest effect.

[0252] 3. Experimental conclusions:

[0253] In a golden hamster hyperlipidemia model induced by a high-fat diet for 7 weeks, oral administration of WS012-A or WS012-B at a dose of 1000 mg / kg once a day for 4 weeks reduced blood lipid levels (TC, TG, LDL-C) and alleviated hepatic fatty lesions (liver weight, liver coefficient, gross liver). WS012-A was the most effective, outperforming the similar high-molecular-weight bile acid binder colesevelam hydrochloride.

[0254] Based on the above experimental results of the present application, it can be concluded that the ammonium salt polymer of the present invention has a good adsorption effect on bile acid both in vitro and in vivo, which is better than colesevelam hydrochloride and other types of ammonium salt polymers WS012-B and WS012-C prepared in this application. BRIEF DESCRIPTION OF THE DRAWINGS

[0255] FIG1 is an infrared spectrum of the ammonium salt polymer WS012-A prepared in Example 1.

[0256] Figure 2 is the ammonium salt polymer WS012-A prepared in Example 1 13 C NMR spectrum.

[0257] FIG3 is a DSC graph of the ammonium salt polymer WS012-A prepared in Example 1.

[0258] FIG4 is an infrared spectrum of the ammonium salt polymer WS012-B prepared in Example 5.

[0259] FIG5 is a DSC graph of the ammonium salt polymer WS012-B prepared in Example 5.

[0260] FIG6 is an infrared spectrum of the ammonium salt polymer WS012-C prepared in Example 6.

[0261] FIG7 is a DSC graph of the ammonium salt polymer WS012-C prepared in Example 6.

[0262] Figure 8 shows the experimental results of the DDC-induced primary sclerosing cholangitis model in mice of Example 15, wherein Normal represents the normal group, Model represents the model group, Odevixibat represents the Odevixibat treatment group, Coleselam represents the coleselam hydrochloride treatment group, WS012-A-Low represents the low-dose treatment group of WS012-A prepared in Example 2, and WS012-A-High represents the high-dose treatment group of WS012-A prepared in Example 2.

[0263] Figure 9 is the experimental results of the ANIT-induced rat primary biliary cholangitis model of Example 16, wherein Normal represents the normal group, Model represents the model group, Odevixibat represents the Odevixibat treatment group, Coleveselam represents the coleselam hydrochloride treatment group, WS012-A represents the WS012-A treatment group prepared in Example 1, and WS012-C represents the WS012-C treatment group prepared in Example 6.

[0264] Figure 10 shows the experimental results of the high-fat diet-induced golden hamster hyperlipidemia model in Example 17, wherein Normal represents the normal group, Model represents the model group, Coleveselam represents the coleselam hydrochloride treatment group, WS012-A represents the WS012-A treatment group prepared in Example 2, and WS012-B represents the WS012-B treatment group prepared in Example 5.

[0265] The specific embodiments and examples provided herein are intended only to illustrate the present invention and do not limit the scope of the claims. Based on the contents disclosed herein, those skilled in the art will readily appreciate equivalent variations of the technical solutions of the present invention, which are also encompassed within the scope of the present invention.

Claims

1. An ammonium salt polymer obtained by polymerizing a monomer of formula (I) and a monomer of formula (II), in: m, n, p, q, r are each independently selected from 1, 2 or 3, The ammonium salt polymer contains monovalent, divalent or trivalent anions as counter ions, and the anions are selected from chloride ions (Cl - ), bicarbonate ion (HCO3 - ), hydrogen sulfate ion (HSO4 - ), acetate ion (C2H3O2 - ), carbonate ions (CO3 2- ), sulfate ion (SO4 2- ), phosphate ions (PO4 3- ) and anions derived from malic acid, citric acid, fumaric acid, and maleic acid.

2. The polymer according to claim 1, wherein m, n, p, q and r are each independently selected from 1 and 2, preferably wherein m, n, p, q and r are all 1.

3. The polymer according to claim 1 or 2, wherein the ammonium salt polymer is obtained by polymerizing a monomer of formula (I) and a monomer of formula (II) in a molar ratio of 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, more preferably 10:1 to 10:4, and most preferably 10:

2.

4. The polymer according to any one of claims 1 to 3, wherein the polymer has a glass transition temperature of about 60°C to about 130°C, preferably about 70°C to 120°C, more preferably about 70°C to about 105°C.

5. The polymer according to any one of claims 1 to 4, wherein the anion contained in the polymer is a bicarbonate ion (HCO3 - ) and / or carbonate ions (CO3 2- ), and wherein m, n, p, q and r are all 1.

6. The polymer according to any one of claims 1 to 5, wherein the polymer has an in vitro bile acid adsorption capacity of 7-10 mmol / g, preferably 7-8 mmol / g.

7. A method for preparing a polymer according to any one of claims 1 to 6, comprising the steps of: (1) acidifying the monomer of formula (I) and the monomer of formula (II) with an acid, preferably, the molar ratio of the monomer of formula (I) to the monomer of formula (II) is 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, further preferably 10:1 to 10:4, and most preferably 10:2; (2) polymerizing the acidified monomer obtained in step (1) to obtain the desired ammonium salt polymer.

8. The method according to claim 7, wherein step (1) is carried out as follows: at a temperature of 30° C. or lower, preferably at a temperature of 15° C. or lower, a monomer of formula (I) and a monomer of formula (II) in a molar ratio of 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, further preferably 10:1 to 10:4, and most preferably 10:2 are added dropwise to a suitable acid; after completion of the dropwise addition, the mixture is stirred at a temperature of 5-30° C., preferably at a temperature of 10-20° C., for a period of time to obtain a mixture of the two acidified monomers.

9. The method according to claim 7 or 8, wherein the acid described in step (1) is selected from hydrochloric acid, sulfuric acid, acetic acid, malic acid, citric acid, maleic acid and fumaric acid.

10. The method according to any one of claims 7 to 9, wherein step (2) comprises the following sub-steps: (2.1) dissolving an initiator in water and adding the mixture to the acidified monomers obtained in step (1) to obtain an aqueous phase mixture; (2.2) in another container, dissolving the emulsifier in an organic solvent to obtain an oil phase mixture, wherein the organic solvent is immiscible with water and has a boiling point higher than the polymerization temperature, and the organic solvent is preferably selected from toluene, n-heptane, cyclohexane, xylene, n-octane, and any mixture thereof; (2.3) adding the aqueous phase mixture to the oil phase mixture to carry out a polymerization reaction to generate the desired ammonium salt polymer; (2.4) performing conventional post-treatment on the reactants of step (2.3) to isolate and obtain the desired ammonium salt polymer; Optionally, (2.5) converting the wet cake in step (2.4) or the ammonium salt polymer separated in step (2.4) into an ammonium salt polymer containing other anions.

11. A method for preparing the ammonium salt polymer of claim 5, comprising the steps of: (1) acidifying a monomer of formula (I) and a monomer of formula (II) in a molar ratio of 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, further preferably 10:1 to 10:4, and most preferably 10:2 with concentrated hydrochloric acid to obtain a mixture of the two acidified monomers; (2.1) dissolving an initiator in water and adding the mixture to the acidified monomers obtained in step (1) to obtain an aqueous phase mixture; (2.2) in another container, dissolving an emulsifier in an organic solvent to obtain an oil phase mixture, wherein the organic solvent is immiscible with water and has a boiling point higher than the polymerization temperature, and the organic solvent is selected from toluene, n-heptane, cyclohexane, xylene, n-octane, and any mixture thereof; (2.3) adding the aqueous phase mixture to the oil phase mixture, and performing a polymerization reaction at a temperature of 40-100° C., preferably 50-85° C., to generate the desired ammonium salt polymer; (2.4) Filter the reactant from step (2.3), slurry the wet cake with methanol or ethanol once, filter, and then slurry the wet cake with purified water and filter, repeating 2-3 times to obtain a wet cake; (2.5) adding the wet filter cake obtained in step (2.4) to a saturated aqueous sodium bicarbonate solution and / or an aqueous sodium carbonate solution, stirring for a period of time, filtering, drying, and crushing to obtain an ammonium salt polymer containing bicarbonate ions and / or carbonate ions.

12. The method according to any one of claims 7 to 11, wherein the initiator is selected from 2,2'-azobisisobutylamidine dihydrochloride (V-50), 2,2'-azabis(2-imidazoline) dihydrochloride (VA-044), ammonium persulfate, potassium persulfate, hydrogen peroxide, and any mixture thereof; and the emulsifier is selected from polyoxyethylene sorbitan fatty acid esters (such as Tween 40, Tween 80, etc.), sorbitan fatty acid esters (such as Span 40, Span 60, Span 80, etc.), and any mixture thereof.

13. The ammonium salt polymer according to any one of claims 1 to 6, for use as a medicament, preferably as a medicament for treating or preventing diseases associated with excessive bile acids, preferably the diseases associated with excessive bile acids are selected from dyslipidemia (e.g., hyperlipidemia), cholestatic liver diseases (e.g., progressive familial intrahepatic cholestasis (PFIC), benign recurrent intrahepatic cholestasis (BIRC), congenital bile acid synthesis disorders, Alagille syndrome (ALGS), neonatal cholestasis caused by citrin protein deficiency, ARC syndrome, primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC), autoimmune hepatitis (AIH), drug-induced cholestasis (DRIC), biliary atresia (BA) and intrahepatic cholestasis of pregnancy (ICP)), bile acid diarrhea and diabetes.

14. A pharmaceutical composition comprising the polymer of any one of claims 1 to 6, and optionally a pharmaceutically acceptable carrier.

15. A method for treating or preventing a disease associated with excessive bile acid, the method comprising administering an effective amount of a polymer according to any one of claims 1 to 6 to a subject in need thereof, wherein the disease is preferably selected from dyslipidemia (e.g., hyperlipidemia), cholestatic liver disease (e.g., progressive familial intrahepatic cholestasis (PFIC), benign recurrent intrahepatic cholestasis (BIRC), congenital bile acid synthesis disorder, Alagille syndrome (ALGS), neonatal cholestasis caused by Citrin protein deficiency, ARC syndrome, primary biliary cholangitis (PBC), primary sclerosing cholangitis (PSC), autoimmune hepatitis (AIH), drug-induced cholestasis (DRIC), biliary atresia (BA), and intrahepatic cholestasis of pregnancy (ICP)), bile acid diarrhea, and diabetes.

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