Ammonium salt polymers, methods for preparing same, and their use as bile acid sequestrants - Patents.com

Abstract

Disclosed is an ammonium salt polymer, a method for preparing the same, and its use as a bile acid chelator. The ammonium salt polymer comprises a monomer of formula (I) and a monomer of formula (II): [Formula 1] JPEG2025528899000016.jpg48131 (wherein m, n, p, q and r are each independently selected from 1, 2 or 3). The ammonium salt polymers contain monovalent, divalent, or trivalent anions as counterions and can be used to treat or prevent dyslipidemia (e.g., hyperlipidemia), cholestasis, bile acid diarrhea, and other diseases.

Description

[Technical Field]

[0001] The present invention relates to the field of polymeric drugs. Specifically, the present invention relates to ammonium salt polymers, methods for preparing the same, and their use as bile acid sequestrants. The ammonium salt polymers are useful in the treatment or prevention of diseases such as dyslipidemia (e.g., hyperlipidemia), cholestatic liver disease, and bile acid diarrhea (BAD). [Background technology]

[0002] Bile acids are important components of bile. Their physiological functions mainly include promoting the digestion and absorption of lipids and inhibiting the precipitation of cholesterol in bile (gallstone formation). Bile acids are also involved in glucose and lipid metabolism. Therefore, bile acids are associated with various diseases, such as dyslipidemia (e.g., hyperlipidemia), cholestatic liver disease, and bile acid diarrhea.

[0003] 1. Dyslipidemia Dyslipidemia refers to the abnormal metabolism of lipoproteins in the human body. A small number of dyslipidemias are secondary dyslipidemias caused by systemic diseases, while the majority of dyslipidemias are primary dyslipidemias caused by inherited genetic defects or their interaction with environmental factors. The incidence of dyslipidemia is high in China and is gradually increasing, which is closely related to the significant improvement in the living standards of Chinese people and the changes in Chinese dietary habits.

[0004] Hyperlipidemia is the most common form of dyslipidemia and is also known as hyperlipoproteinemia. Hyperlipidemia refers to a condition involving abnormally elevated levels of any one or all of the 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 higher than 230 mg / 100 ml and triglycerides higher than 140 mg / 100 ml are collectively referred to as hyperlipidemia.

[0005] The damage caused by hyperlipidemia to the human body is slow, progressive, and systemic, resulting in numerous problems. When blood lipid levels rise over a long period of time, blood lipids and their degradation products gradually accumulate on the walls of blood vessels, forming fibrous tissue, narrowing blood vessel channels and reducing vascular elasticity, ultimately leading to vascular blockage. When lipid plaques are deposited on coronary arteries, they can cause ischemic symptoms in the heart, leading to coronary heart disease; when lipid plaques are deposited in the brain, they can cause cerebral infarction; when lipid plaques are deposited in the lower limbs, they can cause pain and even necrosis of the lower limbs; when lipid plaques are deposited in the orbital plexus, they can cause blindness; and when lipid plaques are deposited in the liver, they can cause fatty liver. Hyperlipidemia is recognized as a risk factor for stroke, coronary heart disease, myocardial infarction, and sudden death. In addition, hyperlipidemia can also induce acute pancreatitis.

[0006] Despite the adverse effects of hyperlipidemia, it is necessary to administer antilipidemic drugs to patients. The mechanisms of action of such drugs mainly include inhibiting cholesterol biosynthesis, reducing the level of cholesterol in the body, increasing HDL levels, and inhibiting cholesterol absorption. For example, statins (e.g., lovastatin, simvastatin, atorvastatin, rosuvastatin) can inhibit hydroxymethylglutaryl-CoA reductase, the rate-limiting enzyme in the biosynthesis of cholesterol in the body, thereby reducing the biosynthesis of cholesterol in the body; phenoxyacetic acid antilipidemic drugs (e.g., clofibrate, fenofibrate) can also interfere with the synthesis of cholesterol in the body, significantly reducing the levels of triglycerides and total cholesterol; nicotinic acid analogs (e.g., acipimox) can increase the level of HDL; and bile acid-binding resins (e.g., cholestyramine, colesevelam hydrochloride) can reduce the level of cholesterol in the body.

[0007] Among the above-listed antilipidemic drugs, bile acid-binding resin is a safe oral drug with good lipid-lowering effect. Bile acids are important components of bile and play an important role in lipid metabolism. During digestion in the body, cholesterol is the only precursor of bile acids. Bile acids produced by the decomposition of cholesterol are secreted into the intestine, where they are absorbed in large amounts 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 and has a major role in adsorbing bile acids, thereby excreting bile acids from the body and preventing their reabsorption. High consumption of bile acids promotes the conversion of more cholesterol into bile acids, thereby achieving the goal of reducing cholesterol in the body.

[0008] Colesevelam hydrochloride is the latest representative of this type of agent and has the chemical name N,N,N-trimethyl-6-(2-allylamino)-1-hexylamine copolymer hydrochloride and the chemical structure:

[0009] [ka]

[0010] It has. Colesevelam hydrochloride not only reduces cholesterol levels in the body but also increases the clearance of low-density lipoproteins from the blood. Colesevelam hydrochloride, which has the trade name WELCHOL, is approved as a bile acid sequestrant for the treatment of primary hyperlipidemia and type 2 diabetes.

[0011] Aging is a major challenge facing many countries today. Therefore, the number of patients with hyperlipidemia is continuously increasing, and more lipid-lowering drugs are needed.

[0012] 2. Cholestatic liver disease Cholestasis refers to a pathological condition in which various factors both inside and outside the liver lead to impaired production, secretion, and excretion of bile, resulting in bile not flowing normally into the duodenum but instead flowing into the bloodstream.Clinical symptoms include itching, fatigue, dark urine, and jaundice.Cholestasis is often asymptomatic in the early stages and is characterized only by elevated serum ALP and GGT levels.Hyperbilirubinemia may occur later in the course of the disease, and in severe cases, liver damage or even death may occur.

[0013] Hepatic biliary diseases with cholestasis as the main symptom, caused by various reasons, are often collectively referred to as cholestatic liver disease.The diagnostic criteria for cholestatic liver disease are usually in accordance with the clinical practice guidelines for the management of cholestatic liver disease developed by the European Association for the Study of the Liver (EASL) in 2009, namely, "ALP > 1.5 × ULN and GGT > 3 × ULN".However, it should be noted that GGT may not be elevated in some specific cholestatic liver diseases, such as PFIC type 1 or 2 and BRIC.

[0014] 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 (BRIC), congenital anomalies of bile acid synthesis, Alagille syndrome (ALGS), neonatal cholestasis caused by citrin deficiency, and ARC syndrome. Acquired cholestatic liver diseases are often caused by infection, drugs, hormones, pregnancy, tumors, bile duct obstruction, and other factors, and 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).

[0015] Currently, the primary drug used to treat cholestatic liver disease is ursodeoxycholic acid (UDCA), which has been proven to reduce cholestasis in a variety of cholestatic diseases and is the standard first-line treatment for PBC. However, UDCA is not effective against most pruritus, being effective in only approximately 50% of PBC patients, and 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 are intolerant to UDCA, only obeticholic acid (OCA) is available. Obeticholic acid is a farnesoid X receptor (FXR) agonist and has side effects such as severe skin pruritus, fatigue, abdominal pain and discomfort, and rash. Therefore, its use is also limited.

[0016] Malalixibat, an apical sodium-dependent bile acid transporter (ASBT) inhibitor, is approved for the treatment of pruritus in pediatric patients aged 1 year and older with ALGS, but has side effects such as diarrhea, abdominal pain, vomiting, fat-soluble vitamin deficiency, liver test abnormalities, gastrointestinal bleeding, and bone fractures. Odevixibat, an ileal bile acid transporter (IBAT) inhibitor, is approved for the treatment of progressive familial intrahepatic cholestasis (PFIC), but has side effects such as liver test abnormalities, diarrhea, abdominal pain, vomiting, and fat-soluble vitamin deficiency.

[0017] The incidence of cholestatic liver disease has recently increased significantly, so there is an urgent need for new and effective therapeutic agents. Currently, peroxisome proliferator-activated receptor (PPAR) agonists elafibranor (GFT505) and seladelpar (MBX-8025) are being tested in clinical trials, and are promising to become new drugs for the clinical treatment of cholestatic liver disease. However, the preliminary results of this clinical trial show that these drugs also have serious side effects.

[0018] In contrast, bile acid-binding resins are not absorbed by the intestine and are therefore relatively safe oral medications. They can adsorb bile acids in the intestine, reduce the reabsorption of bile acids, promote the excretion of bile acids, and reduce cholestasis, thereby showing therapeutic effects in cholestatic liver diseases.

[0019] 3. Bile acid diarrhea Bile acid diarrhea (BAD), also known as "bile acid malabsorption (BAM)," is a chronic diarrhea caused by increased intestinal mucosal permeability, increased intestinal fluid secretion, and increased intestinal peristalsis due to the stimulation of excess bile acids in the large intestine, and the excess bile acids result from abnormal bile acid metabolism. In 1967, Hofmann of the Mayo Clinic first proposed the concept of "bile diarrhea and bile enteropathy." Later, bile acid-associated diarrhea was divided into three types according to their cause: Type I is bile acid malabsorption 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, and it is noted that 25% to 50% of cases of irritable bowel syndrome with significant diarrhea (IBS-D) manifest BAD to some degree; and Type III is bile acid malabsorption induced by other gastrointestinal diseases, including cholecystectomy, bacterial overgrowth in the small intestine, and microscopic colitis.

[0020] Depending on the severity of malabsorption, large amounts of bile acids may pass from the ileum to the colon, inducing several pathogenic effects. Bile acids, especially hydrophobic bile acids, are cytotoxic due to their cleansing properties. Physiological concentrations of bile acids in the colon have been shown to stimulate the propagation of motor waves, increasing the rate of colonic transit and enhancing the urge to defecate. Excess bile acids also induce colonocytes to secrete chloride ions, leading to secretory diarrhea, a major feature of BAD.

[0021] 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 the conventional drug for the clinical treatment of BAD, with a response rate of approximately 70% in several clinical trials. However, its poor taste leads to poor compliance. The new-generation bile acid sequestrant colesevelam hydrochloride, launched in 2000, has been used clinically as a first- and second-line treatment for BAD. However, additional bile acid sequestrants are still needed to provide patients with more treatment options.

[0022] The present invention aims to address the above-listed unmet market needs. Summary of the Invention [Means for solving the problem]

[0023] In a first aspect, the present invention provides a monomer of formula (I) and a monomer of formula (II):

[0024] [ka]

[0025] (In the formula, m, n, p, q, and r are each independently selected from 1, 2, or 3. The present invention provides an ammonium salt polymer obtained by the polymerization of

[0026] The ammonium salt polymer contains a monovalent, divalent, or trivalent anion as a counterion. The anion may be derived from an organic or inorganic acid. Anions derived from inorganic acids include chloride ions (Cl). - ), bicarbonate ion (HCO3 - ), bisulfate ion (HSO4 - ), acetate ion (C2H3O2 - ) monovalent anions preferably selected from; carbonate ion (CO 2- ), sulfate ions (SO4 2- ) divalent anions preferably selected from; preferably phosphate ions (PO4 3- anions derived from organic acids can be derived from malic acid, citric acid, fumaric acid, maleic acid, and the like.

[0027] It will be understood by those skilled in the art that the ammonium cations and anions contained in the polymer act as counterions to each other. In one embodiment of the first aspect, the ammonium salt polymer is obtained from the polymerization of 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, even more preferably 10:1 to 10:4, and most preferably 10:2.

[0028] 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. 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.

[0029] The glass transition temperature of the ammonium salt polymer of the present invention is about 60°C to about 130°C, preferably about 70°C to 120°C, and more preferably about 70°C to about 105°C. Preferably, the infrared (IR) spectrum of the ammonium salt polymer of the present invention exhibits the following absorption bands (cm -1 ) : 2918, 2855, 2359, 2072, 1627, 1529, 1402, 1362, 1200, 1000, 909, 810. More preferably, the IR spectrum of the ammonium salt polymer of the present invention is shown in FIG.

[0030] Preferably, the ammonium salt polymer of the present invention 13 The C solid-state nuclear magnetic resonance (NMR) spectrum has the following lines (ppm): 28.2, 42.9, 50.5, 163.3. More preferably, the ammonium salt polymers of the present invention 13 The C solid-state NMR spectrum is shown in Figure 2.

[0031] The ammonium salt polymer of the present invention has the ability to absorb bile acids in vitro and in vivo. For example, the in vitro bile acid absorption capacity of the ammonium salt polymer of the present invention is 7 to 10 mmol / g, more preferably 7 to 8 mmol / g.

[0032] In a second aspect, the present invention provides a method for preparing the ammonium salt polymers of the present invention, comprising the following steps: General Scheme 1:

[0033] [ka]

[0034] (In the formula, X ― represents the aforementioned monovalent, divalent, or trivalent anion as a counterion to ammonium), the method includes the steps of: (1) acidifying a monomer of formula (I) and a monomer of formula (II) using an acid; and (2) polymerizing the acidified monomer obtained from step 1 to obtain the ammonium salt polymer of the present invention.

[0035] The acidification in step 1 may be carried out by adding the monomer of formula (I) and the monomer of formula (II) dropwise in a molar ratio of 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, even more preferably 10:1 to 10:4, and most preferably 10:2 to a suitable acid at a temperature of 30°C or less, preferably 15°C or less, and stirring for a period of time at a temperature of 5 to 30°C, preferably 10 to 20°C, after the addition, to obtain a mixture of two acidified monomers.

[0036] Suitable acids for use in step 1 may be inorganic or organic, such as hydrochloric acid, sulfuric acid, acetic acid, malic acid, citric acid, fumaric acid, maleic acid, and the like.

[0037] The polymerization in step 2 may be carried out by: 2.1. Dissolving an initiator in water and adding the resulting solution to the acidified monomer mixture obtained from step 1 to obtain an aqueous phase mixture; 2.2. In a separate vessel, dissolving an emulsifier in an organic solvent to obtain an oil phase mixture, wherein the organic solvent is immiscible in 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 and carrying out polymerization to produce the desired ammonium salt polymer; preferably, the polymerization is carried out at a temperature of, for example, 40 to 100°C, preferably 50 to 85°C; 2.4. Subject the reaction product obtained from step 2.3 to conventional workup to isolate the ammonium salt polymer of the present invention. For example, this workup can be carried out by filtering the reaction product, drying, and pulverizing to obtain the ammonium salt polymer of the present invention; preferably, this workup can be carried out by filtering the reaction product, slurrying the wet filter cake once with methanol or ethanol, filtering, further slurrying the wet filter cake with purified water, filtering, repeating the process of slurrying with purified water and filtering 2-3 times, drying, and pulverizing to obtain the ammonium salt polymer of the present invention; Optionally, step 2 includes: 2.5. Converting the wet filter cake obtained from step 2.4 or the ammonium salt polymer obtained from step 2.4 to an ammonium salt polymer of the present invention containing another anion It may further include.

[0038] 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, the anion X in the ammonium salt polymer obtained from Step 2.3 ―is a chloride anion. The polymer can be converted to an ammonium salt polymer containing another anion through step 2.5. For example, it can be converted to a bicarbonate and / or carbonate salt by adding the wet filter cake obtained from step 2.4 or the ammonium salt polymer obtained from step 2.4 to a saturated aqueous sodium bicarbonate or sodium carbonate solution, stirring for a period of time, followed by filtering, drying, and grinding to obtain an ammonium salt polymer, wherein the anion X in the ammonium salt polymer is ― is a bicarbonate and / or carbonate anion.

[0039] The initiator used in step 2.1 is not particularly limited, and may be any suitable initiator. For example, the initiator may be selected from 2,2'-azodiisobutylamidine dihydrochloride (V-50), 2,2'-azobis[2-(2-imidazolin-2-yl)propane]dihydrochloride (VA-044), ammonium persulfate, potassium persulfate, aqueous hydrogen peroxide, and any mixture thereof.

[0040] The emulsifier used in step 2.2 is not particularly limited, and the emulsifier may be any substance, such as a surfactant, having emulsifying properties that are useful for mixing the aqueous phase and the oil phase. For example, the emulsifier may be selected from polyoxyethylene sorbitan fatty acid esters (e.g., Tween 40, Tween 80, etc.), sorbitan fatty acid esters (e.g., Span 40, Span 60, Span 80, etc.).

[0041] In a third aspect, the present invention provides a pharmaceutical composition comprising an ammonium salt polymer of the present invention, and optionally a pharmaceutically acceptable carrier. The pharmaceutical composition may be used as a bile acid sequestrant, particularly for the treatment or prevention of diseases associated with excess bile acids.

[0042] Pharmaceutical compositions may be formulated into solid formulations (including but not limited to capsules, tablets, pills, granules, powders) or liquid formulations (including but not limited to suspensions) for oral administration by methods known in the art.

[0043] For oral formulations, pharmaceutically acceptable carriers may be: a) fillers, such as lactose, dextrose, sucrose, mannitol, sorbitol and cellulose; b) lubricants, such as silica, talc, stearic acid, magnesium or calcium stearate and / or polyethylene glycol; c) binders, such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidone; d) disintegrants, such as starch, agar, alginic acid or its sodium salt, or effervescent mixtures; and / or e) coloring and / or flavoring agents; f) Suspension agents.

[0044] In a fourth aspect, the present invention provides the use of an ammonium salt polymer of the invention in the manufacture of a medicament as a bile acid sequestrant, as well as the use of an ammonium salt polymer of the invention in the manufacture of a medicament for the treatment or prevention of a disease associated with excess bile acid.

[0045] In a fifth aspect, the present invention provides a method for treating or preventing a disease associated with excess bile acids, comprising administering to a subject in need thereof an effective amount of an ammonium salt polymer of the present invention.

[0046] The diseases associated with excess bile acids described herein are selected from dyslipidemia (e.g., hyperlipidemia), cholestatic liver disease (e.g., progressive familial intrahepatic cholestasis (PFIC), benign recurrent intrahepatic cholestasis (BRIC), inborn errors of bile acid synthesis, Alagille syndrome (ALGS), neonatal cholestasis caused by citrin 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. [Effects of the Invention]

[0047] The inventors have surprisingly found that the ammonium salt polymer of the present invention exhibits excellent bile acid adsorption capacity, which is significantly better than that of colesevelam hydrochloride, and its in vivo effect is in good agreement with its in vitro effect.

[0048] The ammonium salt polymer of the present invention is not absorbed into the bloodstream and adsorbs bile acids in the intestine, which are then excreted from the body in the feces, thereby effectively reducing intestinal bile acid reabsorption, improving intrahepatic and extrahepatic cholestasis, preventing the progression of cholestatic liver diseases such as PSC, and alleviating complications such as pruritus. Due to the non-systemic exposure of the ammonium salt polymer of the present invention, toxic side effects caused by small molecules such as odevixibat that enter the bloodstream are avoided. As a result, the ammonium salt polymer of the present invention exhibits good safety, can be administered over long periods of time, and has good patient compliance.

[0049] In addition, the method for preparing the ammonium salt polymers of the present invention is simple. [Brief explanation of the drawings]

[0050] [Figure 1] 1 shows the IR spectrum of the ammonium salt polymer WS012-A prepared in Example 1. [Figure 2]1 shows the 13C NMR spectrum of the ammonium salt polymer WS012-A prepared in Example 1. [Figure 3] 1 shows the DSC curve of the ammonium salt polymer WS012-A prepared in Example 1. [Figure 4] 1 shows the IR spectrum of the ammonium salt polymer WS012-B prepared in Example 5. [Figure 5] 1 shows the DSC curve of the ammonium salt polymer WS012-B prepared in Example 5. [Figure 6] 1 shows the IR spectrum of the ammonium salt polymer WS012-C prepared in Example 6. [Figure 7] 1 shows the DSC pattern of the ammonium salt polymer WS012-C prepared in Example 6. [Figure 8A] 1 shows the results of the DDC-induced primary sclerosing cholangitis mouse model of Example 15, where "Normal" represents the normal group, "Model" represents the model group, "Odevixibat" represents the odevixibat-treated group, "Cholevelam" represents the colesevelam hydrochloride-treated group, "WS012-A-Low" represents the group treated with low doses of WS012-A prepared in Example 2, and "WS012-A-High" represents the group treated with high doses of WS012-A prepared in Example 2. [Figure 8B] As above, the results of Example 15 for the DDC-induced primary sclerosing cholangitis mouse model are shown. [Figure 8C] As above, the results of Example 15 for the DDC-induced primary sclerosing cholangitis mouse model are shown. [Figure 8D] As above, the results of Example 15 for the DDC-induced primary sclerosing cholangitis mouse model are shown. [Figure 9A]1 shows the results of the ANIT-induced primary biliary cholangitis rat model of Example 16, where "Normal" represents the normal group, "Model" represents the model group, "Odevixibat" represents the odevixibat-treated group, "Colesevelam" represents the colesevelam hydrochloride-treated group, "WS012-A" represents the WS012-A-treated group prepared in Example 1, and "WS012-C" represents the WS012-C-treated group prepared in Example 6. [Figure 9B] As in the above, the results of the ANIT-induced primary biliary cholangitis rat model of Example 16 are shown. [Figure 9C] As in the above, the results of the ANIT-induced primary biliary cholangitis rat model of Example 16 are shown. [Figure 9D] As in the above, the results of the ANIT-induced primary biliary cholangitis rat model of Example 16 are shown. [Figure 10A]

[0049] Figure 17 shows the results of the high-fat diet-induced hyperlipidemia golden hamster model of Example 17, where "Normal" represents the normal group, "Model" represents the model group, "Colesevelam" represents the colesevelam 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. [Figure 10B] As above, the results of the high-fat diet-induced hyperlipidemia golden hamster model of Example 17 are shown. [Figure 10C] As above, the results of the high-fat diet-induced hyperlipidemia golden hamster model of Example 17 are shown. [Figure 10D] As above, the results of the high-fat diet-induced hyperlipidemia golden hamster model of Example 17 are shown. [Figure 10E] As above, the results of the high-fat diet-induced hyperlipidemia golden hamster model of Example 17 are shown. [Figure 10F] As above, the results of the high-fat diet-induced hyperlipidemia golden hamster model of Example 17 are shown. DETAILED DESCRIPTION OF THE INVENTION

[0051] Specific Embodiments Embodiment 1. A monomer of formula (I) and a monomer of formula (II):

[0052] [ka]

[0053] (In the formula, m, n, p, q, and r are each independently selected from 1, 2, or 3. An ammonium salt polymer obtained from the polymerization of The counter ion is a monovalent, divalent, or trivalent anion, and the anion is a chloride ion (Cl - ), bicarbonate ion (HCO3 - ), bisulfate ion (HSO4 - ), acetate ion (C2H3O2 - ), carbonate ions (CO3 2- ), sulfate ions (SO4 2- ), phosphate ions (PO4 3- ), and anions derived from malic acid, citric acid, fumaric acid, and maleic acid.

[0054] Embodiment 2. A polymer according to embodiment 1, wherein m, n, p, q and r are each independently selected from 1 and 2, and preferably m, n, p, q and r are all 1. Embodiment 3. The polymer according to embodiment 1 or 2, wherein the ammonium salt polymer is obtained from the polymerization of 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, even more preferably 10:1 to 10:4, and most preferably 10:2.

[0055] Embodiment 4. A polymer according to any one of embodiments 1 to 3, wherein the glass transition temperature of the polymer is from about 60°C to about 130°C, preferably from about 70°C to 120°C, and more preferably from about 70°C to about 105°C.

[0056] Embodiment 5. In the formula, the anion contained in the polymer is bicarbonate ion (HCO - ) and / or carbonate ions (CO3 2- 5. A polymer according to any one of embodiments 1 to 4, wherein m, n, p, q and r are all 1.

[0057] Embodiment 6. A polymer according to any one of embodiments 1 to 5, having an in vitro bile acid absorption capacity of 7 to 10 mmol / g, preferably 7 to 8 mmol / g. Embodiment 7. A method for preparing a polymer as defined in any one of embodiments 1 to 6, comprising: (1) acidifying the monomer of formula (I) and the monomer of formula (II) with an acid, wherein the molar ratio of the monomer of formula (I) to the monomer of formula (II) is preferably 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, even more preferably 10:1 to 10:4, and most preferably 10:2; (2) polymerizing the acidified monomer resulting from step (1) to obtain the desired ammonium salt polymer. A method comprising:

[0058] Embodiment 8. The method according to embodiment 7, wherein step (1) is carried out by adding the monomer of formula (I) and the monomer of formula (II) dropwise at a molar ratio of 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, even more preferably 10:1 to 10:4, and most preferably 10:2 to a suitable acid at a temperature of 30°C or less, preferably 15°C or less, and stirring for a period of time at a temperature of 5 to 30°C, preferably 10 to 20°C, after completion of the addition, to obtain a mixture of two acidified monomers.

[0059] Embodiment 9. The method according to embodiment 7 or 8, wherein the acid used in step (1) is selected from hydrochloric acid, sulfuric acid, acetic acid, malic acid, citric acid, maleic acid and fumaric acid. Embodiment 10. Step (2) comprises the following substeps: (2.1) dissolving an initiator in water and adding the resulting solution to the acidified monomer mixture obtained from step (1) to obtain an aqueous phase mixture; (2.2) in a separate vessel, dissolving an emulsifier in an organic solvent to obtain an oil phase mixture, wherein the organic solvent is immiscible in 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 and carrying out polymerization to produce the desired ammonium salt polymer; (2.4) subjecting the reaction product obtained from step (2.3) to a conventional workup to isolate the ammonium salt polymer of the present invention; for example, this conventional workup may be carried out by filtering the reaction product, drying, and pulverizing to obtain the ammonium salt polymer of the present invention; preferably, this conventional workup may be carried out by filtering the reaction product, slurrying the wet filter cake once with methanol or ethanol, filtering, further slurrying the wet filter cake with purified water, filtering, repeating the process of slurrying with purified water and filtering 2-3 times, drying, and pulverizing to obtain the desired ammonium salt polymer; Optionally, (2.5) converting the wet filter cake obtained from step (2.4) or the ammonium salt polymer obtained from step (2.4) into an ammonium salt polymer containing another anion; 10. The method according to any one of embodiments 7 to 9, comprising:

[0060] Embodiment 11. A method for preparing an ammonium salt polymer as defined in embodiment 5, comprising: (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, even more preferably 10:1 to 10:4, and most preferably 10:2 using concentrated hydrochloric acid to obtain a mixture of two acidified monomers; (2.1) dissolving an initiator in water and adding the resulting solution to the acidified monomer mixture obtained from step (1) to obtain an aqueous phase mixture; (2.2) in a separate vessel, dissolving an emulsifier in an organic solvent to obtain an oil phase mixture, wherein the organic solvent is immiscible in 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 carrying out polymerization at a temperature of 40 to 100°C, preferably 50 to 80°C, to produce the desired ammonium salt polymer; (2.4) filtering the reaction product obtained from step (2.3), slurrying the wet filter cake with methanol or ethanol once, filtering, further slurrying the wet filter cake with purified water, filtering, repeating the process of slurrying with purified water and filtering 2 to 3 times to obtain a wet filter cake; (2.5) adding the wet filter cake obtained from step (2.4) to a saturated aqueous sodium bicarbonate or sodium carbonate solution, stirring for a period of time, followed by filtering, drying, and pulverizing to obtain an ammonium salt polymer containing bicarbonate and / or carbonate anions. A method comprising:

[0061] Embodiment 12. The method according to any one of embodiments 7 to 11, wherein the initiator is selected from 2,2'-azodiisobutylamidine dihydrochloride (V-50), 2,2'-azobis[2-(2-imidazolin-2-yl)propane] dihydrochloride (VA-044), ammonium persulfate, potassium persulfate, aqueous hydrogen peroxide, and any mixture thereof; and the emulsifier is selected from polyoxyethylene sorbitan fatty acid esters (e.g., Tween 40, Tween 80, etc.), sorbitan fatty acid esters (e.g., Span 40, Span 60, Span 80, etc.), and any mixture thereof.

[0062] Embodiment 13. An ammonium salt polymer according to any one of embodiments 1 to 6 for use as a medicament, preferably for use as a medicament for the treatment or prevention of a disease associated with excess bile acids, preferably wherein the disease associated with excess bile acids is selected from dyslipidemia (e.g. hyperlipidemia), cholestatic liver disease (e.g. progressive familial intrahepatic cholestasis (PFIC), benign recurrent intrahepatic cholestasis (BRIC), inborn errors of bile acid synthesis, Alagille syndrome (ALGS), neonatal cholestasis caused by citrin 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.

[0063] Embodiment 14. A pharmaceutical composition comprising a polymer according to any one of embodiments 1 to 6, and optionally a pharmaceutically acceptable carrier. Embodiment 15. Use of a polymer according to any one of embodiments 1 to 6 in the manufacture of a medicament as a bile acid sequestrant, or for the treatment or prevention of a disease associated with excess bile acids, 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 (BRIC), inborn errors of bile acid synthesis, Alagille syndrome (ALGS), neonatal cholestasis caused by citrin 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.

[0064] Embodiment 16. A method for treating or preventing a disease associated with excess bile acids, comprising administering to a subject in need thereof an effective amount of the polymer of any one of embodiments 1 to 6, 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 (BRIC), inborn errors of bile acid synthesis, Alagille syndrome (ALGS), neonatal cholestasis caused by citrin 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.

[0065] Embodiment 17. The invention as described herein above.

[0066] definition As used herein, the term "subject" refers to an animal. Preferably, the subject is a mammal, such as a primate (e.g., human, monkey, chimpanzee), cow, sheep, goat, horse, dog, cat, rabbit, rat, mouse, etc. More preferably, the subject is a human.

[0067] As used herein, the term "treatment" or "treating" refers to the alleviation or cure of a disease or a symptom thereof, or the prevention of the progression of a disease or a symptom thereof. As used herein, the term "prevention" or "preventing" refers to slowing or preventing the development of a disease or its symptoms.

[0068] As used herein, the term "effective amount" refers to the amount of the ammonium salt polymer of the present invention that significantly reduces bile acid levels in the body, preventing the onset of a disease or its symptoms associated with excess bile acid, or ameliorating a disease or its symptoms associated with excess bile acid. The "effective amount" of the ammonium salt polymer of the present invention depends on various factors, including, but not limited to, the type and severity of the disease being treated or prevented, and the patient's general health and response. Generally, the "effective amount" of the ammonium salt polymer of the present invention is 0.5 to 24 g / day for a 70 kg human. However, if desired, the ammonium salt polymer of the present invention may be administered in amounts outside the aforementioned dosage ranges. The effective amount for a particular patient can be readily determined by a physician.

[0069] As used herein, the term "about" indicates that the numerical values ​​given hereinafter may be expanded by 20% or more, preferably 10% or more. For example, "about 100" means 80 to 120, preferably 90 to 110; "about 60°C" means 48 to 72°C, preferably 54 to 66°C. [Example]

[0070] The following examples illustrate the present invention and should not be construed in any way as limiting the scope of the invention. The disclosed data (e.g., amounts, temperatures, etc.) are provided as accurately as possible, but some experimental error and deviation may exist. Unless otherwise stated, temperatures are given in degrees Celsius and pressures are given at or near atmospheric pressure. Unless otherwise stated, all reagents used in the examples are publicly available.

[0071] The abbreviations used herein have the following meanings:

[0072] [Table 1]

[0073] Example 1 Preparation of Polymer WS012-A according to General Scheme 1 1.76 kg of concentrated hydrochloric acid was added to the reaction flask, and 1.46 kg (15 mol) of DAA and 0.41 kg (3 mol) of TAA were added dropwise to the reaction flask, maintaining the internal temperature below 15°C. After the addition was complete, the reaction mixture was stirred at 10-20°C for approximately 0.5-1 hour. 0.146 kg of V-50 and 1.46 kg of water were added to the beaker. After stirring to dissolve, the solution in the reaction flask was added to the beaker, and the mixture was stirred to ensure uniform mixing.

[0074] 8.76 kg of n-heptane and 0.133 kg of Span 60 were added to a reactor and heated to 40-50°C with stirring, with the mechanical stirring speed set at 200-250 RPM. The mass in the beaker was added to the reactor and heated to 50-55°C. The mixture was stirred at 50-55°C for 15-24 hours. After GC analysis of the reaction solution showed that the conversion of the DAA and TAA monomers was nearly complete, the reaction mixture was filtered. The wet filter cake was slurried with approximately 5.84 kg of ethanol for 1-2 hours, followed by filtration. The wet filter cake was then slurried with approximately 18.0-20.0 kg of water for 1-2 hours, followed by filtration. This process of slurrying with purified water and filtering was repeated two or three times. The wet filter cake was added to approximately 30.0 kg of saturated aqueous NaHCO3 solution and stirred at 25-35°C for 18-24 hours, followed by filtration. The process of slurrying and filtering was repeated twice. The wet filter cake was slurried with approximately 18.0-20.0 kg of water for 1-2 hours, followed by filtration. The process of slurrying and filtering was repeated two to three times. The wet filter cake was then rinsed with purified water. The filter cake was air-dried at 45-55°C, then crushed and sieved to yield 1.8 kg of polymer WS012-A as a pale yellow solid.

[0075] Attempts to dissolve the prepared polymer in various conventional solvents, such as water, methanol, N,N-dimethylformamide, dimethyl sulfoxide, showed that the polymer was not soluble in all of these solvents.

[0076] The prepared polymer WS012-A was characterized using a SHIMADZU IRSpirit-T Fourier transform infrared spectrophotometer according to infrared spectrophotometry (Chinese Pharmacopoeia 2020 Edition, Volume IV, General Chapters, 0402). The obtained IR spectrum is shown in Figure 1, which shows the following characteristic absorption bands (cm): -1 ) has: 2918, 2855, 2359, 2072, 1627, 1529, 1402, 1362, 1200, 1000, 909, 810.

[0077] IR spectra showed the presence of structures such as methylene, tertiary ammonium salts, secondary ammonium salts, CN, and carbonate in the product. The polymer WS012-A prepared in this example was detected using a Jnm-600ECZ solid-state NMR spectrophotometer. The test sample was packed into a 3.2 mm ZrO rotor, and the experimental parameters were as follows: a magic angle spinning speed of 15 KHz, a recycle delay time of 5 seconds, 1024 scans at 293 K, and the chemical shift of the carbonyl group of glycine at 176.04 ppm as the external standard. 13 The C solid-state NMR spectrum is shown in Figure 2 and has the following lines (ppm): 28.2, 42.9, 50.5, 163.3.

[0078] 13 C solid-state NMR spectra showed the presence of tertiary carbons, methylene carbons, C–N, and carbonates in the test samples. The prepared product WS012-A was detected by differential scanning calorimetry (DSC). Instrument: METTLER TOLEDO DSC3 differential scanning calorimeter. Analysis method: N2 atmosphere, 50 ml / min. Scanning procedure: Heat from 30°C to 180°C at 10°C / min, hold the temperature for 5 minutes, then cool to 30°C at 10°C / min, hold the temperature for 5 minutes, and heat to 180°C at 30°C / min, recording a second heating curve. The characteristic glass transition temperature was determined from the curve. An aluminum sample tray was used. The obtained DSC curve is shown in Figure 3. The results showed that the glass transition temperature of the polymer WS012-A was 99.5°C.

[0079] Example 2 (Alternative method for preparing polymer WS012-A according to general scheme 1) 101.54 g of concentrated hydrochloric acid was added to reaction flask 1. 82.59 g (0.85 mol) of DAA and 23.33 g (0.17 mol) of TAA were added dropwise to reaction flask 1, maintaining the internal temperature below 15°C. After the addition was complete, the reaction mixture was stirred at 10-20°C for approximately 0.5-1 hour. 5.93 g of VA-044 and 54.0 g of water were added to the Erlenmeyer flask and stirred to dissolve. The aqueous solution was then added to reaction flask 1 and stirred to ensure uniform mixing.

[0080] 580 mL of toluene and 4.1 g of Span 60 were added to reaction flask 2 and heated to 40-50°C with stirring, using a mechanical stirrer set at 300-400 RPM to dissolve the mixture. The mass from reaction flask 1 was added to flask 2 and heated to 50-55°C. The mixture was stirred at 50-55°C for 15-24 hours. After GC detection of the reaction solution indicated that the conversion of DAA and TAA was nearly complete, the internal temperature of the reaction mixture was increased to 80-85°C, and the reaction mixture was stirred at 80-85°C for 1-2 hours, then cooled and filtered. The wet filter cake was slurried in approximately 1 L of water for 0.5-1 hour, followed by filtration. This slurrying and filtering process was repeated 3-5 times. The wet filter cake was added to 500 mL of 10% aqueous NaOH solution and stirred at ambient temperature for approximately 8-12 hours. The reaction mixture was then filtered. The wet filter cake was slurried in approximately 1 L of water for 0.5-1 h and then filtered. The wet filter cake was added to approximately 1 L of saturated aqueous NaHCO3 solution and stirred at 20-30°C for 12-18 h, followed by filtration. These procedures were repeated once or twice. The wet filter cake was slurried in approximately 1 L of water for 0.5-1 h and then filtered. The slurrying and filtration process was repeated two or three times. The wet filter cake was rinsed with purified water. The filter cake was air-dried at 45-55°C, crushed, and sieved to obtain 80 g of polymer WS012-A as a pale yellow solid.

[0081] The product was detected using a SHIMADZU IRSpirit-T Fourier transform infrared spectrophotometer and a Jnm-600ECZ solid-state NMR resonator in Example 1. The results are shown in the IR spectrum of the polymer and13 The C solid-state NMR spectrum showed it to be the same as that of the polymer in Example 1 (spectrum not given).

[0082] In addition, the solubility of the product of this example in various solvents was also determined. The results showed that, like the product in Example 1, the polymer was not soluble in any of the solvents.

[0083] Example 3 (Alternative method for preparing polymer WS012-A according to general scheme 1) 101.54 g of concentrated hydrochloric acid was added to reaction flask 1. 82.59 g (0.85 mol) of DAA and 23.33 g (0.17 mol) of TAA were added dropwise to reaction flask 1, maintaining the internal temperature below 15°C. After the addition was complete, the reaction mixture was stirred at 10-20°C for approximately 0.5-1 h. 5.93 g of VA-044 and 54.0 g of water were added to the Erlenmeyer flask and stirred to dissolve. The aqueous solution was then added to reaction flask 1 and stirred to ensure uniform mixing.

[0084] 600 mL of n-heptane and 8.36 g of Span 60 were added to reaction flask 2 and heated to 40-50°C with stirring, dissolving the mixture with the mechanical stirring set at 300-400 RPM. The mass from reaction flask 1 was added to reaction flask 2 and heated to 50-55°C. The mixture was stirred at 50-55°C for 15-24 hours. After GC analysis of the reaction solution showed that the conversion of DAA and TAA was nearly complete, the reaction mixture was filtered. The wet filter cake was slurried in approximately 1 L of methanol for 0.5-1 h, followed by filtration. The wet filter cake was then slurried in approximately 1 L of water for another 0.5-1 h, followed by filtration. This slurrying and filtration process was repeated two or three times. The wet filter cake was added to 500 mL of 10% aqueous NaOH solution and stirred at ambient temperature for approximately 8-12 hours. The reaction mixture was then filtered. The wet filter cake was slurried in approximately 1 L of water for 0.5-1 h and then filtered. The wet filter cake was added to approximately 1 L of saturated aqueous NaHCO3 solution and stirred at 20-30°C for 12-18 h, followed by filtration. These procedures were repeated once or twice. The wet filter cake was slurried in approximately 1 L of water for 0.5-1 h and then filtered. The slurrying and filtration process was repeated two or three times. The wet product was then 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.

[0085] The product was detected using a SHIMADZU IRSpirit-T Fourier transform infrared spectrophotometer and a Jnm-600ECZ solid-state NMR resonator as described in Example 1. The results are shown in the IR spectrum of the polymer and 13 The C solid-state NMR spectrum showed it to be the same as that of the polymer in Example 1 (spectrum not given).

[0086] In addition, the solubility of the product of this example in various solvents was also determined. The results showed that, like the product in Example 1, the polymer was not soluble in any of the solvents.

[0087] Example 4 (Alternative method for preparing polymer WS012-A according to general scheme 1) 101.54 g of concentrated hydrochloric acid was added to reaction flask 1. 82.59 g (0.85 mol) of DAA and 23.33 g (0.17 mol) of TAA were added dropwise to reaction flask 1, maintaining the internal temperature below 15°C. After the addition was complete, the reaction mixture was stirred at 10-20°C for approximately 0.5-1 h. 5.93 g of VA-044 and 54.0 g of water were added to the Erlenmeyer flask and stirred to dissolve. The aqueous solution was then added to reaction flask 1 and stirred to ensure uniform mixing.

[0088] 480 mL of cyclohexane and 7.17 g of Span 60 were added to reaction flask 2 and heated to 40-50°C with stirring. The mixture was dissolved using a mechanical stirrer set at 300-400 RPM. The mass from reaction flask 1 was added to reaction flask 2 and heated to 50-55°C. The mixture was stirred at 50-55°C for 15-24 hours. After GC analysis of the reaction solution indicated that the conversion of DAA and TAA was nearly complete, the reaction mixture was filtered. The wet filter cake was slurried in approximately 1 L of methanol for 0.5-1 h, followed by filtration. The wet filter cake was then slurried in approximately 1 L of water for another 0.5-1 h, followed by filtration. This process of slurrying and filtering was repeated two or three times. The wet filter cake was added to 500 mL of 10% aqueous NaOH solution and stirred at ambient temperature for approximately 8-12 h. The reaction mixture was then filtered. The wet filter cake was slurried in approximately 1 L of water for 0.5-1 h and then filtered. The wet filter cake was added to approximately 1 L of saturated aqueous NaHCO3 solution and stirred at 20-30°C for 12-18 h, followed by filtration. These procedures were repeated once or twice. The wet filter cake was slurried in approximately 1 L of water for 0.5-1 h and then filtered. The slurrying and filtration process was repeated two or three times. The wet product was then 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.

[0089] The product was detected using a SHIMADZU IRSpirit-T Fourier transform infrared spectrophotometer and a Jnm-600ECZ solid-state NMR resonator as described in Example 1. The results are shown in the IR spectrum of the polymer and 13 The C solid-state NMR spectrum showed it to be the same as that of the polymer in Example 1 (spectrum not given).

[0090] In addition, the solubility of the product of this example in various solvents was also determined. The results showed that, like the product in Example 1, the polymer was not soluble in any of the solvents.

[0091] Example 5 (Preparation of Polymer WS012-B)

[0092] [ka]

[0093] 304.6 g of concentrated hydrochloric acid was added to reaction flask 1. 247.7 g (2.55 mol) of DAA and 69.99 g (0.51 mol) of TAA were added dropwise to reaction flask 1, maintaining the internal temperature below 15°C. After the addition was complete, the reaction mixture was stirred at 10-20°C for approximately 0.5-1 h. 17.79 g of VA-044 and 180 g of water were added to the Erlenmeyer flask and stirred to dissolve. The aqueous solution was then added to reaction flask 1 and stirred to ensure uniform mixing.

[0094] 1740 mL of toluene and 12.3 g of Span 60 were added to reaction flask 2 and heated to 40-50°C while stirring. The mixture was dissolved using a mechanical stirrer set at 300-400 RPM. The mass from reaction flask 1 was added to reaction flask 2 and heated to 50-55°C. The mixture was stirred at 50-55°C for 15-24 hours. After GC detection of the reaction solution indicated the reaction was complete, the internal temperature in the reaction flask was increased to 80-85°C. The mixture was stirred at 80-85°C for 1-2 hours, cooled, and filtered. The wet filter cake was slurried in approximately 4 L of water for 0.5-1 hour, followed by filtration. This slurrying and filtering process was repeated 3-5 times. The wet filter cake was added to 4 L of 10% aqueous NaOH solution and stirred at ambient temperature for approximately 8-12 hours. The reaction mixture was then filtered. The wet filter cake was slurried in approximately 1 L of water for 0.5-1 h and then filtered. The filter cake was air-dried at 45-55°C, crushed, and sieved to obtain a solid that was an ammonium salt polymer with chloride ions as the anion.

[0095] The solid, 55.0 g (0.18 mol) of 1-bromo-6-(trimethylammonium)hexyl bromide, and 2.4 L of methanol were added to a reaction flask, heated to 65-70°C, and stirred at 65-70°C for 24-36 hours. After the reaction was complete, the reaction mixture was filtered. The wet filter cake was slurried in approximately 1 L of methanol for 0.5-1 hour, followed by filtration. The wet filter cake was then slurried in approximately 1 L of water for 0.5-1 hour, followed by filtration. This process of slurrying with purified water and filtering was repeated two to three times. The wet filter cake was added to approximately 4 L of saturated aqueous NaHCO3 solution, stirred at 20-30°C for 12-18 hours, followed by filtration. These procedures were repeated once or twice. The wet filter cake was then slurried in approximately 1 L of water for 0.5-1 hour, followed by filtration. This process of slurrying with purified water and filtering was repeated two to three times. The wet product was then rinsed with purified water. The filter cake was air-dried at 45-55° C. The dried product was crushed and sieved to give 170 g of polymer WS012-B as a pale yellow solid.

[0096] The product of this example was detected using the SHIMADZU IRSpirit-T Fourier transform infrared spectrophotometer described in Example 1. The IR spectrum is shown in FIG. In addition, the solubility of the product of this example in various solvents was also determined. The results showed that, like the product in Example 1, the polymer was not soluble in any of the solvents.

[0097] The product WS012-B prepared in this example was determined using differential scanning calorimetry (DSC) as in Example 1. The resulting DSC curve is shown in Figure 5. The results showed that the glass transition temperature of the polymer WS012-B was 58.8°C.

[0098] Example 6 (Preparation of Ammonium Salt Polymer WS012-C)

[0099] [ka]

[0100] 178.9 g of concentrated hydrochloric acid was added to a 3 L reaction flask. 246.8 g (1.8 mol) of TAA was added dropwise to the reaction flask, maintaining the internal temperature below 15°C. After the addition was complete, the reaction mixture was stirred at 10-20°C for approximately 0.5-1 h. 15.7 g of VA-044 and 45 g of water were added to an Erlenmeyer flask and stirred to dissolve. The aqueous solution was then added to reaction flask 1, and 242.5 g (0.9 mol) of 60% dimethyldiallylammonium chloride aqueous solution was added and stirred to ensure uniform mixing.

[0101] 1.6 L of toluene and 11.8 g of Span 60 were added to a reaction flask and heated to 40-50°C with stirring. The mixture was dissolved using a mechanical stirrer set at 200-300 RPM. The mass in the reaction flask was then added to the reactor. The temperature was increased to 50-55°C, and the reaction mixture was stirred at 50-55°C for 15-24 hours. After GC analysis of the reaction solution indicated that the conversion of TAA was nearly complete, the reaction mixture was filtered. The wet filter cake was slurried in approximately 3 L of methanol for 0.5-1 hour, followed by filtration. The wet filter cake was then slurried in approximately 10 L of water for another 0.5-1 hour, followed by filtration. This slurrying and filtration process was repeated once or twice. The wet filter cake was added to approximately 7.5 L of saturated aqueous NaHCO3 solution, stirred at 20-30°C for 12-18 hours, followed by filtration. These procedures were repeated once or twice. The wet filter cake was slurried with approximately 10 L of water for 0.5-1 h, followed by filtration. The slurrying and filtration process was repeated 2-3 times. The wet product was then rinsed with purified water. The filter cake was air-dried at 45-55°C, crushed, and sieved to yield approximately 200 g of polymer WS012-C as a pale yellow solid.

[0102] The product of this example was detected using the SHIMADZU IRSpirit-T Fourier transform infrared spectrophotometer described in Example 1. The IR spectrum is shown in FIG. The product WS012-C prepared in this example was determined using differential scanning calorimetry (DSC) as in Example 1. The resulting DSC curve is shown in Figure 7. The results showed that the glass transition temperature of the polymer WS012-C was 177.8°C.

[0103] Examples 7 to 13 In Examples 7-13, polymers were prepared using different amounts of each DAA and / or TAA according to the method described in Example 1, and the products were detected using the detection method described in Example 1. The amounts of each reactant and the experimental results in Examples 7-13 are shown below:

[0104] [Table 2]

[0105] The above results showed that no solid polymer was formed when DAA was used as the sole monomer for the reaction in Example 2, but polymers were formed as pale yellow solids in Examples 3 to 13. The products were detected using the DSC method described in Example 1, and the resulting glass transition temperatures are shown in the last column of the table above.

[0106] Example 14 (In vitro bile acid adsorption experiment) 1. Solution Preparation Mixed bile acid solution: 6 mmol of sodium glycocholate, 6 mmol of sodium glycochenodeoxycholate, and 2 mmol of sodium taurodeoxycholate were each contained in 1 L of 0.68% phosphate buffered saline to obtain a mixed bile acid solution (pH 6.8).

[0107] Linear solutions: The mixed bile acid solution was diluted to obtain linear solutions 1 to 5, each containing 0.06, 0.18, 0.3, 0.48, or 0.6 mmol of sodium glycocholate, 0.06, 0.18, 0.3, 0.48, or 0.6 mmol of sodium glycochenodeoxycholate, and 0.02, 0.06, 0.1, 0.16, or 0.2 mmol of sodium taurodeoxycholate in 1 L of purified water.

[0108] Test solution: 100 mg of test sample was accurately weighed into a 250 ml iodine flask, and 100 ml of the mixed bile acid solution was added. The mixture was stirred using a magnetic stir bar in a water bath (37°C ± 0.5°C). 5 ml of the solution was sampled after 5 hours and allowed to stand at room temperature. (After sampling, the solution was allowed to stand at room temperature for at least 5 minutes. The iodine flask was then tilted and shaken to remove any water droplets on the top of the iodine flask. The iodine flask was shaken and rotated for at least 15 seconds.) The sample was filtered. 1.0 ml of the filtrate was placed in a 10 ml volumetric flask, diluted to the mark with purified water, and shaken until homogenous for injection.

[0109] [Table 3]

[0110] 2. Determination of bile acid excretion capacity using the standard curve method A linear regression equation, Y = aX + b, was fitted using the bile acid concentration as the linear solution (unit: mmol / L) as the X-coordinate and the peak area as the Y-coordinate.

[0111]

number

[0112] 3. Experimental Results

[0113] [Table 4]

[0114] The results of the above experiments showed that the ammonium salt polymer WS012-A of the present invention had the highest total bile acid adsorption capacity in vitro, and was superior to the ammonium salt polymer WS012-B, ammonium salt polymer WS012-C, ammonium salt polymer WS012-D, and colesevelam hydrochloride.

[0115] Example 15 Effect of WS012-A on DDC-induced primary sclerosing cholangitis (PSC) in mice 1. Experimental Procedure: After the acclimation period, 36 male C57BL / 6J mice (18–22 g, 6–8 weeks old) were randomly grouped according to their body weight into six experimental groups: normal control group, model control group, active control group 1, active control group 2, low-dose WS012-A group, and high-dose WS012-A group, with six animals in each group. Vehicle (0.5% CMC-Na aqueous solution) was administered to the normal control group and the model control group, 0.3 mg / kg of odevixibat (BiChemPartner) was administered to active control group 1, 1,300 mg / kg of colesevelam hydrochloride (Formosa Laboratories, Inc.) was administered to active control group 2, 300 mg / kg of WS012-A prepared in Example 2 was administered to the low-dose WS012-A group, and 1,000 mg / kg of WS012-A was administered to the high-dose WS012-A group. Modeling was performed according to a known method (Peter Fickert et al., The American Journal of Pathology, Vol. 171, No. 2, August 2007, pp. 525-536). Specifically, except for the mice in the normal control group, the mice in the other groups were fed 0.1% DDC for 14 consecutive days with ad libitum consumption of food and water. The animals were administered once daily for 14 consecutive days under modeling. The specified test sample or vehicle was administered to all groups by gavage at a volume of 20 ml / kg. On the day after the final administration, blood was collected from the animals under anesthesia for the detection of alkaline phosphatase (ALP), total bile acids (TBA), and total bilirubin (TBIL). Feces were collected for 24 hours on days 13 and 14 to detect the TBA content in the feces. The feces were pretreated as follows: 50 mg of feces from each case was weighed, and a 2% aqueous SDS solution was added at a ratio of 1:20 w / v, followed by ultrasonication for 20 minutes and centrifugation at 1000 rpm for 5 minutes. The supernatant was taken for detection.

[0116] All parameters were expressed as mean ± SEM. Pre- and post-treatment animal parameters in various groups were plotted using Graph Pad Prism 8 software. Data were analyzed using SPSS 22.0 statistical software. Parameters were subjected to a test for homogeneity of variance using Levene's test. When variances were homogeneous (P ≥ 0.05), differences between groups were compared using Dunnett's test and least significant difference test in one-way analysis of variance (ANOVA); when variances were heterogeneous (P < 0.05), differences between groups were compared using the Mann-Whitney U test (MW method) in the Kruskal-Wallis H rank sum test (KW method).

[0117] 2. Experimental results:

[0118] [Table 5]

[0119] The results of the experiment in this example are shown in FIG. Figure 8A shows that, compared with the normal group, serum ALP in the model group was significantly elevated (P<0.001); compared with the model group, serum ALP in the odevixibat-treated group was significantly decreased (P<0.001), serum ALP in the colesevelam hydrochloride-treated group was slightly elevated without statistical significance, and serum ALP in both the low-dose WS012-A-treated group and the high-dose WS012-A-treated group was significantly decreased, which exhibited a dose-dependent relationship to some extent, where the decrease in ALP in the high-dose WS012-A-treated group was statistically significant (P<0.05).

[0120] Figure 8B shows that, compared with the normal group, serum TBA in the model group was significantly elevated (P<0.001); compared with the model group, serum TBA in the odevixibat-treated group was significantly decreased (P<0.05), serum TBA in the colesevelam hydrochloride-treated group was not substantially decreased, and serum TBA in both the low-dose WS012-A-treated group and the high-dose group was significantly decreased, which showed a dose-dependent relationship to some extent, where the decrease in TBA in the high-dose WS012-A-treated group was statistically significant (P<0.05).

[0121] Figure 8C shows that, compared with the normal group, serum TBIL in the model group was significantly elevated (P<0.001); compared with the model group, serum TBIL in the odevixibat-treated group was significantly decreased (P<0.05), serum TBIL in the colesevelam hydrochloride-treated group was elevated, and serum TBIL in both the low-dose WS012-A-treated group and the high-dose WS012-A-treated group was significantly decreased, which exhibited a dose-dependent relationship to some extent, in which the decrease in TBIL in the high-dose WS012-A-treated group was statistically significant (P<0.05); and serum TBIL in the low-dose WS012-A-treated group and the high-dose WS012-A-treated group was significantly lower than that in the colesevelam hydrochloride-treated group (P<0.01 and P<0.001, respectively).

[0122] Figure 8D shows that, compared with the normal group, the fecal TBA excretion rate in the model group was significantly decreased (P<0.001); compared with the model group, the fecal TBA excretion rate in the odevixibat-treated group was significantly increased (P<0.001), the fecal TBA excretion rate in the colesevelam hydrochloride-treated group was significantly increased (P<0.05), and the TBA excretion rate in both the low-dose WS012-A-treated group and the high-dose WS012-A-treated group was significantly increased (P<0.05).

[0123] The above results showed that: (1) Compared with the normal group, serum ALP, TBA, and TBIL in the model group were all significantly elevated, and the fecal TBA excretion rate was significantly decreased, indicating that DDC feeding led to significant changes in the physiological indexes of cholestasis in mice and that this model was successfully established.

[0124] (2) Compared with the model group, serum ALP, TBA, and TBIL significantly decreased, and fecal TBA excretion rate significantly increased in the odevixibat [first-line drug for the treatment of progressive familial intrahepatic cholestasis (PFIC)] treatment group, indicating that this drug has a good therapeutic effect on cholestasis.

[0125] (3) Compared with the model group, in the colesevelam hydrochloride-treated group, serum ALP and TBIL increased, which represents an undesirable effect on the treatment of cholestasis; serum TBA did not decrease significantly, indicating that colesevelam hydrochloride had no significant effect on this parameter; and fecal TBA excretion rate increased significantly, which is beneficial for the treatment of cholestasis. Overall, colesevelam hydrochloride did not exhibit a significant improving effect on cholestasis and is not suitable for the treatment of cholestasis.

[0126] (4) Compared with the model group, serum ALP, TBA, and TBIL in both the low-dose and high-dose WS012-A treatment groups were significantly reduced, which exhibited a dose-dependent relationship to some extent, where the reductions in ALP, TBA, and TBIL in the high-dose WS012-A treatment group were statistically significant; and the increase in fecal TBA excretion rate in both the low-dose and high-dose WS012-A treatment groups was statistically significant. Overall, WS012-A exhibited a therapeutic effect on cholestasis, and this therapeutic effect was dose-dependent.

[0127] (5) By comparing the colesevelam hydrochloride-treated group with the low-dose WS012-A-treated group, it can be seen that the effects on these four investigated parameters produced by low-dose WS012-A were better than those produced by colesevelam hydrochloride.

[0128] 3. Experimental conclusion: In a DDC-feeding mouse model of PSC induced by oral administration of WS012-A by gavage at 300 mg / kg and 1000 mg / kg once daily for 14 consecutive days, WS012-A increased the fecal TBA excretion rate, reduced serum ALP, TBA, and TBIL, and alleviated the pathological changes of hepatic cholestasis, and these effects were better than those of colesevelam hydrochloride, which also belongs to the polymeric bile acid sequestrants.

[0129] Example 16 Effect of WS012-A on ANIT-induced acute cholestasis in rats 1. Experimental Procedure: After the acclimation period, 48 male SD rats (180-220 g, 6-8 weeks old) were randomly divided into five experimental groups based on their body weight: normal control group, model control group, active control group 1, active control group 2, WS012-A group, and WS012-C group, with eight animals in each group. Vehicle (0.5% CMC-Na aqueous solution) was administered to the normal control group and model control group, 0.3 mg / kg of odevixibat was administered to active control group 1, 1,000 mg / kg of colesevelam hydrochloride was administered to active control group 2, 1,000 mg / kg of WS012-A prepared in Example 1 was administered to group WS012-A, and 1,000 mg / kg of WS012-C prepared in Example 6 was administered to group WS012-C. The specific test sample or vehicle was administered to all groups by gavage at a volume of 20 ml / kg once daily for 7 days. Modeling was performed according to methods known in the art [Yoshiji Ohta et al., Toxicology 139 (1999) pp. 265-275]. Specifically, except for rats in the normal control group, rats in the other groups were intraperitoneally injected with ANIT at a single dose of 80 mg / kg on the fourth day of administration. On the seventh day of administration (72±1 h after injection of the modeling agent ANIT), the animals were anesthetized, and whole blood was collected from the jugular venous plexus and centrifuged to collect serum for the detection of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and total bile acid (TBA). All parameters were expressed as mean ± SEM. Pre- and post-treatment animal parameters in various groups were plotted using Graph Pad Prism 8 software. Data were analyzed using SPSS 22.0 statistical software. Parameters were subjected to a test for homogeneity of variance using Levene's test. When variances were homogeneous (P ≥ 0.05), differences between groups were compared using Dunnett's test and least significant difference test in one-way analysis of variance (ANOVA); when variances were heterogeneous (P < 0.05), differences between groups were compared using the Mann-Whitney U test (MW method) in the Kruskal-Wallis H rank sum test (KW method).

[0130] 2. Experimental results:

[0131] [Table 6]

[0132] The results of the experiment in this example are shown in FIG. 9A shows that serum ALT levels in the model group were significantly elevated (P<0.001) compared with the normal control group; serum ALT levels in the odevixibat-treated group, colesevelam hydrochloride-treated group, WS012-A-treated group, and WS012-C-treated group were reduced compared with the model group, with the reductions in the colesevelam hydrochloride-treated group (P<0.05), WS012-A-treated group (P<0.01), and WS012-C-treated group (P<0.01) being statistically significant. WS012-A and WS012-C exhibited better effects than colesevelam hydrochloride in reducing serum ALT.

[0133] Figure 9B shows that compared with the normal group, serum AST in the model group was significantly elevated (P<0.001); compared with the model group, serum AST in the odevixibat-treated group, colesevelam hydrochloride-treated group, WS012-A-treated group, and WS012-C-treated group was decreased, where the decrease in the colesevelam hydrochloride-treated group was not statistically significant, while the decrease in both the WS012-A-treated group and the WS012-C-treated group was statistically significant (P<0.05), with the WS012-A group showing the best effect.

[0134] Figure 9C shows that compared with the normal group, serum ALP in the model group was significantly increased (P<0.001); compared with the model group, serum ALP in the odevixibat-treated group, colesevelam hydrochloride-treated group, WS012-A-treated group, and WS012-C-treated group was significantly decreased (P<0.01), with the WS012-A group showing the best effect.

[0135] Figure 9D shows that, compared with the normal group, serum TBA in the model group was significantly increased (P<0.01); compared with the model group, serum TBA in the odevixibat-treated group, colesevelam hydrochloride-treated group, WS012-A-treated group, and WS012-C-treated group was significantly decreased (P<0.05 or P<0.01), and serum TBA in the WS012-A-treated group and WS012-C-treated group was lower than that in the colesevelam hydrochloride-treated group.

[0136] The above results showed that: (1) Compared with the normal group, serum ALT, AST, ALP, and TBA in the model group were significantly elevated (P<0.05~0.001), suggesting that ANIT feeding led to significant changes in the physiological indexes of cholestasis and that the model was successfully established.

[0137] (2) Compared with the model group, serum ALT, AST, ALP, and TBA were reduced in the odevixibat-treated group, colesevelam hydrochloride-treated group, and WS012-A-treated group. The reductions in serum ALP and TBA in the odevixibat-treated group were statistically significant (P<0.01), the reductions in serum ALT, ALP, and TBA in the colesevelam hydrochloride-treated group were statistically significant (P<0.05-0.01), and the reductions in serum ALT, ALP, and TBA in the WS012-A-treated group and WS012-C-treated group were statistically significant (P<0.05-0.01).

[0138] 3. Experimental Conclusion In a rat model of acute cholestatic cholestasis induced by a single intraperitoneal injection of ANIT (80 mg / kg), followed by oral administration of WS012-A or WS012-C by gavage at 1000 mg / kg once daily for 7 consecutive days, WS012-A significantly reduced serum ALT (P<0.01), serum ALP (P<0.01), and serum TBA (P<0.05), and to some extent reduced serum AST; WS012-C also significantly reduced serum ALT (P<0.01), AST (P<0.05), ALP (P<0.01), and TBA (P<0.01). Overall, both WS012-A and WS012-C exhibited cholestasis-ameliorating effects that were better than those of colesevelam hydrochloride, which also belongs to the polymeric bile acid sequestrants, and even better than those of the small molecule drug odevixibat (an IBAT inhibitor).

[0139] Example 17 (Effect of WS012-A on high-fat diet-induced hyperlipidemia in rats) 1. Experimental Procedure: After a 7-day acclimation period, 8 of the 48 male golden hamsters (100-150 g, 6-7 weeks old) were grouped into a blank control group and continuously fed a basal diet, while the remaining animals were grouped into experimental groups and fed a high-fat diet (basal diet + 15% lard + 20% sucrose + 1.2% cholesterol + 0.2% sodium cholate). The animals' condition, including vitality and diet, was regularly observed. After 7 weeks of feeding the high-fat diet, the animals were fasted overnight. The next day, blood was collected from the orbital venous plexus for detection of plasma TC, TG, and LDL-C contents. The golden hamsters were grouped according to the detection results. Thirty-two golden hamsters with hyperlipidemia were screened based on blood biochemical indices and body weight and divided into four groups on average: model control group (model), test sample group 1 (WS012-A prepared in Example 2, 1000 mg / kg), test sample group 2 (WS012-B prepared in Example 5, 1000 mg / kg), and active control group (colesevelam hydrochloride, 1000 mg / kg). The specified test sample or vehicle was administered once daily by gavage at a volume of 20.0 ml / kg for 4 weeks. The blank control group was fed a normal diet, and the other groups continued to be fed a high-fat diet. During the administration period, the hamsters were weighed once a week to observe changes in body weight. Blood was collected before administration and at 1, 2, 3, and 4 weeks after administration for the detection of serum total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C); feces were collected at 2 and 4 weeks (within 24 hours after administration) for the detection of total bile acids (TBA) in the fecal supernatant. After the final fecal collection, the animals were euthanized, and the livers were removed for observation.

[0140] All parameters were expressed as mean ± SEM. Pre- and post-treatment animal parameters in various groups were plotted using Graph Pad Prism 8 software. Data were analyzed using SPSS 22.0 statistical software. Parameters were subjected to a test for homogeneity of variance using Levene's test. When variances were homogeneous (P ≥ 0.05), differences between groups were compared using Dunnett's test and least significant difference test in one-way analysis of variance (ANOVA); when variances were heterogeneous (P < 0.05), differences between groups were compared using the Mann-Whitney U test (MW method) in the Kruskal-Wallis H rank sum test (KW method).

[0141] 2. Experimental results: The results of the experiment in this example are shown in FIG.

[0142] Figure 10A shows that, compared with the normal group, blood total cholesterol (TC) in the model group was significantly elevated before and 1 to 4 weeks after administration (P<0.001). Total cholesterol (TC) increased slowly due to the prolonged high-fat diet induction period. Compared with the model group, TC concentrations in the active control (colesevelam hydrochloride) group were reduced to some extent due to drug administration. However, the reduction in TC in the colesevelam hydrochloride-treated group was statistically significant only after 3 weeks of administration (P<0.01). The reductions in TC at other sampling time points were not significantly different from those in the model group. After 1 to 4 weeks of administration, TC levels in the WS012-A-treated group were significantly reduced (P<0.05-0.001); after 2 to 4 weeks of administration, TC levels in the WS012-B-treated group were significantly reduced (P<0.05-0.001). Compared with the active drug (colesevelam hydrochloride) group, the TC levels in both the WS012-A and WS012-B treatment groups were significantly reduced after 4 weeks of administration (P<0.01). The results showed that both WS012-A and WS012-B exhibited better effects than colesevelam hydrochloride in reducing TC, with WS012-A exhibiting the best effect.

[0143] Figure 10B shows that, compared with the normal group, blood triglycerides (TG) in the model group were significantly elevated before and after 1 to 4 weeks of treatment (P<0.05). Compared with the model group, TG in the colesevelam hydrochloride-treated group was not significantly different after drug treatment; TG in the WS012-A-treated group was lower than that in the model group throughout the entire treatment period. Compared with the model group, TG in the WS012-A-treated group was significantly decreased after 1, 2, and 4 weeks of treatment (P<0.05); TG in the WS012-B-treated group was not significantly different from that in the model group throughout the entire treatment period (P>0.05). Compared with the active control (colesevelam hydrochloride) group, TG in the WS012-A-treated group was significantly lower after 4 weeks of treatment (P<0.01). The above results showed that WS012-A exhibited excellent effect in reducing TG, which was better than that of colesevelam hydrochloride.

[0144] FIG. 10C showed that compared with the normal group, blood low-density lipoprotein (LDL-C) in the model group was significantly elevated before and after 1 to 4 weeks of administration (P<0.001). Compared with the model group, the LDL-C concentration in the colesevelam hydrochloride-treated group was reduced to some extent after drug administration. However, the reduction in LDL-C in the colesevelam hydrochloride-treated group was significant only after 3 weeks of administration (P<0.05), and the reductions in LDL-C at other sampling time points were not significantly different from those in the model group (P>0.05). The LDL-C levels in the WS012-A-treated group were all significantly reduced after 1 to 4 weeks of administration (P<0.05-0.001). The LDL-C levels in the WS012-B-treated group were lower than those in the model group throughout the entire administration period, but the reductions in LDL-C in the WS012-B-treated group were significant only after 1 and 4 weeks of administration (P<0.01-0.001). Compared with the colesevelam hydrochloride group, the LDL-C levels in the WS012-A group were significantly reduced after 1, 2, and 4 weeks of administration (P<0.05-0.001); the LDL-C levels in the WS012-B group were significantly reduced after 1 and 4 weeks of administration. The results showed that both WS012-A and WS012-B exhibited significantly better effects on lowering LDL-C than colesevelam hydrochloride, with WS012-A exhibiting the best effect.

[0145] Figures 10D and 10E show that, compared with the normal group, the liver weight and liver index in the model group significantly increased after completion of drug administration (P<0.001), suggesting that the model was successfully established. Compared with the model group, the liver weight and liver index in the WS012-A and WS012-B treatment groups decreased after completion of drug administration, with the decrease in liver weight and liver index in the WS012-A treatment group (P<0.05) being statistically significant. Compared with the active control colesevelam hydrochloride group, the liver weight and liver index in the WS012-A and WS012-B treatment groups decreased after completion of drug administration, but only the decrease in liver weight (P<0.05) and liver index (P<0.01) in the WS012-A treatment group was statistically significant. The above results showed that both WS012-A and WS012-B were effective in ameliorating high-fat diet-induced hepatomegaly, but only the effect of WS012-A was significantly better than that of colesevelam hydrochloride.

[0146] Figure 10F shows that the livers of animals in the normal control group were bright red and had a smooth surface and soft texture. The livers of golden hamsters in the model control group were significantly increased in size, tended to be white, were generally swollen, exhibited slightly rounded edges, and a brittle texture. The liver tissue of animals in the active control colesevelam hydrochloride group was slightly different from that of the model group, but the difference was small. The livers of animals in the active control group were significantly increased in size, tended to be white, were generally swollen, exhibited slightly rounded edges, and a brittle texture, while no decrease was found in liver weight or liver index. Compared with the model control group and the normal control group, the livers of animals in the WS012-A treatment group and the WS012-B treatment group were bright red and tended to be white at the edges, and their overall size was between that of the model control group and the normal control group. Among the test samples, WS012-A showed the best effect.

[0147] 3. Experimental conclusion: In a 7-week high-fat diet-induced hyperlipidemia model in golden hamsters, oral administration of WS012-A or WS012-B by gavage at 1000 mg / kg once daily for 4 consecutive weeks reduced blood lipid (TC, TG, LDL-C) levels and alleviated liver fatty pathological changes (liver weight, liver index, liver overall observation). Among the tested samples, WS012-A exhibited the best effect, which was better than that of colesevelam hydrochloride, which also belongs to the polymeric bile acid sequestrants.

[0148] Based on the above experimental results of the present disclosure, it can be concluded that the ammonium salt polymer of the present invention exhibited excellent adsorption properties of bile acids in vitro and in vivo, better than that of colesevelam hydrochloride and better than that of the other ammonium salt polymers WS012-B and WS012-C prepared according to the present disclosure.

[0149] The specific embodiments and examples given in this specification are only for the purpose of illustrating the present invention and do not constitute any limitation on the scope defined by the claims. Based on this disclosure, those skilled in the art can clearly understand equivalent variations of the technical solutions of the present disclosure. These variations are also encompassed by the present disclosure.

Claims

1. Monomers of formula (I) and monomers of formula (II): 【Chemical 1】 (In the formula, m, n, p, q, and r are each independently selected from 1, 2, or 3. An ammonium salt polymer obtained from the polymerization of The counter ion contains a monovalent, divalent, or trivalent anion, and the anion is a chloride ion (Cl － ), bicarbonate ion (HCO ３ － ), bisulfate ion (HSO ４ － ), acetate ion (C ２ H ３ O ２ -), carbonate ion (CO ３ ２－ ), sulfate ions (SO ４ ２－ ), phosphate ions (PO ４ ３－ ), and anions derived from malic acid, citric acid, fumaric acid, and maleic acid.

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

3. 3. The polymer according to claim 1 or 2, obtained from the polymerization of a monomer of formula (I) and a monomer of formula (II) in a molar ratio of from 10:0.5 to 10:15, preferably from 10:1 to 10:12, more preferably from 10:1 to 10:10, even more preferably from 10:1 to 10:4, and most preferably from 10:

2.

4. 4. The polymer of any one of claims 1 to 3, wherein the polymer has a glass transition temperature of 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.

5. The anion contained in the polymer is a bicarbonate ion (HCO ３ － ) and / or carbonate ions (CO ３ ２－ 5. The polymer of claim 1, wherein m, n, p, q, and r are all 1.

6. 6. The polymer according to any one of claims 1 to 5, having an in vitro bile acid absorption capacity of 7 to 10 mmol / g, preferably 7 to 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, wherein the molar ratio of the monomer of formula (I) to the monomer of formula (II) is preferably 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, even more preferably 10:1 to 10:4, and most preferably 10:2; (2) polymerizing the acidified monomer resulting from step (1) to obtain the desired ammonium salt polymer. A method comprising:

8. 8. The method of claim 7, wherein step (1) is carried out by adding the monomer of formula (I) and the monomer of formula (II) dropwise in a molar ratio of 10:0.5 to 10:15, preferably 10:1 to 10:12, more preferably 10:1 to 10:10, even more preferably 10:1 to 10:4, and most preferably 10:2 to a suitable acid at a temperature of 30°C or less, preferably 15°C or less, and stirring for a period of time at a temperature of 5 to 30°C, preferably 10 to 20°C, after completion of the addition, to obtain a mixture of two acidified monomers.

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

10. Step (2) comprises the following sub-steps: (2.1) dissolving an initiator in water and adding the resulting solution to the acidified monomer mixture obtained from step (1) to obtain an aqueous phase mixture; (2.2) In a separate vessel, dissolving an emulsifier in an organic solvent to obtain an oil phase mixture, wherein the organic solvent is immiscible in 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 and carrying out polymerization to produce the desired ammonium salt polymer; (2.4) subjecting the reaction mixture obtained from step (2.3) to conventional workup to isolate the desired ammonium salt polymer; Optionally, (2.5) converting the wet filter cake obtained from step (2.4) or the ammonium salt polymer obtained from step (2.4) into an ammonium salt polymer containing another anion; 10. The method of any one of claims 7 to 9, comprising:

11. 6. A method for preparing the ammonium salt polymer of claim 5, comprising: (1) acidifying the monomer of formula (I) and the 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, even more preferably 10:1 to 10:4, and most preferably 10:2 using concentrated hydrochloric acid to obtain a mixture of two acidified monomers; (2.1) dissolving an initiator in water and adding the resulting solution to the mixture of acidified monomers obtained from step (1) to obtain an aqueous phase mixture; (2.2) in a separate vessel, dissolving an emulsifier in an organic solvent to obtain an oil phase mixture, wherein the organic solvent is immiscible in 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 carrying out polymerization at a temperature of 40 to 100°C, preferably 50 to 80°C, to produce the desired ammonium salt polymer; (2.4) filtering the reaction product obtained from step (2.3), slurrying the wet filter cake with methanol or ethanol once, filtering, further slurrying the wet filter cake with purified water, filtering, repeating the process of slurrying with purified water and filtering 2-3 times to obtain a wet filter cake; (2.5) adding the wet filter cake obtained from step (2.4) to a saturated aqueous sodium bicarbonate or sodium carbonate solution, stirring for a period of time, followed by filtering, drying, and pulverizing to obtain an ammonium salt polymer containing bicarbonate and / or carbonate anions. A method comprising:

12. 12. The method according to any one of claims 7 to 11, wherein the initiator is selected from 2,2'-azodiisobutylamidine dihydrochloride (V-50), 2,2'-azobis[2-(2-imidazolin-2-yl)propane] dihydrochloride (VA-044), ammonium persulfate, potassium persulfate, aqueous hydrogen peroxide, and any mixture thereof; and the emulsifier is selected from polyoxyethylene sorbitan fatty acid esters (e.g., Tween 40, Tween 80, etc.), sorbitan fatty acid esters (e.g., Span 40, Span 60, Span 80, etc.), and any mixture thereof.

13. 10. The ammonium salt polymer according to claim 1, for use as a medicament, preferably for use as a medicament for the treatment or prevention of diseases associated with excess bile acids, preferably wherein said diseases associated with excess bile acids are selected from dyslipidemia (e.g. hyperlipidemia), cholestatic liver diseases (e.g. progressive familial intrahepatic cholestasis (PFIC), benign recurrent intrahepatic cholestasis (BRIC), inborn errors of bile acid synthesis, Alagille syndrome (ALGS), neonatal cholestasis caused by citrin 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 a polymer according to any one of claims 1 to 6, and optionally a pharmaceutically acceptable carrier.

15. 10. A method for treating or preventing a disease associated with excess bile acids, comprising the step of administering to a subject in need thereof an effective amount of the polymer of any one of claims 1 to 6, 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 (BRIC), inborn errors of bile acid synthesis, Alagille syndrome (ALGS), neonatal cholestasis caused by citrin 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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